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Retrovirology 2012, Volume 9 Suppl 1 http://www.retrovirology.com/supplements/9/S1 MEETING ABSTRACTS Open Access Abstracts from the 17th International Symposium on HIV and Emerging Infectious Diseases (ISHEID) Marseille, France. 23-25 May 2012 Edited by Alain Lafeuillade Published: 25 May 2012 These abstracts are available online at http://www.retrovirology.com/supplements/9/S1 INVITED SPEAKER PRESENTATIONS I1 Managing antiretrovirals in co-infected patients in 2012 Jürgen Rockstroh University of Bonn, Germany Retrovirology 2012, 9(Suppl 1):I1 In patients with HIV and hepatitis B or C co-infection initiation of early HAART therapy is clearly recommended by the international guidelines reflecting the overall survival benefit with regard not only to HIV but also clinical endpoints of liver disease in the co-infected patient population. In patients with hepatitis C co-infection administration of successful HAART is associated with less inflammation in liver biopsies and hence, over time less fibrosis generation. Obviously, drugs which are dually active against HIV and HBV do not only prevent HIV disease progression, but also decrease the progression of chronic hepatitis B related liver disease. Nevertheless, recent reports on the emergence of non cirrhotic portal hypertension as a rare cause of upper gastrointestinal bleeding in HIV patients most likely following prolonged didanosin exposure have raised questions with regard to potential drug toxicity as sequelae of HAART causing possible damage to the liver and the portal vascular system. Under consideration of metabolic changes including dislipidemia and insulin resistance caused by various components of different drug classes the question also remains in how far risk for fatty liver disease may rise after decades of HAART administration. Therefore, close surveillance programs and monitoring are required to answer the long term safety questions around HAART administration and liver safety. Nevertheless, the clear demonstration of overall increased survival under HAART in patients with concomitant hepatitis and HIV clearly underlines that the benefits of HAART outweighs potential toxicity risks. Also with the knowledge obtained around mitochondrial toxicity observed under D-nucleoside therapy treatment algorithms for co-infected patients now specifically refrain from using these drugs in particular. With the advent of new drug classes with low hepatotoxicity profiles new combinations arise which potentially improved liver safety profile over time. But again, long term data will be needed to eventually decide what are the best HIV treatment options in patients with concomitant liver disease. I2 Clinical use of pharmacogenomics Amalio Telenti University of Lausanne, Switzerland Retrovirology 2012, 9(Suppl 1):I2 The expanding list of antiretroviral drugs, the increasing possibilities in therapy of HCV and other co-infections represent an attractive setting for individualized prescription. However, there has been considerable hype, and limited impact of pharmacogenetics in care. The development of pharmacogenetic tools would be of interest considering the complexity of treatment, the cost, and the expectation of long-term exposure. It would also be desirable given the possibility of serious toxicity, but also treatment intolerance, cumulative toxicity or the unfavorable interaction of drugs and aging, metabolic, cardiovascular and bone disease processes. This presentation will review available tests, the position of agencies on the inclusion of pharmacogenetic information in labels, and a perspective on developments in genomic medicine. I3 Gene therapy to cure HIV? Prospects and realities Gero Hütter Heidelberg University, Württemberg, Germany Retrovirology 2012, 9(Suppl 1):I3 Introduction: HIV-1 can persist in a latent form in resting memory CD4+ cells and macrophages carrying an integrated copy of the HIV genome. Because of the presence of these stable reservoir cells, eradication by antiretroviral therapy is unlikely and in order to achieve eradication, alternative treatment options are required. Materials and methods: Recently, we have described a successful hematopoietic stem cell transplantation in an HIV-1 infected patient transferring donor derived cells with a natural resistance against HIV infection. These haematopoietic stem cells engrafted, proliferated and differentiated into mature myeloid and lymphoid cells. Results: At present the patient is more than five years after allogeneic transplantation without requirement of any antiretroviral treatment. Analyzing peripheral blood cells and different tissue samples including gut, liver, and brain, no viral load or proviral DNA could be detected. Conclusions: There is a degree of optimism that gene therapy in combination with SCT can improve HIV-1 treatment. After nearly 30 years of research and progress HIV infection has become a chronic disease in developed countries. However, the evolution of this infectious agent, due to its rapid mutation, is quite unpredictable. Therefore, strategies to eliminate the virus from the body to achieve cure are desirable. Currently, HIVtargeted gene therapy appears to have great potential as an effective strategy that may eliminate or reduce the need for antiretroviral therapy and achieve elimination of the viral reservoir. © 2012 various authors, licensee BioMed Central Ltd. All articles published in this supplement are distributed under the terms of the Creative Commons Attribution License (http://creativecommons.org/licenses/by/2.0), which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited. Retrovirology 2012, Volume 9 Suppl 1 http://www.retrovirology.com/supplements/9/S1 I4 Update on HIV pandemic Anna Mia Ekstrom Karolinska Institutet, Stockholm, Sweden Retrovirology 2012, 9(Suppl 1):I4 The aim of this presentation is to present the state of the HIV pandemic. The talk will start with an overview of current trends in HIV prevalence and incidence, mode of transmission, and, access to prevention and treatment in various regions of the world. Thereafter, the presentation will focus on three main dilemmas related to the interpretation of HIV prevalence, treatment coverage and recent policy guidelines for HIV treatment and prevention. The first dilemma to be discussed is the difficulty in measuring HIV incidence using current methodologies. HIV prevalence has become a poor estimate of HIV incidence given the varying and rapidly changing access to antiretroviral treatment (ART) as well as retention in ART programs affecting both survival and incidence. The dilemma of selecting an appropriate population group for monitoring HIV trends will also be mentioned. Neither antenatal clinic measurements of point prevalence rates or demographic health surveillance techniques are free of bias. Secondly, the dilemma of using coverage measures to assess the success of scaling up access to ART and prevention of mother to child transmission (PMTCT) will also be discussed. The problem of identifying an appropriate denominator for a lethal disease is one problem. Another one is the high drop-out from both ART and PMTCT programs, making reported coverage estimates based on enrollment, rather than completion, hard to interpret. Finally some recent policies related to HIV treatment and prevention such as Test and Treat, couple testing and the new WHO guidelines for PMTCT will also be briefly discussed in relation to country GDP and health systems capacity. I5 HIV prevention by vaccine Robert Gallo Institute of Human Virology, Baltimore, USA Retrovirology 2012, 9(Suppl 1):I5 Page 2 of 54 successfully demonstrated in 2010 in the iPREX trial. Moreover, this randomized placebo-controlled study showed that men who were adherent in regard to the once-daily use of TDF/FTC were at low risk for acquisition of HIV in comparison to both non-treated controls and men who were not as adherent to their recommended drug intake (44% protection overall but 92% protection among adherent populations). Similar findings were obtained in a different study termed Partners/Prep performed in two developing countries. Yet, despite these positive results, the idea of PrEP remains controversial in some settings, and the following concerns among others have been expressed. 1. The use of antiretroviral drugs in PrEP may deprive some people already infected by HIV of the chance to benefit from these same drugs, due to competition for resources and/or drug supply. 2. The use of PrEP will promote sexual promiscuity and lead to more STDs. 3. Although PrEP has worked in controlled trials, it will be less likely to be successful in real world settings, where adherence and counselling may be less than adequate, 4. PrEP may promote HIV drug resistance in real world settings if ARVs are taken by people who may already be HIV-infected and don‘t know it. 5. Not all PrEP trials have been successful, e.g. VOICE and FEM-PrEP. 6. The daily consumption of ARVs may be toxic. One potential solution to some of these concerns may be the use of PrEP on an intermittent basis, in which ARVs are taken only on the day of anticipated sexual relations and for two days after a sexual encounter. This concept could have the advantage of lowering both costs and drugrelated toxicities. I7 New antiretrovirals Roy Gulick Weill Medical College of Infectious Diseases, New York, USA Retrovirology 2012, 9(Suppl 1):I7 Three major HIV vaccine efficacy clinical trials have now been completed. Two failed, one of the two even increased infection and the third was modestly successful. First: VaxGen, used conventional gp120 protein and like in monkey trials it failed likely due to type specific Abs and perhaps inadequate Ab titans; the second, by the Vaccine Research Center (VRC) and Merck based solely on CMI and predictably failed. Less predictably it actually increased the numbers infected. This was likely due to use as a vector of an adenovirus strain already exposed to a sizeable percentage of people from earlier infections, thereby leading to increased T-cell activation which is accompanied by an increase in CCR5 co-receptor for HIV and consequently to increased susceptibility to HIV infection. The third large efficacy trial involved a novel gp120 delivered by the canary pox virus known as ALVAC made by Sanofi as well as some other HIV genes and boosted by a gp120 containing a herpes virus small sequence known as Gd. This trial was run by Colonel Dr. Nelson Michael and his co-workers in the U.S. Army AIDS research group in collaboration with colleagues in Thailand and resulted in modest success associated with Abs (not CMI) which binds V2 of gp120 and without detectable neutralizing Ab activity. Of great interest to us was the short duration of the Abs resulting in far greater success in the first half year than in the remaining part of the study. This is typical for Abs to gp120, and it is precisely what we have found in our primate challenge experiments with our candidate vaccine, a complex of gp120 with binding region of CD4 which we (A. DeVico, G. Lewis, T. Fouts, and Y. Guan) call the full length single chain (FLSC). I will summarize our rationale for the FLSC, our updated primate results, and our plans for clinical trials in collaboration with Sanofi, N. Michael and his group, and the Gates Foundation. There are 26 approved antiretroviral drugs available in 2012 in 6 mechanistic classes: reverse transcripatase inhibitors (both nucleosides and nonnucleosides), protease inhibitors, entry inhibitors (both fusion inhibitors and CCR5 receptor antagonists), and integrase inhibitors. Current antiretroviral therapy combinations dramatically decrease HIV-related morbidity and mortality. However, despite these advances, some current antiretroviral regimens may be inconvenient, toxic, and/or have suboptimal antiretroviral activity, particularly against drug-resistant viruses. Thus, newer compounds are needed that improve convenience and tolerability, reduce toxicity, and improve antiretroviral activity, particularly against drug-resistant viruses. Additionally, new drugs may better penetrate tissue reservoirs (e.g. genital tract, central nervous system), exploit new targets with new mechanisms of action, or be administered in new formulations. There are a number of HIV investigational drugs in development currently. These include a new pharmacokinetic “boosting” agent, cobicistat (GS-9350) and newer antiretroviral agents in a number of classes, including new nucleoside reverse transcriptase inhibitors, non-nucleoside reverse transcriptase inhibitors, protease inhibitors, entry inhibitors, and integrase inhibitors. Of those in the pipeline, a few compounds are in advanced stages of development: the nucleoside analogue GS-7340, a pro-drug of tenofovir (phase 2); and the integrase inhibitors, elvitegravir (phase 3 completed) and dolutegravir (phase 3). In addition, there are drugs with new mechanisms of action in development, including the CD4 attachment inhibitor, BMS-663068 (phase 2), and the CCR5 antagonist, cenicriviroc (phase 2). Probably the greatest need in the HIV clinic today is compounds that have activity against multidrug-resistant viral strains. Another important need is alternative one-pill, once-daily formulations for both initial and subsequent regimens. However, the clinical use of these newer agents will depend on the results of phase 3 clinical trials, and the timeline for development and availability. I6 Pre-exposure prophylaxis against HIV: pros and cons Mark Wainberg McGill University, Montreal, Canada Retrovirology 2012, 9(Suppl 1):I6 I8 New drug interactions in HIV and HCV David Back University of Liverpool, UK Retrovirology 2012, 9(Suppl 1):I8 The successful use of the co-formulation of Tenofovir (TDF) and Emtricitabine (FTC) to prevent HIV infection among gay men was Drug-drug interactions (DDIs) remain one of the challenges faced by health care professionals involved in patient management and by Retrovirology 2012, Volume 9 Suppl 1 http://www.retrovirology.com/supplements/9/S1 researchers seeking to understand the many different mechanisms that may be involved. While in HIV treatment there has been an understanding that checking for DDIs is part of routine management, the development of directly acting antiviral (DAA) drugs is changing the whole approach to the treatment of HCV. DDIs have not really been high on the agenda while pegylated interferon and ribavirin have been the standard of care for Hepatitis C; however the first generation HCV protease inhibitors while representing a huge advance, also present new treatment challenge of awareness and management of DDIs. In this presentation, some of the key areas of DDIs with a) HIV drugs, b) HCV drugs and c) co infected patients receiving both HIV and HCV drugs will be discussed. While it has long been established that boosted HIV protease inhibitors are likely to interact with other medications metabolised by CYP3A4 (and other enzymes/transporters) to increase drug exposure and that drugs like efavirenz, nevirapine and etravirine are inducers of drug metabolism some of the current challenges are to determine i) the likelihood of interactions where there are no study data to guide us and ii) what happens when a patient switches therapy off an enzyme inducing drug. In relation to the new DAAs, Boceprevir is primarily metabolised by the enzyme aldo-ketoreductase (AKR) but also undergoes metabolism by CYP3A4; telaprevir is metabolised by CYP3A4. In addition, both boceprevir and telaprevir are strong inhibitors of CYP3A4 and are substrates for P-glycoprotein. Co administration of drugs that are highly dependent on CYP3A4 for clearance and for which elevated plasma concentrations are associated with serious or life threatening events means a contraindication with boceprevir and/or telaprevir. However, many other interactions require close monitoring, alteration of drug dosage or timing of administration and here we clearly need help to understand the potential magnitude of an interaction and strategies for patient management. Since targeted drug interaction studies are done in the development programme and then post-licensing there are a limited number of drugs for which there are actual study data. Where there are no data, guidance can only be given based on knowledge of the pharmacology of the respective drugs. This is the approach being used in the resource http://www.hep-druginteractions. org. One particular area of concern is HIV positive patients who are receiving antiretrovirals (ARVs) since they are relatively highly represented among patients with HCV infection, and are at a high risk of DDIs. Recent drug interaction data presented on telaprevir and boceprevir have reinforced the initial suspicions that we are faced with a whole new challenge of managing multiple interactions in co-infected patients. Management of DDIs with these exciting new agents certainly poses a challenge and awareness of the potential for DDIs is fundamental for safe prescribing. Think DDIs for the DAAs. But we are still on a steep learning curve and there are unanswered questions. I9 Animal models of functional HIV cure Guido Silvestri Emory University School of Medicine, Atlanta, USA Retrovirology 2012, 9(Suppl 1):I9 Developing new therapeutic strategies to achieve a functional cure for HIV infection is a central objective for contemporary AIDS research. A key obstacle towards a functional cure for HIV/AIDS is the paucity of information regarding the establishment and maintenance of the persistent reservoirs of latently HIV-infected cells which support the rapid reactivation of virus replication when antiretroviral therapy (ART) is stopped. Given the many and important similarities between HIVinfection of humans and SIV infection of macaques, it is very likely that crucial information on the nature of HIV reservoirs may come from studies conducted in SIV or SHIV-infected macaques that are treated with ART. While this area of AIDS research is still in its early stages, it is important to note that these studies have now become possible due to the availability of antiretroviral regimens that successfully reduce SIV or SHIV replication below detectable limits. Specific advantages of the nonhuman primate models for HIV latency, reservoirs and functional cure include the following: (i) possibility to conduct extensive characterization of the virus reservoirs and pathogenic processes in tissues (including elective necropsy); (ii) performance of in vivo pilot trials of new therapeutic approaches can be conducted in a timely and controlled fashion; (iii) developing and testing of “risky” interventions (i.e., cell Page 3 of 54 depletion experiments, stem cell-based interventions etc) that may pose significant ethical challenges in humans; and (iv) possibility to control for various clinical parameters that are very hard to control in humans (time of infection, duration of ART etc). On the other hand, current limitations to this type of studies are their cost and the lack of standardized research resources (i.e., assays, virus stocks, etc). In this presentation, the potential and limitations of animal models of functional HIV cure will be discussed together with selected examples of therapeutic approaches that could be developed and tested in non-human primates. I10 Molecular control of HIV-1 postintegration latency: implications for therapeutic strategies Carine Van Lint Gosselies Campus, Charleroi, Belgium Retrovirology 2012, 9(Suppl 1):I10 The current antiretroviral therapy HAART is effective and life-prolonging but does not eradicate HIV-1 from infected patients. A reduction of HIV-1 RNA levels in the plasma in HAART-treated individuals to less than 50 copies/ml is frequently achieved but residual viremia persists as detected by ultrasensitive assays. The sources of this persistent viremia are still not fully understood but could arise from ongoing cycles of residual viral replication and/or from the reactivation of viral expression from latently-infected cells. These latently-infected cells contain stablyintegrated, transcriptionally-silent but replication-competent proviruses, thereby representing latent reservoirs of HIV-1. They are a permanent source for virus reactivation and could be responsible for the rebound of plasma viral load observed after HAART interruption. HIV-1 transcriptional repression is crucial to the establishment and maintenance of postintegration latency. Several elements contribute to HIV-1 transcriptional repression including: 1) the site of integration and mechanisms of transcriptional interference, 2) the absence of crucial inducible host transcription factors, 3) the presence of transcriptional repressors, 4) the nucleosomal organization of the HIV-1 promoter, 5) the epigenetic control of the HIV-1 promoter (histone posttranslational modifications, such as acetylation and methylation, and DNA methylation), 6) the sequestration in an inactive form of the cellular positive transcription elongation factor b (P-TEFb), composed of cyclin-dependent kinase 9 (CDK9) and human cyclin T1, 7) the absence of the viral transactivator Tat, which promotes transcription via recruitment to the HIV1 promoter of P-TEFb, histone-modifying enzymes and ATP-dependent chromatin-remodeling complexes required for nucleosomal disruption and transcriptional processivity. The involvement of these elements in postintegration latency depends on the status of activation and differentiation of the heterogeneous CD4+ T cell populations hosting the HIV-1 reservoirs. Further understanding of the epigenetic and non-epigenetic mechanisms regulating HIV-1 latency and reactivation from latency should help devise novel strategies to eliminate latent HIV-1 infection or to restrict the latent pool to a size bearable by the host immune system. I11 A roadmap to a cure Mario Stevenson University of Miami, Leonard M. Miller, Miami, USA Retrovirology 2012, 9(Suppl 1):I11 Highly active anti-retroviral therapy (HAART) effects sustained suppression of viral replication in infected individuals. Despite this, viral replication rapidly resumes if therapy is interrupted. The prevailing view is that viral persistence during HAART is sustained by a reservoir of latently infected, quiescent CD4+ T-lymphocytes, a view that is supported by the apparent lack of viral evolution in the resting CD4+ T cell reservoir and the apparent lack of change in low level plasma viremia when therapy is intensified. A less popular view is that viral persistence in HAART may be sustained by a low level of ongoing or “cryptic” replication, a view that is supported in part by our recent work measuring unintegrated cDNA and specifically episomal cDNA in HAART-treated patients. In order to characterize the reservoirs that persist in the face of HAART, we have been examining the dynamics of HIV-1 in tissue viral reservoirs and the virologic response to therapy in lymphoid tissue. Lymphoid Retrovirology 2012, Volume 9 Suppl 1 http://www.retrovirology.com/supplements/9/S1 tissue was obtained from patients at various intervals after HAART initiation and the virologic response as well as intracellular drug levels in cells from lymphoid tissue was examined. Our analysis indicates that suppressive therapy is limited in its ability to curtail HIV-1 replication in lymphoid tissue and that this may be a consequence of poor intracellular sequestration of some antiretrovirals in cells of lymphoid tissue. These results have implications for strategies aimed at limiting viral persistence in the face of HAART. I12 Towards a cure for HIV: a long road ahead Tae-Wook Chun National Institute of Health, Bethesda, USA Retrovirology 2012, 9(Suppl 1):I12 Plasma viremia can be effectively suppressed and maintained below the limits of detection for extended periods of time in most human immunodeficiency virus (HIV)–infected individuals receiving antiretroviral therapy (ART). However, it has not been possible to eradicate HIV by ART alone, likely due in part to the persistence of various viral reservoirs in lymphoid tissues. In this regard, the existence of latently infected, resting CD4+ T cells carrying replication-competent HIV has posed one of the major challenges to the long-term control or eradication of HIV in infected individuals on ART. Consequently, there has been considerable focus on therapeutic strategies to reactivate the latent viral reservoir using various agents, such as cytokines, histone deacetylase inhibitors, and mitogens, under the assumption that these cells would die due to HIV induced cytopathic effects and antiretroviral drugs would prevent spread of infection. However, such approaches have shown no clinical benefit to date. Moreover, it also has become clear that HIV persists in subsets of CD4+ T cells in blood and lymphoid tissues of infected individuals receiving ART. Recent data from our laboratory will be discussed which will include potential mechanisms of HIV persistence and prospects for eradication and new therapeutic approaches in HIV-infected individuals receiving effective ART. I13 Universal access to anti-HIV therapy Yves Souteyrand HIV/AIDS Department World Health Organization, Geneva, Switzerland Retrovirology 2012, 9(Suppl 1):I13 WHO, in collaboration with UNAIDSs and UNICEF, report annually on progress in the different HIV/AIDS interventions. In 2010, more than 6.6 million people had access to Antiretroviral Therapy (ART), in low-and – middle income countries, representing a 17-fold increase in less than 7 years. This is one of the most successful public health achievements. However, this represents less than 50% coverage of needs. The 2010 global commitment of Universal Access to ART has not been achieved. The presentation will address determinants of progress and of inequalities in access to ART, according to geographical and population criteria. West and Central Africa as well as Eastern Europe and Central Asia are reporting low coverage rates compared to other similar regions. Coverage among children less than 15 years old is much lower than adult coverage (23% versus 51%). Access to ART for intravenous drug users is very low in most countries. In many settings, access for people in rural areas is much more limited than in public area. Closing the gap in access to ART will need implementing huge efforts to reduce inequities in access. A comprehensive approach should include improvement of health delivery systems, decentralization of care, simplification of treatment and laboratory monitoring. It will need also to address essential human right issues, including stigmatization and discrimination of key populations who currently do not have access to testing and counseling and other essential interventions. I14 Primary HIV infection: the French experience Cécile Goujard General Hospital of Bicêtre, Kremlin-Bicêtre, France Retrovirology 2012, 9(Suppl 1):I14 Page 4 of 54 The ANRS PRIMO is a prospective multicenter French Cohort which had enrolled 1433 patients during primary-HIV-1 infection (PHI) since 1996, with the objectives to improve pathophysiological knowledge on PHI, to assess the impact of an early antiretroviral treatment and to contribute to document the epidemiology of new infections in France. Clinical and laboratory data are collected at inclusion, month (M)1, M3, M6, and then every 6 months. All patients are antiretroviral-naïve at inclusion. Patients were included early after infection (median time, 31 days; 26% acute infection); 84% were men. The % of patients infected with non-B subtypes increased in the last decade, while the frequency of resistant viral strains remained stable. At inclusion, median CD4 cell counts and viral HIV load were 518/mm 3 and 5.1 log 10 copies/mL at PHI, with a wide range of values in the whole population (IQR, 372-678/mm3 and 4.5-5.7 log respectively), depending only partially to the time from infection. Median cellular DNA at inclusion was 3.4 log cp/10 6 PBMCs (IQR, 2.9-3.7). Early immunological (CD4 counts) and virological parameters (HIV RNA and DNA) could predict the risk of progression to a CD4 count < 350 cells/mm3 in untreated patients and the chance of spontaneous persistent control of viral replication after infection. 52% of patients initiated an antiretroviral treatment at inclusion. Although a transient treatment during PHI did not translate into a benefit in terms of viral set-point compared to untreated patients, it could lead to long term preservation of CD4 cell counts. Furthermore, we showed that rare treated patients were capable of controlling viral replication after treatment interruption, those patients had lower viral reservoir at inclusion before treatment and at treatment interruption compared to non controllers. Cellular activation was high during PHI and declined substantially thereafter, with lower levels in patients treated early after infection compared to patients treated during the chronic phase. Antiretroviral treatment is now given in less advanced infection and even discussed as a universal approach to prevent HIV transmission. The question of treatment initiation in all patients diagnosed during PHI is relevant. I15 Treating since the beginning? Jean-Pierre Routy McGill University, Montreal, Canada Retrovirology 2012, 9(Suppl 1):I15 Like a pendulum, the therapeutic recommendations to initiate antiretroviral therapy (ART) have changed over the last two decades. Indeed, initiation of ART has swung from an initial “treat hard and early”, to a CD4 T cell count spanning from 200 to 500 and recently to a “as soon as the patient is ready”. Changes on the optimal timing to initiate ART were based on the following 1) Availability of new ART with increased potency, improved tolerance and reduced pill burden; 2) Evidence that controlled viral replication on ART is associated with a dramatic decrease in HIV transmission; 3) New insights on HIV pathogenesis and on the establishment and maintenance of HIV reservoirs. Recent findings on HIV pathogenesis concerning the relationships between CD4 T cell counts, immune activation, non-infectious clinical events and cancers will be presented. Specifically, the influence of CD4 nadir on the quality of long-term immune reconstitution and viral reservoir persistence following ART initiation will be highlighted. We will also focus on the clinical relevance of maintaining certain CD4 T cell subsets like central memory pool for an optimal cytotoxic HIV-specific and vaccination responses. We will also revisit the tissue and cellular localizations of HIV reservoirs according to the time of treatment initiation. Furthermore, ethical considerations on a “patient-centered medicine” for early ART initiation will be discussed. Particularly, the advantages and inconveniences of a life-long treatment on a patient’s quality of life, drug-resistance development, long-term drug toxicity, cost and observance issues will be presented. The very early ART initiation reveals a new frontier 1) Allowing for some patients an ART-free viral control post-drug discontinuation era 2) Selecting optimal patients harboring a reduced HIV reservoir to be invited to participate in clinical trials aiming to HIV eradication. We are entering in a new era where “Treating early to be able to stop early” will be the focus of our future collaborating research efforts to vividly improve the life of HIV-infected people. Retrovirology 2012, Volume 9 Suppl 1 http://www.retrovirology.com/supplements/9/S1 I16 Anti-latency agents to purge HIV reservoirs Santiago Moreno Ramón y Cajal Hospital, Madrid, Spain Retrovirology 2012, 9(Suppl 1):I16 The persistence of latent HIV-infected cellular reservoirs represents the major hurdle to virus eradication with highly active anti-retroviral therapy (HAART), since latently infected cells remain a permanent source of viral reactivation. HIV establishes a persistent infection in CD4+ T lymphocytes (and to a lesser extent in macrophages as well), creating a persistent reservoir consisting mainly of latently infected resting memory CD4+ T cell. Although pre- and post-integration latencies have been described in HIV-1, the reservoir that appears to be the major barrier to eradication is composed of latently infected cells carrying an integrated provirus that is transcriptionally silent. It has been suggested that reactivation of the latent reservoirs could allow effective targeting and possible eradication of the virus. Immunoactivation therapy to reduce the latent pool of HIV by treatment with the anti-CD3 antibody OKT-3 alone or in combination with interleukin-2, substantially failed to significantly decrease the viral reservoir. Non-specific T-cell activation may induce high-level viral replication above a level that can be fully contained by ART, while increasing the susceptibility of uninfected cells. Selective targeting of HIV provirus via agents that induce the expression of quiescent HIV, but have limited effects on the uninfected host cell is an alternate approach to attack latent HIV. Activation from latency to completion of the replication cycle should result in lytic cell death of CD4+ T cells. Multiple mechanisms that contribute to the maintenance of proviral latency could be targeted to activate the latent virus. As examples of potentially useful agents, IL–7 can reactivate HIV–1 in latently infected cells in vitro through the induction of the Janus kinase–signal transducer and activator of transcription (JAK–STAT) signalling pathway. The use of different chemical compounds targeting the PKC signalling pathway (prostratin, bryostatin) has also been proposed as a means of reactivating viral reservoirs. Finally, HDAC blocking is an attractive potential means of inducing broad reactivation of HIV–1 reservoirs, and promising results have been achieved using the HDAC inhibitor vorinostat. I17 Hepatitis C as a metabolic disease Patrice Andre Croix-Rousse Hospital, Lyon, France Retrovirology 2012, 9(Suppl 1):I17 Before liver cirrhosis and hepatocellula carcinoma can develop, the early and long lasting features of hepatitis C are host metabolism modifications with a specific and so far unique metabolic syndrome that may associate insulin resistance, liver steatosis and hypo-betalipoproteinemia. These metabolic perturbations are directly induced by heptatitis C virus (HCV) replication and regress after viral suppression. Symmetrically HCV depends on glucose and lipid metabolism for its replication. HCV induced insulin resistance is both hepatic and peripheral. The insulin receptor pathway is impacted at several steps by viral proteins with probable functional consequences. Mechanism of peripheral insulin resistance remains obscure and the viral signals send to adipose tissue or muscles have to be identified. One major IR metabolic consequence might contribute to the mobilization of free fatty acids from periphery to the liver and to the constitution of liver steatosis. Importantly, HCV modifies with genotype-specific differences, the synthesis, degradation and secretion of lipid in a coordinated fashion to promote the accumulation of neutral lipids. The lipid droplets (LD) that are the lipid storage organelles are mandatory platforms for the assembly of infectious viral particles. In particular localization of core protein and NS5A on LD and mobilization of the LD are essential steps that control viral infectivity. Interestingly, the efficiency of HCV to the LD disposal and use correlates to viral production and may influences the extent of hepatic accumulation of lipids. The most striking association of HCV with lipid metabolism resides in the coincidence of the betalipoproteins and viral particles pathways with the formation of unique hybrid viral particles. Indeed HCV depends on a Page 5 of 54 functional very low density lipoproteins (VLDL) assembly and secretion process to be secreted. HCV also modifies at different degree the VLDL composition forming sub viral particles, which are minimally modified VLDL that bear the viral envelop glycoproteins, or hybrid viral particles known as lipo-viral-particles (LVP), which contain all the viral and VLDL components that are highly infectious. Functions of these hybrid particles on the disease features and progression as well as therapeutic targets remain to be fully characterized. Thus, HCV appears to have developed an original and so far unique way with major clinical consequences to modify and use the lipid metabolism to persist in the host. I18 HCV resistance Philippe Halfon Ambroise Paré Hospital, Marseille, France Retrovirology 2012, 9(Suppl 1):I18 The efficacy of Direct Antiviral Agents (DAAs) is limited by the presence of Resistant associated virus mutations (RAVs) resulting in amino-acid substitutions within the targeted proteins which affect viral sensitivity to these compounds. Six major position mutations in the NS3 HCV Protease (36, 54, 155, 156, 168, and 170), fifteen in the NS5B polymerase (96, 282, 316, 365, 414, 419, 423, 448, 482, 494, 495, 496, 499, 554, 559) and five in the NS5 A region (28, 30, 31, 58 and 93) have now been reported in vitro or in vivo associated with different levels of resistance. The HCV NS3,NS5A, Pol (NNI) mutations occurred quickly (less than 15 days) and longer for Nucleosides Inhibitors in monotherapy and the genetic barrier can be overcome by combination with PegylatedInterferon+Ribavirine in quadruple therapy. There is a long term persistence of HCV NS3 Protease mutations after the end of therapy and it is important to stop the NS3 protease inhibitor early in patients with ongoing replication to avoid the selection of resistant variants with increased fitness and a higher potential of long-term survival. Issues on HCV archived mutations are not solved. Virological failure was more likely in patients with genotype 1a infection than 1b and was associated with the presence of resistant variants. Furthermore, the time taken for resistant HCV variant populations to return to WT is longer for patients with genotype 1a versus genotype 1b. Selection of resistant variants that, in turn, could produce cross-resistance to whole class of drugs with overlapping resistance profiles Combinations of Protease Inhibitors with other class of antiviral with separate modes of action & non-overlapping resistance profile is preferable. Additionally, Ribavirine prevents viral breakthrough in combination with PegylatedInterferon and DAAs and the effect of ribavirin important seems limit the DAA resistance. The resistance profiling does remain a challenge for the next generation of protease, NS5A, non nucleoside inhibitors and probably for nucleoside inhibitors; thus, the lessons from HIV infection and the first clinical proof of IFN-free regimen treatment indicate that combinations of drugs with different mechanisms of action will be an attractive strategy for hepatitis C. I19 What happens after hepatitis C eradication? Stanislas Pol Hepatology Unit, Cochin Hospital, Paris, France Retrovirology 2012, 9(Suppl 1):I19 The treatment of hepatitis C virus infection (HCV) by a combination of pegylated interferon and ribavirin, according to early viral kinetics, leads to a sustained virological response (SVR) in more than 50% of patients with chronic infection. This SVR is a complete recovery of the infection but more than 50% of genotype 1-infected patients do not achieve SVR. A better understanding of the viral cycle, and the characterization of viral enzymes which are potential targets, resulted in the development of new molecules, direct acting antiviral drugs (DAA) targeted against HCV, either specific of genotype 1 (NS3/NS4A protease inhibitors and NS5B polymerase inhibitors) or with a wider spectrum (NS5A or entry inhibitors), and nonspecific antivirals (new interferons, cyclophilin inhibitors). The results of these phase II and III trials which clearly demonstrated a 20 to 30% increase Retrovirology 2012, Volume 9 Suppl 1 http://www.retrovirology.com/supplements/9/S1 in the SVR rate of genotype 1-infected patients, either naive or treatment experienced. These new drugs has now been approved by the end of 2011, after a temporary approval for compassionate use in cirrhotic genotype 1 patients with previous relapse or partial response to the combination therapy and a new turn appears with the “interferon free regimens” which combine different direct acting antivirals. The complete virologic recovery which is achieved with all the antiviral treatments clearly reduces the rates of liverrelated morbidity and mortality but, in the absence of biopsy-proven cirrhosis reversal, the risk of occurrence of hepatocellular carcinoma is still present and requires a regular US follow-up in those patients with extensive fibrosis or cirrhosis for an early screening. In the experienced patients with active replication, new antivirals are mandatory to reduce the risks of complications, including liver transplantation, which is high, especially in patients with rapid fibrosis. In the future, the main limitations of triple therapy will be safety (cutaneous rash or anemia which may be controlled), cost, compliance, viral resistance, and drug-drug interactions that must be avoided by educating patients and physicians. I20 Antiretroviral generics Véronique Andrieu Galenic Laboratory Faculty of Pharmacy, Marseille, France Retrovirology 2012, 9(Suppl 1):I20 The current definition for generic medicinal products is found in Directive 2001/83/EC, Article 10(2)(b), which states that a generic medicinal product is a product which has the same qualitative and quantitative composition in active substances and the same pharmaceutical form as the reference medicinal product, and whose bioequivalence with the reference medicinal product has been demonstrated by appropriate bioavailability studies. The different salts, esters, ethers, isomers, mixtures of isomers, complexes or derivatives of an active substance are considered to be the same active substance, unless they differ significantly in properties with regard to safety and/or efficacy. Furthermore, the various immediate-release oral pharmaceutical forms shall be considered to be one and the same pharmaceutical form. Concept of bioequivalence is fundamental: The purpose of establishing bioequivalence is to demonstrate equivalence in biopharmaceutics quality between the generic medicinal product and a reference medicinal product in order to allow bridging of preclinical tests and of clinical trials associated with the reference medicinal product. In bioequivalence studies, the plasma concentration time curve is generally used to assess the rate and extent of absorption. Selected pharmacokinetic parameters and preset acceptance limits allow the final decision on bioequivalence of the tested products. AUC, the area under the concentration time curve, reflects the extent of exposure. Cmax, the maximum plasma concentration or peak exposure, and the time to maximum plasma concentration, tmax, are parameters that are influenced by absorption rate. In studies to determine bioequivalence after a single dose, the parameters to be analysed are AUC(0-t), or, when relevant, AUC(0-72h), and Cmax. For these parameters the 90% confidence interval for the ratio of the test and reference products should be contained within the acceptance interval of 80.00-125.00%. To be inside the acceptance interval the lower bound should be ≥ 80.00% when rounded to two decimal places and the upper bound should be ≤ 125.00% when rounded to two decimal places. Evaluation of Generic medicinal product is also based on the pharmaceutical quality of the active substance: absence of impurities,.. and of the finished product : reproducibility of the manufacturing process, stability,…. A table of the Antiretroviral drugs and of the Generics available in Europe is presented. I21 Impact of HIV minor species and tropism Karin Metzner University Hospital, Zurich, Switzerland Retrovirology 2012, 9(Suppl 1):I21 Resistance against antiretroviral drugs is one challenge when treating HIV infected subjects. Resistance occurs in consequence of mutations in the HIV Page 6 of 54 genome, thus, viral proteins can no longer be inhibited by antiretroviral drugs or, in case of the CCR5 inhibitor Maraviroc, the virus can also escape through coreceptor switch. Drug-resistant viruses (1) can be rapidly selected leading to virological failure, (2) can persist also in the absence of drugs, (3) can be transmitted, and (4) their presence can limit further treatment options. This is all known in the context of drug-resistant viruses representing the majority of the virus population. Here, the focus will be on minority drug-resistant HIV variants: Virus subpopulations not detectable using standard genotype resistance testing based on population sequencing, thus, representing less than 20-25% of the total virus population. New technologies enable the detection and quantification of minority drug-resistant HIV variants to levels far below 1%. However, their impact on antiretroviral therapy is still controversially discussed. This presentation will provide an overview of the current techniques to quantify minority viral variants, the clinical studies investigating the prevalence and impact of minority drug-resistant HIV variants, and the challenges and potential benefits of clinical implementation. I22 A global virus network and a perspective on viral infections in humans Robert Gallo Institute of Human Virology, Baltimore, USA Retrovirology 2012, 9(Suppl 1):I22 During the flu pandemic of the 1910s, the polio outbreaks of the 1950s, and the AIDS crisis of the 1980s, invaluable time was wasted and a large number of people died or became seriously ill while authorities planned a response, identified and concentrated resources, and developed a plan to prevent others from becoming infected. Each of these 3 pandemics of the last century as well as the smallpox epidemics were caused by viruses. I have noted that often there are no expert responsible medical virologists, and concomitantly I noted a decline in newly trained medical virologists. Without experts, initial responses to viral outbreaks can be needlessly harsh and economically devastating. Fear replaces logical analysis, and entire populations of healthy pigs and birds have been slaughtered simply because they could potentially harbor swine or avian flu strains. These overreactions hurt small farmers and the economies in developing nations, and make crafting a vaccine more difficult. Public health authorities require a reliable source of experts on basic human and animal virology to turn to for rapid answers. A select group of frontline virologists is needed to work quickly to convert preliminary information into advances in the laboratory to protect the blood supply when needed and begin looking for clinical treatments and, ultimately, a vaccine. Organizations involved in the surveillance of viruses, like the World Health Organization and CDC, and those involved in the delivery of care during emergencies, like the Gates Foundation’s GAVI Program, often do not have the manpower for training initiatives and basic virus research which are important parts of addressing any public health threat. The Global Virus Network is uniquely positioned to utilize the expertise of its members to bridge these efforts. The overall aim of the network is to achieve accelerated, innovative solutions to human viral diseases. The GVN also focuses on coordinated virology training for junior scientists and developing scholar exchange programs for recruiting and training the next generation of medical virologists. Today, the GVN is made up of 30 centers and 3 affiliates in 19 countries on 6 of the world’s 7 continents. Its membership includes all 7 medical virologists in the U.S. National Academy of Sciences, and others who are members of their own countries’ national academies. Their collective expertise covers all types of existing, emerging, and re-emerging viruses which affect humans. I23 Human papilloma viruses Denise A Galloway Fred Hutchinson Cancer Research Center, Seattle, USA Retrovirology 2012, 9(Suppl 1):I23 A group of approximately 13 human papillomaviruses (HPVs) are responsible for virtually all cervical cancers as well as the majority of vulvar, vaginal anal and penile cancers, as well as at least half of Retrovirology 2012, Volume 9 Suppl 1 http://www.retrovirology.com/supplements/9/S1 oropharyngeal cancers. Currently two prophylactic vaccines are highly efficacious in preventing HPV 16 and 18 associated disease, and the quadrivalent vaccine also prevents HPV 6 and 11 warts. Currently therapeutic vaccines targeting the viral E6 and E7 oncogenes are being developed. One topic that will be discussed is whether the prophylactic vaccine can have utility in any setting where individuals are already infected. Secondly, this review will discuss the basis of long-term immunity and whether it is afforded by both vaccination and natural infection. Finally, strategies to provide more broad based coverage to other HPV types will be discussed. I24 Implications of influenza resistance Laurence Calatayud Marseille, France Retrovirology 2012, 9(Suppl 1):I24 Antiviral treatment is an important tool in the clinical management of severe or complicated influenza. Two classes of antiviral agents for influenza are available: adamantanes (amantadine and rimantadine), and neuraminidase inhibitors (oseltamivir, zanamivir, peramivir, laninamivir). The currently circulating human influenza viruses, influenza A (H1N1)pdm09, influenza A(H3N2), and influenza type-B, are all resistant to adamantanes, but most are sensitive to neuraminidase inhibitors. From April 2009 to 5 October 2011, a total of 605 cases of oseltamivirresistant infections with influenza A(H1N1) pdm09 virus have been reported to WHO from 32 countries. All resistant strains carry the H275Y substitution in the neuraminidase glycoprotein, which is known to confer a high level of resistance to oseltamivir. Of 468/605 (77%) cases with available clinical information, 133/468 (28%) occurred among patients who were severely immunocompromised. Of the patients who were not immunocompromised, 211/335 cases (63%) occurred after receiving treatment or prophylaxis with antiviral drugs. Four clusters of oseltamivir-resistant viruses with person-to-person transmission have been reported: 2 occurred among severely immunocompromised patients, while 2 were in healthy adults. Most viruses carrying the H275Y substitution remain sensitive to zanamivir. However reports indicate that other virus variants have emerged resulting in a reduced sensitivity to zanamivir and peramivir. Resistance to neuraminidase inhibitors in influenza A (H3N2) and influenza type-B, has been reported rarely, and has usually been associated with prolonged treatment in immunocompromised patients. The prevalence of resistance to neuraminidase inhibitors is still low, and oseltamivir remains the first line treatment recommended for patients with severe influenza or patients at a high risk of developing severe disease. However zanamivir is a therapeutic alternative for patients infected with a virus that is highly suspected to be resistant to oseltamivir, i.e., immunocompromised patients who have received oseltamivir, but still have evidence of persistent viral replication; or people who develop illness despite taking oseltamivir for a post-exposure prophylaxis. In addition appropriate infection control measures should be implemented to prevent spread of the resistant virus. Although prevalence of resistance to neuraminidase inhibitors remains low, reports of person-to-person transmission, and an increased prevalence of resistant viruses in community-based cases in specific regions, emphase the necessity of virological and epidemiological surveillance. Page 7 of 54 Materials and methods: The HIV-1IIIB virus, U373-MAGI-CXCR4CEM, HTLV-IIIB, Monoclonal antibody to HIV-1 gp41 (126-7), HIV-1 gp120 Antiserum (PB1 Sub 2), HIV-1 gp120 Antiserum (PB1), and HIV-1 gp120 Monoclonal Antibody (F425 B4e8) were obtained from the NIH AIDS Research and Reference Reagent Program, Division of AIDS, NIAID. The silver nanoparticles coated with 0.2 wt% PVP were obtained from Nanoamor, Houston, TX. Stock solutions and serial dilution of AGNPs were prepared in RPMI 1640 cell culture media. Cytotoxicity of AgNPs was ascertained in U373-MAGI-CXCR4CEM cells. The cell viability was assessed using a CellTiter-Glo® Luminescent Cell Viability Assay and Glomax Multidirection System (Promega). The neutralizing activity of AgNPs and NABs against HIVIIIB cell-free and cell associated virus was evaluated in an assay involving U373-MAGI-CXCR4CEM cells, AgNPs and NABs. Assessment of HIV-1 infection was performed with the Beta-Glo Assay System using Glomax Multidirection System (Promega). The percentage of residual infectivity after NABs, AgNPs, NABS+AgNPs, or media was calculated with respect to the positive control. The 50% inhibitory concentration (IC50) was defined according to the percentage of infectivity inhibition relative to the positive control. The inhibition data was statistically analyzed with the help of Wilcoxon rank-sum (WilcoxonMann-Whitney test) test. Results: The four NABs used in the study inhibited HIV-1 cell free infection at a dose response manner. They were however largely ineffective against the cell-associated virus. AgNPs alone however were able to inhibit both cell free and cell associated virus infection at a dose dependent manner. AgNPs when mixed together with NABs significantly increased inhibition of Cell associated HIV-1. Conclusions: The addition of AgNPs to NABs has significantly increased the neutralizing potency of NABs in prevention of cell-associated HIV-1 transmission/infection. O2 Search for small molecule activators of latent HIV Romas Geleziunas*, George Stepan, George Wei, Helen Yu, Michael Graupe, Nikos Pagratis, Tiffany Barnes, Tomas Cihlar, Joe Hesselgesser Clinical Virology at Gilead Sciences, Inc., Foster City, USA E-mail: [email protected] Retrovirology 2012, 9(Suppl 1):O2 Reservoirs of HIV that persist during ART represent barriers to eradication of this virus. One well documented reservoir of latent HIV is found in memory CD4 + T-cells. Identifying means to safely eliminate latently infected memory CD4+ T-cells is an important goal that may contribute to a cure for HIV. One approach toward this end is to activate latent proviruses with the premise that viral particles emanating from these cells will cause a cytopathic effect leading to the demise of the host cell. We have optimized and automated a primary cell-based HIV latency assay that can be used for high throughput screening of small molecule libraries in search of HIV activators. Using this assay, we have identified novel histone deacetylase (HDAC) inhibitors fromGilead’s compound collection that activate latent HIV. Analysis of these inhibitors revealed that the magnitude of HIV expression correlated with the breadth of cellular HDAC inhibition. In addition, we have identified a variety of other compounds that activate latent HIV such as kinase inhibitors which may point to novel mechanisms that govern HIV latency. This screening assay has the potential to identify novel molecular targets for drug discovery and new chemical classes that could be optimized to create new drugs to eliminate reservoirs of latent HIV. ORAL PRESENTATIONS O1 Inhibition of cell-associated HIV-1 by silver nanoparticles Dinesh K Singh*, Humberto H Lara Winston Salem State University, Winston Salem Nc, USA E-mail: [email protected] Retrovirology 2012, 9(Suppl 1):O1 Introduction: The glycoprotein gp120 and gp41 of HIV are the main targets for neutralizing antibodies (NABs). It appears that silver nanoparticles (AgNPs) also inhibit HIV-1 targeting same glycoproteins. In this study, we demonstrated that silver nanoparticles are efficient in neutralizing HIV-1 at non toxic concentrations. We also found an additive effect between the four NABs and AgNPs when combined against cell-associated HIV-1 infection in vitro. O3 New transcription regulatory mechanisms of latent HIV LTR Haleh Rafati1†, Yuri Moshkin1†, Tokameh Mahmoudi1*†, Maribel Parra2, Shweta Hakre3, Eric Verdin3 1 Department of Biochemistry, Erasmus University Medical Centre, Rotterdam, The Netherlands; 2Cancer Epigenetics and Biology Program (PEBC), IDIBELL, Barcelona, Spain; 3Gladstone Institute of Virology and Immunology, UCSF, San Francisco, CA, USA E-mail: [email protected] Retrovirology 2012, 9(Suppl 1):O3 Despite the effectiveness of antiretroviral medication, the HIV virus persists in resting memory T cells of infected patients in a latent state, Retrovirology 2012, Volume 9 Suppl 1 http://www.retrovirology.com/supplements/9/S1 providing the main impediment to eradication of the virus. We are interested in identifying the molecular mechanism responsible for the establishment and maintenance of HIV latency and its re-activation. We recently used a cell system reflecting HIV latency in my lab to determine the high resolution nucleosomal landscape of the latent HIV LTR and examine its dynamic changes upon re-activation (Rafati et al., Nov 2011 PLoS Biology). We combined mathematical predictions of nucleosome positioning with a combinatorial biochemical approach based on formaldehyde crosslinking of latent and activated HIV infected cells (using FAIRE, ChIP and high resolution MNase nucleosomal mapping) to define LTR nucleosome positioning and regulation during active and latent HIV infections. We found that BAF, an ATP-dependent chromatin remodelling complex generates a chromatin structure at the LTR that is energetically unfavorable to its intrinsic histone-DNA sequence preferences. Specifically, we find that BAF positions a repressive nucleosome immediately downstream of the HIV transcription start site, abrogating transcription, and in this way contributes to the establishment and maintenance of HIV latency. Our data describe a novel molecular mechanism for the establishment and maintenance of HIV latency, and we identify the catalytic subunit of BAF, the enzyme BRG1, as a putative molecular target to deplete the latent reservoir in infected patients. We will also present preliminary data addressing the role of a novel signalling pathway in derepression of latent HIV, and the effect of small molecules and ligands, which activate this pathway to study reactivation of latent HIV. We anticipate these experiments will further our understanding of HIV transcription regulation and identify both novel cofactors for targeting and molecules with potential to purge HIV latency. O4 Epigenetic modifications of HIV proviral LTRs: potential targets for cure W Doerfler1, S Weber1, K Kemal2, B Weiser2, K Korn1, K Anastos3, H Burger2* 1 Erlangen University, Institute for Virology, Erlangen, Germany; 2Wadsworth Center, New York State Department of Health, Albany, NY USA; 3Albert Einstein College of Medicine/Montefiore Medical Center, Bronx, NY USA E-mail: [email protected] Retrovirology 2012, 9(Suppl 1):O4 Introduction: HIV-1 cure remains elusive despite HAART due to the reservoirs of proviral DNA integrated into the human genome. Efforts to cure HIV-1 therefore need to aim at eliminating proviral DNA from cellular reservoirs. The first epigenetic signal identified in virus infected and uninfected cells has been promoter methylation. Compelling evidence confirms that specific promoter methylation can lead to gene silencing. Previous studies have examined HIV-1-epigenetics mostly in vitro. Materials and methods: We determined methylation patterns in HIV-1 proviral genomes from PBMCs obtained from 21 individuals with a spectrum of disease progression. The CpGs in the long terminal repeats (LTRs) of proviral DNA were investigated by bisulfite sequencing in up to 85 genomic variants per individual. This approach facilitates the study of the full range of CpG methylation and sequence variability of HIV-1 proviruses under conditions of natural selection in human populations. Results: In patients with advanced disease, the HIV-1 proviruses remained essentially unmethylated in their LTRs. In one long-term nonprogresssor, the percentage of methylated proviruses varied from 0-77% at different times after infection. More important and unexpected was the detection of three specific LTR-located CpG dinucleotides that had been selectively mutated to TpAs in >20 out of the 32 samples analyzed. Comparison to 11 HIV-1 LTR sequences in the Los Alamos HIV data base demonstrated that mutations in the sites identified by our study occurred more frequently than at other locations, although the mutations were different from TpAs. Conclusions: These specific CpGs, possibly including their abutting sequences, might indicate weak spots in the proviral genomes whose sacrifice by mutation to TpAs could enhance the HIV-1 potential for longterm proviral survival. These data suggest that the sites of the mutated CpGs occurring at conserved sites may serve as potential targets for therapeutic interventions to eliminate integrated proviruses. (Grants: DFG-DO165/28-1; NIH-UO1-AI35004) Page 8 of 54 O5 Hepatitis C virus fails to activate NF-kappaB signaling in plasmacytoid dendritic cells Ruzena Stranska*, Jonathan Florentin, Clélia Dental, Besma Aouar, Francoise Gondois-Rey, David Durantel, Thomas F Baumert, Jacques A Nunes, Daniel Olive, Ivan Hirsch Centre de Recherche en Cancérologie de Marseille, Marseille, France E-mail: [email protected] Retrovirology 2012, 9(Suppl 1):O5 Introduction: Plasmacytoid dendritic cells (pDCs) respond to viral infection by production of interferon a (IFN-a), proinflammatory cytokines and cell differentiation. The elimination of hepatitis C virus (HCV) in more than 50% of chronically infected patients by treatment with IFN-a suggests that pDCs can play an important role in the control of HCV infection. pDCs exposed to HCV-infected hepatoma cells, in contrast to cell-free HCV virions, produce large amounts of IFN-a. Materials and methods: To further investigate the molecular mechanism of HCV sensing, we studied whether exposure of pDCs to HCV-infected hepatoma cells activates in parallel to interferon regulatory factor 7 (IRF7)mediated production of IFN-a also nuclear factor kappa B (NF-B)dependent pDC responses such as expression of the differentiation markers CD40, CCR7, CD86, and tumor necrosis factor (TNF)-related apoptosis-inducing ligand (TRAIL), and secretion of the proinflammatory cytokines TNF-a and interleukin 6 (IL-6). RESULTS: We demonstrate that exposure of pDCs to HCV-infected hepatoma cells surprisingly did not induce phosphorylation of NF-B or cell surface expression of CD40, CCR7, CD86, and TRAIL, or secretion of TNF-a and IL-6. In contrast, CpG-A and CpG-B induced production of TNF-a and IL-6 in pDCs exposed to the HCV-infected hepatoma cells, showing that cell-associated virus did not actively inhibit toll-like receptor (TLR)mediated NF-B phosphorylation. Conclusions: Our results suggest that cell associated HCV signals in pDCs via endocytosis-dependent mechanism and IRF7 but not via NF-B pathway. In spite of IFN-a induction, cell-associated HCV does not induce a full functional response of pDCs. These findings contribute to the understanding of evasion of immune responses by HCV. O6 HCV full-length genome reconstruction with sequence independent amplification combined with next generation sequencing Barbara Bartolini*, Emanuela Giombini, Isabella Abbate, Ubaldo Visco-Comandini, Gianpiero D’offizi, Gabriella Rozera, Marina Selleri, Giuseppe Ippolito, Maria Rosaria Capobianchi Virology at National Institute for Infectious Diseases L. Spallanzani, Rome, Italy E-mail: [email protected] Retrovirology 2012, 9(Suppl 1):O6 Introduction: HCV genome variability is related to both disease progression and treatment response. De novo high-throughput pyrosequencing was used to obtain full length HCV genome characterization directly from clinical samples. Material and methods: Plasma samples from 3 HCV-infected subjects were analyzed (two patients with subtype 1b, one patient with subtype 2a/ 2c; viral load: 6.0 x 106,20.8 x 106 and 7.3 x 106 IU/ml viral load, respectively). All samples were analyzed in a single run, using sample-specific barcoding adapters. Data were generated with a modified sequence-independent single primer amplification followed by 454 sequencing (GS-FLX Roche, Titanium version), using the shotgun approach. The reads were assembled using cap3 program; HCV contigs were identified using BLAST against full HCV genome database. Reads of HCV contigs were used for genome reconstruction with gs Mapper (Roche software). Results: A total of 297,493 reads were obtained (average length 267 bp). Using a minimum read length cut off of 40 nt with >90% identity and >40% overlapping, BLAST analysis classified a total of 27,107 reads (10,682 from patient 1, 11,920 from patient 2, and 4,505 from patient 3) as HCVspecific. In all patients, genome reconstruction was achieved for more than 98 % of the entire HCV genome. The mean coverage was 315, 307 and 142 reads per site for patients 1, 2 and 3, respectively (overall mean coverage: Retrovirology 2012, Volume 9 Suppl 1 http://www.retrovirology.com/supplements/9/S1 253 reads per site). Within-patient variability was calculated, resulting in E1 and E2 as the most variable structural genes in all patients, as expected. Conclusion: The present study describes a unifying approach for HCV full genome sequencing, based on sequence-independent amplification combined with next generation sequencing. This may represent a relevant innovation, since so-far HCV full genome direct sequencing was based on genotype-specific multiple primer approach and conventional sequencing. The possibility of performing simultaneous analysis of pooled samples may represent a further advantage for cost saving.High coverage allows to analyze virus variability along the entire genome providing important information on possible viral variants which could impact on clinical and therapeutic outcome. O7 Raltegravir switch improves hepatitis C transaminitis in HIV-1 and hepatitis C (HCV) co-infected individuals Muge Cevik*†, Gurmit Singh†, Laura Dickinson, Andrew Scourfield, Marta Boffito, Mark Nelson Chelsea and Westminster Hospital, London, UK E-mail: [email protected] Retrovirology 2012, 9(Suppl 1):O7 Introduction: HCV is one of the most relevant co-morbidities seen in HIVinfected individuals as evidenced by the negative impact that HIV exerts on the course of HCV infection. Despite remarkable results on HIV infection alone, the impact of highly active antiretroviral therapy (HAART) on liver disease in co-infection remains unknown. We sought to explore the impact of Raltegravir (RAL) on amino transferase (ALT) in HIV/HCV co-infected individuals. Methods: HIV-infected individuals co-infected with HCV within the last 5 years receiving non-integrase inhibitor containing HAART with a subsequent switch to RAL-containing HAART were identified from a retrospectively maintained outpatient database. Patient demographics were extracted. Biochemical, virological and immunological parameters were collated and individuals received pegylated interferon with ribavirin were excluded. ALT levels at switch and post switch were compared using Kruskal-Wallis test. Spearman’s Rank correlation was used to assess the relationship between ALT and HCV-RNA. Results: Twenty seven HIV-HCV co-infected individuals were identified between January 2007 and January 2012 and seven individuals were excluded. Median age was 44 years (range: 31-68). Five had acute and fifteen had chronic HCV infection during the switch. Twenty (100%) had HIV-RNA-1. Conclusion: In our study, RAL had a favourable effect on the liver up to 24 weeks after switch in HIV/HCV infected individuals. O8 Insect cell endocytosis of chikungunya virus adapted to Aedes albopictus, a mosquito recently introduced into southern France Christian Devaux*, Eric Bernard, Bernard Gay, Nathalie Chazal, Laurence Briant Infectious Diseases at Cpbs (Centre d’Études d’Agents Pathogènes et Bio. Santé), Umr5236 Cnrs, Um1, Um2, Montpellier, France E-mail: [email protected] Retrovirology 2012, 9(Suppl 1):O8 Introduction: Since the first isolation of chikungunya virus (CHIKV) more than 50 years ago in Eastern and Central Africa, CHIKV epidemics have been repeatedly recorded from various countries. The 2005-2006 outbreak in Reunion Island, was characterized by a genome microevolution in the E1 envelope glycoprotein gene (E1-A226V mutation) that enhances CHIKV fitness for Aedes Albopictus vector. More recently, CHIKV caused explosive outbreaks in India and propagated to temperate areas in Southern Europe, including France in 2010. Along with Aedes albopictus colonization of new geographical areas and climate change facilitating vector proliferation, the epidemic risk for “tropical infectious diseases” represents a real threat for naïve populations. It is therefore important to better understand the replication cycle of CHIKV in cells from Aedes albopictus. Materials and methods: CHIKV strains (the African reference strain of CHIKV 37997; the LR-OPY1 (E1-226V) variant isolated from Reunion Island and the LR-OPY1V226A bearing the reverse E1-V226A mutation were Page 9 of 54 tested for replication in the C6/36 Ae. albopictus cell line. Experiments were performed to assess the role of clathrin and dynamin-dependent endocytic pathways implication of endosomal pH acidification and requirement for membrane cholesterol in CHIKV infection of mosquito cells. Result and conclusions: Our data indicate that CHIKV uses a clathrindependent, caveolae-independent pathway to infect Aedes albopictus cell cultures and requires membrane cholesterol as well as a low-pH environment for entry. These features, especially membrane cholesterol requirement, are modulated in some extent by the E1-A226V mutation. Altogether, our data provide the first information regarding the pathways used by CHIKV to infect Aedes albopictus cells and points the consequences of recent genome microevolution on these entry routes. O9 The adequacy of encephalitis surveillance for emerging infectious diseases in Australia David N Durrheim1*, Clare Huppatz1, Beverley Paterson2 1 University of Newcastle, Wallsend, Australia; 2Hunter Medical Research Institute, University of Newcastle, Wallsend, Australia E-mail: [email protected] Retrovirology 2012, 9(Suppl 1):O9 Introduction: The acute encephalitis syndrome has heralded the emergence of multiple virulent pathogens in Australia, including Murray Valley encephalitis, and recently Hendra virus and Australian bat lyssavirus. In Australia, the encephalitis syndrome in humans is not notifiable. The burden of Australian encephalitis hospitalisations and deaths was not known; encephalitis aetiology and trends had not been investigated at population level; and the adequacy of neurological review in hospitalised encephalitis cases to exclude emerging pathogens had not been investigated. Materials and methods: A series of studies was conducted to better understand the status of encephalitis surveillance in Australia. Firstly, Australian Bureau of Statistics mortality and population data for the period 1979–2006 were obtained and cause of death data were extracted using ICD-9 (1979–1998) and ICD-10 (1999–2006). Secondly, rates of hospitalisation for patients with encephalitis in Australia’s most populous state, New South Wales (NSW), from January 1990 through to December 2007 were reviewed, with encephalitis-associated hospital stays extracted using ICD-9-CM (1990– 1998) and ICD-10-AM (1999–2007) from the NSW Department of Health Inpatient Statistics Collection library. Finally, a retrospective clinical audit was performed, of all adult encephalitis admissions between July 1998 and December 2007 to the three hospitals with adult neurological services in northern NSW. Case notes were examined for evidence of relevant history taking, clinical features, physical examination, laboratory and neuroradiology investigations, and outcomes. Results: Between 1979 and 2006 there were 1,118 encephalitis-associated deaths in Australia, with an average annual death rate of 2.3 per million population. The aetiology of 576 deaths was unknown and the proportion of deaths due to ‘unknown’ encephalitis increased from 47.0% between 1979 and 1992, to 57.2% from 1993 to 2006. Encephalitis was the primary discharge diagnosis for 5,926 hospital admissions in NSW with an average annual hospitalisation rate of 5.2/100,000 population. Toxoplasma encephalitis and subacute sclerosing panencephalitis showed notable declines. The proportion of patients hospitalised with encephalitis and no identified pathogen (69.8%, range 61.5%–78.7%) was stable during the study period. Amongst patients admitted with an encephalitis diagnosis in the NSW regional neurological hospital network, treating clinicians suspected a specific causative organism in 18.9% of cases and a cause was confirmed by laboratory testing in 12.1% of cases. However, only 14.9% were tested for flaviviruses and tests for specific locally occurring zoonotic encephalitis viruses were only conducted in 0.0-6.8% of cases. Conclusions: The non-notifiable status of human encephalitis in Australia and the high proportion of cases, including deaths, with no known aetiology may conceal emergence of novel pathogens. Unexplained encephalitis should be investigated, and encephalitis hospitalisations should be subject to statutory notification to facilitate prompt public health investigation and action as necessary. The utility of hospital sentinel surveillance, and standardised diagnostic and testing algorithm is currently being explored. Retrovirology 2012, Volume 9 Suppl 1 http://www.retrovirology.com/supplements/9/S1 O10 Dengue virus-pandemic influenza virus co-infection results in enhanced influenza virus replication through inhibition of apoptosis Thomas G Voss*, Mei-Chun Chen, Gena J Nichols, Somanna K Naveen, Benjamin T Bradley, Robert W Cross Tulane University, School of Medicine, New Orleans, USA E-mail: [email protected] Retrovirology 2012, 9(Suppl 1):O10 Introduction: Dengue fever and Dengue Hemorrhagic fever in humans are the result of infection with Dengue virus, a mosquito-borne member of the Flaviviridae. In 2009, the appearance of a novel, swine origin, pandemic (H1N1) influenza A virus in humans resulted in identification of patients co-infected with dengue and influenza with enhanced clinical disease. To elucidate potential mechanism(s) of enhanced pathogenesis observed during human Dengue/Influenza co-infection, we examined the effects of co-infection in cells (A549) and also in animals. Materials and methods: A549 cells were infected with Dengue virus or pandemic influenza A virus. Virus replication was measured by plaque assay and immune fluorescence. Apoptosis was measured by multiple assays including TUNEL, Annexin V expression, and Casapse 3 expression. Ferrets were challenged with Dengue for 24 hours prior to influenza challenge. Clinical disease was monitored for 10 days post Dengue challenge. Nasal aspirates were collected and tissues harvested for virology, immunology, and histology studies through the 10-day in life period. Clinical chemistry and hematology were also measured on infected ferrets. Results: In A549, co-infection enhances influenza virus replication and reduces dengue virus replication compared to singly infected A549. Influenza-specific inhibition of dengue replication was dependant on multiplicity of infection (moi) and timing of influenza infection with dengue inhibition detected when influenza infection occurs up to 48 hours post Dengue infection. Co-infected cell apoptosis was reduced suggesting a mechanism for increased influenza virus loads. In ferrets co-infected with dengue and pandemic influenza, influenza virus loads and clinical disease signs compared to influenza infection alone were observed. In the lungs of co-infected ferrets, apoptosis was reduced, confirming in vitro results. Our results indicate a potential pathogenic interaction between dengue and influenza viruses that models human co-infection cases. O11 Potent suppression of HIV viral replication by a novel inhibitor of Tat Guillaume Mousseau1, Mark A Clementz1, Wendy N Bakeman1, Nisha Nagarsheth1, Michael Cameron2, Jun Shi3, Phil Baran3, Rémi Fromentin4, Nicolas Chomont4, Susana T Valente1* 1 Department of Infectology, The Scripps Research Institute, Jupiter, Florida, USA; 2Department of Molecular Therapeutics and Translational Research Institute, The Scripps Research Institute, Jupiter, Florida, USA; 3Department of Chemistry, The Scripps Research Institute, La Jolla, California, USA; 4Vaccine and Gene Therapy Institute, Port St. Lucie, Florida, USA E-mail: [email protected] Retrovirology 2012, 9(Suppl 1):O11 Background: Tat, the HIV Trans-Activator of Transcription is a potential antiviral target. Tat binds to the 5’ terminal region of HIV mRNAs stembulge-loop structure called the Trans-activation Responsive (TAR) element and activates transcription from the HIV promoter. Plasma viremia stubbornly persists in HIV-1 infected subjects despite receiving HAART, suggesting that residual levels of viral production originate from an integrated form of the HIV genome that is continuously transcribed at low levels. As current antiretrovirals (ARVs) fail to inhibit transcription from integrated viral genomes or viral production from stable cellular reservoirs, novel classes of ARVs are needed to inhibit this process. Results: Cortistatin A is a steroidal alkaloid isolated from the marine sponge Corticium simplex. Here, we show that its analog didehydroCortistatin A (dCA) inhibits Tat-mediated trans-activation of the integrated HIV provirus by binding specifically to the TAR-binding domain of Tat. dCA reduces cell-associated viral RNA and capsid p24 antigen production in acutely and chronically infected cultured and primary cells, at a half maximal effective concentration (EC50) of 0.7 pM to 2.5 nM, depending on the multiplicity of infection (MOI). dCA reduces both transcriptional initiation/elongation from the viral promoter and alters the nucleolar Page 10 of 54 localization of Tat. Termination of dCA treatment does not result in immediate virus rebound as the HIV promoter is transcriptionally silenced. dCA inhibits both HIV-1 and HIV-2, and displays high bioavailability. dCA added to a combination of ARVs mediates a statistically significant reduction in viral replication from primary CD4 + T cells isolated from viremic patients compared to the ARVs alone, and abrogates low-level virus replication from CD4 + T cells isolated from aviremic patients undergoing HAART treatment. Conclusions: With a therapeutic index of over 8000, dCA defines a novel class of HIV anti-viral drugs endowed with the ability to decrease residual viremia during HAART, and should be considered as a promising drug to be included in therapeutic eradication strategies. POSTER PRESENTATIONS P1 In vitro study of the trans-epithelial crossing of HIV-1 through the female genital mucosa and of the role of epithelial cells in the selection of CCR5-tropic virus R Terrasse*, O Delezay, A Brunon-Gagneux, L Heyndrickx, H Hamzeh-Cognasse, B Pozzetto, T Bourlet Gimap Ea3064, Saint-Étienne, France E-mail: [email protected] Retrovirology 2012, 9(Suppl 1):P1 Object: HIV heterosexual transmission mainly occurs by exposition of female genital mucosa with infected male seminal secretions. The notion of compartmentalization of HIV in the male genital tract is well established by the demonstration of the existence of several viral populations between blood and semen, especially in the gp160 viral envelope. The objectives of the present work were (i) the study of the role of genital epithelial cells in the heterosexual transmission of HIV and (ii) the in vitro modeling of this crossing by using chimeric viruses (pBrNL4.3-eGFP/dsRedExpress) that express the gp160 glycoprotein isolates from patients’semen. Methods: Seminal samples that showed a positive viral load were selected from 50 semen specimens obtained from 39 HIV-infected patients followed at the University-Hospital of Saint-Etienne. Chimeric viruses were constructed by cloning a pBrNL4.3 vector with a viral envelope containing a GFP or a dsRedExpress fluorochrome. HEC genital epithelial cells were infected by CXCR4-tropic (LAI) or CCR5-tropic (BaL) HIV-1 viruses (0.001 MOI per cell during 24h) in different conditions (presence or absence of AZT or proinflammatory cytokines). Results: Two days after infection, intracellular viral RNA was detected for both virus strains (LAI and BaL) either in the presence or in the absence of AZT. By contrast, the detection of proviral DNA (LTR) was observed only for the CXCR4-tropic variant (500 ± 100 copies / 106 cells); the addition of AZT abolished this infection. Neither intracellular viral proteins nor extracellular viral RNA could be detected in infected cells. Conclusion: These results confirm the selective sequestration of CXCR4tropic viruses by epithelial cells. Complementary analyses are ongoing for determining the exact role of these cells in HIV-1 heterosexual transmission and as a viral latency reservoir that could be reactivated under inflammatory conditions. A confocal microscopy imaging using the chimeric viruses will complete these preliminary results. P2 MiRNA profile in CD4 positive T cells from HTLV-2 and HIV-1 monoand co-infected subjects E Pilotti1*†, C Casoli1†, MV Bianchi1, F Bignami1, Francesca Prati2 1 GEMIB Laboratory, Parma, Italy; 2Hospital Santa Maria Nuova, Reggio Emilia, Italy E-mail: [email protected] Retrovirology 2012, 9(Suppl 1):P2 Introduction: The HTLV-2/HIV-1 co-infection has been shown to be associated with a delayed progression to AIDS (Turci et al. 2007). Casoli et al. (2007) have demonstrated the key role played by CCL3L1 chemokine, whose expression is induced by HTLV-2 infection, in slowing down HIV-1 disease. MiRNAs (miRNAs) are small non-coding RNAs that regulate fundamental cellular processes. Since HTLV-2 creates a cellular environment favourable to itself and adverse to HIV-1, it is supposable that host miRNAs profile can be modulated by HTLV-2 to this aim. Here, we investigated the expression profile of miRNAs in HTLV-2 and HIV-1 Retrovirology 2012, Volume 9 Suppl 1 http://www.retrovirology.com/supplements/9/S1 mono-infected, in double infected, and HIV-1 exposed uninfected (MEU) subjects. Materials and methods: CD4+ T cells were purified from blood samples of 7 LTNP HTLV-2/HIV-1 co-infected, 5 HIV-1MEU, 7 HTLV-2/HIV-1MEU, 10 viremic and 7 LTNP HIV-1 mono-infected individuals and 10 healthy donors as controls. The expression profile of 377 miRNAs was obtained by real-time quantitative PCR and 2-ΔΔCt method, by which the PCR signal of miRNAs transcript in CD4+ T-lymphocytes from infected subjects were related to that of healthy donors. By real time PCR, each cohort of subjects was also tested for the expression of miRNA processing enzymes, Dicer and Drosha. Results: Analyzing these miRNA identified by the comparison of the two viral infection, 4 miRNAs (329, 337-5p, 379, and 503) were down-regulated and 6 miRNAs (34a, 125a-3p, 155, 203, 449a, and 502-5p) were up-regulated in both conditions, suggesting a retroviral exposure signature. HTLV-2/HIV1MEU subjects are characterized by a miRNA profile similar to that of healthy donors, while a strong up-regulation of miRNA profile marked subjects infected by o exposed to HIV-1. Dicer and Drosha expressions seem to explain the miRNAs changes observed. Conclusions: These findings enable us to better understand the potential role of miRNAs in the development of resistance to HIV-1 infection by HTLV-2. P3 Production and expression of recombinant anti-V3 scFvs from HIV-1 clade C infected Indian patient Rajesh Kumar*†, Raiees Andrabi†, Ashutosh Tiwari, Somi Sankaran Prakash, Naveet Wig, Durgashree Dutta, Anurag Sankhyan, Lubina Khan, Subrata Sinha, Kalpana Luthra All India Institute of Medical Sciences, New Delhi, India E-mail: [email protected] Retrovirology 2012, 9(Suppl 1):P3 Introduction: Neutralizing antibodies are an important component of the humoral immune response directed against viral infections So far the few available anti HIV-1 broadly neutralizing antibodies have a limited breadth and potency against clade C viruses. More than 50% of the HIV-1 infections worldwide belong to clade C. Clade C is the most prevalent subtype in India. Materials and methods: A human scFv phage display library was constructed from the peripheral blood mononuclear cells of an HIV-1 Clade C infected Indian patient, having a good titre of serum neutralizing antibodies. Diversity of the scFv library was checked by sequencing and DNAfingerprinting analysis of randomly selected clones from the preselected library. One round of biopanning was done against V3 peptide of clade C and clade B. Single chain fragments were checked for their soluble expression in E. coli HB2151 and purfied using Ni+2 affinity colums. Specificity of the soluble scFvs was checked by indirect ELISA. Single chain fragments were checked for their stability in different agents like 30%DMSO. 4M NaCl, pH 2-11. Results: 50 clones were randomly selected after biopanning and they were checked for their binding to V3C and V3B peptides. In phage ELISA 15 out of 50 clones showed binding to both the V3 peptides. A 32kDa band was observed in polyacrylamide gel electrophoresis as predicted. The expressed product was confirmed by Western blot analysis using anti His Tag antibody. Specificity of the purified scFvs was confirmed by their binding to V3 peptides and no reactivity against other unrelated peptides. Further these scFvs displayed a stable binding to V3 peptides in different denaturing agents. Conclusions: This is the first study to generate human anti-V3 scFvs against HIV-1 clade C. Further characterization of these scFvs for their neutralization potential will help identify unique and shared epitopes responsible for neutralization of clade C and non clade C viruses. P4 Genetic variability and high proportion of HIV-1 BF1 recombinant strains among vertically infected children in São Paulo, Brazil Ana Carolina Soares de Oliveira*, Antonio Charlys da Costa, Vanessa Pouza Martinez, Maria Teresa Maidana, Giret Regina Celia de Menezes Succi, Ester Cerdeira Sabino Esper, Georges Kallas, Sabri Saeed Sanabani São Paulo Institute of Tropical Medicine, São Paulo, Brazil E-mail: [email protected] Retrovirology 2012, 9(Suppl 1):P4 Introduction: The enormous genetic variability of human immunodeficiency virus type 1 (HIV-1) continues to present a major challenge for vaccine Page 11 of 54 design and frustrate efforts to halt the epidemic. A proper understanding of this phenomenon is a prerequisite for proper epidemiology, genetic diagnosis, and successful drugs and vaccines. In this study, we undertook a detailed molecular epidemiological investigation on HIV-1 vertically-infected children born from 1993 to 2008 in the state of São Paulo, Brazil. Material and methods: HIV-1 proviral DNA was extracted from the peripheral blood mononuclear cells of 48 participants. The near full-length genomic (NFLG) and partial fragments were determined by overlapping nested PCR and direct sequencing. The data were phylogenetically analyzed. Results: Of the 48 samples (median age 11.8 years, range 4-20.6 years) studied, 3 (6.2%) NFLGs and 39 (81.2%) partial fragments were successfully subtyped. Of the successfully subtyped sequences, 20 (47.6%) were subtype B sequences, 17 (40.4%) BF1 recombinants, and 5 (11.9%) subclade F1. Two of the partial BF1 chimeric isolates shared an identical recombination structure. Predictions of viral tropism using the computer program geno2pheno [co receptor] for phenotype prediction were determined for 15 subjects. X4 or X4 dual or mixed-tropic viruses were seen in 3 (20%) of participants and the V3 sequences of 12 patient virus strains (80%) were predicted to be R5-tropic virus. Conclusions: Our data provided evidence of unexpectedly high proportion of BF1 recombinants viruses transmitted from the first mother-to-child since the earliest days of the epidemic to the present time in Brazil. These findings offers additional insights to understanding the diversity of HIV-1 strains currently circulating in Brazil, with future implications for diagnosis, therapy, and efficient vaccine development. P5 Next generation of antiretroviral agents targeting the RNA binding site of the HIV-1 cellular cofactor DDX3: an innovative therapeutic approach Giovanni Maga1*, Anna Garbelli1, Marco Radi2, Federico Falchi2, Alberta Samuele1, Stefania Paolucci3, Fausto Baldanti3, Fabrizio Manetti2, Sandra Beermann4, Ursula Dietrich4, Maurizio Botta2 1 Instituto di Genetica Molecolare, IGM-CNR Via Abbiategrasso 207, I-27100 Pavia, Italy; 2Dipartimento Farmaco Chimico Tecnologico, University of Siena,Via Alcide de Gasperi 2, I-53100 Siena, Italy; 3Molecular Virology Unit, Foundation IRCCS Policlinico S. Matteo, piazzale Golgi, I-27100 Pavia, Italy; 4Georg-Speyer-Haus Institute of Biomedical Research, Paul-Ehrlich-Str. 42-44, 60596 Frankfurt, Germany E-mail: [email protected] Retrovirology 2012, 9(Suppl 1):P5 Introduction: Efficacy of currently approved anti-HIV drugs is hampered by mutations of the viral enzymes, leading invariably to drug resistance and chemotherapy failure. Recent data suggest that cellular co-factors also represent useful targets for anti-HIV therapy. We have recently provided evidence for the possibility to block HIV-1 replication by targeting its cellular cofactor DDX3. Material and methods: Molecular modeling and in silico technologies were applied to rationally design small molecules specifically targeting the RNA binding site of human DDX3. Biochemical studies of mutated DDX3 enzymes were also used to identify additional potential drug binding sites. Results: Optimization of compounds identified by application of a highthroughput docking approach afforded a promising lead compound which proved to inhibit both the helicase and ATPase activity of DDX3 and to reduce the viral load of peripheral blood mononuclear cells (PBMC) infected with HIV-1. A novel interaction site has been also identified in DDX3, which, when blocked, can reduce viral replication, representing an additional target for small molecules inhibitors. Conclusions: We have identified the first inhibitors of HIV-1 replication targeting the RNA binding site of the cellular cofactor human DDX3. These compounds may offer superior selectivity over the ATP-competitive inhibitors previously developed. In addition, a novel RNA interacting motif specific to DDX3 has been identified, opening new venues for HIV-1 drug development. P6 Monoclonal antibodies that recognize important functional elements of the HIV-1 integrase enzyme Richard G Maroun1*, Farah Ammar1,2, Zeina Hobaika1, Loussinée Zargarian2, Serge Fermandjian2 1 Unité de Biochimie, Faculté des Sciences, Université Saint-Joseph, CST-Mar Roukoz, Beirut, Lebanon; 2ENS de Cachan, CNRS, Cachan, France E-mail: [email protected] Retrovirology 2012, 9(Suppl 1):P6 Retrovirology 2012, Volume 9 Suppl 1 http://www.retrovirology.com/supplements/9/S1 Aim: HIV integrase (IN) is a privileged target for antiviral treatments. These induce the emergence of resistant strains, prompting the search of new drugs. To better understand the relationships between structure and function of IN and identify new anti-HIV inhibitors we prepared antibodies recognizing the IN a4 helix that binds viral DNA ends and contributes to the integration process and antibodies recognizing a loop in between the a4 and a5 helices which participates to the binding of LEDGF a protein that helps IN to anchor viral DNA. Materials and methods: Polypeptide K159 (sequence 147-175 of IN) was injected to mice. Several hybridomas producing monoclonal antibodies (Mabs) were obtained Mabs were characterized by ELISA and blotting techniques using peptide fragments, IN and viral DNA sequences. Results: We prepared two Mabs (Mab-a4 and Mab-loop) exhibiting high affinities against the antigenic peptide K159 and IN. An epitope mapping showed that Mab-a4 interacted with N-terminal segment (147-163) and Mab-loop with the C-terminal (164-175). Mab-a4 blocked the interaction of IN with viral DNA end, while the loop segment 164-175 recognized by the Mab-loop constitutes a strong epitope also found in African seropositive patients. Spectroscopic studies of the antibody-antigen complexes are under progress. Crystallization of the Fab moiety of Maba4 has been recently obtained. Conclusions: We showed that the important immunogenic properties demonstrated by the a4 helix and the loop 164-175 coincided with their important functional properties in IN. We wish to collect details on the interactions and the energies stabilizing these complexes and compare them with those stabilizing the complexes with their biological targets (DNA, LEDGF). Finally at a medical level, these Mab could be used as valuable tools for HIV diagnostics in ELISA or western blot assays. P7 The orally bioavailable allosteric CXCR4 HIV-1 entry inhibitor AMD11070 Simon Fricker1, Renee Mosi2, Virginia Anastassova2, Jean Labrecque2, Rebecca Wong2, Renato Skerlj1, Gary Bridger1, Dana Huskens3, Dominique Schols3* 1 Genzyme Corporation, Framingham, MA, USA; 2Formally of AnorMED Inc., Langley, BC, Canada; 3Rega Institute for Medical Research, University of Leuven, Leuven, Belgium E-mail: [email protected] Retrovirology 2012, 9(Suppl 1):P7 In order to enter and infect human cells HIV must bind to the CD4 receptor in addition to either CXCR4 or CCR5. AMD11070 was the first orally available small molecule inhibitor of CXCR4 to enter the clinic. Here, we report in detail the molecular pharmacology of AMD11070 which is a potent inhibitor of X4 HIV-1 replication in various CD4 + T cell lines, CXCR4-transfected cell lines and in PBMC (IC50 values of 14 ± 3 nM). In addition, AMD11070 potently inhibited cell fusion between a CHO-K1 cell line expressing viral gp120 and the P4-R5 MAGI cells which express CD4 and CXCR4 with an IC50 of 1.5 ± 0.3 nM. No antiviral activity was observed with AMD11070 against CCR5-using (R5) HIV-1 replication. Using CD4 + T cell lines that endogenously express CXCR4 we demonstrate that AMD11070 is an antagonist of CXCL-12 (SDF-1a)-ligand binding (IC50: 12.5 ± 1.3 nM), inhibits CXCL-12mediated signaling (IC50: 9 ± 2 nM) and that it inhibits CXCL-12mediated chemotaxis (IC50: 19 ± 4 nM). AMD11070 does not inhibit chemokine-induced Ca2 + -signaling in cells expressing CXCR3, CCR1, CCR2b, CCR4, CCR5 or CCR7, demonstrating the compound selectivity for the CXCR4 receptor. In addition, AMD11070 is able to inhibit the SDF-1beta isoform interactions with CXCR4 and N-terminal truncated variants of CXCR4 with equal potency as to the wild type CXCR4 receptor. These data indicate that AMD11070 is an allosteric antagonist of CXCR4. A proof-of-concept clinical trial has shown that AMD11070 can reduce the viral load of X4 HIV-1 in HIV-1-infected persons. Together these data further support to the potential beneficial role of orally bioavailable CXCR4 inhibitors as a therapeutic option for HIV/ AIDS treatment. Page 12 of 54 P8 Human beta-defensins induce APOBEC3G expression by interacting with chemokine receptors, protecting highly susceptible cells from HIV infection Alfredo Garzino-Demo*, Mark K Lafferty, Jennifer Bharucha, Lingling Sun, Walter Royal, Suzanne Gartner, Wuyuan Lu Institute of Human Virology, U. Maryland School of Medicine, Baltimore, USA E-mail: [email protected] Retrovirology 2012, 9(Suppl 1):P8 ß-defensins are antimicrobial peptides secreted by epithelial cells that can bind to cellular receptors. CCR2 and CCR6 are the two cellular receptors known to bind human ß-defensin (hBD) 2 and -3. Both of these receptors are of crucial relevance in HIV infection. CCR6 is expressed, often in concert with CCR5, on cells that are highly susceptible to HIV infection: memory T cells, Th17 cells, a4ß7+ cells, and defects in CD4+CCR6+ cells have been associated with faster AIDS progression. CCR2 is expressed on monocytes and macrophages, cells that are reservoirs of HIV infection and that are known to mediate central nervous system damage. Our studies show that hBD2, hBD3, and CCR6 ligand MIP-3a/CCL20 inhibit HIV infection via CCR6 by increasing expression of the antiviral protein APOBEC3G. This increase is due to a transcriptional mechanism mediated by intracellular signaling. hBD2 also inhibits HIV replication in macrophages that express CCR2. Our findings suggest novel therapeutic and preventive approaches that exploit CCR6 and CCR2-mediated intracellular signaling to inhibit HIV infection in highly susceptible cells. P9 Three-dimensional modeling of DCIR and identification of new drugs blocking HIV-1 attachment and propagation Caroline Gilbert*, Arezki Azzi, Alexandra A Lambert, Sheng-Xiang Lin, Geneviève Allaire, Karianne P St-Gelais, Michel J Tremblay Laval University, Québec, Canada E-mail: [email protected] Retrovirology 2012, 9(Suppl 1):P9 Introduction: The HIV-1 pandemic continues to expand while no effective vaccine is yet available. Finding new therapeutic targets and drugs is therefore crucial. We have previously shown that the dendritic cell immunoreceptor (DCIR), a C-type lectin receptor expressed in dendritic cells (DCs), acts as an attachment factor for HIV-1 to DCs and contributes to HIV-1 transmission to CD4+ T lymphocytes (CD4TL). Directly involved in HIV-1 infection, DCIR is expressed in apoptotic or infected CD4TL and promotes trans-infection to bystander cells. The aim of the present study is to characterize the extracellular domain of DCIR and to test chemical inhibitors of HIV-1 attachment thereto. Results: We present the first three-dimensional model of DCIR structure. Based on this structure, several inhibitors were selected to target viral interaction with the carbohydrate recognition domain and the EPS motif. Preliminary screening using Raji-CD4-DCIR cells identified two inhibitors that decreased HIV-1 attachment and propagation. These inhibitors did not affect the proliferation of peripheral blood mononuclear cells. Conclusions: The results of this study thus suggest structures for novel molecules capable of blocking HIV-1 transmission by DCs and CD4TL. P10 Nevirapine-associated liver toxicity and hypersensitivity reactions in a cohort of HIV-1-infected patients,clinical analysis Sylvie Jonckheere*†, JC Yombi†, Leila Belkhir, Anne Vincent, Bernard Vandercam Medecine Interne Infectiologie at Centre Refrence St Luc Ucl, Bruxelles, Belgium Retrovirology 2012, 9(Suppl 1):P10 Introduction: Antiretroviral drug-related liver injury is a common cause of morbidity and treatment discontinuation in HIV-infected patients. Nevirapine is incriminated as one of the liver toxicity inducer especially in Retrovirology 2012, Volume 9 Suppl 1 http://www.retrovirology.com/supplements/9/S1 patients with high CD4-cells count. The purpose of our study was to analyze the role of CD4 cell count at treatment initiation and that of several co-factors (Hepatitis C or Hepatitis B virus co-infection, concurrent use of protease inhibitors) on the incidence of liver toxicity and hypersensivity reactions induced by Nevirapine in our HIV1-infected patients. Material and method: We analyzed retrospectively a cohort of 930 HIV-1 infected patients. Patients, who were started on NVP between 1998 and 2003, regardless of CD4-cells count, were included. We assigned patients to two groups: (A) group with high CD4-cells counts (women with CD4 cells ≥ 250 cells/mm³ and men with ≥ 400 cells/mm³), and (B) group with low CD4-cell counts. Liver toxicity is considered severe when at least a grade 3 toxicity is observed (WHO classification). Results: In total 108 patients were included. Eight (7.40%) and 15 (13.9%) patients interrupted treatment because of severe liver toxicity and hypersensitivity reactions respectively. There was no overlap between these two groups, and hypersensitivity reactions tended to occur sooner (22 vs 45 days respectively). HCV and HBV co-infection rates were 7.4% and 8.33%. Severe liver toxicity was seen in 15.74% of patients. Comparing group A and B, rates of severe liver toxicity were 15.68% and 17.30% respectively. There was no significant difference. In a multiple linear regression model, we found viral hepatitis C co-infection to be the only independent risk factor in the occurrence of liver toxicity (p<0.006). Conclusion: In our study the rate of severe liver toxicity due to NVP was high. HCV co-infection was an independent risk factor for liver toxicity, contrary to CD4 cell counts at treatment initiation. These findings are in keeping with recent published data. A careful analysis of the literature shows that hypersensitivity reactions due to NVP are strongly correlated with high CD4 cell counts. The limitation of our study is the low number of patients included. P11 Immunological response after initiation of second line anti-retroviral therapy in HIV patients Basavaprabhu Achappa*†, Keerthi Pillai†, John T Ramapuram, Satish B Rao, Deepak R Madi, Unnikrishnan Kasturba Medical College, Manipal University, Mangalore, India E-mail: [email protected] Retrovirology 2012, 9(Suppl 1):P11 Introduction: Treatment with second line ART is initiated when the first line therapy fails. There is less experience with the immunologic response for second-line ART for adults. Hence this study was done to find out the immunological response after initiation of second line ART by analysis of CD4 counts. Methods: This retrospective study was conducted in a tertiary level hospital attached to a medical college that caters to a large number of HIV positive patients. The study population for this analysis included all HIV positive individuals who were undergoing second line ART treatment. The data was collected using semi-structured pre-tested proforma from the hospital records of HIV positive individuals. The immunological response after the initiation of second line ART was analyzed using the CD4 cell counts taken at intervals of 3 months and 6 months after the initiation. Results: Out of the 32 patients studied, 27(84.4%) were males and only 5 (15.6%) were females. Mean age of the patients was 40.56±6.78 years. The mean CD4 value at initiation was 152.35±142.89 cells/μL, which significantly increased to 324.43 ±163.65 cells /μLby 3 months after initiation (p value= .000) and to 348.21± 253.57 cells /μLby 6 months after initiation.Around 91.3% of patients had a baseline CD4 T cell count <350 cells L-6. After 3 months of therapy, 65.2% of patients and after 6 months 46.2% had a baseline CD4 T cell count <350 cells /μL.The mean weight at initiation was 50.548± 11.37 kg, which very significantly increased to 53.30± 11.1 kg by 3 months of therapy (p = .001, table 1) and to 54.63 ± 10.29kg at the end of 6 months. Conclusion: The CD4 counts increase very significantly within the first 3 months of initiation of second line therapy.The rise in CD4 count between 3 months and 6 months is not as statistically significant as the earlier one. Also, there is significant gain in weight within 6 months of initiation of second line therapy. Page 13 of 54 P12 Immunological effect of ten-year c-ART in treatment-naive and pretreated HIV-1 patients in Bulgaria Ivaylo Elenkov1†, Maria Nikolova2*†, Ivanka Radeva1, Margarita Yankova1, Nina Yancheva1 1 Specialized Hospital for Active Treatment of Infectious and Parasitic Diseases, Sofia, Bulgaria; 2Clinical Immunology at National Center of Infectious and Parasitic Diseases, Sofia, Bulgaria E-mail: [email protected] Retrovirology 2012, 9(Suppl 1):P12 Introduction: Highly effective combination anti-retroviral therapy (c-ART) has been applied in Bulgaria since 1999. The aim of the present study was to compare retrospectively the long-term immunological effect of cART in treatment-naïve and pre-treated HIV-1+ patients. Patients and methods: The study included HIV-1+ patients (n=56) that have started c-ART between March 1999 and December 2001, have been on continuous treatment, with good adherence, death being the only reason for ART stop. Of them, 27 had a history of irregular pre-treatment with AZT or AZT/LMV for an average of 4.6 yrs (Group A), and 29 were ART-naïve (Group B). CD4 absolute counts (AC) were determined by single-platform flow cytometry (BD Biosciences). Viral load (VL) was measured by RT-PCR (Roche). Comparisons were performed by unpaired t-test (SPSS 17.0). Results: The demographic characteristics of groups A and B did not differ significantly: mean age (yrs): 34 vs. 35; male to female ratio: 9 vs. 7, respectively. Baseline CD4 AC (cells/ml) and VL (log HIV RNA copies/ml) were comparable: mean 124 vs. 119, and 5.1 vs. 4.6, respectively, (p>0.05 for both comparisons). In the long term, suppression of viral replication was observed in both groups: mean VL at 5 yrs 3.7 vs. 3.1 for groups A and B, respectively, (p>0.05). However, treatment-naïve patients (group B) had a better immune recovery than group A, and the difference became significant in the long term: mean CD4 AC 177 vs. 252 after 6 months of c-ART, (p>0.05), 391 vs. 240 at 2 yrs (p Conclusion: Similarly to other studies, (SHM Monitoring report, 2009), a more complete and lasting long-term immunologic response to c-ART was observed in treatment – naïve patients. According to us, a previous sub-optimal and irregularly applied ART regimen, may promote the selection of gradually outgrowing drug-resistant viral strains, compromising therapeutic efficacy in the long run. P13 Use of new targets (D-Mannose receptor, sodium channel voltage dependent) in a new effective, low cost HAART. Validation with the presentation of a clinical case Adrien Caprani1*, Guy MK Tran2, Laurent Roudiere3 1 Alternative Therapies-Basic Science at Association Positifs, Vidauban, France; 2 Clermont-Ferrand University, Hospital Hotel-Dieu, Public Health, ClermontFerrand, France; 3Hopital Pitie Salpetriere, Paris, France E-mail: [email protected] Retrovirology 2012, 9(Suppl 1):P13 Background: Neglected data more or less recent, showed that the voltage-dependent sodium channel (Tran MKG) and the Mannose receptor [Bandivdekar AH 14°ISHEID Toulon 2006 (PP 2.14); J Acquir. Immune.Defic.Syndr.2008,Virology.2008] are involved in the transmission of HIV. In particular, the mannose Receptor seem essential for contamination since in a discordant couple, the uninfected male partner does not own this receptor (14° ISHEID, Bandivdekar A.H.). From these facts we have experienced a patient of 68 years, HIV + since 28 years, under active antiretroviral therapy (Epivir, Reyataz, resveratrol) (another presentation at this conference), the following chemotherapy: Epivir (150mg 2daily), resveratrol (500mg 2daily), D-Mannose (1 g 3daily), Omacor (1 g 2daily). Indeed it is known that omega-3fatty acids bind the Na+ channel. Resveratrol has an anti TaT activity (Zhang HS, 2009) and a synergy with nucleoside analogues ((HerediaA, 2008.) Methods: Measurements of viral load, CD4 and CD8 and other usual blood parameters were followed during 9 months, every month. Results: Our results show that over a period of 9 months the patient remains undetectable and CD4 count increases significantly from the one Retrovirology 2012, Volume 9 Suppl 1 http://www.retrovirology.com/supplements/9/S1 of the previous therapy (557+/-43 vs 478+/- 35). Besides, it seems that the CD4/CD8 ratio tended to increase (0.61 vs 0,56 ). Moreover strong activation of the immune system almost always observed among patients HIV+ is standardized at this patient (CD3+/HLADR+ 7%) and the NK strongly increase (24%). Conclusion: Our results show the feasibility of HAART including Epivir, resveratrol, D-mannose, and omega3 fatty acids. In addition control of dyslipidemia induced by orthodoxic anti retrovirals should be unnecessary. These results paves the way for clinical trials with effective, low toxicity and low cost agents. Moreover the fact that mutations on the Mannose receptor and sodium channel (cell structures) are unlikely, make the appearance of resistance to such therapies unlikely. P14 Polymorphims of innate immunity genes influence disease progression in HIV-1 infected children Riccardo Freguja*, Ketty Gianesin, Marisa Zanchetta, Francesco Carmona, Sandro Malacrida, Osvalda Rampon, Carlo Giaquinto, Anita De Rossi Researcher at University of Padua, Padua, Italy E-mail: [email protected] Retrovirology 2012, 9(Suppl 1):P14 Introduction: Toll-Like Receptors (TLRs) and Defensins play a crucial role in host’s innate immune response. Genetic variations in Defensins and TLRs may affect host-virus interactions and impact HIV1 disease progression, particularly in infants who acquire immune infection when adaptive immune response is still under development. Methods: The study was performed in 95 perinatally HIV1 infected children followed since birth. The endpoint was the onset of disease (stage C) or initiation of highly active antiretroviral therapy. The median (interquartile) followup from birth to endpoint was 87 (46-134) months. Single nucleotide polymorphisms (SNPs) on Beta-defensin1 (DEFB1 -44C>G;-52G>A), and TLR9 (1174G>A;1635A>G) genes were identified with TaqMan allelic discrimination assay. The probability of acquiring disease was calculated with Kaplan-Meier method. Hazard ratios and their 95% confidence interval (95% CI) based on the Cox proportional hazards model were estimated to test the association between genotypes, haplotypes and risk of stage C. Results: TLR9 1635AG genotype was associated with rapid disease progression with both KaplanMeier (p=0.008) and Cox analysis (p=0.009), while DEFB1 -44CG genotype was associated with slower disease progression with both KaplanMeier (p=0.020) and Cox analysis (p=0.024). Notably, TLR9 [G;G] haplotype, previously associated with a higher risk of mother-to-child transmission of HIV1 (MTCT), was also found to be associated with rapid disease progression (p=0.033). In addition, DEFB1 [G; G] haplotype, found to be protective against MTCT, was associated (p=0.016) with a better clinical outcome in HIV1 infected children. Conclusions: Overall, these findings support the role of innate immunity in pediatric HIV1 pathogenesis. Specific SNPs in DEFB1 and TLR9 genes may affect the functional ability of their encoded proteins to modulate innate immunity, thus contributing to the variability of clinical outcome in HIV1 infected children. P15 Emergence of IFN-alpha TRAIL-expressing killer pDCs (IKpDCs) as a consequence of a crosstalk with NK cells. Influence of HIV-1 infection and implication of HMGB1 Marlène Bras*, Héla Saidi, Pauline Formaglio, Marie-Thérèse Melki, Marie-Lise Gougeon Institut Pasteur, Paris, France E-mail: [email protected] Retrovirology 2012, 9(Suppl 1):P15 Background: Plasmacytoid dendritic cells (pDCs) mainly contribute to antiviral immunity through recognition of viral components resulting in the production of type-I interferon (IFN), a powerful innate antiviral cytokine. IFN-alpha production by pDCs is promoted by a cross-talk with NK cells, that triggers in return the cytotoxicity of NK cells. Given the essential role of pDCs and NK cells in viral control, we addressed the Page 14 of 54 question of the impact of HIV on NK-pDC cross-talk, and the consequences on viral innate immunity. Methods: pDCs and NK cells were negatively sorted from PBMC of healthy donors. NK cells were kept either unstimulated (rNK) or activated with PMA/ionomycine for 2 hrs (aNK). pDC were either uninfected or infected with R5-HIV-1 BAL (pDCHIV) and cocultured with NK cells at different ratios for 24 h. The fate of both cell types was studied by multiparametric flow cytometry combined to Multianalyte Profiling technology. Results: HIV-1-infection of primary pDCs induced their maturation, characterized by the expression of maturation markers (HLA-DR, CD80, CD83, CD86) and the homing receptor CCR7. In addition, HIV-1 triggered the emergence of IFN-induced TRAIL expressing killer pDCs. The crosstalk of pDCHIV with aNK cells strongly increased the differentiation of pDCs into killer pDCs. Interestingly, the alarmin HMGB1, secreted by pDC upon HIV-1 infection, seems to be important in this cross-talk, since its modulation altered pDC maturation and the emergence of IKpDCs. At high concentrations of HIV-1, pDCs were able to activate rNK cells, as assessed by CD69 expression, and to induce IFN-gamma and TNF-alpha expression as well as perforin degranulation by aNK cells. Conclusion: We report for the first time that NK-pDCHIV crosstalk potentiates the emergence of TRAIL-expressing IFN-alpha-producing pDCs, and also triggers beta-chemokines synthesis and NK cell killing activity in a HMGB1 dependent manner. Overall these data suggest that the dialogue of HIV-infected pDCs with NK cells favors the emergence of both killer pDC and cytotoxic NK cells and promotes host innate immunity through the activation of potent antiviral effectors. P16 Naive CD8+ T cells from ART respond to primary vaccination against autologous HIV-1 antigen Kellie N Smith1*, Robbie B Mailliard2, Weimin Jiang2 1 University of Pittsburgh School of Medicine, Department of Molecular Virology and Microbiology, Pittsburgh, USA; 2University of Pittsburgh Graduate School of Public Health, Department of Infectious Diseases and Microbiology, USA E-mail: [email protected] Retrovirology 2012, 9(Suppl 1):P16 Introduction: Antiretroviral therapy (ART) decreases HIV-1 viremia and AIDS-associated mortality. Despite this, HIV infected patients are unable to clear virus during treatment interruption due to insufficient cytotoxic T cell (CTL) activity against the autologous reservoir. It is unclear if naïve T cells from patients on ART can respond to immunotherapies that induce CTL specific for their own, unique virus. Unfortunately, late-evolving virus and the ART reservoir contain escape epitope variants that confer a lack of CTL control. We hypothesize that a dendritic cell (DC)-based immunotherapy during ART can induce CTL capable of eliminating the autologous reservoir, despite their failure to do so during natural infection. Materials and methods: We use a naïve T cell flow cytometry panel to evaluate changes in the naive CD4+:CD8+ T cell ratio before seroconversion, during untreated infection, and after ART in an HIV infected subject. We then use this panel to isolate naive CD4+ and CD8+ T cells from this patient during ART and from HIV negative donors. These purified naive T cells are then used in an in vitro model of dendritic cell (DC) vaccination at their in vivo ratios to induce primary IFNg-producing CTL against autologous HIV-1 Gag, Env, and Nef peptide antigens derived from ART. Results: Although partial immune reconstitution occurs during ART, we observed a disproportionate recovery in the naïve CD4+:CD8+ T cell ratio compared to pre-infection. Despite this, we show that naïve CD4 + and CD8+ T cells from ART, when primed at their skewed in vivo ratio against late-acquired, “escape” epitope variants, differentiate into IFNg-producing CTL comparable to those induced in pre-seroconversion T cells. Additionally, we show that primary CTL responses induced during ART are comparable to those observed in HIV negative donors. Figure 1. Conclusion: These data indicate that, despite a disproportionate recovery in the naive CD4+:CD8+ T cell ratio, DC vaccination of naïve T cells from ART can induce CTL specific for autologous “escape” HIV-1 variants, and that these naive T cells can respond to primary vaccination at a level similar to pre-infection. These data support the use of DC immunotherapies in HIV infected patients on ART. Retrovirology 2012, Volume 9 Suppl 1 http://www.retrovirology.com/supplements/9/S1 Page 15 of 54 Figure 1(abstract P16) P17 Universal mapping of humoral immune response using a versatile high-content and high-density peptide microarray Ulf Reimer1*, Nikolaus Pawlowski2, Janina Seznec2, Tobias Knaute2, Paul von Hoegen2, Holger Wenschuh2, Dan H Barouch2 1 Jpt Peptide Technologies, Berlin, Germany; 2Division of Vaccine Research, Beth Israel Deaconess Medical Center, Harvard Medical School, Boston, MA 02115, USA E-mail: [email protected] Retrovirology 2012, 9(Suppl 1):P17 Background: Humoral immune responses are often the hallmark of efficient vaccines. The recent RV144 vaccine trial has turned attention to the stimulation of humoral immune response as a potential mode of action for HIV vaccines. Therefore, detailed monitoring of antibody reactivities in patient specimens before and after vaccination is crucial. The determination of these reactivities on a sub-protein level provides information on the site of antigen/antibody interaction. In contrast to assays relying on whole antigens such as ELISA, peptide microarrays are efficient tools to deliver such information. Besides, complex peptide libraries can cover HIV sequence diversity, a special challenge provided by this virus. Materials and methods: Based on the sequence database of LANL a complex peptide library of more than 6500 peptides was generated. The peptides were synthesized and printed onto glass slides. Initial incubations with serum samples of non-human primates from vaccination studies were performed and evaluated. Results: The peptides span the immunogenic regions of the HIV proteome including full-length ENV, NEF and fractions of GAG, POL, TAT, REV and VIF and allow an overall coverage above 50% of all HIV sequences. Each clade (A, B, C, D, G, CRF1 and CRF2) is represented by at least one sequence. Additional sequences were added to improve coverage. Retrovirology 2012, Volume 9 Suppl 1 http://www.retrovirology.com/supplements/9/S1 Experimental data for serum samples from vaccination trials allow the identification of antibody reactivities following vaccination. The representation of different clades allows a detailed evaluation of specificity for the raised antibodies. Conclusion: High-density high-content peptide microarrays can tackle the tremendous sequence diversity of HIV and deliver information on clade-specific antibody response. This enables monitoring of humoral immune response in HIV patients independent of geographical origin and to study a broad range of different vaccines. The results can shed light on the underlying protective mechanisms of vaccinations. P18 Evaluation of the accuracy of primary CD4 gating in Senegalese individuals coinfected with HIV and tuberculosis Abdoul Aziz Diallo*, Aliou Niang, Géraldine Daneau, Joséphine Khady Badiane, Makhtar Camara, Abdoul Almamy Hane, Luc Kestens, Souleymane Mboup, Tandakha Dieye Laboratory of Bacteriology, Dakar, Senegal E-mail: [email protected] Retrovirology 2012, 9(Suppl 1):P18 Background: The classic multiparameter measurement of CD4+ T cells (standard method) by flow cytometry is complicated and expensive for resource-limited countries. Simpler and less expensive methods like primary CD4 gating have been described but must be evaluated in patients where an overestimation of CD4 can be measured due to monocytes. To address this issue, we compared primary CD4 gating with the standard method in individuals coinfected with HIV and tuberculosis where the risk of CD4 T cell overestimation exists. Methods: Ninety eight patients were recruited, including 32 individuals infected with HIV alone (control group) and 66 individuals coinfected. Each fresh blood sample was analyzed within 6 hours with the fACSCalibur cytometer (Becton Dickinson) in the laboratory of Bacteriology-Virology of Le Dantec hospital in Dakar, Senegal. For each sample, 2 Trucount tubes were used: one for the standard method containing anti-CD3 FITC, antiCD4 PE and anti-CD45 PerCP monoclonal antibodies, and the other tube for the primary CD4 gating method containing only the anti-CD4 PE. Linear regression and Bland-Altman tests were used for statistical analysis. Results: The correlation of absolute CD4 T cell counts obtained by primary CD4 gating and the standard method was high for the HIV control and HIV-TB coinfected groups (R2 = 0.9897 and 0.9795, respectively). The mean bias was 15 cells/μl for the control group and 16 cells/μl for the coinfected group. For the interval < 200 cells/μl, the correlation is R2 = 0.9978 and 0.8327 (mean bias for both less than 7 cells/μl); for 200 - 500 cells/μl, the correlation was R2 = 0.8112 and 0.9624 (mean bias for both less than 2 cells/μl); for > 500 cells/μl, the correlation was R2 = 0.9809 and 0.9841, (mean bias for both less than 72 cells/μl). Conclusions: Primary CD4 gating is an affordable and accurate method for CD4 counting. With one antibody, it has the potential to be an effective alternative to complex panels for resource-limited countries. However, because of the intensive manual data analysis required, a highly skilled operator is necessary. P19 A high dimensional immune monitoring model of HIV-specific CD8 T cell responses accurately identifies subjects achieving spontaneous control of viral replication Zaza M Ndhlovu*, Lori B Chibnik, Jacqueline Proudfoot, Seanna Vine, Ashley McMullen, Kevin Cesa, Donna Marie Alvino, Alicja Piechocka-Trocha, Philip L de Jager, Daniel E Kaufmann, Bruce D Walker Ragon Institute of Mgh, Mit and Harvard, Charlestown, USA E-mail: [email protected] Retrovirology 2012, 9(Suppl 1):P19 Introduction: A major challenge in HIV vaccinology is the development of appropriate immune monitoring models to determine vaccine efficacy. Studies of HIV-specific CD8 T cells (CTL) suggest that these responses play an important role in infected individuals capable of spontaneous viral control (HIV elite controllers) and that they will likely play a role in Page 16 of 54 immune interventions. However, no single CTL assay is uniquely associated with the controller phenotype. Method: We compared functionality of HIV-1-specific CTLs in individuals with spontaneous viral control and subjects with treated or untreated progressive infection. A model integrating multiple features of epitopespecific CTL responses that delineate HIV controllers from subjects with treated or untreated progressive infection was built. Results: Area Under the Receiver Operating Characteristic (ROC) Curve showed that proliferative capacity, absolute early cytokine production and kinetics of cytokine secretion were all associated with HIV control. However, only integrated modeling of these different dimensions of data allowed reaching the remarkable 90% accuracy, which was validated in separate cohorts. Conclusions: Our results suggest that while the search for a common determinant of protective immunity remains elusive, combining parameters generated by various well-established assays in models that can be iteratively refined may have important applications for predicting disease outcome and for immune monitoring of HIV-1 vaccine trials. P20 The levels of apostosis markers in different HIV infected patients groups Ilze Eksteina*†, Valentina Sondore†, Baiba Rozentale, Andrejs Ivanovs, Inga Januskevica, Gunta Sture , Ludmila Viksna Infectologist at Infectology Center of Latvia, Riga, Latvia E-mail: [email protected] Retrovirology 2012, 9(Suppl 1):P20 Introduction: HIV-1 infection is characterized by a progressive loss of CD4+ T cells. The role of apoptotic processes was identified recently, but a limited information is available so far. The aim of this study was to compare levels of apoptosis markers - cytokeratin 18 neoepitope (CK18) and cytochrome C (CC) in different HIV infected patient groups. Methods: There were 69 HIV infected patients enrolled in the study and divided into four groups according to CD4+ T cell count and presence of opportunistic infections (OI): 19 patients with CD4+ T cell count above 200 c/mcl without OI, 15 patients with CD4+ T cell count below 200 c/mcl without OI, 7 patients with CD4+ T cell count above 200 c/mcl with OI, 28 patients with CD4+ T cell count below 200 c/mcl with OI. Opportunistic infections included tuberculosis, cryptococcosis, CMV infection, PCP. The serum levels of cytokeratin 18 neoepitope and cytochrome C were determined. Comparisons between groups were made using paired T- test. Results: CC levels were not significantly different between groups with CD4+ cell count above and below 200 c/mcl (with opportunistic infections 0,5>p>0,4, without opportunistic infections p=0,5). Levels of CC were not significantly influenced by presence of opportunistic infections (with CD4+ cell count above 200 c/mcl p=0,6, with CD4+ cell count below 200 c/mcl p=0,7). We found significant diference of CK18 levels between group without opportunistic infections and CD4+ cell count above 200 c/mcl (210,58 ±26,98 u/l) and group without opportunistic infections and CD4+ cell count above 200 c/mcl (132,95±14,09 u/l), p=0,02, as well as between group without opportunistic infections and CD4 + cell count below 200 c/mcl (132,95±14,09 u/l) and group with opportunistic infections and CD4+ cell count below 200 c/mcl (174,56±20,83 u/l), 0.02>p>0.01. Conclusion: The results obtained from our study demonstrate elevation of levels of apoptosis serum markers early in HIV infection which anticipate further decrease of CD4 cell count. P21 Immune reconstitution inflammatory syndrome (IRIS) in HIV positive patients initiated on antiretroviral therapy (ART) Basavaprabhu Achappa*, Deepak Madi Unnikrishnan, B Anand Venugopal Kasturba Medical College Hospital, Mangalore, India E-mail: [email protected] Retrovirology 2012, 9(Suppl 1):P21 Background: IRIS is defined as ‘occurrence or manifestation of new opportunistic infections or existing opportunistic infections within six weeks to six months after initiating antiretroviral therapy with increase in CD4 count’. The objective of this study was to determine profile of IRIS in Retrovirology 2012, Volume 9 Suppl 1 http://www.retrovirology.com/supplements/9/S1 HIV Positive patients after initiation of ART and correlation of IRIS with CD4 count. Methods: This was a case control study done on HIV positive patients newly initiated on ART at KMC hospital, Mangalore. IRIS was diagnosed based on National Aids Control Organization Guidelines. Cases were defined as those who developed IRIS after initiation of ART and controls who did not develop IRIS. Results: 40 cases and 80 controls were studied between May 2008 and May 2010, who were newly initiated on ART during this period. The cases and controls were compared based on age, sex, and initial CD4 count, final CD4 count, duration of ART, ART regimen, opportunistic infections and their relationship to CD4 count were analyzed. Mean age of patients who developed IRIS was 36 years. 75% of these patients were men and remaining 25% were females. Initial mean CD4 count was 135 and CD4 count at development of IRIS was 239. The mean duration of ART following which IRIS developed was 4 months. 65% of patients were on Lamivudine+Stavudine+Nevirapine regimen, 25% were on Zidovudine +Lamivudine+Nevirapine, 7.5% were on Stavudine+Lamivudine+Effavirenz, 2.5% were on Zidovudine+Lamivudine+Effavirenz. Most common opportunistic infection occurring as IRIS was pulmonary tuberculosis followed by tubercular lymphadenitis and pneumocystis jheroveci pneumonia. Other opportunistic infections seen were oesophageal candidiasis, isospora duodenitis, CMV Retinitis, cryptococcal meningitis, herpes zoster. Tuberculosis accounted for 52.5% of cases of IRIS. Conclusion: Most common IRIS was tuberculosis followed by pneumocystis. Tuberculosis as IRIS occurred 2.5 months after initiation of ART. There was weak correlation between low CD4 count at ART initiation and occurance of IRIS. P22 A review of probiotics studies in HIV research suggests improved immunological presentation and preservation of viral host restrictive factors of TH17 in HIV patients M Selbovitz*, Keller, Miller, Moore, Farmer, Bray Health Action, Bronx, USA E-mail: [email protected] Retrovirology 2012, 9(Suppl 1):P22 Background: Recovery of gut mucosal immune system is slow and incomplete during HAART therapy, leading to elevated inflammation rates, increased mitochondrial damage and the pathogenesis of replicatively competent escape mutations. A recent study of Ganeden BC30, Bacillus coagulans GBI-30, demonstrated safety, increased CD4+ counts in patients on HARRT, and a significant increase in CD69 and maturation of dendritic cells in vitro. Lactobacillus rhamnosus GR-1 and Lactobacillus reuteri RC-14 demonstrate efficacy in treating naïve HIV patients against gastrointestinal and urogenital infections . Restoration of viral host restrictive factors of Th17 cell lines in GALT may prevent the evolving viral diversity, provoking increased CD4 presentation and regulation of inflammatory cytokine levels responsible for increased viral replication. Methods: Metagenomic sequencing analysis has shown alternations in intestinal microbiota in HIV patients. Measurements of immunological parameters including Serum cytokine levels and total serum IgE levels (CD4 lymphocyte count, CBC, levels of TNFα, NFAT, IL-12, IL-10, and G-CSF [Kim, et. al. 2006]) were measured in studies reviewed here. Results: A recent study of Ganeden BC30, bacillus coagulans GBI-30, 6086 demonstrated safety, and an increase in %CD4, a significant increase in CD69 activation and maturation of dendritic cells in HIV patients on HARRT and improvement in GI function. Lactobacillus rhamnosus GR-1 and Lactobacillus reuteri RC-14 have demonstrated efficacy treating naïve HIV patients gastrointestinal and urogenital infections and prevent diarrhea and increased CD4 T-lymphocyte percentages in HIV patients. Conclusions: Intestinal microbiota are integral to the homeostasis and functioning of immune cells. The loss of intestinal flora by HIV infection is severely detrimental to the recombination of CD4 cells. Restoring proper biodiversity in the gut by safe, efficacious probiotics demonstrates promise in increasing Th17 cells by restoring GALT. GanedenBC30 may down regulate TNF-a and other inflammatory cytokines and mitigate ARV-related SAEs. Page 17 of 54 Data suggests that replenishment of Th17 CD4 cells in the gut mucosa during HAART correlates with improved function of the gut mucosal immune system and its function. Studies by NIH of probiotics to lower microbial translocation and immune activation in HIV-infected adolescents. P23 Alpha-1-proteinase inhibitor regulates CD4 lymphocyte levels and is rate limiting in HIV-1 disease Cynthia L Bristow*, Mariya A Babayeva, Michelle Labrunda, Michael P Mullen, Jose Cortes, Ronald Winston Weill Cornell Medical College, New York, USA E-mail: [email protected] Retrovirology 2012, 9(Suppl 1):P23 Introduction: Adult stem cell migration through human hematopoietic tissue requires the chemokine CXCL12 and its receptor CXCR4. In addition, human leukocyte elastase (HLE) plays a key role. When HLE is located on the cell surface (HLECS), it acts not as a proteinase, but as a receptor for a1proteinase inhibitor (a 1 PI, a1antitrypsin). Binding of a 1 PI to HLECS forms a motogenic complex. We previously demonstrated that a 1 PI deficiency attends HIV-1 disease. Here we investigate the mechanism and therapeutically address the a1PI deficiency of HIV-1 infection. Materials and methods: Blood was collected from 30 HIV-1 uninfected and 39 HIV-1 infected adults. Residual sera was obtained from 20 HIV-1 uninfected chimpanzees, 2 chimpanzees pre- and 42 months post-HIV-1 challenge, 12 HIV-1-immunized macaques, and 3 SHIV-infected macaques. Three HIV-1 infected individuals received a1PI augmentation therapy. Results: In HIV-1 uninfected individuals, CD4+ lymphocytes were correlated with the combined factors a 1 PI, HLECS+ lymphocytes, and CXCR4+ lymphocytes (r2 = 0.91, p < 0.001, n = 30), but not CXCL12. In contrast, in HIV-1 individuals with >220 CD4 cells/μl, CD4+ lymphocytes were correlated solely with active a1PI (r2 = 0.93, p < 0.0001, n = 26). The monoclonal antiHIV-1 antibody 3F5 present in HIV-1 patient blood bound and inactivated human a1PI. Chimpanzee a1PI differs from human a1PI by a single amino acid which lies within the 3F5-binding epitope. Unlike human a1PI, neither chimpanzee nor macaque a 1 PI bound to 3F5, nor was a 1 PI depleted following HIV-1 challenge, consistent with the normal CD4+ lymphocyte numbers of HIV-1 infected chimpanzees. The presence of IgG- a1PI immune complexes correlated with decreased CD4+ lymphocytes in HIV-1 individuals, and a1PI augmentation quadrupled the number of immunocompetent CD4+ lymphocytes with no untoward effects. Conclusions: An autoimmune component of HIV-1 disease was identified and was overcome therapeutically. Results identify an achievable vaccine modification with the novel objective to protect against AIDS as opposed to the current objective to protect against HIV-1 infection. P24 CD4.CD8 ratio decrease in AIDS, explained by a molecular mimicry between African HIV-1 Nef and Notch-1. Nef as a target for vaccine and NF-Kb inhibitors (salicylate, resveratrol,curcumin, epigallocatechine-3-gallate) Guy MK Tran1,2*, Adrien Caprani2, Laurent Gerbaud1 1 Clermont-Ferrand University, Hospital Hotel-Dieu, Public Health, ClermontFerrand, France; 2Association POSITIFS, Paris cedex 18, France E-mail: [email protected] Retrovirology 2012, 9(Suppl 1):P24 Background: The AIDS hallmark is the simultaneous fall in CD4 and rise in CD8 T lymphocytes. Interestingly, this very pathognomonic but unexplained decrease of CD4/CD8 ratio is also characteristic of a member of the EGF family, Notch-1 function (Fowlkes BJ, 2002). Calenda V (1994) found that Nef hampered drastically bone marrow progenitors cells functionality. African HIV-1 strain NDK (Spire B, 1989), which induced a fulminant AIDS killing the patient in only 15 days, decreases dramatically CD4 counts. Nef is the most abundant HIV-1 protein in infected cells (85% of mRNA). Nef is a superantigen, its action is amplified 10,000 times compared to a common antigen. Objective: We found previously Notch-1 in the LTR (Long Terminal Repeat) of another retrovirus [Mouse Mammary Tumor Virus (MMTV)] (Tran Retrovirology 2012, Volume 9 Suppl 1 http://www.retrovirology.com/supplements/9/S1 MKG, Eurocancer, Paris, 1999). As Nef is located also in HIV-1 LTR, we looked for Notch-1 in Nef. Methods: Amino Acid (AA) alignment between Epidermal Growth Factor (EGF) family members (including Notch-1) and Nef (Los Alamos HIV sequences Database, 2002). Results: Nef COOH-terminus of HIV-1 clade D African strains (from Congo Democratic Republic, Chad, Tanzania, Uganda, South Africa, Kenya,…), but not from other parts of the world (other non-D clades), was a perfect molecular mimetic of Notch-1: They shared a heptapeptide (7 AA) SRLAFEH. The homology between Nef (Poon AFY, 2009) and Notch (BLASTP on mouse Notch-1) chimera was 67 AA long with 4 His, 1 Cys and 1 Trp (highly significant): Nef : GWCFEVEEDTEGET NSLLHPISQHGMEDPERQVLVWRFNS RLAFEHKARLMHPEFYKNC Notch : GWLLD…FEQDSEGETNSLPHLISQHAL ANPEMQALA-HGKSRLAFEHQVRLSHLPVANNC It included the Nef LL and ED doublets precisely implicated in CD4 down-regulation and EE in b-COP recruitement(Benichou S.1994). Conclusions: This opens new avenues for a vaccine targeted to NefNotch specific to Africa, a continent devastated by AIDS and tuberculosis (in South Africa, about 60% HIV-1 infected patients had also tuberculosis). P25 Abstract withdrawn Retrovirology 2012, 9(Suppl 1):P25 Abstract withdrawn: P26 Role of functional avidity in HIV-specific memory CD8 T cell effector functions Tiffany Lemon*, Donna Alvino, Zaza Ndhlovu, Bruce Walker Louisiana State University - Dept of Biological Sciences, Baton Rouge, USA E-mail: [email protected] Retrovirology 2012, 9(Suppl 1):P26 CD8+ T cells provide protective antiviral defense in HIV-1 infection. Although studies demonstrate which effector functions are employed, knowledge of the underlying mechanisms is lacking and inconclusive. Here, we investigate the functional avidity of CD8 + T cells, based on cytokine secretion and proliferation, to compare the effective antigen concentration required to induce each response. Our preliminary data Figure 1(abstract P26) Page 18 of 54 suggests that the functional avidity of CD8+ T cells differs based on the effector function used for measurement, indicating that the requirements for activation differ within a single CD8+ profile. We also found that elite controllers, individuals who control the virus without antiretroviral treatment, require lower avidity interactions than chronic progressors. These and future results will help to determine optimal doses of antigens for the induction of effective responses in new vaccine formulations. P27 Prevalence of, risk factors for, and oxidative stress associated with Toxoplasma gondii antibodies among asymptomatic blood donors in Egypt Manar S Azab*, Nashwa K Abousamra, Mohammad H Rahbar, Doaa M Elghannam, Douaa Raafat Departments of Parasitology, and Clinical Pathology, Faculty of Medicine, Mansoura University, Mansoura 35516, Egypt Divisions of Epidemiology and Biostatist, Egypt Retrovirology 2012, 9(Suppl 1):P27 Background: Since existing therapies are not fully effective, and no Toxoplasma gondii vaccine is available, efforts to reduce toxoplasmosis transmission are crucial to reducing the impact of this disease. Objectives: To evaluate the seroprevalence of, risk factors for, and oxidative stress associated with T.gondii antibodies in asymptomatic blood donors in northeastern Egypt in a cross-sectional study. Methods: Blood donors were recruited (169 men and 61 women) from blood banks, Mansoura University Hospital, Egypt. We interviewed blood donors about sociodemographic characteristics and potential risk factors for T. gondii infection using a structured questionnaire. A venous blood sample was taken to document their T. gondii antibody status using enzyme-linked immunosorbent assay (ELISA). Also, serum level of malondialdehyde (MDA) and activity of glutathione peroxidase (GSH-Px) and tocopherol fractions (a, g, δ) was assessed. Results: Overall, 155 (67.4%) of 230 blood donors were positive for anti-T. gondii IgG antibodies and 24 (10.4%) of them were also positive for anti-T. gondii IgG avidity antibodies, which is high compared to many countries. Univariate logistic regression analysis showed an association between T. gondii seropositivity and area of residence, blood type, older ages, level of education, contact with cats, professional contact with farm animals, agricultural activities, washing hands before meals, eating unwashed vegetables, drinking raw milk, eating luncheon or shawerma. In a multivariate logistic regression analysis, eating luncheon or shawerma showed a strong significant association with T. gondii antibodies. T. gondii-seropositive blood Retrovirology 2012, Volume 9 Suppl 1 http://www.retrovirology.com/supplements/9/S1 donors had significantly higher MDA level paralleled with significant decrease in the level of GSH-Px and tocopherol fractions compared with T. gondii negative blood donors. Conclusion: This study highlights that T. gondii is prevalent among healthy blood donors in northeastern Egypt, and that there is a need to introduce T. gondii screening in the blood donation scheme. P28 Near full-lenght genome characterization of a newly derived unique recombinant form AG HIV-1 circulating in Siberia Pavel Borisovich Baryshev, Natalya Matveevna Gashnikova*, Vladislav Viktorovich Bogachev State Research Center of Virology and Biotechnology VECTOR, Koltsovo, Russia E-mail: [email protected] Retrovirology 2012, 9(Suppl 1):P28 Background: Genotyping of HIV-1 variants isolated from patients of Siberian region (Russia) in 2006-2010 revealed a sharp increase of 02_AG recombinant variant proportion among HIV-1 genetic variants, circulating in the Siberia, from 2% in 2006 to 52% in 2010. The majority of HIV 02_AG variants isolated in Siberia form a separate branch of phylogenetic tree. The objective of this analysis was to study structure of the near fulllength genome sequence of HIV-1 strains recombinant form AG rapidly spreading in Siberia. Material and methods: Sample 10.RU.6637 was collected in 2009 from a 28-year-old man, infected in 2008 through heterosexual contact. Three regions of the viral genome were independently amplified using nested PCR from the cDNA to contain a nearly full-length genome of HIV-1. Sequencing of all amplicons was performed by using cycle sequencing and dye termination on an automated sequencer, DNA sequences were assembled using Sequencher software. Jumping profile hidden Markov model (jpHMM) program was used to analyze the subtype assignment of all sequences retrieved. The software Simplot v3.5.1 was initially used to perform bootscanning analyses of a query sequence against a set of other sequences. Phylogenetic trees were constructed with the program PhyML v.3.0 using a maximum likelihood approach. Results: Sequence 10.RU.6637 HIV-1 from Siberia was submitted to Genbank under accession number JN230353. Points of recombination were determined and uniqueness of 10.RU.6637 HIV-1 genome structure was shown through in-depth genetic analysis. Recombinant breakpoint analysis of 10.RU.6637 HIV-1 genome sequence revealed that it was recombinant form between CRF 02_AG and sub-subtype A1. We will designate this variant 02_AG/IDU A HIV-1, to reflect its close relationship to the CRF 02_AG and A1 (IDU A) subtype from Russia. Conclusion: The majority of HIV 02_AG variants circulating in Siberia represented of a newly derived unique recombinant form 02_AG/IDU A HIV-1. Wider spread of HIV-1 URF02_AG/IDU A is possible in the territory of Russia in the coming years. P29 Comparison of HIV-1 viral load based on RNA or reverse transcriptase activity in patients with suspected viral load underestimation Beatrice N Vetter*, Cyril Shah, Jürg Böni, Jörg Schüpbach Swiss National Center for Retroviruses, Institute for Medical Virology, University of Zürich, Switzerland E-mail: [email protected] Retrovirology 2012, 9(Suppl 1):P29 Introduction: In HIV diagnostics, viral load (VL) measurement is based on viral RNA. Occasionally, untreated patients present with a low VL of ≤1000 copies/ml, in which case the VL may be underestimated. In Switzerland, patients with suspected underestimation of VL by nucleic acid testing (NAT) are offered VL assessment by the Product Enhanced Reverse Transcriptase (PERT) assay. Here, we compared VL measurement by these two methods in order to assess the frequency and magnitude of NATbased VL underestimation. Material and methods: We compared VL by PERT and NAT for three different patient groups: (1) newly diagnosed patients with suspected Page 19 of 54 underestimation of VL by NAT (n=19); (2) patients receiving VL monitoring by PERT based on previously confirmed underestimation (n=28), and (3) a reference group of untreated, subtype B-infected patients (n=16). The output of both assays was copies/ml viral RNA. For the PERT assay, conversion to copies/ml was based on a reference correlation of NAT VL and RT-activity (qPCR). Results: In newly diagnosed patients approximately 4% have a suspected VL underestimation by NAT (≤1000 copies/ml). PERT results were available for 19 of 59 of such newly diagnosed patients (32.2%). The median difference (log copies/ml) between PERT and NAT VL for this group was 1.36, compared to 0.92 for PERT-monitored patients and -0.004 for the reference group. In 74% of newly diagnosed and 68% of PERT-monitored patients the VL by PERT was ≥5x higher compared to NAT (reference group: 0%). Correlation between PERT and NAT was at R2=0.02 for newly diagnosed patients, 0.63 for PERT-monitored patients and 0.89 for the reference group. Patient groups (1) and (2) both comprised a mixture of subtypes, including subtype B. Conclusions: This analysis confirms that VL underestimation still occasionally occurs, even with the improved contemporary VL tests. Causes include sequence variations leading to impaired primer/probebinding during cDNA amplification. As inadvertent VL underestimation may lead to further infections or inappropriate treatment decisions, a sequence-independent test, like the PERT, remains valuable for confirming a low VL. P30 Molecular epidemiology and drug resistance prevalence of strains from newly diagnosed HIV-1 patients in Northern Greece during 2009-2010 Zoe Antoniadou, Ioanna Kousiappa*, Johana Hezka, Lemonia Skoura, Simeon Metallidis, Pavlos Nikolaidis, Nicolaos Malisiovas, Leondios G Kostrikis University of Cyprus, Nicosia, Cyprus E-mail: [email protected] Retrovirology 2012, 9(Suppl 1):P30 Introduction: As part of a continuing effort to monitor the molecular epidemiology of HIV-1 in northern Greece, in this study we determined the genetic diversity and the prevalence of drug resistance transmission among HIV-1 strains isolated from 94 newly-diagnosed untreated consenting patients in the period 2009 to 2010. Materials and methods: Peripheral blood mononuclear cells (PBMC’s) and plasma were collected from the patients at the AIDS National Reference Laboratory of Northern Greece. Plasma RNA encoding partial pol(protease and reverse transcriptase) was amplified using TRUGENE HIV-1 genotyping assay (Siemens). Antiretroviral drug resistance prevalence was estimated by using the HIVseq program (HIV Drug Resistance Database, Stanford University). PBMC’s DNA encoding partial env(gp120) V3-loop region was amplified by nested PCR and sequenced using an in-house tropism assay at the Department of Biological Sciences, University of Cyprus. Phylogenetic analysis was performed and trees were constructed for each region by using neighbor-joining with Kimura two-parameter method by means of MEGA v5.0 software. Results: The media age of patients is 32 years with the percentage of male infection at 89.4%. The main risk groups were homosexual contact (73.4%) and heterosexual contact (17.0%). The phylogenetic analysis indicated B and A1 as the dominant subtypes, 47.8% and 41.5% respectively, followed by subtype C, CRF02_AG (3,2% each), CRF04_cpx (2,1%) and subtypes F1 and G (1,1% each). Twenty-one clusters showed epidemiologically linked HIV-1 patients. Resistance mutations to protease inhibitors were found in three individuals, while high resistance associated mutations to reverse transcriptase inhibitors (NRTI’s/NNRTI’s) were shown in sixteen patients. Tropism testing indicated most of the HIV-1 strains as R5-tropic (96%). Conclusions: These newly found data demonstrate a heterogeneous epidemiological status of HIV-1 in northern Greece during 2009-2010, with subtype B and A1 being the dominant subtypes in relation to the other subtypes. The prevalence of antiretroviral resistance mutations is high among the newly diagnosed untreated patients (17%) in comparison with other European countries (10%). Retrovirology 2012, Volume 9 Suppl 1 http://www.retrovirology.com/supplements/9/S1 P31 Frequency of subtype B and F1 dual infection in HIV-1 positive, Brazilian men who have sex with men Sabri Saeed Sanabani*, Ana Carolina Soares de Oliveira, Antonio Charlys da Costa, Mariana Melillo, Sauer Katia Cristina Bassichetto Solange, Maria Santos Oliveira, Priscilla Ramos Costa, Claudia Tomiyama, Helena Tomoko Iwashita Tomiyama, Ester Cerdeira, Sabino Esper, Georges Kal Instituto de Medicina Tropical de São Paulo, São Paulo, Brazil E-mail: [email protected] Retrovirology 2012, 9(Suppl 1):P31 Introduction: Co-infection, otherwise known as superinfection, with 2 or more HIV-1 isolates, has been documented frequently. However, in Brazil, few data are available regarding the frequency of HIV-1 co-infection. Because various HIV vaccination studies are in progress, it is important to understand often inter- and intra-subtype co/superinfection occurs in different HIV-infected high risk groups. In this cross-sectional study, we report the frequency of subtype B and F1 co-infection in a clinical group of 41 recently HIV-1 infected men who have sex with men (MSM) in São Paulo, Brazil. Material and methods: Proviral HIV-1 DNA was isolated from subject’s peripheral blood polymorphonuclear leukocytes that were obtained at the time of enrollment. Each subject was known to be infected with a subtype B virus as determined in a previous study. A small fragment of the integrase gene (nucleotide 4255-4478 of HXB2) was amplified by nested PCR using subclade F1 specific primers. The PCR results were further confirmed by phylogenetic analysis. Viral loads (VL) data were extrapolated from the medical records of each patient. Results: In the 41 samples from MSM who were recently infected with subtype B virus, in five patients it was possible to detect subclade F1 proviral DNA, which represents a co-infection rate of 12.2%. In subjects with dual infection the median VL was 5.3 X 104 (range, 1.5 X 104 - 12.5 X 104 copies/ml), whereas in MSM that were infected with only subtype B virus the median VL was 3.8 X 104 copies/ML (range < 400 - 39.3 X 104 copies/ ml) (p > 0.8). Conclusions: This study indicated that subtype B and F1 co-infection occurs frequently within the HIV-positive MSM population as suggested by large number of BF1 recombinant viruses reported in São Paulo, Brazil. We conclude that the co-infection is a potentially important event that significantly contributes to HIV-1 genetic variability with serious implications for diagnosis, drug treatment and optimal vaccine development. P32 A new unique recombinant HIV-1 revealed in Belarus VF Eremin*, EL Gasich, SV Sasinovich Briem, Minsk, Belarus E-mail: [email protected] Retrovirology 2012, 9(Suppl 1):P32 Material and methods: Blood plasma, EIA, western blot, RT-PCR, sequencing, SeqScape, BioEdit, Mega4.1, statistica 6.0, software. Results: In April 2010 we have performed resistance tests of plasma sample obtained from patient Mos, 6 years old girl, born from HIVinfected mother. The phylogenetic analysis of the DNA fragment of patient Mos had shown that sample has been clustered with HIV-1 subtype A on gene pol, but was different from other analyzed samples, subtype A consensus IDU-A and reference sequences (AF004885). The Mos isolate is the most similar to AF413987 from Ukraine (subtype A) the p-distance was 0.066. The comparison of sequences from gag gene p17/ p24 region of Mos isolate with reference sequences HIV-1 of subtype A demonstrates that average p-distance was 0.129, and with reference sequences of subtype B was 0.075. Average p-distance on gag gene (the Mos isolate) with CRF03_AB (AF414006.1, Belarus and AF193276.1 CRF03_AB KAL153) was 0.121 if compared with 0.013 p-distance between reference sequences. The analysis of Mos isolate sequences on V3 loop gp120 gene env region HIV-1 has shown that average p-distance with reference isolates subtype B was 0.323, and with A subtype was 0.155. Average p-distance sequence of Mos isolate with reference isolates AF414006.1 and AF193276.1 (CRF-03_AB) was 0.308. Page 20 of 54 Conclusion: Thus, it has been shown that Mos isolate is a unique recombinant form, but differs in genome structure from the one described earlier CRF03_ AB (AgagBpolBenv). The new recombinant HIV-1 has the following structure: BgagApolAenv. Sequences of new HIV-1 unique recombinant in gag, pol and env genes were submitted to EMBL/ Genbank/DDBJ under accession numbers: FR775442.1, FN995656.1, FR775443.1. P33 Molecular epidemiology of HIV-1 subtype B in the Basque Country (Spain) Juan Angel Patiño, Miguel M Thomson, Fernando González-Candelas* Csisp-University of Valencia, Valencia, Spain E-mail: [email protected] Retrovirology 2012, 9(Suppl 1):P33 The goal of this work was to study the HIV-1 subtype B epidemic in the Basque Country (Spain). For this, we used HIV samples submitted for genotypic testing of anti-retroviral resistance mutations from 2005 until 2008. Consequently, 2115 HIV-1 sequences comprising protease and retrotranscriptase (PR/RT) coding regions were analyzed. HIV transmission groups were identified by phylogenetic analysis. The 10 largest such groups were subsequently subjected to Bayesian phylogenetic and coalescent reconstructions, using a relaxed molecular clock model. The results obtained show that these groups have been long-standing: most of them were originated in the late 70s or early 80s, and none after the year 2000. Most of these groups comprise both intravenous drug users (UDIs) and people who got infected through unprotected heterosexual sex (HTs). MSM clades were also represented in the sampled population. By comparing different demographic models, it was concluded that all the transmission groups are growing in an exponential manner. Time between infections was significantly lower in MSM groups than in those mainly containing IDUs (P-value < 0.05 in the Mann Whitney test). In conclusion, this work suggests that in the near future the HIV-1 subtype B epidemics in the Basque Country will be characterized by a growth of the existing transmission groups. Due to the high diversity of these clusters, it is necessary to design campaigns for HIV prevention focused on the different risk groups. P34 Clinical consequences of persistent low level viremia Toby Dyner*, Virginia Cafaro, Valby Chow Hiv Disease at Shared Perspectives On Therapies (Spot), San Francisco, USA E-mail: [email protected] Retrovirology 2012, 9(Suppl 1):P34 Introduction: Work done by this group nearly 7 years ago, evaluated a cohort of patients who, for a variety of reasons, elected to remain on a virologically failing ARV regimen. We re-evaluated these patients to determine whether their earlier years living with low level viremia impacted their future treatment options. In the absence of a cure and with the goal of maintaining HIV infection as a chronic, yet manageable disease, it is important to understand the consequences of persistent immune system activation. Materials and methods: Two community based HIV practices with nearly 1000 patients evaluated a subset of 25 patients on stable ARV regimens for >24 months who refused treatment change despite the presence of low level viremia. Patients were counseled regarding the need for change. They acknowledged a variety of reasons for their refusal including fear of change and fear of the unknown; a comfort with their current regimen; fear of “burning through options”, etc. Charts were reviewed for CD4 counts, HIV bDNA levels, HIV related OIs or malignancies, as well as other co-morbidities. Results: As new HIV medications became available, patients agreed to switch regimens. 19 of 25 patients were started on regimens which resulted in virologic suppression below the level of quantification (BLQ). 1 patient died of an MI and 1 died of a neuroendocrine tumor and 4 patients were lost to follow-up. Given the potency of the new regimens that were constructed, those patients who switched were able to suppress without significant difficulties. Retrovirology 2012, Volume 9 Suppl 1 http://www.retrovirology.com/supplements/9/S1 Conclusions: With the new paradigm of test and treat despite CD4 level, we can expect to be treating many more patients for longer periods of time. The issues of adherence and “pill fatigue” are well known and contribute to patients’ potential inability to maintain fully suppressive regimens for long and sustained periods of time. Our experience of patients who refused to switch regimens despite low levels of virus and yet, when ready, were able to fully suppress with a regimen change, is encouraging. P35 The dual (activating/suppressive) effect of extracellular TatHIV-1 is driven by the infalmmatory microenvironment of infected lymphoid foci Hélène Le Buanec1,2,3, Thomas Sené1,2,3, Armand Bensussan1,2,3, Robert Gallo4, Daniel Zagury5* 1 INSERM U976, F-75475, Paris, France; 2Université Paris Diderot, Sorbonne Paris Cité, Laboratory of Immunology, Dermatology & Oncology, UMR-S 976, F-75475, Paris, France; 3Service de dermatologie, Hopital Saint Louis, F-75010, Paris, France; 4Institute of Human Virology, University of Maryland Baltimore, Maryland, USA; 5Neovacs SA, Paris, France E-mail: [email protected] Retrovirology 2012, 9(Suppl 1):P35 It has been shown that HIV-1 infects activated but not resting CD4 + T cells [1] and that CPE induced by viral replication together with the immunosuppressive effect triggered by extracellular Tat protein [2] account for the decrease of CD4+ T cell count in infected patients. In lymphoid foci, dependent on the level of viral infection, the stromal microenvironment surrounding immune cells could include, together with extracellular Tat [3] and circulating antiviral IFN-a, inflammatory innate factors such as ATP and derivatives released by CPE-derived dead cells. We show that, according to its concentration and the presence of inflammatory factors (IFN-a, ATP and ATP-derivatives), Tat protein may exert either an activation with enhanced production of IL2 or an immune suppression of stimulated CD4+ T cells subpopulations. The double-edged sword of Tat activity on CD4+ T cells could account for its immunopathogenic effects both at the early stage of infection (by allowing CD4+ T cells activation and viral replication) and at late stages (by inducing immuosuppression, source of opportunistic infections). Indications for targeting Tat protein by therapeutic vaccines in subgroups of HIV-1 infected patients will be discussed. References 1. Zagury D, et al: Long-term cultures of HTLV-III–infected T cells: a model of cytopathology of T-cell depletion in AIDS. Science 1986, 231(4740):85. 2. Viscidi RP, et al: Inhibition of antigen-induced lymphocyte proliferation by Tat protein from HIV- Science. 1989, 246(4937):1606. 3. Ensoli B, et al: Release, uptake, and effects of extracellular human immunodeficiency virus type 1 Tat protein on cell growth and viral transactivation. J Virol 1993, 67(1):277. P36 Genotype characterization of human papillomavirus in women infected and uninfected with HIV in Ouagadougou, Burkina Faso Florencia W Dkigma*, Djeneba Ouermi, Tani Sagna, Charlemagne Ouedraogo, Cyrille Bisseye, Moctar Zeba, Simplice D Karou, Virginio Pietra, Jean-Baptiste Nikiema, Jacques Simpore Centre de Recherche Biomoléculaire « Pietro Annigoni » CERBA/LABIOGENE, Ouagadougou, Burkina Faso E-mail: [email protected] Retrovirology 2012, 9(Suppl 1):P36 Introduction: Our study aims to compare the prevalence and genotypes of HPV in HIV-positive and negative women in Ouagadougou, where HIV and HPV prevalence are respectively estimated at 4.0% and 24%. Materials and methods: The study involved 410 women: 205 HIVpositive followed by two sites of HIV/AIDS care in the eastern outskirts of Ouagadougou, matched by age with 205 HIV-negative consulting the gynaecological services in the same area. HPV genotyping was done by PCR followed by reverse hybridization on nitrocellulose strips with the kit “STAR HPV Blot” (Diatech®, Italy). Page 21 of 54 Results: Prevalence of HPV was: 25.4% among HIV-negative women and 59.0% among HIV-positive women (p <0.01). Means of age was respectively 33.6 (SD±8.6) versus 33.4 (SD±6.4). Prevalence of HPV subtypes at low risk of oncogenicity (6,11,LR) among HIV-positive women was 9.8% and 9.3% in the control group (p=ns). Prevalence of HPV subtypes at high risk of oncogenicity was significantly(p<0.01) higher among HIV-positive women for subtypes 50’S (22.0% versus 9.3%, OR=2.8 ; CL95% 1.5-5.1), 18 (20.0% versus 2.9%, OR=8.3 ; CL95% 3.2–23.0), 30’S (17.6% versus 1.5%, OR=14.3 ; CL95% 4.1–60.1) and HR ( 7.3%, versus 1.0%, OR=8.0 ; CL95% 1.7–52.1) while no significant difference was observed for high risk subtypes 16 and 45. Co-infections by two or more subtypes at high risk were detected in 27/205 (13.2%) HIV-positive women and 3/205 (1.5%) HIV-negative women (p<0.01; OR 10.2; CL95% 2.9-43.5). Among HIV-positive women, prevalence of oncogenic subtypes was significantlycorrelated with a lower CD4 count (P = 0.05). Conclusions: HIV-positive women are at high risk of coinfection by HPV oncogenic subtypes. This study confirms the need to integrate the screening of cervical cancer in HIV care protocols in Burkina Faso. Further investigations should be continued for the establishment of vaccine that matches all genotypes circulating in the country. P37 Tungiasis (jigger infestation) in Rural Kenya, an emerging infectious disease Nicholas N Njau*, Peter Wanzala, Marion Mutugi, Liana Ariza, Jorg Heukelbach Kenya Medical Research Institute, Center for Public Health Resear, Nairobi, Kenya E-mail: [email protected] Retrovirology 2012, 9(Suppl 1):P37 Objective: To describe the prevalence of tungiasis (jigger flea infestation) and associated risk factors in a sentinel group (children 5-12 years of age) in rural Central Kenya. Methods: A cross-sectional study was carried out in Murang’a South district during high transmission season (dry season, August – September 2009). A total of 385 randomly selected households were visited. Children were examined for presence of tungiasis, and a questionnaire was administered to collect demographic, behavioral and environmental data. Results: Prevalence of tungiasis was 57% (218/385; 95% CI=51.7%-61.6%). Itching (89.1%) was the most common associated symptom, followed by pain upon pressure (67.3%), sleep disturbance (58.2%) and walking difficulties (53%). In multivariate logistic regression analysis the following independent factors were identified to be associated with tungiasis: living in houses with an earthen floors (adjusted OR=3.84; 95%IC: 2.09-7.06), walking barefooted (OR=3.28; 1.78-6.04), having a common resting place outside the house (OR=2.36; 1.01-5.51) and presence of rats on the compound (OR=1.69; 1.03-2.75). Conclusion: Tungiasis is an emerging neglected disease found in Africa. It is highly endemic in rural Central Kenya and associated with considerable morbidity. The disease is associated with poverty. Modifiable risk factors were identified that should be the focus of sustainable and effective control measures. P38 Impact of macrophages on Balamuthia mandrillaris virulence properties using human brain microvascular endothelial cells in vitro Abdul Matin*, Khalid Mehmood, Suk-Yul Jung Institute of Biomedical and Genetic Engineering, Islamabad, Pakistan E-mail: [email protected] Retrovirology 2012, 9(Suppl 1):P38 Introduction: Balamuthia amoebic encephalitis (BAE) is a serious human disease almost always leading to death. An important step in BAE is amoebae invasion of the bloodstream, followed by their haematogenous spread. Balamuthia mandrillaris entry into the central nervous system (CNS) most likely occurs at the blood–brain barrier (BBB) sites. Macrophages are thought to be the first line of defense in many infectious diseases and are present in high numbers during infections. Retrovirology 2012, Volume 9 Suppl 1 http://www.retrovirology.com/supplements/9/S1 The objective of the present study was to determine the impact of cytokines and macrophages on the virulence characteristics of B. mandrillaris in vitro. Materials and methods: In vitro, B. mandrillaris were used to demonstrate the effects of cytokines and macrophages on the physiological and morphological characteristics of amoeba. Using human brain microvascular endothelial cells (HBMEC), which constitutes the BBB, adhesion and cytotoxicity assays were performed. To investigate the engulfing property and proteolytic activity of the amoeba, phagocytosis and zymography assays were conducted respectively. Results: It was observed B. mandrillaris exhibited >90 % binding and >70 % cytotoxicity to HBMEC which was further enhanced in the presence of cytokines and macrophages. It has also been observed that cytokines TNF-a and TGF-b significantly increased the B. mandrillaris numbers in the presence of macrophages. It is important to note that amoebic numbers were more than doubled in the presence of cytokines and macrophages within 24h. We have shown in the past the bacteria uptake by B. mandrillaris is limited which is further significantly inhibited in the presence of cytokines during phagocytosis assays. Zymography assays revealed that cytokines and macrophages have no inhibitory effect on proteolytic activity of B. mandrillaris. In addition the activated macrophages did not show any vital effects on amoebic virulence properties. Conclusion: Overall we described for the first time that cytokines and macrophages has no inhibitory effects on the virulence properties of B. mandrillaris in vitro. P39 Investigation of measles outbreak-Herena and Dawe-Serer Districts of Bale Zone, Oromia Region, Ethiopia, February 2011 Abyot Bekele Woyessa*, Tesfaye Deti, Amanuel Yadata, Ashenafi Kenna, K Addisalem, M Yesuf, M Abebe Ethiopian Health and Nutrition Research Institute, Ethiopia, Addis Ababa, Ethiopia E-mail: [email protected] Retrovirology 2012, 9(Suppl 1):P39 Introduction: An estimated 10 million cases and 164,000 deaths from measles occur worldwide each year. On 08 Feb 2011 Bale Zonal health department reported suspected measles outbreak. We investigated to identify the etiology of the outbreak and undertake appropriate prevention and control interventions. Materials and methods: Patient observation was made and active-cases were searched house to house. Medical registration-books were assessed and suspected measles cases were identified from 24-Nov-2010 to 15Feb-2011 in Herena and Dawe-Serar districts using the following case definitions: maculopapular rash with fever ≥38.5°C with coryza, conjunctivitis or cough or epidemiologically linked by contact with laboratory confirmed outbreaks in neighboring districts. Immunization coverage and vaccine-storage facilities were assessed. Descriptive analysis was conducted using Epi-Info version3.5.1. Results: A total of 329 suspected measles cases and 30 community deaths (case fatality rate (CFR) 9.1%) were reported of which 159 (48.3%) were from Dawe-Serar and 170 (51.7%) were from Herena. CFR was 25/ 159 (15.7 %) in Dawe-Serar and 5/170 (2.9%) in Herena and higher among females than males (12.2% vs. 6.1%). All deaths and 140/329 (42.6%) of the cases were not vaccinated against measles. Vaccination coverage was 45.4% in Dawe-Serer and 54% in Herena. The attack rate was highest among those 15 years of age. About 5/7 (71%) refrigerators used for vaccine-storage were not functional. Prior to investigation period, 110/159 specimens from 14 districts of the zone were tested positive for measles-IgM. Conclusions: An outbreak of suspected measles occurred in 2 districts affecting primarily those <5 years of age. Low-vaccination coverage and non-functional cold storage likely contributed to the outbreak. Undertaking supplementary vaccination activities, enhancing routine vaccination coverage and improvement in cold chain operation and maintenance need to be emphasized in the districts to reduce measles incidence. Page 22 of 54 P40 Seroepidemiological feature of Q fever among sheep in Northern Iran Ehasn Mostafavi*, Saber Esmaeili, Mahin Shahdordizadeh, Hadi Mahmoudi, Hamid Liriayii, Fahimeh Bagheri Amiri Department of Epidemiology at Pasteur Institute of Iran, Tehran, Iran E-mail: [email protected] Retrovirology 2012, 9(Suppl 1):P40 Introduction and aims: Q fever is a zoonosis caused by Coxiella burnetii, which infects various hosts, including humans and animals. As Q fever is considered an important factor in public health, and there is little epidemiological information on the status of the disease in various parts of Iran, this study has been carried out to evaluate the seroepidemiology of Q fever among sheep in the province of Mazandaran, northern Iran. Materials and methods: In this study, samples from sheep were collected from western, central and eastern regions of Mazandaran in 2010-2011. Serum samples were analyzed by ELISA test. Results: In this study, 253 serum samples were collected. The infection rate with Q fever was 23.7%. The chi-square test showed a significant statistical relationship between central (33.8%) and eastern (27.2%) regions compared to western regions (8.5%). There was no significant difference between the three groups of sheep with respect to age. No significant statistical relationship was seen between infection rate and age and gender. Discussion: The infection rate of coxiella burnetii in this study, is similar to the results of other research carried out in various parts of the country. With respect to the fact that there is a higher infection rate in the eastern and central regions of the province, compared to the western region, and also in imported animals from Afghanistan, the hypothesis that the disease is spreading from eastern boundaries becomes more probable. It is recommended that complementary research be carried out on other animals, on high-risk persons and on ticks, in order to reveal the status of the disease in the province. P41 Molecular detection of Rickettsia and Borreliosis pathogens in the areas of Boumerdes and Tizi Ouzou (Algeria) Afif T Chaouche*, I Bitam, K Amara, I Yahiaoui Microbiologie at Université, Tlemcen, Algeria E-mail: [email protected] Retrovirology 2012, 9(Suppl 1):P41 Aim: Mediterranean spotted fever, and the Lyme disease are emerging infectious diseases with significant impact on public health, a study of their detection methods and transmission modes is very useful. As vector diseases, this study consist of finding the agents in question on ticks (vectors of these diseases) in the area of Boumerdes and Tizi-ouzou, coastal areas in central Algeria, and checking the transmission of these germs from generation of ticks to another. Materials and methods: The investigation launched from December 2009 to June 2010, has raised ticks on dogs and cows in these areas, and after the identification of genus and species of the 182 collected ticks, 56 of them were analyzed by PCR followed by electrophoresis. The detection of Rickettsia was also done by the Gimenez stain from a drop of hemolymph of a tick collected on a slide. A breeding of ticks has been launched to track their life cycle and eventual transmission of Rickettsia by ovarian trans-staidly. Results: 11 of 56 ticks were found carrying Rickesttsia conorii conorii, (the bacteria responsible for Mediterranean spotted fever), and no ticks carrying Borrelia Burgdorferi (bacteria that causes Lyme disease) was found. After analyzing the ticks of the first and second generation, the presence of these bacteria has been found. Conclusion: The results of this investigation allow confirm the presence of Rickettsial diseases in the regions of Boumerdes and Tizi Ouzou, but also the transmission from one generation of germs on ticks, the risk of transmission to humans is certain in case of tick bite, from which the necessity of the vector control. 1. Sahibi H. & Rhalem A. (2007) : Tiques et maladies transmises par les tiques chez les bovins au Maroc, MADER/DERD N°151 2. Doudier B., Pages F., Parola P., Socolovschi C., Tiques et maladies Retrovirology 2012, Volume 9 Suppl 1 http://www.retrovirology.com/supplements/9/S1 transmises à l’homme en Afrique, Médecine Tropicale, Med Trop 2008, 68, 119-133 3. Bitam I., (2008) : Rickettsioses associées aux arthropodes, la revue medicopharmaceutique N°48-3éme trimester, p38-39. P42 Risk factors of Trichomonas vaginalis in women attending central Sexually Transmitted Diseases Clinic Sri Lanka Sathyadevi Herath*, Deepika Fernando, Saman Jayasinge Ministry of Health, Sri Lanka, Colombo, Sri Lanka E-mail: [email protected] Retrovirology 2012, 9(Suppl 1):P42 Introduction: Trichomonas vaginalis is one of the common infections among women attending Sexually Transmitted Disease Clinics (STD) of Sri Lanka. Yet majority with symptoms don’t attend STD clinics and treated syndromically. In this scenario this study was carried out to identify risk factors of Trichomonas vaginalis in women inorder to review treatment by signs and symptoms. Methods: Three hundred and fifty new female clinic attendees were recruited. Participants were interviewed on sociodemographic data, sexual history, symptoms, knowledge on STI/HIV, and condom use. Laboratory specimens were collected for theroutinescreening ofSTD diagnosis including Trichomoniasis. Results: Mean age of the sample was 32.8 years (SD ± 9.27). More than half (223; 64%) were married. Approximately 53% had completed Grade 10. Almost 76% tested positive being in 21-45 years and 20% of positives were unmarried. Trichomoniasis prevalence was 7.2% (25 out of 346). Pruritus, vaginal discharge and vulvovaginal soreness, were significantly higher amongst positives (P< 0.05 for all). Educated women had higher risk (OR= 3.0; CI=1.28-7.26) of infection. Trichomoniasis was less common among women engaged in sex work (OR= 0.3; 95% CI=.0.14-0.85), reported multiple sexual partners (OR = 0.02; 95% CI= 0.073-0.408) and women reporting extra marital relationship (OR = 0.3%, 95% CI= 0.123-0. 733). Conclusions: These findings are the reflection of female clinic attendees involving in commercial sex trade and high use of condoms amongst sex workers. It further emphasizes that the primary prevention activities are widespread especially among MARPs. Findings also suggest that non use of condoms among women in monogamous relationship may contribute for Trichomoniasis thus HIV acquisition among housewives in Sri Lanka. P43 A retrospective analysis among male and female infants EID results in Cross River State, Nigeria Onovo Amobi1,2 1 University Of Calabar, Nigeria; 2Monitoring and Evaluation Officer at ProHealth International, Calabar, Nigeria E-mail: [email protected] Retrovirology 2012, 9(Suppl 1):P43 Background: Early definitive diagnosis of HIV infection in infants is critical to ensuring that HIV-infected infants receive appropriate and timely care and treatment. The purpose of this study is to investigate the possible determinant of EID test results among male and female infants in south-south region of Nigeria. Methods: A retrospective study was conducted in July, 2011 among male and female infants receiving PMTCT intervention for Early Infant Diagnosis in PHC’s at four different LGA’s of Cross River namely: Akamkpa, Calabar South, Odukpani and Biase respectively. Relevant data of the HIV infection status to male and female infants, whose samples were collected and diagnosed using PCR, was obtained from the National PMTCT-EID register. The data was analyzed using Cross-tabulation. Results: About 42.9% male infants and 57.1% female infant’s blood samples were collected using the DBS technology for diagnosis by PCR assay. The age distribution of the infants ranged from 2 - 11 months with the mean age of 5 months. EID samples (19.0%) diagnosed tested HIV positive and 81.0% tested HIV negative by PCR. 7.1% (Male) and 9.5% (Female) infants tested HIV positive by PCR. 35.7% (Male) and 47.6% Page 23 of 54 (Female) infants tested HIV negative by PCR. There was a significant association between infant’s ages at 9 months, 10 months and 11 months with the EID test results (Standardized residual of 3.7, 1.9, and 2.6 respectively). The test of model of gender as predictor was statistically significant for female infants (Standardized residual of 1.6) and a Pearson chi-square which appeared statistically significant (P=0.006). Conclusion: From our study, there is a significant association between EID test results and specific ages of male and female infants with gender as a perfect predictor. P44 In vitro antifungal susceptibility of Candida albicans isolates from oral cavities of patients infected with human immunodeficiency virus in Ethiopia Nasir Tajure Wabe*, Jemal Hussein, Sultan Suleman, Kedir Abdella Jimma University, Jimma, Ethiopia E-mail: [email protected] Retrovirology 2012, 9(Suppl 1):P44 Summary objective: Oral Candidiasis is the most common HIV related oral lesion. Most patients are infected with a strain originally present as a commensal of the oral cavity. The chronic use of antifungal agents, in the treatment of candidiasis mainly in HIV/AIDS patients leads to the selection of strain resistant to this therapy. The objective of this study was to evaluate the in vitro susceptibility of Candida albicans to commonly used antifungal agents in Ethiopia. Methods: In vitro susceptibility tests were performed using the broth microdilution method following the National Committee for Clinical Laboratory Standards (NCCLS) M27-A guidelines. Data were then analyzed using SPSS for windows version 16.0. Tests of proportions were done with Chi-Square, and a p value of <0.05 was considered as statistically significant. Results: A total of 42 oral C.albicans isolates from HIV-infected patients were included in this study. Forty one (97.7%) of all isolates were determined fully susceptible to amphotericin B, 40 (95.3%) to nystatin, and 39 (92.9%) to ketoconazole and miconazole. On the other hand, the isolates showed highest rates of resistance against fluconazole (11.9%) relatively. There was little difference in the antifungal susceptibilities of C. albicans isolated from patients who had a history of previous antifungal therapy compared with those who had not received antifungal treatment. Conclusion: The in vitro antifungal susceptibility testing of C.albicans in this study showed relatively high resistance to commonly used azoles. As with the prescribing of any antimicrobial agent, the use of a systemic antifungal drug must be justified. Efforts must be maintained to avoid inappropriate or unnecessary prescribing of these antifungal. P45 Reproductive Tract Infections (RTI) among married women in Sri Lanka Sathyadevi Herath1*, Pushpa Fonseka2, Sujatha Samarkoon1 1 National Sexually Transmitted Diseases, AIDS Control Program, Ministry Of Health, Sri Lanka; 2University of Sri Jayawardenepura, Sri Lanka E-mail: [email protected] Retrovirology 2012, 9(Suppl 1):P45 Introduction: Feminization of HIV epidemic, and increasing in HIV infection among house wives is seen in most of the world. Yet, community prevalence data on RTI are sparse in Sri Lanka, and little is known about rates among community women. The objective was to describe the prevalence and risk factors of RTI in women aged 22–49 years, living in highly populated poor urban settlements of Sri Lanka. Methodology: A community-based cross-sectional study was conducted among 770 married women living in 116 urban slums of Colombo city, Sri Lanka. Participants were interviewed on sociodemographic data, sexual history, knowledge on STI/HIV, and condom use. Laboratory specimens were collected for thediagnosis of RTI. Prevalence was calculated with corresponding 95% confidence intervals (CI). Analyses of risk factors were carried outseparately for the outcomes of sexually transmitted infections: chlamydia, gonorrhoea, trichomoniasis; and endogenous infections:bacterial vaginosis (BV) and candida. Retrovirology 2012, Volume 9 Suppl 1 http://www.retrovirology.com/supplements/9/S1 Results: Ninety three percent of women had single life time partner, only 9% was previously screened for STI/HIV. Condom use was mainly decided by the male partner and none of them used condom for prevention of STI/HIV. Endogenous infections were relatively common [BV 8.6% (95% CI: 6.6.10.6)candida 6.8 % ( 95%CI: 5.0-8.6)], and sexually transmitted infections (STI) were infrequent (1.1%-95% CI: 0.34-1.86). Of the risk factors investigated none of the factors were associated with RTI on multivariate analysis. Conclusion: Married women in this community had a low prevalence of RTI and risky sexual behavoiurs were infrequent. Most of the population burden of RTI is attributedto endogenous infections. However, education and outreach screening facilities are needed to reduce the stigma, embarrassment and lack of knowledge related to STI/HIV in order to facilitate screening and condom use. P46 Investigation of acute febrile illness outbreak- Asyaita and Dupti districts, Afar Region, Ethiopia, February 2011 Abyot Bekele Woyessa*, Worknesh Ayele, Abdi Ahimed, A Nega Ethiopian Health and Nutrition Research Institute, Ethiopia, Addis Ababa, Ethiopia E-mail: [email protected] Retrovirology 2012, 9(Suppl 1):P46 Introduction: Acute Febrile Illnesses (AFIs) due to different etiologic agents are the most common causes of morbidity and mortality in developing tropical and subtropical countries. Afar region reported unidentified AFI outbreak on 10-Aug-2011. We investigated to identify etiologic agent, risk factors and to recommend prevention and control measures. Methods: Unmatched case control study was employed. Study subjects (57 cases and 57 controls) were obtained and interviewed. Cases were defined as any person with fever ≥ 38.3C°, headache, pains in joints, muscles and back, anorexia and weakness. Medical records were reviewed and suspected AFI cases were identified from 07-Aug-2011 to 11-Sep2011in Asyaita and Dupti districts. Active cases were searched house to house. Blood-samples, blood-serums and throat-swabs were collected and analyzed for heamoparasites, bacterial-pathogens, hemorrhagic fevers and respiratory viruses at national and CDC Kenya laboratories. Environmental scanning was performed. Odd-Ratio (OR) in 95%Confidence-Interval (CI) was calculated using Epi-Info version-3.5.1. Results: A total of 12816 suspected AFI cases with no death were identified. Of the cases 9107(71%) were male and 3709 (29%) were female. Attack-Rate (AR) was 8.7% (11.5% in male and 5.4% in female) and 13.8% among 15-44 age-groups. On bivariate analysis factors associated with illness were living with sick family member (OR: 2.8; 95%CI: 1.3-6.2), contact with patient (OR: 3.8; 95%CI: 1.5-9.6) and drinking deep-well water (OR: 2.7; 95% CI: 1.2-5.8). However, on multivariate analysis only having contact with patient (OR: 4.1; 95%CI: 1.5-11.3) was associated with illness. Twenty-five specimens were tested negative for malaria parasites, Salmonella species, brucella species, dengue fever, yellow fever and rift valley fever. Conclusions: An outbreak of suspected AFI occurred in 2 districts affecting primarily males and older age. Having contact history with patient was risk factor to contract the illness. Etiologic agent and source of the outbreak was not identified yet. Large-scale investigation is recommended. P47 Acanthamoeba castellanii. antibody prevalence among diverse tribal Pakistani population Abdul Matin*, Muhammad Ismail, Khalid Mehmood Institute of Biomedical and Genetic Engineering, Islamabad, Pakistan E-mail: [email protected] Retrovirology 2012, 9(Suppl 1):P47 Introduction: Acanthamoeba is opportunistic protozoan pathogen and is known to be one of the most ubiquitous organisms that can produce Page 24 of 54 keratitis and rare but fatal encephalitis. Infections due to Acanthamoeba have increased over the year, which is due to presence of Acanthamoeba in the natural environment and have a direct contact with human in everyday life and is responsible for human diseases. Given the free-living nature of the organisms, it is anticipated that we encounter Acanthamoeba during our normal life. The aim of the present study was to investigate anti-Acanthamoeba antibodies in Pakistani healthy population to combat this pathogen in normal situation. Materials and methods: Acanthamoeba isolation from environmental sources (water, soil and air samples) was done using plating assay. Acanthamoeba identification from environmental samples was based on the morphology of cyst and trophozoite forms by non-nutrient agar plates seeded with E. coli K12 and PCR amplification with a genus specific primer pair. The presence of anti-Acanthamoeba sIgA in mucosal secretions of tribal Pakistani population (saliva were obtained from healthy individuals) were determined using Enzyme-linked immunosorbent assays (ELISA). Results: Acanthamoeba was successfully isolated from the water sources of Pakistan during this study. ELISA demonstrated the presence of Acanthamoeba-specific sIgA in mucosal secretions of in different age groups and both genders. A total of 524 samples of 45 tribes, were collected from different age groups ranged from 15 to 60 years. The overall prevalence was 78.8% in males and 73.8% in females. No significant difference was observed between genders. The high level of anti-Acanthamoeba antibodies was observed among the people in 25-30 years of age. Furthermore the prevalence of antibodies was observed high in tribal population of Khyber Pakhtunkhwa province as compared to rest of the country. Conclusion: Here, we for the first time isolated Acanthamoeba from the natural environment of Pakistan and presented the prevalence level of anti-Acanthamoeba secretory IgA antibody in mucosal secretions of the normal Pakistani population. P48 Study of intestinal parasitic infections associated with HIV infection in Douala, Cameroon Leopold G Lehman, Lafortune Kangam*, Eveline Nguepi, Marthe-Lilianne Mbenoun, Charles F Bilong Bilong University of Yaoundé I, Douala, Cameroon E-mail: [email protected] Retrovirology 2012, 9(Suppl 1):P48 Aim: Gastrointestinal infections are common in people living with HIV. Diarrhea occurred in 30 to 90% of them. However reports on the prevalence of gastrointestinal parasites (GIP) and HIV infection are very few in Cameroon. The purpose of this study was to assess the GIP prevalence in the context of increasing availability of highly active antiretroviral therapy (HAART). Materials and methods: A prospective study was conducted from January to December 2011 wherein stool and blood samples were collected from 201 patients in 3 hospitals of Douala. The stool samples were stained with Kinyoun and Safranin to identify coccidian oocysts. Saline preparations were used to identify ova, cysts and larva. Preparations were observed under UV light microscope CyScope® (Partec Görlitz, Germany). CD4 + T lymphocytes were counted with a flow cytometer CyFlow® (Partec Görlitz, Germany). Chi-square test was used for statistical analyses and P-value <0.05 was considered significant. Results: The global prevalence of intestinal parasites was 27.8%. Seventeen (48.6%) out of 37 patients with diarrhea and 38 (23.2%) out of 164 without diarrhea were parasitized. The most frequent parasites were Candida spp. (13.2%), Cryptosporidium spp. (7.4%) and Entamoeba histolytica/dispar (3%). A significant correlation (p=0.002) was observed between the presence of parasites and diarrhea. The highest parasite counts (p=0.035) and diarrhea (p<0.0001) were found in patients with CD4 + < 200 cells/μl. Srongyloïdes stercoralis, Trichuris trichuira and Isospora belli were only found in diarrheal sample. Conclusions: The overall prevalence of GIP is decreasing in Douala, probably due to the growing avaibility of HAART. This study highlights the importance of looking for intestinal parasite in HIV patient with low immunity presenting with diarrhea in Douala, which is not the case in our hospitals. Retrovirology 2012, Volume 9 Suppl 1 http://www.retrovirology.com/supplements/9/S1 P49 Epidemiology of Hepatitis Delta Virus infection in HIV-infected individuals in Taiwan Hsi-Hsun Lin*, Susan Shin-Jung Lee, Ming-Lung Yu, Bo-Sean Hu, Shiou-Haur Liang, Wen-Chien Ko, Jaw-Ching Wu, Fan-Ceng Zheng, Chung-Hsu Lai, Jin-Long Lin E-Da Hospital, I-Shou University, Kaohsiung, Taiwan, Province of China E-mail: [email protected] Retrovirology 2012, 9(Suppl 1):P49 Introduction: HIV-infected individuals are at higher risk for acquiring HDV. We sought to study the prevalence, genotypes, and associated risk factors causing HDV infection in HIV-infected individuals from an area with high prevalence of hepatitis B virus infection. Materials and methods: A multicenter study of 341 (22.1%) HBsAg+ from 1543 HIV-infected patients was conducted from 2005 through 2011. Blood samples were collected and analyzed for the presence of antibody to HDV and to determine the genotype of HDV. Results: The overall prevalence of HDV infection among HBsAg+ carriers was 54.8% (187/341). However, the prevalence among different risk group was distinct. The prevalence of HDV was 73.6%, 13.5%, and 9.2% among HIV-infected IDUs, heterosexual, and MSM, respectively. The main circulating HDV subtypes in our study were genotype IV (60.5%), genotype II (27.6%), and genotype I (11.8%). Multivariate logistic regression analysis revealed that the major risk factor associated with HDV infection was injection drug use, following by HCV infection, HBsAg titer >=250 IU/mL, and duration of injection drug use. A significant increase of cumulative seroprevalence of HDV with duration of IDU from 1 to 15 years was observed (OR: 1.20, 95% CI: 1.09-1.32, P<0.01). Conclusions: Our study demonstrated high prevalence of HDV infection among HIV-infected IDUs. Effective strategies are needed to prevent injection drug use and to educate ongoing IDUs about the avoidance of practices that lead to infection with HIV, HCV, and HDV. P50 Genetic variability of Hepatitis C Virus in Moroccan population Ikram Brahim*, Abdelah Akil, El Mostafa Mtairag, Régis Pouillot, Abdelouhad El Malki, Richard Njouom, Pascal Pineau, Sayeh Ezzikouri, Soumaya Benjelloun, Salwa Nadir, Rhimou Alaoui Faculty of Sciences Ain Chock Casablanca, Casablanca, Morocco E-mail: [email protected] Retrovirology 2012, 9(Suppl 1):P50 Hepatitis C virus (HCV) evolution is a highly dynamic process. There is little information about molecular epidemiology of HCV isolates in Morocco, an area known for an intermediate prevalence of HCV infection. The primary aim of this study was to determine the subgenotype distribution of HCV strains in patients with chronic HCV infection in Morocco and an eventual association between HCV subgenotypes and liver cancer. The secondary aim was to estimate the prevalence of amino acid substitutions in the HCV core region in treatment-naive patients from Morocco and an eventual association between amino acid substitutions and liver cancer. Serum samples from a total of 185 anti-HCV positive patients were included in this study (81 males and 104 females). The identification of HCV genotype and subtype was respectively performed by sequencing of the 5’UTR and core regions and phylogenetic analysis of the NS5B region. HCV demographic history was inferred using a Bayesian Monte Carlo Markov chain analysis. Of the 174 patients with detectable viremia, the core and the NS5B regions were amplified in 152 (87.4%) and 141 (81.0%) patients respectively. Phylogenetic analysis based on NS5B region revealed that most HCV strains were classified into subtypes 1b (75.2%) followed by subtypes 2i (19.1%), 2k (2.8%). Subtypes 2a, 1a, and 4a were found in a single patient. HCV subtype 1b had an even higher prevalence in liver cancer cases (84.4% vs 67.5% in chronic hepatitis, P= 0.031). Using a Bayesian approach, the mean date of appearance of the most recent common ancestor was estimated to be 1910 for HCV-1b and 1854 for HCV2i. Based on core region, mutations at R70Q or L91M were detected in more than one fourth of patients infected with HCV 1b. Page 25 of 54 P51 Molecular epidemiology and genetic analysis of HCV infection in Cyprus within an intravenous drug user study cohort Demetris C Iacovides*, Johana Hezka, Natasa Savvopoulou, Athos Chrysanthou , Leontios G Kostrikis University, Nicosia, Cyprus E-mail: [email protected] Retrovirology 2012, 9(Suppl 1):P51 Introduction: The HCV genome is highly heterogeneous, owing to the high mutation rate of RNA viruses. There are seven HCV genotypes, with numerous subtypes and viral quasispecies. The prevalence and distribution of HCV genotypes varies globally, and strains show highly variable sensitivity to available therapeutics. Thus, genetic and epidemiological studies of hepatitis C infection are significant, especially within high-risk cohorts where viral evolution can be rapid and unpredictable. Materials and methods: We collected blood samples from intravenous drug users (IVDU) and performed ELISAs to determine HCV positivity. RNA extraction, reverse transcription PCR and DNA sequencing were performed in the Core-E1 and NS5B regions. Strain subtyping was performed using the Oxford HCV subtyping tool v2.0. Phylogenetic analysis was done by aligning and comparing the sequences of both regions to reference strains from the Los Alamos database, using the neighbor-joining method and the Kimura two-parameter distance estimation approach in MEGA v4. The reliability of the phylogenetic clustering was evaluated using bootstrap analysis with 1,000 replicates, and bootstrap values above 70 were considered sufficient for subtype assignment. Results: 21 out of 64 research subjects (32.8%) were positive for HCV, a percentage much lower than the global average (60%) amongst IVDUs. All individuals were infected with genotypes 3a (62%), or 1b (32%), and no unique recombinants were identified. Interestingly, none of the subjects were infected with the 1a strain, a genotype that is highly associated with intravenous drug use, and is also prevalent in the general Cypriot population. In contrast, 38% of the subjects were infected with strain 1b, which is fairly uncommon amongst IVDUs. Finally, we observed three small clusters within the IVDU group, suggesting possible sharing of injecting equipment. Conclusions: HCV infection within IVDUs in Cyprus is polyphyletic, with high genetic heterogeneity, as seen by the limited clustering within this group. Absence of large clusters also suggests that sharing of injecting equipment is uncommon. HCV prevalence amongst IVDUs is significantly lower than the global average, and only circulation of subtypes 1b and 3a is observed, in contrast to the general population where all genotypes are present. P52 Living with hepatitis C, multiple issues to face Mathilde Coudray*, Elisabete de Carvalho Sida Info Service, Montpellier, France E-mail: [email protected] Retrovirology 2012, 9(Suppl 1):P52 Aims: Since 2010, viral hepatitis has been declared a public health priority, although it remains little known to the general population. In France, 220.000 people are positive to the hepatitis C virus (HCV) and live with its chronic form, but limited data exists. This study was undertaken to assess the current face of the epidemic in France, and to identify the profiles of people living with HCV and their specific challenges. Methods: The survey consisted of a questionnaire accessible from November 2010 to February 2011 on the Hépatites Info Service website, a French hepatitis helpline. Callers meeting the criteria could also complete the questionnaire with a professional counselor on the toll-free line. After 3 months, 165 questionnaires had been collected. Results: Participants’ backgrounds are diverse with equal female to male ratio, and an average age of 50. Women are older and more likely to have been infected by blood transfusion prior to 1991 or by medical procedure with early stage diagnosis. Men, however, frequently presume to have been infected through intravenous drug use with late stage diagnosis. For all participants, HCV treatment presents a major challenge. Almost all (96%) undergoing or having concluded treatment emphasized difficult Retrovirology 2012, Volume 9 Suppl 1 http://www.retrovirology.com/supplements/9/S1 side effects: 26% interrupted treatment early and 69% took sick leave. Inadequate health insurance, solitude or addiction problems and coping with the social stigma of HCV infection multiply general difficulties. Conclusion: Living with HCV has major and multiple physiological and psychological consequences, in part treatment-associated. Economic and social precarity and solitude amid a context of discrimination combine to increase the issues confronting HCV positive people. The study has highlighted the diversity of profiles and problems which need to be integrated into a comprehensive care approach. P53 Factors associated with hepatitis C in residents of Sao Paulo, Brazil Norma Farias*, Umbeliana Barbosa de Oliveira, Débora Moraes Coelho, Iára de Souza, Claudia Afonso Binelli Epidemiology at Secretaria de Estado Da Saúde de São Paulo- Brazil, Sao Paulo, Brazil E-mail: [email protected] Retrovirology 2012, 9(Suppl 1):P53 Introduction: Hepatitis C represents a global public health problem. The aim of the present study was to describe the epidemiological profile and to assess exposure variables associated with hepatitis C in residents of Sao Paulo, from the database of viral hepatitis at the National Databank of Major Causes of Morbidity. Material and methods: We analyzed 24,140 cases of hepatitis C notified in residents of the State of Sao Paulo, among the 46,969 bank records of viral hepatitis between 2007 and 2010. Suspected cases of hepatitis C have been confirmed by the presence of HCV RNA using reverse transcriptionpolymerase chain reaction (RT-PCR). The variables selected from the notifications files were sex, age, race, education, number of sexual partners, history of sexually transmitted diseases, HIV/AIDS, intravenous drugs use, inhaled or crack, injecting drugs, tattooing/piercing, acupuncture, blood/ products transfusion, surgical, dental treatment, hemodialysis, transplantation and accidents with biological material. Factors associated with HCV infection were identified with univariate and multivariate Poisson regression and confidence intervals of 95%. Results: The detection rate of hepatitis C was about 15/100000 inhabitants. People aged 50 and over (PR=2,11;95% CI:1,96-2,27), history of blood transfusion (PR=1,41;95% CI: 1,34-1,48), intravenous drugs use (PR=1,33;95% CI: 1,25-1,42), inhalable drugs use or crack (PR=1,27( 1,20-1,35), HIV/AIDS (PR=1,20;95% CI:1,13-1,28), surgical treatment in the past (PR=1,16;95% CI:1,11-1,21) were the main factors independently associated with infection by hepatitis C virus. Conclusions: These findings reinforce the importance of preventing hepatitis C in vulnerable populations such as drug users and the implementation of drugs misuse related harm reduction programs targeted these segments. P54 Low reduction of HCV-RNA level at week 4 in HIV-1 infected patients with acute HCV Infection A Vincent, S Jonckheere*, B Kabamba, JC Yombi, D Wilmes, L Belkhir, B Vandercam Internal Medicine - Infectious Dieases at Cliniques Universitaires Saint Luc, Bruxelles, Bruxelles, Belgium E-mail: [email protected] Retrovirology 2012, 9(Suppl 1):P54 Introduction: The epidemic of acute Hepatitis C-virus infection (AHC) is an on-going issue, especially among MSM patients. Spontaneous HCV clearance rates have been estimated to be between 5-40%, with lower rates among HIV-1 co-infected patients. High rates of sustained virological response have been observed (60-80%) when patients are treated within 24 weeks of AHC diagnosis. Moreover, a spontaneous 2log reduction in HCV-RNA might be a good predictor of spontaneous HCV-RNA clearance in HIV-1 co-infected patients. The aim of this study is to analyse the incidence of 2-log-HCV-RNA spontaneous reduction in AHC in HIV-1 infected patients. Materials and method: We reviewed medical files of 930 HIV-1-infected patients from our Reference Centre. All patients diagnosed with AHC in Page 26 of 54 Table 1(abstract P54) Demographic data (data shown in numbers (%) or median) Population Male sex 9 (100%) Age (median) 44 y BMI (median) Transmission risk-factor; MSM 23 9 (100%) Ethnicity (Caucasian/Black) 9 (100%) HIV results CD4-cells count (median) 565 HIV-RNA (copies/mL) (median) <37 HAART+ 9 (100%) HCV Genotype 1a 7 (78%) Genotype 4 2 (22%) HCV-RNA To (copies/mL) 1049740 HCV-RNA T+4weeks (copies/mL) 552042 - 2 log at T+ 4w 3 (33%) - 2 log at T+ 4w according to genotype 3 (100%) genotype 1a / 0 genotype 4 Clinical features AST (IU/L) T0 (median) 357 ALT (IU/L) T0 (median) ALT (IU/L) T+4w (median) 654 131 2010-2011 were included. Diagnosis of AHC was based on seroconversion and ALT elevation. HCV-RNA PCR data was collected at time of and on week 4 after diagnosis. Results: Eleven patients were included, but only 10 met our diagnostic criteria. One patient was excluded because of missing data. Demographic and clinical data are summarised in Table 1. All patients were MSM. All but 2 patients had genotype 1a HC-virus. On week 4, 3 patients presented with a spontaneous 2log HCV-RNA reduction. All were diagnosed with genotype 1a AHC. Conclusion: Although completed on a small number of patients, this audit suggests that only a low proportion (33%) of HIV-1 infected patients presenting with AHC will meet a favourable 2log-HCV-RNA-drop criteria. This is slightly lower than rates reported by NEAT (i.e. 48%). It implies that these co-infected patients should benefit from a close followup in order to start treatment within 24 weeks of AHC diagnosis, whenever it is needed and available. P55 The results of a study on the prevalence of HIV, HCV and HBV genotypes in some regions of Ukraine Yurchenko, Tetiana Stepchenkova*, Iryna Karnets, Kenneth Ashworth, Tetiana Cheusova Kiev Regional Aids Center, Kiev, Ukraine E-mail: [email protected] Retrovirology 2012, 9(Suppl 1):P55 Background: Ukraine occupies a leading position in Europe on HIV, HCV and HBV infections. 197083 cases of HIV infection and 2000000 cases of viral hepatitis are officially reported among Ukrainians. Specialists of the Kiev regional AIDS center and other regional AIDS centers of Ukraine with the assistance of the “EW BIOPHARMA” company (by order of “Aldima SA”) are conducting a study on genotyping HIV, HCV and HBV. Methods and test-systems: - ELISA testing (Genscreen ULTRA HIV AgAb; Monolisa HCV Ag-Ab ULTRA; Monolisa HBs Ag Ultra) Retrovirology 2012, Volume 9 Suppl 1 http://www.retrovirology.com/supplements/9/S1 Page 27 of 54 - PCR testing (Versant HIV-1 RNA 3,0(bDNA); RMS CAP CTM HCV PCR; Abbott Real Time HCV; AmpliprepCOBAS TaqMan HCV Test; AmpliSense HCV Monitor FRT; RMS CAP CTM HBV PCR) - Genotyping (Trugene HIV-1 Genotype; Versant HCV v2,0 (Lipa); AmpliSense HCV- genotype FL;Quest HBV_Genotype) Duration of a study - August 2011 - present time. All patients gave their voluntary consent to involvement and publication of the results. The study includes patients with HIV, or HCV, or HBV. Coinfected patients, which was confirmed by ELISA testing, were excluded. 200 samples have been tested during 4 mounths of 2011. Among them: - HIV-58 - HCV-121 - HBV-21 In the genotyping of 58 samples of HIV patients were revealed: - M,A -21 (36,2%) - M,B -8 (13,8%) - M-1 (1,7% - no genotype - 28(48,3%) (undetectable viral load) In the genotyping of 121 samples of HCV patients were revealed: - 1a-2(1,6%) - 1b-51(42,1%) - 2a-1(0,8%) - 2ac-1(0,8%) - 3a-35(28,8%) - 4-1(0,8%) - no genotype - 30(25,1%) (undetectable viral load) In the genotyping of 21 samples of HBV patients were revealed: - A-3(14,2%) - C-1(4,7%) - D-11(52,4%) - no genotype - 6(28,7%) (undetectable level of viral load) Conclusion: Among the samples of plasma of HIV patients, the most common genotype is -M,A (36,2%) Among the samples of plasma of patients with HCV, the most common genotype is 1b -(42,1%) Among the samples of plasma of patients with HBV, the most common genotype is D-(52,4%) P56 Changes in immune activation in the T Cell compartments of HIV HCV coinfected patients during PEG IFN RBV treatment Amélie Menard*, Corinne Brunet, Véronique Obry, Patrick Dukan, Sylvie Brégigeon, Olivia Faucher, Estelle Balducci, Anne-Suzel Ritleng, Françoise Dignat-George, Isabelle Poizot Martin Haematology-Aids Unit Hôpital Sainte-Marguerite, Marseille, France Retrovirology 2012, 9(Suppl 1):P56 Chronic activation of CD8 T-cell compartment is critical during HCV-HIV-1 coinfection. The objective of this study was to evaluate the impact of pegylated-interferon (PEG-IFN) in combination with ribavirin (RBV) on immune activation in HCV-HIV-coinfected patients. T cell phenotype (CD8+CD38+,CD8+DR+) was quantified using flow cytometry analysis,. Measurements were performed at day one of treatment (Baseline, BL), then at week (W)12,W24,W48 and W24 posttreatment. HVC viral load was measured using a PCR (COBAS TaqMan 48; Roche), exhibiting a limit of detection at12 IU/ml. Statistical analysis was performed with SPSS 17.0. 11 pts (64% of males; median age 47.4 [45.1-51]) with a median follow up for HCV infection of 14.7y [11.7; 19.1]) were evaluated. All were treated for HIV infection (PI- based regimen: 63.6%) with an HIV-VL < 40copies/ml. Median CD4 and CD8 T cells count at BL was 886/mm3 [671; 1008] and 825/mm3 [530; 1843], respectively. HCV genotype was 1 for 63.6%, 3 in 27.3% and 4 in one pt. HCV VL at BL was 5.9 [4.6; 6.7] (log UI/ml). Up to now, 2 pts stopped HCV treatment at W2 and W4, 9 pts have reached W24, and 5 of them are between W24 and W48. The results at W12 and W24 are presented in the table. We observed a significant decrease of the number of circulating total lymphocytes and CD4T cells in absolute value (p=0.008), but a significant increase in the percentage of CD4+ T-cells and a significant decrease in the percentage of CD8+ T cells at W24. HCV VL was negative for all of them. These preliminary results show that the immune system hyperactivation driving by HCV disease can be reduced with a control of HCV replication. However, we observed a discrepancy in the evolution of CD8+CD38+ and CD8+DR+ expression at W12 which remains at W24.These results have to be confirmed with the next measurement performed at W48 and W24 post treatment. P57 HBV and HCV genotypes distribution on the territory of Belarus EL Gasich, VF Eremin*, SV Sasinovich, MG Tulinova Briem, Minsk, Belarus E-mail: [email protected] Retrovirology 2012, 9(Suppl 1):P57 Materials and methods: 236 HCV and 158 HBV plasma samples collected during 2004-2011 have been investigated. EIA, PCR, RT-PCR, sequencing, SeqScape, BioEdit, MEGA4.1, statistica v.6 software have been used. Results: Of 158 HBV HBsAg positive specimens 17 (10,8%) were from patients with acute hepatitis B and 141 (89,2%) from chronic hepatitis B. Among surveyed were 83 (43,6%) women and 92 (56,4%) men at the age from 15 till 90 years. Middle age has made 44,5±17,8 years. From 158 surveyed 11 persons have been HBV+HBC co-infected and 1 with HBV +HCV+HIV. HBV genotypes has been defined at 61 (34,9%) patient. The phylogenetic analysis of HBV preS fragment has shown that in 52 (85,3%) cases come to light D genotype (D1 – 32.9%; D2 - 26,2%, D3 – 26.2%), in 7 (11,5%) – A (A2) and in 2 (3,2%) – C (C2) genotype. Of 236 HCV infected patients prevailed 1b (53,8%, n=127) and 3a (28,8%, n=58) genotypes. At 30 (12.7%) cases have been revealed 1a genotype and 2a and 2b genotypes have been revealed in 8 (3,4%) and 3 (1,3%) cases accordingly. Conclusion: In the territory of Belarus a genetic variety of HBV and HCV genotypes, caused both circulation before the brought viruses, and new drifts from neighbouring countries, basically from Russia and Ukraine is observed. P58 Rapid cell-free CD4 enumeration using whole saliva Cynthia L Bristow*, Mariya A Babayeva, Rozbeh Modarresi, Carole P McArthur, Santosh Kumar, Charles Awasom, Leo Ayuk, Annette Nhinda, Paul Achu, Ronald Winston Weill Cornell Medical College, New York, USA E-mail: [email protected] Retrovirology 2012, 9(Suppl 1):P58 Introduction: The determination of CD4 counts in patients with HIV/AIDS is of paramount importance clinically to determine when to initiate antiretroviral therapy (ART). ART slows disease progression, reduces viral load and significantly reduces HIV transmission. Currently, the only methods Table 1(abstract P56) BL W12 p W24 p CD4+ T Cell (%) 27.0 [24.5; 37.1] 37.5 [26.8; 46.0] 0.008 43.0 [30.5; 47.3] 0.011 CD8+ T Cell (%) 46.0 [30.4; 51.0] 43.2 [25.9; 49.7] 0.110 37.4 [26.5; 46.9] 0.028 DR+CD8+ T Cell (%) 11.0 [6.4; 19.5] 5.2 [2.8; 9.9] 0.017 2.4 [1.4; 7.4] 0.008 CD38+CD8+ T Cell (%) 14.7 [6.8; 21.5] 43.2 [28.9; 48.9] 0.008 36.2 [15.7; 47.1] 0.015 Retrovirology 2012, Volume 9 Suppl 1 http://www.retrovirology.com/supplements/9/S1 for obtaining CD4 counts use flow cytometry technology. Unfortunately, the latter technologies require fresh cells, reliable and stable electricity, a high degree of technical training for laboratory personnel, expensive instrumentation, and calibrated pipetting which is often unavailable in rural areas or in resource-limited environments. A rapid, inexpensive, point-of-care method would overcome a critical barrier to HIV prevention because it would have a major impact on the cost and health outcomes of patients as well as the frequency of HIV transmission, all of which markedly impact global economy. A commonly used cost-saving and time-saving laboratory strategy is to calculate, rather than measure certain blood values. For example, LDL levels are calculated using measured levels of total cholesterol, HDL, and triglycerides. Thus, identification of cell-free correlates that directly regulate the number of CD4 + T cells could provide an accurate method for calculating CD4 counts due to the physiological relevance of the correlates. We have recently determined that a 1 proteinase inhibitor (a 1 PI, a 1 antitrypsin) participates in regulating the number of CD4+ T cells in blood. Materials and methods: Stimulated saliva was collected from 20 female and 11 male HIV-1 subjects attending clinic for routine care in Cameroon. The a1PI Index was calculated as the ratio of a1PI activity versus protein content in saliva and compared to CD4 counts determined by the standard method, flow cytometry. Results: The a1PI Index in saliva correlated with CD4 counts determined by flow cytometry (r²=0.91, p<0.0001. n=31). An algorithm was developed (the a-test) based on the a 1 PI Index. The precision of the a-test was approximately 26 CD4 cells/ml, and the accuracy of the a-test was approximately 95%. Conclusions: The a-test is physiologically relevant to CD4 counts and can be performed using saliva thereby providing a noninvasive, accurate and precise point-of-care method for monitoring CD4 counts in endemic regions with no instrumentation at a cost-per-test that is less than a dollar. P59 Disclosure of HIV status outcome of regular counseling in a cohort of patients attending HIV clinics Livingstone Ssali1*, Francis Wasagami1, Agnes Kateeba1, Sophie Nantume1, Andrew Kiboneka2 1 The Aids Support Organisation, Taso, Kampala, Uganda; 2Kampala International University Teaching Hospital, Kampala, Uganda E-mail: [email protected] Retrovirology 2012, 9(Suppl 1):P59 Background: The African continent bears the greatest burden of HIV/AIDS in the world. Efforts by scientists to find a vaccine for curing the pandemic have proved futile to date. The prevalence in Uganda stands at 6.4% in Adults and 0.7 % in Children, and about 1.1 million Ugandans are living with HIV/AIDS. The AIDS Support Organization started in 1987. Non disclosure of HIV sero status affects uptake of HIV/AIDS health services, increases stigma and discrimination. Methodology: A retrospective cohort study was done to review records of patients newly registered between January and December 2007 from Management Information System. We analyzed records of patients who had not previously disclosed their HIV/AIDS Sero status at the time of entry into TASO HIV/AIDS clinic. These patients routinely received counseling services for a period of 36 months to assess their disclosure status. Results: Out of 1413 patients counseled, 117(8.3%) patients had not disclosed their HIV sero status, 27(23%) patients were sexually active. During the first individual counselling sessions, patients were given information on condom use 19%, septrine prophylaxis 18%, sexually transmitted infections 15%, family planning 12%, and antiretroviral therapy 9%, safe water 8%, abstinence 5%, life skills 4%, prevention of mother to child transmission 4%, Tuberculosis 4%, voluntary counselling and testing 2%, and faithfulness 1%. Significant number of these sessions focused on opportunistic infections 39%, disclosure 15%, antiretroviral therapy 14%, drug therapy 9%, STD and HIV prevention 5%, nutrition 4% and welfare 4%, discordance 2%, sexuality and abstinence 3%. Other indirect interventions included HIV prevention sensitization through formation of peer support groups, drama sensitizations, group counseling and health talks during clinics. After 36 months of follow-up, 65-56% of 117 patients had disclosed their HIV sero status. Disclosure of HIV status is statistically associated with the number of counseling sessions (p=0.008). Average Page 28 of 54 number of counseling sessions was 6 sessions. Patients who had not disclose after 36 months recorded an average of 3 counselling sessions. Revealing of HIV sero status is statistically associated by sex, more female reveal their HIV Sero status in as short life span compared to males (p=0.002). Conclusions: The number of counseling sessions someone receives is associated with supported disclosure. Female patients reveal their HIV sero status in a shorter time span compared to males. Integration and more frequent provision of counseling services to patients in HIV/AIDS care and treatment enables them to make informed decisions regarding disclosure of their HIV sero status to their family members, sexual partners, friends and others. This has created support systems to clients and therefore reducing further spread of HIV. P60 Economic modeling in HIV for maraviroc in France in treatment experienced patients. Results from the ARAMIS 2011 model Nicolas Despiegel, Felicitas Kuehne, Monique Martin*, Ahmed Shelbaya Vp Uk French Heor Operations at Optuminsight, Uxbridge, UK E-mail: [email protected] Retrovirology 2012, 9(Suppl 1):P60 Introduction: To update an existing and previously published economic (micro-simulation model) in HIV (ARAMIS) to reflect current treatment patterns and to evaluate the cost-effectiveness of maraviroc (MVC) in France. Materials and methods: A systematic literature review was carried out in PubMed to identify all articles published in the past 5 years to provide new data to update the existing model. A total of 1964 abstracts were identified on opportunistic infections (OIs), health consequences (effects on health over the longer term e.g. cancer), costs, quality of life, adherence, resistance and efficacy of treatments. In addition, current guidelines were identified and reviewed. New data were included for OIs, costs, treatments and LT health consequences. Treatments focussed on maraviroc, etravirine (ETR) and raltegravir (RAL) including optimised background therapy (OBT) for treatment-experienced patients at the model’s start. All relevant trials with these agents were identified, data were extracted and used in a metaanalysis (results not provided here) which provided relative efficacy data at 48 and 96 weeks. Treatment algorithms were updated based on guidelines and expert opinion. Costs were at the 2011 level based on official sources. There were insufficient data on adherence to include this in the model. Results: The updated version of the ARAMIS model indicates that MVC compared to RAL or ETR over a life time is associated with more QALYs (a difference of 0.037 and 0.134 respectively) but higher total costs (a difference of €1,439 and €4,766, respectively). The incremental cost effectiveness ratio for MVC compared to RAL or ETR is €39,300 and €35,700. Assuming a threshold of €50,000 MVC can be considered costeffective compared to RAL and ETR. Life expectancy with MVC was similar to RAL (0.37 month difference) and higher than ETR (1.58 months difference). Conclusion: MVC is a cost-effective treatment option for CCR5 tropic treatment-experienced patients in France. P61 Project control- evaluation of a brief HIV counseling video to improve risk reduction behavior of teenagers Yvette Calderon1*, Cheng-Shiun Leu2, Ethan Cowan1, Jillian Nickerson3, Christopher Brusalis3 1 Albert Einstein College of Medicine, Bronx, NY, USA; 2Columbia University, New York, NY, USA; 3Jacobi Medical Center, Bronx, NY, USA E-mail: [email protected] Retrovirology 2012, 9(Suppl 1):P61 Background: This study compared the effectiveness of a brief theorybased, youth-friendly HIV counseling video series with the standard practice (an HIV counselor) in improving risk reduction behavior among teens recruited in an urban Emergency Department (ED). Methods: A two-armed randomized controlled trial was conducted on a convenience sample of 203 non-critically ill, sexually active individuals aged 15-21 in an urban emergency department. Participants in the Retrovirology 2012, Volume 9 Suppl 1 http://www.retrovirology.com/supplements/9/S1 control (counselor) group received HIV information and counseling from a counselor while those in the intervention (video) group watched a series of youth-friendly counseling videos tailored to patients’ stages of change. All participants completed pre- and post-intervention measures on three mediating variables hypothesized to reduce unsafe sexual behavior: condom intention, condom outcome expectancy, and condom selfefficacy. HIV testing was optional for both arms. Results: 203 patients were enrolled and randomized, 102 in the video group and 101 in the counselor group. The groups were similar with respect to age, gender, race, ethnicity, and sexual history. The video intervention performed as well as in-person counseling at improving several condom use measures. The mean difference between groups (video-counselor) in improvement over time (from pre- to postcounseling) in condom self-efficacy was 0.26, CI(0.03,0.50), in male outcome expectancy was 0.15, CI(0.02,0.28), and in female outcome expectancy was 0.20, CI(-0.01,0.40). Participants in the video group improved their condom use intention score significantly more than those in the counselor group, with a mean difference between arms for change over time of 1.02, p-value = 0.01, CI(.24,1.80). The intervention effect on condom intention score did not differ by gender or ethnicity. Conclusions: The use of theory-based, youth-friendly video can be a valid means to provide post-test counseling education and prevention messages within an urban ED. The theory-based prevention messages can improve specific mediators representing risk reduction behavior among teenagers immediately following the intervention. P62 Relationship between health locus of control and sexual risk behaviour Enejoh Aromeh Victor1*, Karick Haruna2 1 Umsom - Institute of Human Virology, Nigeria, Fct Abuja, Nigeria; 2 Department of Psychology University of Jos Nigeria, Nigeria E-mail: [email protected] Retrovirology 2012, 9(Suppl 1):P62 Background: Knowledge of the means of transmission and prevention of HIV/AIDS has been rated as the most important foundational factor for prevention. However studies have also shown that this knowledge does not always translate into reduced sexual risk behaviour (SRB). Perceived control over health status (“internal” locus of control) or attribution of health status to chance or fate (“external” health locus of control), HLC, is a psychological construct that has been shown to impact health outcomes. This study investigated the relationship between HLC and SRB. Hypothesis tested was, “individuals with an external locus of control will engage in more SRB compared with internals”. Methods: A cross sectional survey design was employed with 9 secondary schools in Jos,Plateau StateNigeria. Sample comprised of 361 students in senior secondary school. Health Locus of Control Scale measured attitude regarding perceived control over personal health, with individuals having either an internal or an external HLC while SRB was assessed using the Brief HIV Screener (BHS). Data was analyzed using the SPSS17.0. Descriptive statistics were computed while Analysis of variance was used to determine difference in SRB by HLC categories. Results: Respondents were 169 (46.8%) males and 192 (53.2%) females with a mean age of 16.9, age range 12-24 years. Mean HLC score was 38.54. When grouped into categories, 178 were internals while 183 were externals. Mean score on the BHS was 1.58, with a significant effect of gender. F (1,359) = 8.136, P < 0.01. Research hypothesis was supported; there was a statistically significant effect of HLC on SRB F (1,359) = 4.15, P < 0.05. Conclusion: Respondents who attributed their health status to chance or fate (external locus of control) significantly scored higher on SRB compared with internals. Prevention programs targeted at adolescents should also aim at internalizing their health locus of control. P63 Physical activity particpation and cardiovascular fitness in people leaving with HIV. A one-year longitudinal study S Fillipas*, FM Cicuttini, AE Holland, CL Cherry Physiotherapist at the Alfred, Melbourne, Australia E-mail: [email protected] Retrovirology 2012, 9(Suppl 1):P63 Page 29 of 54 Introduction: Physical activity (PA) and cardiovascular fitness (CVF) are beneficial for HIV-infected individuals, however long-term effects are unknown. This study aimed to document long-term habitual PA and CVF in stable, HAART-treated individuals with HIV, explore relationships to body composition, body image and cardiovascular disease (CVD) risk and evaluate PA determinants. Materials and methods: This was a 12-month prospective, longitudinal cohort study. Eighty ambulant HIV-infected adults (aged ≥18 years) on HAART were recruited from The Alfred Hospital Infectious Diseases clinic and local HIV community clinics. PA was reported using the International Physical Activity Questionnaire and CVF assessed using the Kasch Pulse Recovery Test. Results: 19-37% participants reported suboptimal PA levels at each study visit, while PA and CVF were largely stable over the study period. Higher CVF was associated with better body composition and this association persisted over time (p=0.05). Greater total energy expenditure was associated with improved body image (r=-0.325, p=0.027) but notCVD risk. At baseline, the proportion of subjects in a permanent relationship was higher among active versus inactive participants [47.7% versus 13.3% (p=0.032)]. A similar trend was seen at six and 12 months. Conclusions: In this stable, HAART-treated HIV–infected cohort a suboptimal level of PA participation was observed. CVF was associated with improved body composition, suggesting HIV- infected individuals should be encouraged to improve and maintain CVF. Increasing PA levels were associated with improved perceived body image, supporting use of PA to improve this aspect of psychological well being. Being in a permanent relationship was associated with higher PA levels suggesting that social isolation may be a risk factor for inactivity in those with HIV. Further work, including larger cohorts and longer follow up is needed to explore factors that influence PA and CVF in HIV. This study found benefits for both longterm PA and CVF for chronic HIV-infection however intervention studies are required to define the benefits obtainable for improving long-term PA uptake and CVF in this population. P64 Consistency between clinician and patient perspectives on ARV treatment commencement and switching examined Jeffrey Grierson*, Marian Pitts, Rachel Koelmeyer la Trobe University, Melbourne, Australia E-mail: [email protected] Retrovirology 2012, 9(Suppl 1):P64 Introduction: In the context of multiple HAART treatment modalities it is critical that common understandings regarding the motivators and barriers to treatment commencement and switching are shared by both prescribing clinicians and PLHIV. Materials and methods: We conducted an online survey of 254 people living with HIV (PLHIV) in Australia and structured interviews with 18 clinicians (HIV S100 prescribers). PLHIV had a median age of 47.5 years. Overall, 87.4% of respondents were currently taking ARV; 5.5% had taken ARV in the past but not currently, and 7.1% had never taken ARV. Clinicians were a mix of high caseload experienced practitioners and newer low caseload clinicians. Results: When we examined the motivations for and barriers to treatment commencement, PLHIV identified concerns about potential side effects and the psychological consequences of acknowledging potential physical decline. Clinicians were most likely to assess readiness in terms of ability to maintain adherence. PLHIV relied on the clinician initiating and advising treatment commencement. Clinicians identified process of clinical assessment, ARV education and adherence training as the antecedents of treatment initiation. Similarly with switching treatments, PLHIV identified concerns about side effect profiles, clinical markers and the potential limiting of treatment options. Clinicians identified resistance patterns, regimen potency and adherence concerns as primary motivators for treatment change. PLHIV generally expected any discussion of switching to be initiated by the clinician. Clinicians discussed the process in terms of resistance testing and patient education around treatment options. Both PLHIV and clinicians identified the relationship between them as the critical component of the processes of commencing and changing ARV Retrovirology 2012, Volume 9 Suppl 1 http://www.retrovirology.com/supplements/9/S1 treatment. Clinicians were keen to involve PLHIV in the decision-making process and PLHIV had a high degree of trust in their clinicians’ knowledge and judgement. Conclusions: Treatment discussions need to occur frequently. There is evidence of resistance on the part of patients to change unless they are experiencing significant side effects. Clinicians may focus unduly on adherence while PLHIV are more concerned about side effects and disease progression. The valued relationship between clinician and PLHIV provides a fertile basis for improved treatment discussion. P65 Drivers of treatment change in PLHIV- psycho-social factors are more important than considerations of adherence Marian Pitts*, Jeffrey Grierson, Rachel Koelmeyer la Trobe University, Melbourne, Australia E-mail: [email protected] Retrovirology 2012, 9(Suppl 1):P65 Introduction: There are increasing options available for the effective management of HIV. Treatment regimens have become simpler and more manageable since the introduction of HAART fifteen years ago. In this context it is important to understand how commonly PLHIV switch treatment s and how the decision to change treatment is negotiated between clinician and patient. Materials and methods: We conducted an online survey of 254 people living with HIV (PLHIV) in Australia. PLHIV had a median age of 47.5 years. Overall, 87.4% of respondents were currently taking ARV; 5.5% had taken ARV in the past but not currently, and 7.1% had never taken ARV. Results: Two-thirds of respondents had changed ARV at some point: 14% in the last 12 months. Respondents last changed treatment a median of 3 years ago. Most respondents (90%) had not used more than six combinations in their lifetime. Of those who had changed treatment, the most significant driver of the decision was the advice of the physician (76%) the second most important was the side effects of the previous regimen (49%). Fatigue/ loss of energy was the most prevalent side effect experienced by 74% of all respondents. It was also rated as the most bothersome on the ACTGHIS scale. Aspects associated with adherence were the lowest rated drivers of treatment change. Only 7% of respondents identified problems with taking doses at the correct time, and less than 5% reported missing doses. Given that the doctor’s advice was the most important driver of treatment change we elicited the important characteristics of the clinician- patient relationship using free text. The doctors’ interpersonal skills and personality was most frequently identified (by 62% of respondents). This was more than double the next most frequently mentioned characteristic (competence- 37%). Open ended contributions from participants also highlighted the importance of open communication and a robust relationship with the clinician. Conclusions: A focus on adherence and neglect of relational, psychosocial and somatic factors in assessing motivations for treatment switching is likely to compromise robust patient-clinician relationships. P66 A multidisciplinary staff for the optimisation of therapy in HIV-infected patients treated for cancer Julie Daigre1*, Meriem Mendjel1, Sylvie Bregigeon2, Catherine Tamalet3 1 APHM La Timone, Laboratoire de Pharmacocinétique et Toxicologie, Marseille, France; 2APHM Sainte Marguerite, Service d’Immunohématologie clinique, CISIH, Marseille, France; 3Pole des Maladies Infectieuses et tropicales Clinique et Biologique, Fédération de Bactériologie Hygiène Virologie, Marseille, France E-mail: [email protected] Retrovirology 2012, 9(Suppl 1):P66 Introduction: With the improvement of HAART, life expectancy of HIVinfected patients sharply increased. The incidence of cancerous diseases is therefore increasing in this population. This raises the problem of drugdrug interactions between HIV treatment and cancer chemotherapy poorly Page 30 of 54 understood because few data are available. However, the potential risk of interactions is important because of the involvement of similar metabolic enzymes and transporters between these drugs and due to the inhibitory and/or inducer effects on different CYP450 isoforms of the PI and NNRTI. These interactions may both lead to inefficiency and/or an increased risk of toxicity of the chemotherapy which can be life-threatening for the patient. Materials and methods: We set up since January 2010 amultidisciplinary staff “AIDS-cancer-transplantation” to optimize the management of HIVinfected patients diagnosed for a tumour disease. Demographic, virological and immunological characteristics and therapeutic decision issued from the staff for these patients have been retrospectively analyzed. Results: As of today, data of 28 patients have been evaluated. A therapeutic adjustment had to be proposed for 14 patients (50%), mainly corresponding to a modification of the antiretroviral therapy (11/14). The new cART therapeutic option was defined according to HIV resistance profile, ART history and after additional biological analyzes if requested. In few cases (4/14), the cancer chemotherapy was adjusted according to the different therapeutic options proposed. The main reasons for such therapeutic adjustment were, for 57% (8/14), related to a metabolic interaction through the CYP3A4 because of the presence of a boosted PI or NNRTI and for 36% (5/14) to an increased risk of nephrotoxicity due to the concomitant administration of tenofovir with a potent nephrotoxic anticancer drug. For all patients, opportunistic infections prophylaxis (PCP) and screening for PCR CMV have been prescribed according to French recommendations. Conclusion: The implementation of the multidisciplinary staff “AIDScancer-transplantation” highlights a high frequency of the risk of drug interactions between antiretrovirals and antitumoral agents. This allows us to optimize the management of HIV patients treated for malignant diseases by apprehending the risk of drug interactions. P67 Correlates of HIV Stigma in prison guards and case workers in a national prison system Carmen E Albizu-García*, Cynthia Pérez-Cardona, Salvador Santiago-Negrón School Public Health University Puerto Rico, San Juan, Puerto Rico E-mail: [email protected] Retrovirology 2012, 9(Suppl 1):P67 HIV is over-represented in correction facilities. Health organizations have provided a framework for effective response to HIV/AIDS in prisons yet many countries lack prison-based HIV prevention and treatment. Prison workforce’s fears of HIV contagion and stigma have been identified as barriers to appropriate responses. We explore the magnitude of stigma towards HIV in correctional officers and case workers in prison system across workforce categories, sex, and HIV knowledge. A self-administered, anonymous questionnaire was completed by 95 case workers and 214 correctional officers from 26 of the 39 penal institutions in Puerto Rico (PR) selected multi stage random sample design. Stigma towards HIV was assessed with a Spanish measure of stigma towards HIV among health professionals containing 18 items measured with 6 categories agreement scale. Predictor variables include demographics, work experience in corrections, and HIV knowledge. The study was approved by the IRB. We report the scale’s reliability and the weighted mean and standard deviation of responses to the stigma measure for the total sample and by workforce category, sex, and HIV knowledge. Of the 309 respondents 58% were male and 68% report 5 or more years iof employment. Scale reliability as measured with Cronbachs alpha was 0 .81. The values of the stigma towards HIV measure ranged from 18-108, with higher scores corresponding to greater stigma. The mean of the stigma score for the total sample was 69.5 (SD 15.6). The means did not differ significantly when compared for correctional officers vs. case workers or by sex, yet an inverse relationship was found between stigma and HIV knowledge. Stigma scores fell within the second highest quartile suggesting that HIV stigma is high in this sample. The availability of a culturally sensitive instrument in Spanish contributes to work in the field and the findings underscore the need to address stigma towards HIV in the prison workforce in this and other contexts. Research should be conducted on the effect of HIV knowledge gains on stigma reduction in this population. Retrovirology 2012, Volume 9 Suppl 1 http://www.retrovirology.com/supplements/9/S1 P68 Knowledge, risk perceptions and attitudes of nurses towards HIV in a tertiary care hospital in Mangalore, India Anand Venugopal*, A Basavaprabhu, B Unnikrishnan Radiology at Kasturba Medical College Hospital, Mangalore, India E-mail: [email protected] Retrovirology 2012, 9(Suppl 1):P68 Introduction: Infectious diseases like HIV are on rise in developing countries like India which puts heavy burden on health care needs. Nurses have a key role and spend considerable time taking care of HIV positive patients admitted in hospitals. Hence a study was conducted in our hospital to have an insight into their knowledge about HIV, their apprehensions while taking care of such patients and their attitudes and willingness to take care of them. Methods: It’s a cross sectional study done among 200 nurses of KMC Hospital, Mangalore. They were given validated questionnaire comprising of 67 items which included knowledge of spread of HIV, universal precautions, risk perceptions and their attitudes towards HIV positive patients. Their responses were analysed using SPSS software. Results: Of the 200 Nurses selected, 152 completed the questionnaire. Regarding knowledge of HIV transmission, the correct response for widely advertised modes of transmission were higher-sexual contact 97.4%, vertical transmission 88.8%. However, 11.2% did not know about mother to child transmission and 28.9% about transmission by breast feeding. 90.1% felt HIV could be transmitted by sharing of plates and 83.6% felt by mosquito bites. 93.4% knew about universal precautions and 78.3% routinely practiced it. 80.3% were aware of post exposure prophylaxis.79.2% described caring HIV Positives as rewarding,86.5% were willing to assist operations on hiv patients and 84.9% were willing conduct deliveries. 13.4% felt they have right to refuse caring HIV patients and 97% felt surgical patients need to be routinely tested for HIV. Conclusions: This study demonstrates that there are deficiencies in knowledge about HIV and false beliefs exist among nurses about spread of HIV. There is need to improve awareness about HIV and HIV patient care through training programmes to clear misconceptions amongst Nurses so that HIV positive patients are not discriminated against and are treated without discrimination. P69 Problems of HIV positive tuberculosis patients’ health care in Latvia Ludmila Viksna*, Inga Januskevica, Valentina Sondore, Baiba Rozentale, Ilze Eksteina, Andrejs Ivanovs, Tatjana Kolupajeva, Vija Riekstina Department of Infectology and Dermatology at Riga Stradins University, Riga, Latvia Retrovirology 2012, 9(Suppl 1):P69 Introduction: The number of patients with HIV/TB alliance has increased recently. There were 1055 TB/HIV cases registered in European Union (6% out of 17 650 TB cases tested for HIV) in 2010, among them 71 in Latvia (9.5% out of all TB cases in 2010, in comparison with 0.5% in 2000). The treatment for TB/HIV cases is provided in Latvia according to WHO recommendations, including DOTS strategy. However the TB treatment results for HIV positive cases are concerning due to increasing resistance to HIV/AIDS drugs and to TB drugs. Methods: The treatment results for 7761 new smear and/or culture positive pulmonary TB cases were analyzed, 234 among them were HIV positive. In HIV/AIDS naive patients in general the resistance is found in 5.3% of cases. In treated patients the resistance to different groups of drugs is detected in 41% of cases: to all -3%, to NRTI – 14%, to NRTI+NNRTI – 9%, to NRTII+PL – 6%, to NNRTI – 6%, to NNRTI+PI – 1%, to PI – 2%. Multidrug resistance (MDR) was diagnosed in 14% of TB cases. Molecular biological, immunofluorescence, bacteriological and bacterioscopic methods were used for detection of initiating agents. The TB treatment results for HIV positive and HIV negative cases from the period 2000-2007 were compared with the results reported in 2010. Data for 2000-2007 was grouped together due to small number of HIV positive cases within the period. Results: The treatment success for HIV positive cases was lower in both periods (60-61%) in comparison with HIV negative cases (78-76%). The level of MDR TB cases among HIV positive patients was higher (15%) than Page 31 of 54 in HIV negative patients (8%) during the years 2000-2007, but similar in both groups (9-10,6%) in 2010. Despite improvements in HIV/AIDS treatment since 2000, the death rate among HIV positive cases in 2010 was higher (18%) than during the years 2000-2007 (11%). Conclusions: The resistance to HIV/AIDS drugs is a factor influencing the TB treatment results. P70 One body, one test, two lives.. patient centered strategy to increase HIV testing in pregnant women and their partners Larisa Kudryashova-Hernandez Ancillary HIV Services at Neighborhood Health Services Corporation, Plainfield, USA E-mail: [email protected] Retrovirology 2012, 9(Suppl 1):P70 Background: NHSC, an urban community-based health center in New Jersey, USA, provides prenatal services, labor/ delivery to 750 uninsured/ impoverished/minority women annually. Given that NJ has the third highest number of HIV women in USA and the highest number of HIV children, early HIV detection/ intervention in pregnant women become paramount. NHSC historically struggled with sub-optimal OB HIV testing rates (60%) and needed to make radical program changes to comply with CDC recommendations to ensure HIV testing is offered to 100% pregnant patients. Methods: PDSA (Plan-Do-Study-Act) was conducted to test a new HIV testing approach: HIV Counselors are located in OB department; HIV counseling/ Rapid testing is done at OB registration; daily registration schedules are available to HIV Counselors; HIV results become part of OB records upon result availability; educational DVDs are utilized in patient areas to increase awareness/ interest. Results: Per PDSA-improved strategy, NHSC sustained 100% compliance with CDC recommendations over the last three years. Rapid HIV testing and Rapid-on-Rapid positive result confirmation allow for smooth/timely transition from HIV testing to care/treatment for newly diagnosed HIV pregnant patients/partners. Conclusions: The collected/analyzed data suggests that coordinated, patient-centered approach helps to: identify HIV positive pregnant patients in the first/second trimesters; immediately connect them to prenatal/HIV care to minimize vertical HIV transmission; provide prevention/treatment for partners including prevention for positives. P71 Scaling up HIV treatment-Karnataka, India experience Salma Fahim1*, Suresh Shastri1, Reynold Washington2 1 Karnataka State Aids Prevention Society, Bangalore, India; 2Karnataka Health Promotion Trust, Bangalore, India Retrovirology 2012, 9(Suppl 1):P71 Background: Karnataka a high HIV prevalence state in south India is home to 10 percent India’s HIV infected. The Government of India sponsored HIV treatment program was initiated on 1st April 2004. A systematic approach to scale up of the ART services is followed in Karnataka where Government is the lead agency to implement the program and all the donors/NGOs compliment the program. Methods: A donor supported technical consultant was identified and located within the State AIDS Society. A logistic Management Information System is established to track procurement, distribution and supplies. Human resource recruitment, training and retention at the ART centres are decentralized to the district AIDS prevention and control units (DAPCU). Centralized classroom based training is complemented with field based onsite training and mentorship. A Google group is created for technical updates. DAPCU conducts coordination meetings to geographically distribute and allocate responsibility to all field level workers in HIV prevention and care programs to minimize loss to follow up. Results: By Sept 2011, 189,179 persons living with HIV (PLHIV) are registered at ART centres and 64,104 are currently on ART. This was possible thanks to a scale up in the numbers of ART centres from 17 to 44 and CD 4 testing machines from 5 to 32 during the period 2008-2011. The proportion of PLHIV detected at Integrated Counseling and Testing Centres and registered in ART centres rose from 46% to 96% in this period. 122 link ART Retrovirology 2012, Volume 9 Suppl 1 http://www.retrovirology.com/supplements/9/S1 centres are established to decentralize drug distribution for those stable on ART. Lost to follow up among those on ART reduced from5.5% to 3.5%. However, death rates among those on ART remain high at around 17 %. The commonest cause of death is TB (21%), while unknown remains high (26%). Conclusions: Despite a rapid scale up, loss to follow up of those initiated on ART has been significantly reduced and stabilized. This was possible because of coordination between government and civil society partners. The high death rates indicate the need for better integration between HIV and TB programs, strengthening clinical competencies, laboratory diagnostic facilities for opportunistic infections and operations research. P72 Acute retroviral syndrome mimincs dengue in Singapore, a dengue endemic country JH Tan*, A Verrall, S Archuleta Infectious Diseases at National University Singapore, Singapore, Singapore E-mail: [email protected] Retrovirology 2012, 9(Suppl 1):P72 Introduction: The differential diagnosis for dengue-like illnesses is broad. However, World Health Organization (WHO) recommended confirmatory testing may not always be feasible. Acute human immunodeficiency virus type-1 (HIV-1) presents similarly to dengue and is an important diagnosis that may be missed when dengue confirmatory testing is not performed. Methods: Cases of acute HIV-1 in adults >15 years diagnosed between January 2010 and December 2011 were identified through HIV service records at our institution. Medical records were reviewed for admitting diagnosis, clinical and laboratory features. Results: Eight patients with acute HIV-1 were identified in the study period, seven of whom were clinically diagnosed as dengue and met the WHO dengue case definition. The duration of illness and clinical features were indistinguishable from dengue: fever, myalgias, nausea and rashes. Five had thrombocytopenia and three had leukopenia. During the same period, emergency physicians at our hospital assigned a dengue diagnosis using prespecified ICD-9 based coding for 276 persons over 15 years old. The ratio of acute HIV-1 cases to those with presumed dengue was 0.025. Conclusion: Acute HIV-1 infection can mimic symptoms of dengue fever. This case series adds to the single case report in the published literature. The ratio of 0.025 is concerning as not all patients with presumed Figure 1(abstract P73) Page 32 of 54 dengue undergo confirmatory testing. This high rate of acute HIV-1 among dengue-like presentations needs further prospective investigation. Physicians in endemic areas like Singapore should test for acute HIV-1 in cases of presumed dengue where dengue confirmatory testing is negative. P73 Integral HIV-STI diagnosis at the first VCT visit as strategy to prevent late ART initiation in the HIV-AIDS program of Mexico City (HIVPMC) L Juárez-Figueroa*, A González-Rodríguez, E Rodríguez-Nolasco HIV Laboratory at HIV-Aids Program of Mexico City, Mexico City, Mexico E-mail: [email protected] Retrovirology 2012, 9(Suppl 1):P73 Introduction: HAART and laboratory monitoring of PLWHIV are available in México. Nevertheless, lack of opportune detection impeded early ART initiation, thus favoring HIV transmission and increased incidence of morbidity and mortality due to AIDS conditions before ART starts. Methods: On 2010 HIVPMC started faster HIV/STI diagnosis at first VCT visit combining a rapid HIV test and parallel blood analysis with automated HIV 4th generation/STI serology (Abbott Architect). Since fall 2011, initial CD4 counting (Becton Dickinson FacsCalibur) in HIV+ clients, also done at the first visit, favoured a rapid HIV infection staging. On 2012 HIV viral load analysis (Abbott RT-PCR), required in Mexico for ART initiation, was added at the first VCT visit. Results: The integral HIV/STI diagnosis at the first VCT visit reduced dramatically the time elapsed before, between the first HIV detection with a rapid test or ELISA and the completion of laboratory studies necessary for starting ART. The attendance of vulnerable groups to VCT at Condesa Clinic scaled up with 51% detection increment during 2011. A 60% of new detected PLWHIV did not return for follow up and treatment as shown by the national HIV data base SALVAR. Figure 1 shows the distribution of new infections on 2011 (UNAIDS model). Conclusions: Integral HIV/STI diagnostic self promoted the HIV VCT increasing VCT demand while reducing desertion of number new patients. This model should be expanded to HIV clinics in 31 Mexican States. Depending on the size of the population to be serviced a combination of serial HIV rapid tests, ELISA and simplified point of care CD4 counting could be used. Retrovirology 2012, Volume 9 Suppl 1 http://www.retrovirology.com/supplements/9/S1 The quantity of non-returning patients highlights the need of education oriented to people at risk of HIV infection while individual post-test counseling/accompanying could be also individually addressed. P74 Cadillac care for the poor... addressing local health emergency around STDs and HIV in an urban community-based medical home Larisa Kudryashova-Hernandez Ancillary HIV Services at Neighborhood Health Services Corporation, Plainfield, USA E-mail: [email protected] Retrovirology 2012, 9(Suppl 1):P74 Background: NHSC, an urban community-based health center in Plainfield, Union County, NJ, USA, provides services to 25,000 uninsured/minority/ impoverished patients. Plainfield consistently ranks first and second among the 29 Union County municipalities for Syphilis, Gonorrhea and Chlamydia. Plainfield ranks second for the numbers of HIV/AIDS. There is, therefore, a dire need to address the existing health emergency around STDs and its correlation with HIV. Methods: NHSC incorporates a coordinated, proactive, patient-centered approach to integrating STD screening/prevention with primary care in a medical home environment. Risk assessments/screenings are done by clinicians. Those identified suspicious for STD or with STD symptoms/ diagnosis receive on-the-spot HIV counseling and Rapid testing. Patients receive immediate treatment intervention for suspected STDs. There are ongoing prevention efforts including development of Risk Reduction Plans agreed upon/signed by patients. Results: Resulting from the integrated STD/HIV prevention approach: 100% of patients presenting with symptoms/suspicion for STDs received HIV counseling/Rapid testing; 100% received prevention education and free condoms; 100% had Risk Reduction Plans developed and agreed upon; 19 persons were identified STD and HIV positive and were immediately linked to care. Conclusions: Integration of STD/HIV prevention with outpatient care under the umbrella of Early Intervention Services allowed to: identify extend of STD/HIV correlation; allow for a seamless one-stop shop preventiontreatment service delivery model; improve patient awareness of on-site prevention/treatment resources. P75 Access to HIV treatment and care in Armenia S Grigoryan*, A Mkrtchyan National Center for Aids Prevention, Yerevan, Armenia E-mail: [email protected] Retrovirology 2012, 9(Suppl 1):P75 Introduction: ART has become available in Armenia since 2005. To evaluate progress in the response of the National Programme to PLHIV’s needs since last evaluation time, UNAIDS country office requested WHO/Europe HIV/ AIDS treatment and care expert for a country mission. The purpose of the mission was to evaluate access of PLHIV to HIV/AIDS treatment and care and other related services. Materials and methods: Prior to the mission some background documents had been reviewed. During the mission key-informant interviews were conducted with a range of stakeholders. Also, a focus group discussion was conducted with PLHIV representing two NGOs. Visits were made to a number of institutions and activities in Yerevan, the capital. Results: As of end 2010, cumulative 971 HIV cases were registered by the National Center for AIDS Prevention (NCAP), 231 persons died. Of 740 PLHIV 542 (73%) were seen for HIV care in 2010. Armenia has accepted a higher threshold of ART initiation (CD43), as recommended by recent WHO guidelines. There is no waiting list for starting ART. By the end 2010, 250 PLHIV (46% of seen for care) were on ART. Adherence to ART is pretty high. Of all 294 PLHIV ever started ART, only 44 (14.9%) dropped it out. However, more than a half of them (25) died, due to late presentation. About a quarter of newly identified HIV patients had CD43at the time of diagnosis. Conclusions: NCAP has demonstrated high coverage with HIV care and ART. PLHIV are satisfied with the range and quality of services they receive. However, low level of CD4 cell count in more than a half newly diagnosed Page 33 of 54 HIV cases (57.7% PLHIV had CD4 <350cells/mm3 at time of diagnosis) in combination with the highest proportion of PLHIV identified due to clinical symptoms compared to other reasons for HIV testing (39.8%), indicates late presentation and raises concern of the health system’s inefficiency to diagnose HIV at early stages and thus to early enroll in HIV care. The expert has made appropriate recommendations to improve early HIV detection for optimal care and patients’ survival. P76 MMT of HIV positive patients in Georgia Khatuna Todadze*, Eka Kavtiashvili Research Institute On Addiction, Tbilisi, Georgia E-mail: [email protected] Retrovirology 2012, 9(Suppl 1):P76 Introduction: The main route of HIV transmission is injective drug use in Georgia (58-60%). The most popular injective narcotics are opioids. Although prevalence of HIV among drug users is only 1-3%, the high number of IDUs and high prevalence of hepatitis C (from 65 to 80% according to the different studies) in this population could be the predictor of HIV increase. Methadone maintenance treatment (MMT) has been implementing throughout the country since 2005 as one of the important strategies to decrease drug related risky behavior, increase adherence to ARV treatment among HIV+ persons and improve the physical and psycho-social status of the patients. Materials and method: 42 randomly selected HIV positive drug users undergoing MMT with intensive psychological counseling have been studied for 3 years. They received ARV therapy before inclusion in MMT at least 6 month. Risky behavior, quality of life, level of depression, anxiety and other data were measured before starting MMT and after 3, 6, 12, 18 months. The illegal use of psychotropic-narcotics was checked through random urine-testing 3 times per patient per month. Results: The study showed significant improvement of patients’ status. The remarkable decrease of depression and anxiety was observed (dynamic of average scores of depression - 24, 14, 14, 13, 14 and anxiety-46, 40, 40, 41, 39). Life quality increased in comparison with the starting data (76, 85, 86, 88, 93). The positive answers on psychotropic-narcotics were observed in 6.7% on average and even those patients didn’t admit any kind of injectionrelated risky behaviors. Conclusions: The analyses of data showed that combination of MMT, ARV and psychological counseling significantly improves the physical and psycho-social status of HIV positive IDUs, improves life quality and treatment adherence , dramatically decreases use of illegal psychotropic-narcotic drugs and decreases the risk of spread of HIV and other blood-transmitted diseases among injecting population in Georgia. P77 AIDS patients family knowledge and behavior toward their patients before and after counseling Behnam Honarvar Communicable Diseases at Shiraz University of Medical Sciences, Shiraz, Iran E-mail: [email protected] Retrovirology 2012, 9(Suppl 1):P77 Background: This study was aimed to measure the effect of ongoing counseling program at behavioral counseling center (BCC) of Shiraz, southern Iran on changing the knowledge, attitude and behavior of AIDS patients’ family members toward them. Methods and materials: 125 members of HIV/AIDS patients’ family were interviewed individually by filling questionnaire before and after performing counseling for them. The findings were analyzed in SPSS. Results: The age of the participants was 40±13 years. Forty four percent had spousal relationships with their patients. Their knowledge about the main routes of HIV transmission were 9.76±2.59 and10.64±0.88 before and after counseling, respectively (P=0.028). Supportive behaviors of families toward their patients reached to 79 % after counseling compared with 44% before that (P=0.004). Belief to isolate the patients and the practice of this approach at home dropped from 71% to 15% and from 29% to 7% after counseling, respectively (P<0.05). In 30% of participants fear of getting HIV from patients was not changed by counseling, and 24% of patients’ spouses Retrovirology 2012, Volume 9 Suppl 1 http://www.retrovirology.com/supplements/9/S1 did report to avoid protected sex with their HIV infected husbands even after taking part in the counseling program (P>0.05). Conclusion: Ongoing counseling for HIV/AIDS patients’ families at BCC of Shiraz did advance their knowledge about AIDS and improved their attitude and behavior toward their patients. However, in some aspects such as the removal of fear about HIVspread in the family or the change of the patients’ wives attitude to have protected sex with their HIV infected husbands, the counseling program did not show remarkable success. P78 Pregnancy and contraception - the perspective of HIV-positive and negative women Chinedu O Oraka*, Tochukwa S Egbunu, Samuel C Ani Build Africa Research Capacity, Lagos, Nigeria E-mail: [email protected] Retrovirology 2012, 9(Suppl 1):P78 Objective: To understand pregnancy intentions and contraception knowledge and use among HIV-positive and negative women in the prevention of mother-to-child transmission (PMTCT) program. Design: A cross-sectional survey of 236 HIV-positive and 162 HIV-negative postpartum women interviewed within 12 months of their expected delivery date in a public-sector health facility providing PMTCT services. Methods: Bi-variant analyses explored fertility intentions, and family planning knowledge and use by HIV status. Multivariate analysis identified socio-demographic and service delivery-related predictors of reporting a desire for additional children and modern family planning use. Results: HIV-positive women were less likely to report wanting additional children than HIV-negative women (8 vs. 49%, P < 0.001), and although a majority of women reported discussing family planning with a health worker during their last pregnancy (HIV-positive 79% vs. HIV-negative 69%, P = 0.0), modern family planning use remained low in both groups (HIV-positive 43% vs. HIV-negative 12%, P < 0.001). Condoms were the most commonly used method among HIV positive women (31%), whereas withdrawal was most frequently reported among HIV-negative women (19%). In multivariate analysis, HIV-negative women were 16 times more likely to report wanting additional children and nearly 85% less likely to use modern family planning. Women who reported making two or less antenatal care visits were 77% less likely to use modern family planning. Conclusion: Our results highlight success in provision of family planning counseling in PMTCT services. As family planning use was low among HIVpositive and negative women, further efforts are needed to improve uptake of modern methods, including dual protection, in the PMTCT settings. P79 The relation between hemispheric lateralisation and measures of immune competence and adherence in Human Immunodeficiency Virus Type 1 (HIV-1) Rachel C Sumner*, Alexander V Nowicky, Andrew Parton, Carolien Wylock, Renata Cserjesi, Patrick Lacor, Yori Gidron Brunel University, Manchester, UK E-mail: [email protected] Retrovirology 2012, 9(Suppl 1):P79 Introduction: Communication from the brain to the immune system is influenced by hemispheric lateralisation (HL). Left-HL is immunopotentiating, right-HL is immunosuppressive. Only one study has examined the effects of HL on the progression of HIV (Gruzelier et al., 1996). That study included a small sample with very little control over third variables. The present study tested whether left HL predicted higher CD4 and CD8 levels, statistically controlling for confounders. Methods: Employing two neuropsychological assessments of HL (line bisection task and Zenhausern’s Hemispheric Preference Test), 69 HIV-1+ patients were followed prospectively. Numerous exclusion criteria and confounder assessments were employed (e.g., age, sex, mode of contraction, medication adherence) to provide a more rigourous and controlled analysis. Results: The present work corroborated the theory of asymmetrical influence on HIV immunity by HL via a moderator: ethnicity. The main Page 34 of 54 analysis of the research findings did not attain statistical significance in the whole group of patients. However, among African patients, left-HL predicted better immunity, while no such relationship was seen in European patients, independent of confounders. Further observations were made between HL and HIV-relevant behaviours. Left HL was related to higher number of sexual partners in Europeans. A near-significant relationship was observed between left HL and longer periods between HIV clinic attendances in Africans. Conclusions: The present study adds new information concerning a moderating factor of the HL-immunity relationship in HIV. As expected, left-HL predicted higher CD4 and CD8 counts, but only in African patients. Further, the added methodological and statistical control employed, extend the validity of the HL-immunity relationship. Moreover, the present study has uncovered behavioural implications of HL in HIV disease. Potential explanations for neurobiological pathways in the relationship between HL and immunity are discussed. P80 Antiretroviral therapy (ART) adherence and correlates to non-adherence among people on ART in Estonia Anneli Uuskula1, Kaja-Triin Laisaar1*, Mait Raag1, K Rivet Amico2, Anjali Sharma3, Jack DeHovitz3 1 Department of Public Health, University of Tartu, Tartu, Estonia; 2Center for Health Intervention and Prevention, University of Connecticut, Storrs, CT, USA; 3Division of Infectious Diseases, State University of New York Downstate Medical Center, Brooklyn, NY USA E-mail: [email protected] Retrovirology 2012, 9(Suppl 1):P80 Introduction: There is limited data on antiretroviral therapy (ART) adherence among patients in Eastern Europe, despite the high incidence of HIV infection and the growing number of HIV-infected individuals who are being prescribed ART. We conducted a study to measure rates of adherence to ART and factors associated with non-adherence among patients receiving care at an outpatient HIV clinic in Estonia. Materials and methods: The study was based on cross-sectional data from a convenience sample of 161 patients receiving outpatient HIV care. Data were obtained via interviewer administered surveys and data abstraction from clinical records. Adherence was measured from 3-day patient selfreport. Results: Among the 161 participants (mean age 33 and 55% male / 45% female), two thirds (64%) had been infected with HIV through intravenous drug use. Most (74%) were co-infected with hepatitis C (HCV). Perfect adherence over the last 3-days was commonly reported [87% (95% CI 80 − 92%)], with non-perfect adherence associated with greater concerns about the potential negative consequences of taking ART [AOR 5.8 (95% CI 1.3 − 45.7)] and fewer antiretroviral medications (ARVs) in one´s current regimen (2 or fewer different ARVs vs 3 or more different ARVs: AOR 17.0 (95% CI 3.7 – 97.6). Conclusions: Self-reported ART adherence in this sample of Estonian HIVinfected patients in clinical care was similar to rates observed in Western Europe and other developed countries. The results suggest that adherence education and support could be most beneficial, if specifically targeting the development of positive beliefs, reduction of negative expectations towards ART and when helping patients manage negative treatment experience, particularly with regimens including multiple ARVs. P81 HIV-AIDS stigma and discrimination in health care sector in Belarus Vera Ilyenkova1*, Svetlana Kunitskaya2, Irina Eramova3 1 World Health Organisation, Regional Office for Europe, Communicable diseases, Minsk, Belarus; 2Belarusian State Medical University, Public Health, Minsk, Belarus; 3World Health Organisation, Regional Office for Europe, Communicable diseases, Copenhagen, Denmark E-mail: [email protected] Retrovirology 2012, 9(Suppl 1):P81 Background: Stigma and discrimination are barriers to effective HIV treatment and care in Belarus. Results of the Stigma Index Survey conducted among people living with HIV (PLHIV) reveal that 40.5% of respondents Retrovirology 2012, Volume 9 Suppl 1 http://www.retrovirology.com/supplements/9/S1 experienced disclosure of their diagnosis and confidentiality breach by health care workers (HCW); 15.5% were refused medical care. Methods: Public health department of Belarusian State Medical University and NGO “Fialta” investigated the extent and possible reasons of HIVrelated stigma in health sector, by conducting a survey on knowledge, misconceptions, attitudes and motivations regarding HIV/AIDS among 40 HCW not routinely involved in HIV-care. This was followed by two one-day sensitizing workshops addressing Belarusian law on HIV/AIDS, emphasizing medical rules/regulations, patients’ rights and confidentiality, stigma and discrimination, including sensitizing role plays. Results: Though the sample size was small, research findings indicate needs to address levels of stigma and discrimination among HCW in Belarus: a quarter of respondents are not willing to provide services to PLHIV at all; more than half are inclined to violate patients’ rights and test patients for HIV without informed consent; majority revealed some negative stereotypes in regards to PLHIV that lead to discrimination outside their professional duties (for example, changing HIV-positive hairdresser, advising their children to reduce contacts with HIV-positive schoolmate); about 2/3 would isolate PLHIV if they are not their friends or relatives. Possible reasons from workshops’ findings: HCW don’t realize what stigma is and don’t know their actions/attitudes are discriminative; lack of knowledge about HIV-infection resulted in fear of contact with PLHIV; negative stereotypes regarding PLHIV exist among population as they “traditionally” represent risk-groups (injection drug users, commercial sex workers, etc.), thus, “they are immoral and dangerous”. Conclusions: Introducing stigma-reducing interventions (sensitizing workshops, educational briefings) in health sector would help to improve the situation. Conducting in-depth interviews among HCW is necessary to analyse stigma-related issues more thoroughly. P82 Decentralizing treatment services with link ART centres- experience from Karnataka, South India Salma Fahim1*, Suresh Shastri1, Bharat Rewari2 1 Karnataka State Aids Prevention Society, Bangalore, India; 2National AIDS Control Organisation, India Retrovirology 2012, 9(Suppl 1):P82 Background: Prior to 2007-08, ART services in Karnataka state, India were delivered through ART centres, located mainly in medical colleges, and tertiary and district hospitals. This led to high rates of defaults, increased travel time and cost and loss of working hours. To make treatment services more accessible to PLHIV, link ART centres (LACs) were developed at subdistrict primary and secondary care levels, co-located with HIV voluntary counselling and testing centres. Methods: After a thorough needs assessment based on existing ART and HIV testing data which included load in ART centre, HIV prevalence, distance and accessibility, sites were identified, health personnel trained, and patients sensitized about the centres. Results: Currently there are 122 LACs attached to 29 nodal ART centres in the state. The LACs provide services to 5,498 PLHIV.Default rate has come down from 3.5 % in ART centres to less than 0.5% in LACs.The average patient travel distance has declined from 70 to 30 kms, saving travel cost and time. Waiting times for refilling prescriptions have declined from four hours to one hour, which has improved patient adherence. Health care staff at primary and secondary level care centres are gradually taking the lead in HIV care and treatment service delivery. Conclusions: The establishment of LACs at primary and secondary care levels has helped improve drug adherence and service access. They save travel cost and time, and more importantly, help strengthen primary and secondary health care services, which is beneficial for program sustainability. P83 An assessment of cultural values contributing to antiretrovial therapy adherence among patients on antiretroviral therapy adherence among patients Kaona AD Frederick*, Sikaona Lenganji, Miti Esnart, Siziya Seter Mwengu Social and Health Research Centre, Ndola, France E-mail: [email protected] Retrovirology 2012, 9(Suppl 1):P83 Page 35 of 54 Background: The study was conducted in 2007, in the Northern Province of Zambia, to assess the impact of cultural values contributing to ARV non-compliance among elderly patients afflicted by AIDS. Providing care for sick child, loss of income by the family, caring for the orphans, seeking treatment were studied. Parental experiences and impact of caregiving during the time of child illness that led to poor compliance are examined. Methods: A community-based survey was conducted in three randomly selected catchment areas of Nakonde, where 250 out of 682 patients receiving ARVs within the twelve months period, were recruited through the District’s Health Management Board Zonal Health Centre. All patients were interviewed using a pre-tested structured questionnaire, consisting of: Sociodemographic characteristics, Socio-economic factors, Knowledge about HIV transmission and prevention. Cultural beliefs regarding ARVs in Zambia and the social taboos surrounding treatment and actual drug adherence were observed. Results: Most male AIDS patient respondents tended to be older and more educated than the female patient respondents. Overall, 32.6% of the patients stopped taking their medication. There were 49.1% of the females and 23% of the males, who reported that AIDS patients stopped taking their medication within the first 4 months of commencing treatment. Age, marital status and educational levels were not significantly associated with compliance. The major factors leading to non-compliance included patients beginning to feel better (55% and 48.6%), lack of knowledge on the benefits of completing a course (25.7%), running out of drugs at home (25.4%) and ARVs drugs too strong (20.1% and 20.2%). There was a significant difference [OR = 1.66, 95% CI 1.23, 2.26] in HIV knowledge, with more males than females reporting shaking hands as a means for HIV transmission, after adjusting for age, marital status and educational levels. Conclusion: This study established that 32% of AIDS patients failed to comply with AIDS drug taking regimen once they started feeling better. Providing means and knowledge for primary caregiver to administer effective drugs and ARVs or to prevent and treat opportunistic infections, TB and pneumonia, would help reduce caregivers distress. P84 Concomitant sexually transmitted diseases in patients with newly diagnosed HIV in Sri Lanka SD Dharmaratne*, K Buddhakarale National STD AIDS Control Program, Sri Lanka E-mail: [email protected] Retrovirology 2012, 9(Suppl 1):P84 Introduction: Diagnosing and treating STI are important both for the health of HIV infected persons, their sexual partners and for HIV prevention efforts. STI following an HIV diagnosis or STI co-infection can serve as a surrogate indicator of continued risk behaviors. Objective: To determine the prevalence of concomitant STI in newly diagnosed HIV positive patients. Method: Data was extracted from randomly selected individual clinical notes of patients who are attending the Central HIV clinic, Colombo. Results: A total of 187 subjects were included to the study of which 106 (57 %) were males and 80 (43 %) were females. The mean age of the sample was 37 years (SD =9.3) and the median CD4 count at diagnosis was 320 cells/μl. Routine STI screening had been done only in 90.3% (169) of the sample. Among the screened subjects for STIs, over one in every four (28%) had an STI. Ulcerative and non-ulcerative STIs were present in 18.2% and 10% of subjects respectively. Syphilis was positive in 9.5% (N=16), symptomatic Herpes simplex infection in 7.7% (N=13), non gonoccocal urithritis /cervicitis in 6.5% (N=11) and symptomatic genital warts in 3% (N=5). There was a statistically significant association between the number of life time partners and STI prevalence (P=0.011) and sexual orientation and STI prevalence (P=0.005). In contrast gender, marital status and age did not show any statistically significant association with STI prevalence. Conclusion: Significant numbers of HIV patients are co-infected with at least one ulcerative or non-ulcerative STI at the time of their HIV diagnosis which enhances HIV transmission. Therefore, careful and continued screening and management of STIs in HIV positive patients is an important HIV prevention intervention. Retrovirology 2012, Volume 9 Suppl 1 http://www.retrovirology.com/supplements/9/S1 P85 Surgical dental implants in people living with HIV-AIDS A Sparaco, M Ghezzi*, G Donati, K Andriella, A Montebello, C Luraghi, G Romanoni, V Rania Dentistry at Luigi Sacco Hospital, Milan, Italy E-mail: [email protected] Retrovirology 2012, 9(Suppl 1):P85 Introduction: The introduction of HAART has considerably improved life expectancy and quality of life of people living with HIV/AIDS (PLWHA). If in the first years HIV infection was considered an absolute contraindication to implantology, it is now possible to employ implants positioning which allows a more complete functional and aesthetic rehabilitation of the oral cavity also in these patients; howewer there is still fear and prejudice in using this technique in PLWHA. We present the experience of Department of Dentistry of Luigi Sacco Hospital of Milan in over twenty years of implant surgery to evaluate the possibility of using implantology techniques in PLWHA without exposing them to greater risks of developing infections both during and after surgical intervention. Materials and methods: The study considers a consecutive series of 21 HIV-positive patients (for a total 80 implants) and 91 HIV-negative patients as control group (for a total 245 implants), treated in the dental clinic of the Luigi SaccoHospitalfrom 1998 to december 2011. The pre-surgery phase included a collection of anamnesis, clinical examination data, diagnostic radiology evaluation and assessment of blood tests. We used the surgical technique of submerged fixture implants and mobile and fixed prostheses for the final prosthetic rehabilitation. Results: Several failures occurred in both groups and were attributable to local factors related to receiving bone or to exceedingly invasive surgical techniques. We detected no lesions in the oral cavity in these subjects concomitant with the plants loss nor changes in their overall health conditions. Conclusions: Patients enrolled in this study presented both functional and aesthetic dental problems. The comparison between the success/failure rates in the two groups shows that implant surgery can be employed without risk for the patient and with success rate comparable to the general population, nevertheless it is important to assess the level of immune competence of the patient. Finally, the prosthetic rehabilitation of the oral cavity, in addition to the clear local benefit, has an important psychological effect on patients and on your quality of life. P86 Improve the quality of counseling for HIV in the district of Guédiawaye Sakho D Maty*, Seck Karim, Ouattara Baly, Gaye Alioune, Sylla Bintou Médecin Spécialiste du Vih at District Sanitaire de Guédiawaye Synergie pour l’Enfance, Dakar, Senegal E-mail: [email protected] Retrovirology 2012, 9(Suppl 1):P86 Objectives: HIV counseling and testing for several years working in the district of Guédiawaye. A situation analysis has revealed shortcomings in the organization of the service given the many other activities of the health center. This is why the medical team decided to use the collaborative model of quality improvement that is based on the implementation of a package of changes and the measurement of indicators of improvement. Methodology: It consisted of a situational analysis of the Board Search, the development of process maps and identify targets for improvement. Improvement targets were selected to make the results of all HIV tests to clients within two hours, to provide the reference of all patients positive for medical care. The changes have been to extend the opening hours of the laboratory, to assign a specific staff for testing and to ensure continuous availability of a pool of consultants. Better communication was established and the customer’s circuit has been reduced. Results: From January to December 2010, the proportion of tests performed in two hours has risen by an average of 35% to 99%, also the proportion of patients actually referred for medical care rose from an average of 33% to 97%. Page 36 of 54 Conclusions and recommendations: The process of improving not only affects the indicators of improvement but the effects on teamwork and a consideration of the basic needs of patients. P87 Estimated numbers of cases with HIV from 1990 to 2010 in Fars province by capture recapture method M Marzban*, H Joulaee, P Kazeroni Epidemilogy at HIV Reseach Center, Shiraz University of Medical Science, Shiraz, Iran E-mail: [email protected] Retrovirology 2012, 9(Suppl 1):P87 Background: HIV is potential risk in different countries of the world. According to UNAIDS, estimated HIV-infected people in Iran 4 times more than the cases have been recorded. Full details of the cases with HIV is one of the most essential tool for planning. The aim of the study is to estimate numbers of infected patients during the selected time. Method: All information contained in three sources hospital, voluntary counseling and testing center (VCT) and prison from 1990 to 2010 were used. Then by record linkage identify common cases finally log-linear methods were fit for eight models. Result: 5167 cases entered to the study. The proportion of men 10 times women also most patients belong to the age group 15 to 44 years old. The main source of recorded cases is voluntary counseling and testing center by 3347 patients. As expected the lowest source is the common cases between prison and hospitals. annually average of 550 patients have been added. The lowest estimate belong to the common cases between prison and hospital but most of them are the presence of three sources. Estimated number of cases in the province are approximately 18914. Conclusion: Low coordination between the various components of the health system, along with social issue such as stigma and discrimination of HIV positive patients is the most important weakness in information systems. Therefore constitutional arrangements for providing high quality information is essential for HIV-infected cases and should be the priorities of health policy investment. P88 TB and HIV among hill tribe marginalized vulnerable population, Thailand Tawatchai Apidechkul School of Health Science, Mae Fah Luang University, Chiang Rai Province, Thailand E-mail: [email protected] Retrovirology 2012, 9(Suppl 1):P88 Introduction: Since 1982, Thailand had been reported 372,874 cases of HIV/AIDS, and 98,153 deaths. The north of Thailand has been report as the highest prevalence areas. There were almost 600,000 hill tribe populations live there as a marginalized and vulnerable people under lacked of access to health care and limited education. Most of them emigrated from China last 200 years ago. Chiang Rai Province is the most favorite living area of hill tribe people. Materials and methods: The retrospective cohort study design aimed to investigates the TB and HIV situation among hill tribe marginalized and vulnerable population. The systematic data collection with the completed questionnaire was conducted in the 12 hospitals, Chiang Rai Province. All questionnaires had been tested for reliability and validity before use. Survival and Cox’s regression were analysis. Results: Of 629 cases of TB reported during 2009-2011 form 12 hospitals were recruited into the study. 60.7% were male 23.8% aged 51-60 years old, and followed by 41-50 years old (20.2%) (min=1, max=93). Of 84.6%were pulmonary TB and extra pulmonary 15.4%, 44.6%recieving AFB testing, 77.4%new cases. The results of treatment found that 22.7% were cure, 28.6%complte, 4.3%defults, 8.1%death, and 1.9% failure. Prevalence of HIV/ AIDS among hill tribe TB cases was 17.2%. Of 88.1% had treatment on CAT1, and 4.6%CAT2, and 2.3%CAT4. Male had greater pulmonary TB (pvale=0.044), and HIV+ than female (p-value=0.023). Survival analysis found that being male (p-value=0.01), non-HIV (p-value<0.01), and CAT1 had Retrovirology 2012, Volume 9 Suppl 1 http://www.retrovirology.com/supplements/9/S1 greater success treatment. Cox’s regression found that only aged 11-20 years old had related to success treatment (HR=2.11, 95%CI=1.05-4.26). Conclusion: Active screening program and increasing the rights of access to care are immediate needed for hill tribe vulnerable people for coping TB problem in Thailand. P89 HIV epidemic in Bulgaria Simon Shamas Merical University of Sofia, Sofia, Bulgaria E-mail: [email protected] Retrovirology 2012, 9(Suppl 1):P89 Background: To identify the groups of highest risk of infection and understand the HIV dynamic in Bulgaria. Methods: By analyzing data presented by ‘’The National Center of Infectious and Parasitic Diseases’’ about HIV/AIDS reported cases between 1986 and 2011. Results: During the period from 1986 to 2011 the number of registered patients is 1438 people. Last year the number of registered cases was 166, from which 81% are men. The annual number of officially registred HIV-infections increased from 49 in 1986 to 127 in 2007. We observe a development of HIV concentrated epidemics among the highest risk groups. Between 1986 and 2011 109 infections were due to Injecting drug use (IDU), the prevalence rate in this group was increased from 0.97% in 2004 to 7.29% in 2011. During the same period in the group of homosexuals the prevalence rate was increased from 0.99% to 1.6%. Romanies people are considered as a risk group because of their low social and economic status. Their prevalence rate reaches 2,9%. The newly diagnosed HIV-infections among prisoners in bulgarian jails increased 7,4 times between 2004 and 2007. In 2010 the lowest age limit of infection reached 15. Discussion: In Bulgaria HIV prevalence among the general population is below the national average for the countries of the EU. However, the country is facing a serious challenge in the form of rapid development of concentrated epidemics among some groups at higher risk. In order to protect the people there are some steps that should be taken as in motivating the groups of highest risk to use the services of voluntary free testing and counseling for HIV, rapid expansion of testing services through a network of Cabinets for anonymous and free counseling and assuring the access to knowledge about HIV/AIDS. P90 Procalcitonin as a marker of bacterial sepsis in immunocompromised patients Lana Gatserelia*, Lali Sharvadze, Marine Karchava, Nino Babridze, Tengiz Tsetvadze, Natia Dvali, Lela Dzigua, Nika Chxartishvili Head of Virology Lab. at Infectious Diseases, Aids and Clinical Immunology Research Center, Tbilisi, Georgia E-mail: [email protected] Retrovirology 2012, 9(Suppl 1):P90 Introduction: Procalcitonin (PCT) is a recently described marker of severe sepsis. It was decided to assess the value of PCT as a marker of secondary infection in patients infected with HIV in Georgian AIDS Center. Materials and methods: PCT plasma levels were measured by quantitative assay BRAHMS-Biomérieux using the VIDAS analyser in a prospective study in 135 HIV-infected individuals: 87 asymptomatic and 48 with lever or suspected secondary infections. Results: The baseline plasma level of PCT was (0.5 ng/ml +/- 0.5), even in the latest stages of the disease, and did not differ from the values of healthy subjects (0.54 ng/ml +/- 0.1). EDTA-treated whole blood was collected from patients before starting specific antimicrobial therapy. No elevation of PCT level was detected in HIV-infected patients with evolving secondary infections including PCP (n = 4), cerebral toxoplasmosis (n = 5), viral infections (n = 9), mycobacterial infections (n = 4), localized bacterial (n = 13) and fungal infections (n = 4), and in various associated infectious and non-infectious febrile events (n = 15). All these plasma values were lower than 2 ng/ml. In contrast, high PCT plasma levels were detected in one HIV-infected patient with a septicaemic influenza infection (17 ng/ml) and another one with a septicaemic Pneudomonas Page 37 of 54 aeruginosa infection (46 ng/ ml), PCT values decreased rapidly under appropriate therapy. Conclusions: We found that PCT is a specific marker of bacterial sepsis in HIV-infected patients, as no increase in other secondary infections could be detected in those patients. A rapid determination of PCT level could be useful to verify or refute bacterial sepsis for a better management of febrile HIV-infected patients. P91 The medicalization of disease as a factor of abandonment and dissatisfaction with HIV treatment services Carmen Rodríguez Reinado*, Teresa Blasco, Hernández Jesus, Nzang Esono University of Huelva, Huelva, Spain E-mail: [email protected] Retrovirology 2012, 9(Suppl 1):P91 Introduction: Medicalization of the disease is a social phenomenon not exclusive of industrialized societies. In countries with medium and low human development indexes and developing economies, this process is prevailing due to the expansion of the biomedical model. This is the case of many African countries and more concretely in Equatorial Guinea. Methodology: Qualitative research, based on the application of semistructured interview (30) as the technique for gathering information. Ambit: Bata, Equatorial Guinea. Purposeful sampling: homogeneous type by subgroup; 1. HIV-positive people who abandoned diagnostic and treatment services. 2. HIV-positive people who are utilising HIV treatment services. Location: General Hospital and Outpatient Treatment Centers. Unit of analysis: utilization of diagnostic and treatment services. Method of analysis: Grounded Theory Method. Data triangulation: internal by two observers and theoretical. Results: - Drugs are a central topic on the discourse about the disease. - Regardless of the population profile, all the informants share a medicalized view of HIV and its treatment - In the asymptomatic phase of HIV, avoiding to prescribe drugs reinforces the process of non-acceptance of the disease leading the patient to question his positive diagnosis. - Within the profile of HIV-positive respondents still in treatment, medication is the element of the care process that takes on more importance. Avoiding to prescribe drugs is an element of dissatisfaction with health services. - In the profile of HIV-positive respondents who abandoned HIV diagnostic and treatment services, no prescription drugs was one of the reasons for abandonment. Conclusions: The medicalization of the disease has affected the social construction of HIV as a disease and is one of the reasons for abandonment and dissatisfaction with the care received in diagnostic and treatment centers. It therefore represents a factor for intervention and to modify in order to reduce the rates of abandonment for such services. P92 Group psychotherapy for HIV patients. A different approach Michele Battuello*, Paolo Roma, Giovanna Celia University of Rome Sapienza, Italy. Sant’Andrea Hospital, Rome, Italy E-mail: [email protected] Retrovirology 2012, 9(Suppl 1):P92 Introduction: HIV is often related to psychological distress, after the diagnosis and the beginning of HAART. Brief Psychological approaches are important but they give only support, enhancing the relationships of the patients. In many cases these relationships are disfunctional too. Psychotherapies don´t focus on the indidvidual´s autonomy but on the support from the others. Objectives: A brief-grouppsychotherapy focused on major objectives of each person but with the main common objective to enhance the psychological individual indipendence and to promove a self- maturation; understanding the disfunctional dynamics realized in the past that are enhanced by the HIV-status, to promove individual indipendence with the objective of focusing the good and valid relationships and change the disfunctional ones and to allow the person to be able to open to the world again. Retrovirology 2012, Volume 9 Suppl 1 http://www.retrovirology.com/supplements/9/S1 Method: A brief group psychotherapy, supportive but mostly expressive. a small group 3 male, 1 female: were choosen for group psychotherapy after 2-3 individual meetings. 16 psychotherapy meetings, weekly, lenghth 1 hour 40-45 minutes. Results: First time patients worked on their disfunctional affective part, that was pre-hiv. In a second time they worked promoting their possibility of indipendence to find their lost self esteem. in the last time they focused to improve their relationships where possible but mostly to think that they can go out alone from the darkness after HIV diagnosis. Conclusions: Quality of life of HIV patients can be focused on changing their disfunctional parts, first enhancing the process of indipendence and individual esteem of the person. P93 Late detection of schizophrenia patient VIH-VHC Karine Bartolo*, Nathalie Labrune, Isabelle Jaquet Chu Sainte Marguerite, Marseille, France E-mail: [email protected] Retrovirology 2012, 9(Suppl 1):P93 Introduction: After several years, some patients on TSO, became custumers of BZD (40 tablets per day). During the maintenance with these patients, we can notice that this consummations are rythmed by periods where there is a new outbreak of productives symptoms. Auditory and visual hallucinations with influence syndrom, persecution syndrom, tense boarder, social isolation, peculiarity contact, mutilations to-himself, disjointed thought. These symptoms can’t be withdrawal symptoms or a decrease of their consummation. Methode: Case report: HDM: He is man, 41 years, co-infected patient (VIH and VHC). ATCD: He’s poly-addict with BZD in first line drug. Symptoms: - persecution - tense boarder - suicidal ideas - decrease motivation - decrease memory - violent acting Treatment and observance: He was having TSO from ten years (buprenorphine 16 mg per day) and a large number of hospitalisation before starting the psychiatric disease treatment. When the diagnosis was established, the treatment LAAA is administrated and the patient progressively gave up the BZD consummation. The symptoms progressively disappeared. The only time he was hospitalised is because the withdrawal of BZD was too violent. Diagnosis: The diagnosis of schizophrenia with deficit disorders was imput to this patient when the symptoms are stand out ,specialy , when he had no drugs: the BZD, taken in exes. Results: We observe that the number of hospitalisations has decreased from 2 since the begining of the new long acting atypical antipsychotics (LAAA). Conclusion: We must make a difference between psychiatrics symptoms and associated drugs withdrawal clinic signs when we consider poly-drug users who are partially stabilized by a TSO. P94 HIV pre-exposure prophylaxis (PrEP)- knowledge and attitudes among a New York City emergency department patient population Yvette Calderon1*, Jason Leider1, Ethan Cowan1, Christopher Brusalis2, Joanne Mantell3, Theo Sandfort1 1 Albert Einstein College of Medicine, Bronx, New York, USA; 2Jacobi Medical Center, Bronx, New York, USA; 3Columbia University, New York, New York, USA E-mail: [email protected] Retrovirology 2012, 9(Suppl 1):P94 Background: HIV Pre-exposure Prophylaxis (PrEP), in which HIV-negative individuals receive antiretroviral medications to prevent HIV acquisition, has shown potential as a means to reduce HIV incidence among high-risk Page 38 of 54 persons. The acceptability of PrEP among at-risk persons will strongly impact the effectiveness of PrEP. This study aimed to assess knowledge and attitudes towards PrEP within a demographically-mixed community with high HIV prevalence. Materials and methods: A cross-sectional study was conducted with a convenience sample of Emergency Department (ED) patients at two New York City municipal hospitals. Eligible participants completed an anonymous written survey about knowledge and acceptability of PrEP. Means and standard deviations were calculated for continuous variables and proportions for categorical variables. Standard bivariate methods were used to compare acceptability and knowledge by race, ethnicity and gender. Results: 474 ED patients agreed to participate. The study population was 40.9% male, 40.7% Latino and 38.2% non-Hispanic Black. 7 participants (1.5%) self-identified as MSM. Mean age was 35.3, SD +/- 13.1 years. 66.4% reported inconsistent condom use and 78.4% had previously had an HIV test. 13.3% reported knowledge of either the term “PrEP” or the use of antiretroviral medications to prevent HIV acquisition. More people indicated they were unlikely or extremely unlikely to use PrEP (40.1%) than indicated they would likely take PrEP if available (32.2%). Many (27.7%) were unsure if they would or would not take PrEP. 44.4% thought that individuals would stop using condoms if on PrEP, while 27.0% thought that individuals would continue using them. Some participants (28.4%) incorrectly thought that PrEP needed to be taken only prior to sex. There were no differences in knowledge or acceptability of PrEP by gender. Latinos were more likely (17.6%) than blacks (8.8%) and others (12.4%) to report knowledge of PrEP. Conclusions: Potential providers of PrEP must consider limitations in acceptability to this HIV prevention strategy. Future administration of PrEP must incorporate patient education to ensure user understanding of the technology and its potential limitations. P95 HIV experts on the decision to use early ART for prevention in France: are we there yet? Bertrand Lebouché1, Kim Engler1*, Joseph Josy Lévy2, Gimore1, Bruno Spire3, Willy Rozenbaum4, Jean-Pierre Routy1 1 Mcgill University Health Centre, Montreal, Canada; 2Université du Québec à Montréal, Canada; 3INSERM-SESSTIM UMR912, Marseille, France; 4Hôpital Saint-Louis, Paris, France E-mail: [email protected] Retrovirology 2012, 9(Suppl 1):P95 Introduction: The finding that successful antiretroviral therapy (ART) can almost eliminate the risk of heterosexual HIV transmission is the scientific breakthrough of 2011 according to Science Magazine. This potential of ART has generated novel approaches to prevention including “Test and Treat” (T&T). Our qualitative study, drawing on the perspectives of French HIV experts, aims to better understand if, and how, a T&T approach might be applied in France and to generally learn more about concerns raised by prevention with early ART. Materials and methods: In 2011, 19 French HIV experts participated in a semi-structured interview on implementing T&T in France. Expertise was typically defined on the basis of contribution to the 2010 French ART guidelines (Rapport Yeni 2010). Participants’ HIV expertise included clinical care, epidemiology, virology, and community activism. Interviews lasted an hour, on average, and explored opinions on T&T, who, how and whether to test and treat, and what public health discourses and evaluations should accompany it. Analyses of interview content pertaining to who or whether to treat early for prevention are presented here. A content analysis of the transcribed interviews was supported by the software Atlas.ti version 5.2. Results: The decision to treat earlier than current guidelines recommend (CD4 >500) was generally associated with uncertainties and involved weighing the risks and benefits, whether potential or known, primarily in terms of patient health, the risk of transmission, patient choice, population health and/or cost. Perspectives on each of these aspects varied as did experts’ position on treating early for prevention. Conclusions: Despite ART-based HIV prevention’s status as evidence based medicine (EBM), French experts had uncertainties about early preventive treatment of HIV-infection which often translated into a weighing of benefits and risks to varying effect. Our findings suggest fostering a culture of dialogue and debate between care providers, community organizations Retrovirology 2012, Volume 9 Suppl 1 http://www.retrovirology.com/supplements/9/S1 and recipients of care on the complex considerations of implementing strategies of ART as prevention in France can help translate EBM into improvements in health and new HIV guidelines towards the eradication of HIV. P96 The research on female partners of injecting drug users-gender aspect and risk of HIV Miljana Grbic*, Verica Lela Ilic, Sladjana Baros, Vesna Ciprus, Jelena Tadzic, Rade Grbic, Gordana Jurican, Milan Parlic, Svetomir Samardzic Unaids Focal Point for Serbia at Undp, Belgrade, Serbia E-mail: [email protected] Retrovirology 2012, 9(Suppl 1):P96 The goal of this research is to assess whether there is an increased risk of HIV infection based on gender among female partners of IDUs and to what degree is this risk present. The research has been conducted using quantitative and qualitative methodology. For the purpose of collecting the research data among the female partners of IDUs and male partners who are IDUs, a qualitative methodology was used (in-depth interviews) complemented with a quantitative methodology (survey). Within the research 50 male injecting drug users (IDUs) have been reached and 50 steady female partners of the injecting drug users, regardless of the fact whether they themselves use drugs or not. Research findings indicate the following: Gender aspect is very important in initiation into drug use. Girls and women often start with drug use practice with their partner. Out of 19 IDU males who are in a relationship with a female partner who is an IDU, eight male subjects reported that their female partners had the first contact with drugs through them. On the other hand, out of 50 male participants, only one had the first experience with drugs during a relationship with a woman IDU. Sharing of injecting equipment is still present . In most cases a male partner has priority in distribution and taking drugs. Traditional understanding and acceptance of gender roles is reflected in attempts to explain or “justify” a man’s violent behaviour with the withdrawal crises or need for drugs. Sex work is one of the frequent activities for procuring money, or sex services are offered to drug dealers in exchange for drugs. The decision on purchasing and using a condom is mostly left to a male partner, and women and girls are not enough empowered to impose the use of a condom. Men are more prone to changing partners. Women prove their devotion to a partner by agreeing to risky behaviour (for ex. intercourse without a condom) or even by willingness to get infected with HIV. They tend to remain in a relationship with a person who is HIV positive, even when that person has infected them. In conclusion, the research findings have to a great degree confirmed gender specific risks for women that have been stated in the professional literature: unequal power relations and often weaker economic position of a woman. Women are in a weaker position when they make a decision about when, with whom and under what conditions they would have a sexual intercourse. The research findings, although they are not representative due to the size and sampling method, are certainly indicative – confirm to a great degree the thesis about the specific vulnerability of women and girls, both related to situations that lead to HIV infection, and related to availability and specific content of services offered to women living with HIV. P97 Impact of educational program on knowledge, attitude and preventive behaviors related to HIV STIs in female sex workers in Shiraz south Iran Mahmood Amini Lari*, Minoo Ali pour Sakha, Farbod Ebadi Far Azra, Parvin Afsar Kazerooni, Mehrab Sayadi Shiraz HIV/AIDS Research Center, Shiraz, Israel E-mail: [email protected] Retrovirology 2012, 9(Suppl 1):P97 Background: Female sex workers (FSW) have been well identified as core groups that play an important role in the effluence of sexually transmitted diseases (STD) and HIV. The purpose of present study was to assess the impact of educational intervention on promoting STIs /HIV related knowledge, attitude and preventive behaviors among female sex workers In Shiraz Iran in 2009. Page 39 of 54 Methods: In this quasi-experimental study with a pre-test -post-test design which was conducted from February to July 2009, 80 female sex workers were selected from 5 Shiraz Drop in Centers using the classified random sampling technique. At the beginning FSWs´ HIV/STI related information was assessed in a safe place by interview, using standard questioners as pre-test and then educational intervention program (lecture, face to face education, pamphlet, educational movie, role playing health educational competition) have performed. After 2 months post test was administered to evaluate the effect of interventional program. Results: The average age of the participants was 32.62± 9.11 and the average number of participants´ partners was 4 (range 1-15).After the educational intervention, there was a significant difference between the pre-test and post-test knowledge scores ,it means that the mean score of general knowledge related to HIV/STI increased from 13.7±0.95 to 19.47±11.62 (P < 0.01). Similarly, there was a significant improvement in the sex workers´ attitude and their sexual preventive behaviors such as safe sex and condom use (P < 0.01). Conclusion: According to results of this study, the educational program was successful in increasing and promoting the HIV/AIDS-related knowledge and attitudes of the participants and enhancing their HIV sexual risk behaviors. More interventional studies should be developed for other high risk population. P98 Influx of uncommon HIV-1 strains from Eastern Europe and identification of a new unique recombinant strain among young Cypriot MSM in Cyprus Ioanna Kousiappa*, Yiota Lazarou, Katerina M Othonos, Johana Hezka, Leondios G Kostrikis University of Cyprus, Nicosia, Cyprus E-mail: [email protected] Retrovirology 2012, 9(Suppl 1):P98 Introduction: The polyphyletic picture of HIV-1 infection in Cyprus is a fact, as new variants and unique recombinant forms were found in the recent past. As part of a growing effort to monitor any changes in the molecular epidemiology of HIV we studied two notable cohorts of the known HIV-1 population diagnosed in 2010 to 2011. Near full-length genome sequencing and phylogenetic analysis was carried out to determine the heterogeneity among HIV-1 strains isolated from patients, 10 originated from eastern European countries (mostly Romania) and 8 young Cypriots (<25 yrs), all men who have sex with men (MSM). Materials and methods: Sequence of the near full-length genome was amplified by RT-nested PCR from all HIV-1 seropositives and sequenced. Detailed phylogenetic and bootscanning analyses were performed by MEGA v5.0 to determine phylogenetic associations and subtype assignments. To explore putative recombination patterns in the sequences we performed a bootscanning analysis using Simplot, version 3.5.1. Results: Phylogenetic analyses of the obtained viral sequences showed genetic diversity. In the eastern European cohort, subtype F1 was the dominant subtype (40%), followed by subtype C (20%), A1, A2, CRF02_AG, and CRF03_AB (10% each). In the young MSM cohort subtype B was the main subtype (50%), followed by subtype A1 (25%), CRF01_AE (12.5%) and one HIV-1 isolate that was not classified in any known subtype or recombinant form (12.5%). Complete recombination analysis revealed that this isolate had a new recombinant pattern, comprising segments of subtypes A1 and B, and is distinct from any reported recombinant. Conclusions: These findings exhibit an influx of infrequent HIV-1 genetic forms from eastern European countries in Cyprus, and a stable circulation of B and A1 subtype among the young Cypriot MSM cohort. A unique recombination event between A1 and B subtypes has occurred and the parental strains seem to be formerly characterized Cypriot MSM patients. For the first time, these data show an impact on the evolutionary progress of HIV-1 epidemic of the island. The significance of this study along with the earlier variable epidemiological status of HIV-1 infection in Cyprus reflects the contribution to HIV classification, and the important implications for HIV-1 disease control and surveillance. Retrovirology 2012, Volume 9 Suppl 1 http://www.retrovirology.com/supplements/9/S1 P99 HIV risk in an urban American population Josephine F Wilson Boonshoft School of Medicine, Wright State University, Kettering Oh, USA E-mail: [email protected] Retrovirology 2012, 9(Suppl 1):P99 Introduction: Investigations of specialized populations in the United States have revealed multiple risk behaviors for HIV infections. Identification of significant risk factors for specific populations enables tailoring interventions to populations. The present study examined risk behaviors in a general urban population in order to determine the need for HIV interventions in subgroups of the general population. Materials and methods: HIV testing tied to a small remuneration was offered to adults in a small city in Ohio. Testing was conducted in churches, public health sites, and in a van that was parked at various public housing sites. A total of 3,290 individuals were tested for HIV. At the time of testing, demographic and risk behavior data were collected. Data were analyzed using logistic regression to determine the populations most at risk for HIV infection in this large sample of poor urban residents. Results: The sample tested ranged in age from 18 to 85 years, was 80.2% Black and 18.8% Caucasian, and 43.6% male. Altogether, 0.4% of the sample tested was HIV positive. In this largely African American sample, 77.3% admitted to condom use, 4.9% had sex with an intravenous drug user (IDU), 4.2% used intravenous drugs, 2.1% had sex with a MSM, and 11.4% had sex with 3 or more partners in the past 30 days. Logistic regression indicated significant associations between a positive HIV test and having sex with a MSM, no condom use, being an IDU, having sex with an IDU, and having sex with someone who is HIV positive. Odd ratios were calculated for each of the identified significant risk factors. Conclusions: These data indicate where intervention efforts are needed in our community. Studies of this sort enable public health administrators to conserve valuable prevention funds by targeting interventions to populations at greatest risk. This study revealed that, in this largely African American population, men and women were equally at risk for HIV, thus mandating the development of HIV interventions for men and women in this urban population. P100 Uptake of PMTCT sites for increasing accessibility of services in prevention of mother to child HIV transmission program in Rwanda, January 2005 June 2010 Ange Anitha Irakoze*, Placidie Mugwaneza, Sabin Nsanzimana, Jennifer Mbabzi, Jean Pierre Nyemazi, Eric Remera Hiv Division, Former Trac Plus, Kigali, Rwanda E-mail: [email protected] Retrovirology 2012, 9(Suppl 1):P100 Background: In Rwanda, the Prevention of Mother to Child Transmission of HIV program (PMTCT) started in 1999 as a pilot project; a positive evaluation conducted 1 year later lead to national scale-up. The integration of the PMTCT program into existing antenatal care (ANC) services was done gradually. Nationally, HIV testing is routinely offered to all pregnant women at the time of enrollment in ANC. Political commitment and involvement of key stakeholders in program implementation; public awareness campaigns, especially for pregnant women; and the involvement of local authorities, care providers, and community health workers have greatly contributed to the scale-up of the PMTCT program in Rwanda. Methods: We analyzed routinely reported facility-level data on HIV testing among pregnant women in the national PMTCT program from January 2005 to June 2010. Results: The number of PMTCT sites increased from 209 (46%) at the end of 2005 to 382 (84%) sites in June 2010 (209 in 2005, 234 in 2006, 285 in 2007, 342 in 2008, 377 in 2009 and 382 June 2010) out of 452 health facilities providing ANC services. Overall, 1,623,079 women received ANC services; of these, 1,554,387 (95.76%) accepted HIV testing. The acceptance rate of HIV testing among pregnant women in ANC increased from 89% in 2005 to 98,3% in June 2010. Of women tested, 99,4 % received their HIV test result. HIV prevalence among pregnant women tested decreased from 4.8% in 2005; to 4.4% in 2006; to 3.8% in 2007; to 2.9% in 2008 and 2.6% in 2009 and 2010. Page 40 of 54 Conclusions and recommendations: These results demonstrate that many efforts were done regarding the increasing of PMTCT sites in the line of scale-up of the national PMTCT program in Rwanda, as indicated by increases in number of sites offering PMTCT services, in uptake of HIV testing for pregnant women. Despite the high uptake of PMTCT sites, it is still below the national target of 100% of coverage of PMTCT sites, thus enhanced efforts to increase availability of PMTCT services are warranted. P101 A decade of investment, HIV prevention research and development funding from 2000 through 2011 K Fisher, E Donaldson*, LM Green, T Harmon, P Harrison, R Lande, M Warren Avac - Global Advocacy for Hiv Prevention, New York, USA E-mail: [email protected] Retrovirology 2012, 9(Suppl 1):P101 Introduction: Since 2004, the HIV Vaccines and Microbicides Resource Tracking Working Group has employed a comprehensive methodology to track resource trends in R&D for biomedical HIV prevention options, including HIV vaccines, microbicides, PrEP, treatment as prevention, vertical transmission prevention and adult voluntary medical male circumcision. Materials and methods: Data were collected on annual disbursements by public, private and philanthropic funders for product development, clinical trials and trial preparation, community education and policy advocacy efforts in order to estimate annual investment in HIV prevention R&D. Investment trends were assessed and compared by year, prevention technology type, type of funder and geographic location. Results: Since 2000, there has been significant growth in funding support for HIV prevention research and development. However, in 2011 HIV prevention research began to face increased funding pressures as governments worldwide decreased or flat-lined budgets in many areas of global health and as philanthropic donors worked to revise their investment strategies. Competing funding priorities affected funding for HIV prevention R&D. Still, despite those various funding pressures, the field of HIV prevention research progressed significantly in 2011, with new findings that advanced the field and promising new trials underway. Conclusions: Monitoring funding trends for HIV prevention research is particularly important at this time of critical juncture between economic uncertainty and the point at which the scientific community has articulated a much clearer pathway to the end of the HIV epidemic. Monitoring funding provides the fact base for policy advocacy around spending levels and allocations that will sustain investments in the research required to build on the success of recent trials; bring novel HIV prevention candidates into the pipeline; and support the follow-on clinical trials needed to assure the safety, immunogenicity, efficacy and acceptability of new HIV prevention products. P102 Sexual behavior and reproductive health among HIV infected adolescents in RBC, IHDPC, Clinic during 2011 therapeutic holidays Ange Anitha Irakoze*, Diane Tuyishimire, B Ami Bugingo, Sabin Nsanzimana, M Josee Maliboli, Melanie Muhizi, Angelique Nkuliza, Simon Niyonsenga, M Gasana, Ciprien Baribwira, Jackson Sebeza, Laetitia Umulisa Rbc, Ihdpc, Hiv Division.Former Trac Plus, Kigali, Rwanda E-mail: [email protected] Retrovirology 2012, 9(Suppl 1):P102 Background: Access to SRH services for adolescents in general and for HIV infected specifically is still challenging , resulting in low level of awareness , unsafe sex practices, risk of pregnancies, STI , contamination or super infection. Objectives: To determine the patterns of sexual behavior, reproductive health among HIV infected adolescents followed at IHDPC/RBC HIV clinic. Methods: During 2011 therapeutic holidays for HIV adolescents followed in RBC/IHDPC/HIV Clinic, 151 of them (73 girls and 78 boys) were assessed about sexual behavior and SRH using a modified HEEADS assessment selfadministrated questionnaire and interviews. Parental permission was sought. Results: The mean ages (+/-SD) of the girls and boys were 16.3 +/- 0.165 years. About disclosure: 8.7% of the adolescents were not willing to disclose their HIV status to their sexual partners, yet 76% of interviewed have a partner. Retrovirology 2012, Volume 9 Suppl 1 http://www.retrovirology.com/supplements/9/S1 About safe sex: Overall, 8.4% of adolescent are sexually active all above 15years (4.4% of the girls and 12% of the boys). Boys are three times more sexually active than girls and 66.7.1% of sexually experienced boys had used condoms. Among sexually experienced girls only 33.7% used condoms, surprisingly 96.6% of interviewed declared not willing to have protected sex. About Contraception and pregnancy: among sexually experienced girls 36.7%, had ever used contraception and prevalence of adolescent pregnancy was 2.6%. Information about sexuality: 81.12% of adolescents have ever learnt about sexuality (51.7 % aged 15-19 years), 49.6% can discuss sexual issues, among them 55% are girls and 45%; are boys. Conclusion: In this adolescent cohort, sexual activity seems to debut after 15 years old, safe sex practice is not optimal, and contraception among sexual actives girls is low and information level about SRH need to be improved, underling the urgent need of implementing SRH program for HIV adolescents. P103 Gendered sexual risk patterns and polygamy among HIV serodiscordant couples in Uganda Sarah Khanakwa*, Moses Ngolobe, David Moore, Robert Mwesigwa, Josephine Birungi, Rachel King, Kate Shannon Aids Support Organization (Taso) Uganda, Jinja, Uganda E-mail: [email protected] Retrovirology 2012, 9(Suppl 1):P103 Background: Multiple sexual partnerships and HIV sero-discordant relationships are among the most at-risk for HIV transmission. Polygamy is a common form of multiple-partnered relationships in Eastern Uganda. We investigated the association between HIV risk patterns and polygamy among HIV sero-discordant couples at The AIDS Support Organization in Jinja, Uganda Methods Participants were enrollees in a prospective cohort of HIV sero-discordant couples, the Highly Active Antiretroviral therapy as Prevention (HAARP) Study at TASO Jinja. Descriptive and bivariate analyses to compare sexual risk patterns among HIV sero-discordant men; in polygamous as compared to single-spouse relationship. Results: Polygamous Vs Single-spouse couples ≥2 wives 1 wife P value N = 241 56 185 Male HIV+ve 38 (68%) 99 (54%) 0.065 Male-controlled sexual decision making 34 (61%) 66 (36%) 0.001 Male-controlled condom use 33 (59%) 51 (28%) <0.001 Condom last time had sex 45 (80%) 128 (69%) 0.086 Financial support 45 (80%) 152 (82%) 1.00 HIV positive partner on ART 24 (37%) 88 (48%) 0.143 Median age (IQR) 44 (39– 50) 43 (37– 50) 0.451. Conclusion: This study demonstrates continued gendered risks for women in HIV sero-discordant relationships in sub-Saharan Africa. In particular, men with 2 or more wives are more likely to make decisions about when to have sex or when to use a condom. However, we found no differences in condom use at last sex by polygamy status. P104 Relying on injection drug users to prevent HIV in Ukraine - follow-up results of peer-driven interventions Oleksandra Datsenko*, Pavlo Smyrnov, Robert Broadhead International Hiv-Aids Alliance-Ukraine, Kyiv, Ukraine E-mail: [email protected] Retrovirology 2012, 9(Suppl 1):P104 Introduction: Conventional harm reduction (HR) projects overlook IDUs’ capabilities in offering them services but no active roles to play in preventing HIV. In contrast, “peer-driven interventions” (PDIs) offer IDUs rewards to educate and recruit peers for services. All IDU-recruits receive the opportunity to play both roles. Methods: In 2010, the International HIV/AIDS Alliance-Ukraine implemented PDIs in 12 Ukrainian cities that relied entirely on IDUs to access and teach IDUs who either had: (a.) never received HR services, or (b.) former PDI/HR-respondents eligible for a one-year follow-up (FU) intervention. IDU-recruiters were trained to administer two completely different bodies of prevention information: one to new recruits, the other to FU-recruits. Recruits from both groups were administered an 8-point knowledge test (KT) at their appointment that measured how well their recruiters educated them. Results: In 6 months of operation, the 12 PDIs recruited 8,115 IDUs: 2,782 (34%) new recruits Page 41 of 54 5,333 (66%) PDI-FU-recruits - New recruits: 87% scored 7 or higher on the first KT - FU-recruits were administered both KTs at the follow-up appointment. 86.4% scored 7 or higher on the 1st KT 89.6% scored 7 or higher on the 2nd KT (One year earlier, only 81.5% of the FU-group scored 7 or higher, which underscores how the PDI’s repetitive features improved respondents’ retention rate.) Conclusion: The PDIs documented that IDUs can play active roles in preventing HIV by recruiting IDU-peers who have never received HR services, or IDUs eligible for FU intervention. They are also able to deliver two entirely different bodies of prevention information. Compared to HR projects that relied on traditional staffs of salaried outreach workers, the PDI proved to be far more powerful and cost-effective model. AllianceUkraine now plans to broaden its investment in PDIs even more heavily by further expanding projects targeting IDUs, but also female sex workers and homeless/runaway street children. P105 Awareness and attitude of the general public toward HIV AIDS in coastal South India - a community based crossectional study B Reshmi*, B Unnikrishnan, P Mithra, T Rekha Manipal College of Allied Health Sciences, Manipal, India E-mail: [email protected] Retrovirology 2012, 9(Suppl 1):P105 Background: Acquired Immunodeficiency Syndrome (AIDS) is one of the most dreaded entities that modern medicine has ever had to tackle. Adult HIV prevalence in India is approximately 0.36%. HIV-related stigma and discrimination remains an enormous barrier to effectively fighting the HIV and AIDS epidemic, There are several reasons for the stigma toward PLWHA among the general population, one of them could be inaccurate information about the transmission of HIV; creating irrational behavior and misperceptions of personal risks. Objective: To assess the awareness and attitude of the general public toward people living with HIV/AIDS (PLWHA) in Mangalore, a city in Coastal Karnataka. Methods: The study population included 630 individuals aged 18 years and above. The information was collected using a semi structured pre-tested questionnaire. Statistical package SPSS version 11.5 was used, Chi-square test was conducted and P< 0.05 was considered as statistically significant. Results: About one-third of the study population thought that one could get infected by merely touching an HIV positive individual. Approximately 45% stated that they would dismiss their maid on finding out her HIV positive status. About 54% were willing to undergo the HIV test. The respondents with less than secondary school education had a discriminatory attitude toward HIV positive people, with regard to them deserving to suffer, dismissing a HIV positive maid, hesitating to sit next to a HIV positive person in the bus, divorcing the infected spouse, and willingness to get tested for HIV, which was found to be statistically significant. Conclusion: Stigma among the general public was mostly due to fear of contracting the illness. Stigma does exist to significant degrees among the educated people, which was suggested by about 45% of the participants being willing to undergo the HIV test. P106 Sexual behavior and perceived risk of HIV AIDS among returnee labor migrants from Overseas in Nepal Sushma Dahal*, Paras Kumar Pokharel, Birendra Kumar Yadava School of Public Health and Community Medicine, BPKIHS, Dharan, Nepal E-mail: [email protected] Retrovirology 2012, 9(Suppl 1):P106 Introduction: Nepal is a popular country of origin for labour migrants. Migrant workers are risk group for HIV. In Nepal studies on labour migrants have mainly focused on those going to India. Not enough studies have focused on sexual behaviour of migrants going to overseas. Materials and methods: A cross sectional study was done among 110 returnee male labour migrants in Nepal who were interviewed about their sexual behaviour while in overseas and their perceived risk of HIV/AIDS. Recruitment agencies were selected purposively to identify returnee migrants. Snowball technique was also used to trace some migrants in houses and hotels. Retrovirology 2012, Volume 9 Suppl 1 http://www.retrovirology.com/supplements/9/S1 Results: Saudi Arabia, Malaysia, UAE and Qatar were found to be the popular countries of destination. Among respondents returning from Gulf countries, 41.6% had sex with paid/unpaid partner which was 66.7% for those from Non-Gulf countries. Among 21 respondents who didn’t always use condom, 15 were from Gulf countries and 6 were from Non-Gulf countries. Respondents for whom difficulty in finding condom was the reason for non use were from Gulf countries. Co-working female friends were the non spousal unpaid partners for majority of the respondents. There was not much difference in risky sexual behaviour of migrants based on law of destination country regarding sex work and respondent’s habit of drinking alcohol. AIDS was perceived to be a very dangerous killer disease by more than 2/3rd of the respondents; even though, more than 40% perceived themselves to be at some risk and more than 75% perceived their friends to be at some risk of HIV. Only 7% had ever heard and used VCT service. Conclusions: Labour migrants going to overseas are at risk of HIV. HIV related awareness raising activities should focus on migrants. HIV prevention programs in destination and origin country should target both male and female labour migrants. P107 Controlling the spread of Hiv among long haulage workers in Nigeria Evans Benjamin*, Taiwo Kelvin Igie, Monday Udoh Foundation Aid Solution for Talent Empowerment and Development, Ikeja Lagos, Nigeria E-mail: [email protected] Retrovirology 2012, 9(Suppl 1):P107 Introduction: The study investigated the effect of the transport industry on the transmission and spread of HIV/AIDS. Transport workers are highly mobile and spend long intervals away from the comforts of their homes. They are often involved in risky sexual behaviours that make them vulnerable to HIV infection, and so constitute carriers in the spread of the pandemic. Method: The study entailed interviewing more than 1,000 long haul drivers and workers with the objective of inducing frank talk to assess their sexual habits enroute their long hauls. The limitation of this study was their insistence on anonymity to avoid adverse effects on their social status and marriage stability. Result: More than 80% of interviewees had more than 20 female friends stationed at the villages on highways across the country. 60% of the promiscuous group knew about condoms but never used them. To them, what was the use doing it if you could not have the real thing. Sadly, some of the interviewees stated they had no other pleasurable indulgences in life other than sex, and if they were to die doing the only thing they enjoyed then who is complaining. Conclusion: The NURTW was advised: Create rest stations along the nation’s highways, with lodging, canteen, games, TV/Video sets, and other recreational facilities for drivers and motor boys. Provide GSM phone facilities for workers on long distance engagements to allow them keep in touch with their families. Organize seminars to educate workers on the implications, prevalence and management of HIV/AIDS. Provide medical test and care facilities at those rest stations for the quiet testing of workers for HIV/AIDS and dispensing of necessary drugs to sufferers. P108 Comparing knowledge, attitudes and sexual practices of Female Commercial Sex Workers (FCSW) and the general female population in Brazil Célia Landmann Szwarcwald*, Ana Roberta Pati, Pascom Paulo, Roberto Borges de Souza Júnior Fundação Oswaldo Cruz, Rio de Janeiro, Brazil E-mail: [email protected] Retrovirology 2012, 9(Suppl 1):P108 Introduction: The aim of this study was to compare knowledge and risk behavioural practices among female commercial sex workers (FCSW) and the general female population in Brazil. Page 42 of 54 Material and methods: Information on 2523 FCSW were collected in a RDS study carried out in 10 Brazilian cities in 2009. A method for estimating proportions and their variances was proposed, which takes into account the dependency structure of observations. Both the inverse of network size and the size of the city were considered in the estimation of RDS weights. The 2008 Behavioural Surveillance Study provided information of the general female population. The sample design of the BSS was a typical three stage-selection (census tracts, households and individuals), stratified by macro-region. In the statistical analysis, tests of proportions were performed taking into consideration each study sample design. Results: FCSW are less educated than the general female population aged 18 to 64 years. Knowledge of HIV transmission is always worse among FCSW, even after controlling for educational level. As to sexual practices, the onset of sexual activity is much earlier among FCSW and 38% reported sexual abuse at least once during lifetime. Although protected sexual practices showed a slightly more favourable scenario among FCSW, the proportion who would never fail to use a condom was very low (23%). Higher use of drugs among FCSW was another point of note. In relation to prevention measures, the coverage of gynaecological exam in the last three years was much higher in the general female population even though STI signs were more frequently reported among FSW. Similar proportions of HIV testing in the previous 12 months were found. Conclusions: The results showed that there are still shortcomings in preventive care among FCSW in Brazil. Measures specifically aimed at increasing knowledge of HIV transmission, coverage of gynaecological exam and annual HIV testing among FCSW could contribute to the reduction of STI transmission in the Brazilian population. P109 Condom use among long-term intimate partners using drugs baseline results from a randomized trial in Ukraine Liudmyla Shulga1*, Tatiana Andreeva2 1 ICF International HIV AIDS Alliance in Ukraine, Kiev, Ukraine; 2National University of Kyiv Mohyla Academy, Ukraine E-mail: [email protected] Retrovirology 2012, 9(Suppl 1):P109 Introduction: Ukraine faced changes in HIV transmission routes and in 2007 sexual transmission exceeded parenteral way. In response to situation change couple’s counseling for IDUs was introduced. Data presented in the abstract are part of the randomized trial to test couple’s counseling as a mean of HIV prevention among IDUs. Methods: Baseline data were collected in June - September 2011. 560 IDU couples in 10 cities were surveyed and screened for HIV and Hepatitis C. Participants were recruited using respondent-driven sampling method. Results: Age of respondents varied between 30 and 33 years old with women being about 3 years younger compared to men. On average, duration of relationship was 3.7 years. Only 15% of respondents were married, while majority (64.9%) lived in a civil marriage; 20% declared having close intimate relationships. At the same time women tended to consider living together as “marriage” while men differentiated between these two categories. Men had regular jobs more often compared to women (23.8 vs 15.8%), as well as odd jobs (53% vs. 31% respectively). Data showed that there was no difference observed between men and women in sexual activity with regular partners. But men reported having more occasional sexual partners (3 partners/30 days), and women were more often engaged into commercial sex (19 partners/30 days). Condom use with occasional (80%) and commercial (88%) sex partners took place more often compared to sexual intercourse with regular partners (44%). Condom use during anal and oral intercourse was much lower and varied between 8% with regular partners and 25% with occasional partners. The most common reasons for avoiding condom use were sensitivity decrease (28%). Conclusions: Intimate partners are often not engaged into sexual relationships, even if they live together; drug use is sees as a more intimate process. Sex education for IDUs should account for nature of intimate relationsips; focus on reasons of condom non-use; and promote protected anal and oral intercourses. Education should also teach to understand risks related to own or partner’s extramarital sex. Retrovirology 2012, Volume 9 Suppl 1 http://www.retrovirology.com/supplements/9/S1 P110 HIV situation in the Greater Mekong Sub-Region bordering Thailand Tawatchai Apidechkul Professor at Mae Fah Lung University, Chiang Rai Province, Thailand E-mail: [email protected] Retrovirology 2012, 9(Suppl 1):P110 Introduction: Greater Mekong Sub-region (GMS) is composing of the border of Thailand-Myanmar, Republic Lao, and China with various of geographic and cultures and more than 200 million people live there. The migration of population is simple scenario at these areas. Last few decades Thailand has been reported of the highest prevalent of HIV/AIDS particularly in the northern of Thailand. The study aimed to investigate the effect of HIV/AIDS at the areas of GMS. Materials and methods: This retrospective cohort study design aimed to investigates the HIV situation among the people who immigrated into Thailand in GMS. The systemic data extraction from the medical records from 19 hospitals which located in the border of Thailand during 19902009 was analyzed. The instruments had been detected in validity and reliability. Chi-square test was used for identifying the statistical significant at the alpha=0.050. Results: Totally 1,303 cases had been detected. Of 84.50% were still alive, 57.27% were male. Myanmar were 56.52%, 26.44% were unknown, and 11.10% were Republic Lao, 5.24%were Chinese, and 1.00% was Cambodian. Of 37.53% were aged 21-30 years old, 33.15% were aged 3140 years old, and 12.89% were aged 41-50 years old. Of 38.94%were agricultural, 34.56% were employee, 4.15%were student, and 3.46% were young children. Of 51.65% were full bone AIDS, and 28.32 were symptomatic AIDS. Of 92.31% had infected by sexual intercourse, 6.06% were mother to child, and 1.63% were IDU. Distribution of age by sex was statistical difference (p-value<0.001). Being female had a longer live than male (p-value=0.002). The live status was different according to nationality (p-value<0.001) and risk factor (p-value<0.001). Conclusion: The cooperation between countries to control the HIV spreading is immediately need for the GMS region especially the free trade market in 2015. P111 Risk behaviors and reasons for not getting tested for HIV among men who have sex with men in Peru Magaly M Blas*, Isaac E Alva, Robinson Cabello, Cesar Carcamo, Ann E Kurth Cayetano Heredia Peruvian University, Lima, Peru E-mail: [email protected] Retrovirology 2012, 9(Suppl 1):P111 Introduction: Men who have sex with men (MSM) account for the greatest burden of the HIV epidemic in Peru. Given that MSM are frequent users of the Internet, understanding the risk behaviors and the reasons for not getting tested among MSM who surf the Internet may improve the tailoring of future online behavioral interventions. Methods: From October 2007 to April 2008, we conducted an online survey among users of seven Peruvian gay websites. Results: We received 1,481 surveys, 1,301 of which were included in the analysis. The median age of the participants was 22.5 years (range 12-71), 67% were homosexual, and the remainder was bisexual. Of survey respondents, 49.4% had never been tested for HIV and only 11.3% were contacted in-person during the last year by peer health educators from the Peruvian Ministry of Health and NGOs. Additionally, 50.8% had unprotected anal or vaginal sex at last intercourse, and a significant percentage reported a condom broken (22.1%), slipped (16.4%) or sexual intercourse initiated without wearing a condom (39.1%). The most common reasons for not getting tested for HIV among high-risk MSM were “I fear the consequences of a positive test result” (n = 55, 34.4%), and “I don’t know where I can get tested” (n = 50, 31.3%). Conclusions: A small percentage of Peruvian MSM who answered our online survey, were reached by traditional peer-based education programs. Given that among high-risk MSM, fear of a positive test result and lack of awareness of places where to get tested are the most important reasons for not taking an HIV test, Internet interventions aimed at motivating HIV Page 43 of 54 testing should work to reduce fear of testing and increase awareness of places that offer free HIV testing services to MSM. P112 Social and contextual factors that influence HIV risk behaviors among indigenous MSM in the Peruvian Amazon Isaac E Alva*, E Roberto Orellana Universidad Peruana Cayetano Heredia, Lima, Peru E-mail: [email protected] Retrovirology 2012, 9(Suppl 1):P112 Introduction: Men who have sex with men (MSM) face a significantly higher risk of HIV infection than the general population around the globe. In Peru, HIV prevalence among MSM range from 14% to 23%, with Lima, the capital, and port cities in the Amazonian region being the most affected. Recent studies found that indigenous MSM who leave their villages for cities along the Amazon River and its tributaries, engage in high risk behaviors such as high alcohol consumption and unprotected sex with mestizo (non- indigenous) MSM. This study examined social and contextual factors associated with risky behaviors among indigenous MSM in the Peruvian Amazon. Materials and methods: During a 5-month period in 2009-2010, we purposively recruited indigenous MSM. The study took place in several port cities throughout the Amazon region. Semi-structured in-depth interviews were conducted with indigenous men, who consented to voluntarily participate in the study. Results: We interviewed 34 MSM with an average age of 26 years. They represented 8 different ethnic groups. In most situations, when family and community members learned about the participants’ sexuality, discrimination and violence ensued. Participants reported being beaten up by their relatives. Sometimes, community councils were held to decide on their fate. Council decisions ranged from forcing the person to hard (manly) labor, to undertake traditional medicine treatments, to expulsion. Participants saw their migration to the city as an escape from oppressive forces in the community. In the city, many reported being in abusive relationships with other men. Lacking appropriate education and technical skills, many participants engaged in sex work as a way of making a living. Conclusions: A great deal of discrimination, isolation and lack of social support was experienced by most participants. Besides individual-level interventions, HIV prevention programs should take these factors into account and design programs that increase social support, enhance community building and reduce stigma. P113 Once a man tests, the partner tests as well. A comparison by gender for HCT and STD clinic attendance Maria Nambira*, Lydia Mwolobi Monitoring and Evaluation Specialist at African Medical and Research Foundation, Kampala, Uganda E-mail: [email protected] Retrovirology 2012, 9(Suppl 1):P113 Introduction: Since its outbreak in 1986, Uganda has registered a tremendous decline in HIV prevalence rates from 15% in 1991 to 5% in 2002 though up again to 6.4% in 2006 (2006 UDHS). Currently, an estimated 100,000 new HIV infections occur annually. In recent years, the uptake and practice of preventive behavior have declined, particularly among men. Currently, almost 40% of people with HIV are not diagnosed until they already have developed AIDS. That can be up to 10 years after they first became infected with HIV. Finding out whether a person is infected with HIV is the first step to improving their health and that of their partners and their families. The study aimed to assess the contribution, role of men in the fight of HIV/AIDS through HCT. Methods: The project monitored all individuals attending STD and ART clinics. The project sought to check whether when requested, males brought their female counterparts for testing and vice versa during the September 2010 to Sept 2011 at Luwero health centre IV in Luwero district, Uganda. Clinical data was used. Results: In the reporting period, we estimated that 1,230 people were attending ART clinics and about 3,200 people attending STD clinics. More Retrovirology 2012, Volume 9 Suppl 1 http://www.retrovirology.com/supplements/9/S1 males once tested brought more spouses for testing. The numbers were much less vice versa. For every 1 man tested at least 3 women were tested. This was also recorded in polygamous relationships. Clinic Type Males Females Total Attendants Attendants Who Brought Spouses Males Females STD 1,091 2,109 1,230 567 (52%) 111 (5.3%) ART/HCT 383 847 3,200 146 (38%) 57 (6.7%). Conclusion: HCT needs to emphasized in the struggle to reduce HIV incidence and males contribute to this trend setting. Due to the poor health seeking behavior of males compared to females, low HCT is done. The impact of HCT can be greater when men are targeted to attend ART and STD clinics since they have a bigger ability to bring their spouses to test as well. Health facilities should design ways to attract more males for HCT and STDs. P114 Effectiveness of PMTCT Programme at Mogwase Health Centre, South Africa Iryna Chaparanganda University of KwaZulu Natal, South Africa E-mail: [email protected] Retrovirology 2012, 9(Suppl 1):P114 Introduction: Today, HIV/AIDS-related conditions are a major contributor to childhood morbidity and mortality in South Africa. The PMTCT Unit, Antenatal Clinic (ANC) at Mogwase Health Centre (MHC), North West (NW) Province in South Africa runs a prevention mother-tochild transmission (PMTCT) programme aimed at reducing the number of HIV-infected babies born to HIV-positive mothers. The PMTCT Unit also undertakes polymerase chain reaction (PCR) test for surrounding clinics. Materials and methods: The abstract analyses the effects of PMTCT programme within the 2010 period. 626 (100%) pregnant women booked during this period were tested for HIV with pre- and posttesting counselling. 200 (31.9%) of them were tested positive. 117 (58.5%) of the HIV-positive women received dual therapy (AZT from 14 weeks of pregnancy (before 01.04.2010 – from 28 weeks) and sdNVP at onset of labour) and 83 (41.5%) were initiated on highly active antiretroviral therapy (HAART) (CD4 ≤350 cells/mm3 (before 01.04.2010 - ≤200 cells/mm3) or stage 4) – 200 (100%) in total. 200 (100%) of HIVpositive mothers also were counselled on safe infant feeding at least once. Results: 133 HIV-exposed babies were born at the clinic, and 130 (97.7%) of them received nevirapine syrup whilst 3 (2.3%) did not receive and transferred to hospital. PCR tests around 6 weeks were done on 161 babies inclusive of those from other clinics. 4 (2.48%) infants got infected with HIV through MTCT of which, 1 (25%) was transferred from another clinic; 1 (25%) was from a late booked mother, and 2 (50%) from unbooked mothers. Conclusion: Although more effort is needed to encourage prevention of unplanned pregnancies, early pregnancy testing, and early booking at ANC site PMTCT programmes can be effectively implemented in rural clinics in South Africa, reducing the number of HIV-infected babies born to HIV-positive mothers. Source: ANC register. MHC, NW Province, South Africa. PMTCT register. MHC, NW Province, South Africa. Dispensing book. MHC, NW Province, South Africa. PCR register. MHC, NW Province, South Africa. P115 High incidence of occupationnal blood exposures (OBE) in the health care workers sector of low income countries, using the example of bangui, central African Republic (CAR) HD Mossoro-Kpinde*, CD Mossor-Kpinde, E Gbangba-Ngai, CG Kamalo Bangui, Central African Republic E-mail: [email protected] Retrovirology 2012, 9(Suppl 1):P115 Introduction: CAR has been heavily affected by HIV (6.2%) and hepatitis B (15%) and C (3%), but has not yet developed a prevention plan against OBE, even though its health care staff, already low in numbers, is Page 44 of 54 overwhelmed by a massive patient load.This study aims to assess the current OBE situation and develop a national plan for the management of these accidents. Methods: A preliminary cross-sector study was conducted in 2009 amongst 3 health care facilities groups in Bangui. The parameters being studied were collected using a standard form including serological status for HIV, HBV and HCV, vaccination against hepatitis B, incidents of OBE and their subsequent management. Outcomes: Three hundred members of the health care staff were included in the study. 9.2% had been vaccinated against hepatitis B. Thirty six percent (36%) had already been tested for HIV, with 7.3% of the tests performed within the last three months. Fifty four percent (54%) cited an incident of OBE within the last six months. Sixty eight percent (68%) of these were from accidental needle stick injuries. At the time of the accident, 39.9% knew their HIV serological status, and 22% their HBV status. Three percent had been vaccinated against hepatitis B. Three per cent (3%) of the accidents received subsequent care. The post-OBE care management did not cover hepatitis B. Conclusion: There is a high prevalence of OBEs in the sites studied. The number of health care staff receiving subsequent care is low. As this study was limited to Bangui, it could be interesting to conduct an exhaustive evaluation throughout CAR. Meanwhile, given the current results and the aim of safeguarding the over-stretched pool of health care staff from OBEs, efforts are required to strengthen staff capacities, manage OBEs and improve hospital hygiene in the sites studied. P116 Achieving zero new HIV infection, unsafe sexual practices of out of school border youths AO Sekoni*, AT Onajole College of Medicine, University of Lagos, Lagos, Nigeria E-mail: [email protected] Retrovirology 2012, 9(Suppl 1):P116 Background: Young people aged 15 to 24 account for more than 50 percent of all new HIV infections worldwide, majority are those who engage in unsafe sex (unprotected casual sex and multiple sex partners), unsafe injection drug use, exposure to contaminated blood and blood products or unsterilized skin piercing procedures. Border towns have an admixture of vulnerable population including uniform personnel, out of school youths, traders, drivers, commercial sex workers and migrants as well as risky sites such as bars, hotels, brothels and truck parking areas. Methodology: This cross sectional study was carried out among border youths to assess knowledge and practices of safer sex and use of HCT services. One in two systematic market stall sampling was used in the border market between Nigeria and Republic of Benin to select participants for the study, in each of the selected stalls all the youths 15 to 24 years were interviewed using a validated structured questionnaire until sampling size of 120 was achieved. Results: The mean age was 19years, majority were female (62%) & single (73%). Half of them had at least secondary school education, 22% had no formal education, 93% were Nigerians, 25% live on their own, while 36% were financially responsible for themselves. Two third (68%) did not know what safer sex means although 43% have had sexuality education, mean age at sexual debut was 16yrs, 64% have had sex while 63% were still currently sexually active. The main reason for having sex was to have fun (56%), more than a quarter engage in multiple sexual partnerships, 61% use condoms, among which only 15% were consistent users and 18% have accessed HCT services. Those who consume alcohol were more likely to be sexually active as well as have multiple sexual partners. Conclusion: The out of school border youths in this study engage in risky sexual behavior that can put them at risk of HIV infection, uptake of HCT (a prevention strategy) is poor. P117 Piloting work of outreach nurses in Harm Reduction projects in Ukraine Natalya Dvinskykh*, Sergiy Botvin HIV AIDS Alliance in Ukraine, Kyiv, Ukraine E-mail: [email protected] Retrovirology 2012, 9(Suppl 1):P117 Retrovirology 2012, Volume 9 Suppl 1 http://www.retrovirology.com/supplements/9/S1 In Ukraine, despite growing sexual way of transmission of HIV, drug use remains the driving force of the epidemic. Prevalence of HIV among IDU is the highest compared to other risk groups - 21.6%, according to biobehavioral study conducted in Ukraine in 2009. However IDU often do not have access to most of the needed medical services because of stigma, discriminative attitude, and lack of resources. To address these issues and to bring basic medical services to the places and at a time convenient for drug isers, a project “outreach nurses was piloted in 2011. The aim of the project was to introduce work of medical professionals (nurses) as a part of Harm Reduction Program. The project was led by All-Ukrainian Harm Reduction Association, supported by HIV/AIDS Alliance in Ukraine and implemented in 5 regions of Ukraine. 3 nurses from each region were trained on two workshops: Harm Reduction Basics and VCT. The nurses provided basic medical services and consultations at mobile clinics, fixed NEPs, outreach routs and clients’ houses. They changed bandages, applied ointments, checked body temperature etc., as well as provided information on various health improvement issues and referred to other services. The intervention proved to be extremely popular among IDU 1732 clients were reached by the services during the project’s work. The most popular services included: veins treatment, bandaging, anticeptics dissemination, and medical consultations, including HIV counseling. Those nurses who had not worked in Harm Reduction programs before reported that their attitude changed a lot during the project implementation: from fear to being ready to help and being proud on the ability to help people in need. The new services also gained the most positive feedback from the clients. The project piloting showed that work of outreach nurses can be useful for any Harm Reduction program and can improve quality of its services. There are several factors that are important for the success of the intervention. Among them: adequate training for outreach nurses, ‘nurse social worker’ team work, project’s cooperation with local medical institutions. P118 Accessing and educating female sex workers in Ukraine via a peerdriven intervention Oksana Matiyash*, Pavlo Smyrnov, Robert Broadhead International HIV-AIDS Alliance-Ukraine, Kyiv, Ukraine E-mail: [email protected] Retrovirology 2012, 9(Suppl 1):P118 Introduction: A peer-driven intervention (PDI) for female sex workers (FSWs) was implemented in harm reduction projects in two Ukrainian cities. The goal was to recruit and interview 500 FSWs in 6 months who had never received services before, and to measure how well FSWrecruiters educated FSWs in a body of fresh HIV prevention information. Recruiters were rewarded by earning tickets to win special prizes in a weekly lottery. Methods: A PDI relies on respondent educating and recruiting peers for services. All recruits also get to serve as peer-educator/recruiters. Each recruit’s score on an 8-item knowledge test, measuring how well the recruiter educated her, determined how many lottery tickets the recruiter earned. A weekly lottery was held each week offering prizes to 4 lucky winners. Results: The two PDIs in 6 months recruited 532 and 437 FSWs never seen before, 2-3 times more new FWSW-recruits than the number recruited 6 months prior to the PDIs’ start-up. Both projects held 23 lotteries with an average attendance of 8 FSWs. Very significant differences in levels of drug- and sex-related risk behaviors were found between FSW drug-users and non-users, heavily shaped by education, knowledge, and other social variables, suggesting more targeted types of intervention. Conclusion: The FSW-respondents were eager to serve as peereducators/recruiters. The lottery proved to be a cost-saving and effective reward system that was highly motivating. The PDI offers harm reduction projects an entirely new model for accessing and educating FSWs, as well as a new method for targeting special sub-populations of FSWs. Page 45 of 54 P119 A qualitative study on HIV positive women experience in PMTCT program in Indonesia Martiani Oktavia1*, Anita Alban2, Prisca AC Zwanikken3 1 IMPACT Program with Padjadjaran University and Hasan Sadikin Hospital Bandung, Indonesia; 2University of Copenhagen, Denmark; 3Royal Tropical Institute, The Netherlands E-mail: [email protected] Retrovirology 2012, 9(Suppl 1):P119 Introduction: Indonesia has one of the fastest growing HIV epidemics in South-East Asia, which was largely driven by injecting drug users (IDUs). A projection model suggests that there will be a shifted of HIV epidemic from contaminated needles among IDUs to their sexual partners via heterosexual contact. At present, women are accounted 25% of all reported AIDS cases cumulatively. Despite the growing need for prevention mother to child transmission (PMTCT) of HIV is emerging, coverage of intervention for HIV test and ARV prophylaxis among HIV pregnant women are still low. This study calls for more client-oriented PMTCT program based on women’s need and demand in a changing HIV epidemic. Material and method: Mixed study design, which consists of a literature review and a qualitative study with in-depth interview among HIV positive women with history of PMTCT (purposive sampling). Results: Nine women aged 25 to 33 years old were selected as respondents for qualitative study. Majority of them had contracted HIV from their spouse, who formerly injected drugs. All of them perceived low or no risk factor for HIV. HIV positive women valued high acceptance of HIV testing in primary health care with conditions; ensure confidentiality and quality of counselling. Lack of information about PMTCT and unintended pregnancies presumably correlated with late initiation of ARV prophylaxis among HIV infected pregnant women. Despite almost all of the respondents were expressing no intention to have more children, there was unmet need for contraception. Stigma and discrimination remain exist in various forms; fear of being isolated/separated from friends and family, sub-optimal treatment in hospitals by healthcare workers. Conclusions: Findings from this study provide a basis for establishing PMTCT program responsive to the need and demand of women as subject of intervention. Comprehensive interventions need to be integrated into existing health systems and utilize resources at the locals disposal. A successful and sustainable PMTCT of HIV program requires a close collaboration with stakeholders e.g. governmental institutions, nongovernmental organisations, and civil society representatives. P120 Perceptions of sexual practices among the old people in Sub-Saharan African largest City, Nigeria Odor King*, Isaac Olaseha Public Health at University of Ibadan, Abuja, Nigeria E-mail: [email protected] Retrovirology 2012, 9(Suppl 1):P120 Introduction: Most studies on sexual behaviour in Nigeria focus on young people and adults, limited attention is paid to elderly people. Hence there is dearth of information about elderly persons’ reproductive health challenges and involvement in risky sexual activities. Aim: This study examined the perceptions of sexual practices among the elderly in Ibadan, Nigeria. Methods: The study was cross sectional in design, 400 elderly persons aged 65 years and above were selected using a three-stage sampling technique. Main outcome measures Both qualitative (FGD) and quanti-tative (Questionnaire) methods of data collection were used to collect relevant data on participants, sexual perception, practices and problems. The FGDs were recorded and analysed using the thematic approach, while the data from the questionnaires were analysed using descriptive and Chi-square tests. Results: The participants’ mean age was 71.8 (± 6.7) years. Slightly more than half, (50.5%) were males. Few (18.3%) had sex two years preceding the study. A total of 30.0% of the participants had had extramarital sex since they attained the age of 65 years. Among this subgroup, very few (7.3%) used condom. Half (50.1%) of the respondents were of the perception that condom was not meant for the elderly. Moreover, majority Retrovirology 2012, Volume 9 Suppl 1 http://www.retrovirology.com/supplements/9/S1 Page 46 of 54 (68.8%) were of the perception that sex with virgin could boost immunity against STIs/HIV. Lack of interest for sexual intercourse (59.5%) was the reported main sexual problem of the respondents. Moreover, FGD participants were unanimous in their opinion that sexual dysfunction was due to ageing. Conclusion: Many of the elderly were involved in risky sexual practices. Therefore, health education intervention programmes such as training on safe sex practices and counselling services are needed to address the problem. homosexual-bisexual and people with no risk factor. The information area of interest for people-users about the method of transmission of infectious agents of STDs (60%) (figure 1). Conclusions: These data would seem to suggest the need for interventions aimed at prevention of STDs populations of youth, as well as information strategies able to reach and engage the female target. The value added of telephone counseling for STDs is to provide the person-user cognitive tools, to help you avoid risky behavior and enable empowerment processes aimed at protecting the health of the individual and his community. P121 Modelling HIV modes of transmission in Iran Maryam Nasirian*, Ali-Akbar Haghdoost*, Fardad Doroudi, Mohammad Mehdi Gooya, Abbas Sedaghat, Eshagh Dortaj Rabbori Kerman University of Medical Science, Kerman, Iran E-mail: [email protected] Retrovirology 2012, 9(Suppl 1):P121 P123 Motivating men who have sex with men to get tested for HIV Magaly M Blas*, Luis Menacho, Isaac E Alva, Roberto Orellana Cayetano Heredia Peruvian University, Lima, Peru E-mail: [email protected] Retrovirology 2012, 9(Suppl 1):P123 Introduction: There is inadequate information on high-risk populations even though prevention programmes are the mainstay of the national response to the HIV epidemic in these populations. We used the mode of transmission (MOT) model to understand the sources of new HIV infections and use this information for programme planning. Material and methods: We systematically searched published and grey literature to find the best values for the input parameters required by the MOT model. The data were discussed by a group of national experts before being fed into the MOT model. Using the Monte Carlo technique, we computed the 95-percent uncertainty level (UCL) for the outputs of the MOT. Results: The MOT model estimates that 9136 new HIV infections will occur in Iran in 2010 (UCL 6831-11757). Fifty-six percent (UCL 47.7–61.6%) of new infections were among injecting drug users and 12 percent (UCL 9.5–15%) among their sexual partners. The major routes of direct and indirect HIV transmission in Iran are unsafe injection (68%) and sexual contact (34% heterosexual and 10% same-sex). If current coverage for safe injection among IDUs increased from 80 to 95 percent, the number of new HIV infections in this group would decrease by almost 75%. Conclusion: IDUs remain the key population at highest risk of HIV infection in Iran, so programme coverage for IDUs and their spouses needs to be increased. And as the sexual transmission of HIV increasingly contributes to the pool of new infections, serious measures are required to reduce sexual transmission of HIV among the relevant key populations. P122 STDs help line and prevention Filippo Maria Taglieri*, Pietro Gallo, Anna Colucci, Anna Maria Luzi, Rudi Valli, Francesca Botta, Eleonora Lichtner I.S.S., Rome, Italy E-mail: [email protected] Retrovirology 2012, 9(Suppl 1):P122 Introduction: Sexually Transmitted Diseases (STDs) are a public health problem worldwide, each year about 340 million people, aged between 15 and 49, who contract an STDs (WHO). The Research Unit psycho-sociobehavioral, Communication, Training – U.O. RCF (Department of Infectious Diseases, Parasitic and Immunomediated – National Institute of Health in Italy ) initiated in 2010 under a project funded by the Ministry of Health, a specific STDs prevention activities through the intervention of telephone counseling. This action is already used by researchers at the U.O. RCF since 1987 for the prevention of HIV infection and AIDS. Materials and methods: The data collected, anonymously, during the telephone counseling intervention were entered and stored in a data-entry software that has allowed, through statistical analysis, to define some personal characteristics of those users who choose the telephone to get information on STDs, and to identify their information needs. Results: In the period 21 June 2010 - June 20, 2011 U.O. RCF received 2,017 phone calls during which it as focused attention on STDs. The 91.1% of the people-users are male, 77.3% were on no more than 39 years. The greatest number of calls coming from the North Italy (46%), followed by the Centre (30%), South (18%) and the Islands (6%). Data analysis can distinguish three main groups of people-people: heterosexual, Introduction: Although men who have sex with men (MSM) have the highest HIV prevalence in Peru, they are underserved by traditional preventive programs. Interestingly, in Peru the Internet and cell-phones have emerged as a convenient tool to reach this population. Methods: From October 2010 to February 2011, we conducted eight focus groups with gay and non-gay identified MSM, and eight in-depth interviews with key informants in order to identify key features and preferences to be used to tailor culturally-appropriate behavioral messages that could be delivered through Internet and cell-phones to motivate MSM to get tested for HIV. Results: Participants reported that in order to motivate HIV testing among MSM, interventions need to be based on motivational messages that encourage participants to overcome the fear of getting tested. Messages should increase the HIV risk perception (of participants who do not consider themselves at risk) by eliciting risky situations usually experienced by MSM. Messages should emphasize the confidentiality, professionalism and respect of the personnel conducting the counseling and testing. A thorough explanation of the process of HIV testing, including information about the type of information will be collected, types of tests that will be available (rapid or ELISA), level of pain participants may feel, time to get the results back, and cost of the testing should be included. Additionally, detailed information about the steps participants have to follow if they test positive or negative should be provided. Messages should also contain detailed information about the venue where the test will be conducted in terms of type of clients who attend, location, hours of operation and personnel. Finally, stigmatizing and stereotyped messages or images about “being gay” should not be included, as they act as deterrents for getting tested. Conclusions: Interventions aimed at motivating HIV testing among MSM should include motivational messages that reduce the fear of getting tested and increase the risk perception of participants. They should also market the venue where the testing will be conducted, the professionals who will perform the tests, and the test itself. Stigmatizing messages or images should be avoided. P124 The HIV infection and telephone counseling, the experience of Italian National Institute of Health Anna Maria Luzi, Anna Colucci, Filippo Maria Taglieri*, Pietro Gallo, Rudi Valli, Francesca Botta, Eleonora Lichtner I.S.S., Rome, Italy E-mail: [email protected] Retrovirology 2012, 9(Suppl 1):P124 Introduction: In the context of HIV prevention interventions, counseling has proved a valuable operational tool. This method is characterized by the application of knowledge, personal qualities, skills (active listening, self-awareness, empathy) and communication techniques (reformulation, clarification, investigation skills). The interview give the person counselinguser to make choices and changes in situations perceived as difficult to deal with his problems in an active way and its difficulties. The above constitutes the methodological basis of HIV / AIDS Helpline telephone counseling done by the National Institute of Health, established in 1987. Retrovirology 2012, Volume 9 Suppl 1 http://www.retrovirology.com/supplements/9/S1 Page 47 of 54 Materials and methods: The intervention of telephone counseling provided by AIDS HELP LINE Italian is structured in three phases: initial, intermediate and final. The actions that characterize the three phases of the telephone counseling are as follows: - Accommodating the person / service user and present the Service - To focus the problem and find a shared goal - Provide information scientifically accurate, current and customized - Propose and agree on possible solutions - Summarise and check what has emerged and what has been agreed - Saying goodbye properly, make themselves available for further contacts and enter into the relationship The Italian AIDS Helpline uses a data-entry software that allows you to collect data on calls received. The data are handled anonymously and analyzed in aggregate form. Results: People - users in over 24 years, have turned to the AIDS Helpline Italian ISS are 671,823, of these 73.9% were males aged between 20-39 years (78.0%), reside in Central Italy, declare themselves heterosexual (55.1%) and raise questions regarding information on the procedures for transmitting HIV (27.2%) and ITER diagnostics (25.3%). Conclusions: The intervention of telephone counseling conducted by the AIDS Helpline Italian while placing no longer in a state of emergency and as a social alarm in the ‘80s, however, the data show that a significant number of people are taking advantage of telephone counseling intervention to express their information needs, clarify doubts and get information about the psycho-social-health services on national territory involved in the prevention, diagnosis and treatment of AIDS. Introduction and objectives: AIDS is an acquired syndrome of immunity damage caused by body immunity damaging virus. As our population is young and consciousness raising has an essential role in prevention of a disease and also the young people are the main victims of it, so it is necessary to determine student’s knowledge and attitude as a symbol of young population to understand their ability in campaign against the AIDS and distinguish appropriate educational programs. Materials and methods: This semi experimental study was performed on 150 engineering students of Islamic Azad University of Zanjan Branch. Samples were chosen by random systematic method. The data collection tool was a questionnaire concluding three parts of demographic characteristics, knowledge level, and attitude questions. After performing the pre-test, the educational program was held during three weeks and then the post–test was held. The data was analyzed by SPSS software and the scores which were achieved by samples before and after the educational program were compared using t-test and paired t-test. Results: This study showed that 64/6 % were male and 36/4 were female. The knowledge rate of the students about risk factors of the disease before and after intervention were 16/03 2/04 and 18/02 1/44 respectively. The mean and standard deviation of attitude score of the students also increased from 90/16 10/6 to 96/60 10/33 after the education. Conclusions: Findings of this research show the positive effects of education on knowledge and attitude of the students, and providing education and appropriate background in educational environment seems necessary for employing effective behaviors. Keywords: AIDS, Attitude, Education, knowledge. P125 Level of knowledge on risks to HIV and AIDS among secondary school students in the Kisumu District JT Ongwara1*, Obadha Odenyo2 1 Maseno University, Private bag Siriba Maseno, Kenya; 2Institute of Tropical Medicine, Kisumu, Kenya, Kisumu, Kenya E-mail: [email protected] Retrovirology 2012, 9(Suppl 1):P125 P127 Faculty members of Islamic Azad University of Zanjan’s knowledge regarding AIDS and preventing it Vida Sadeghzadeh*, Eshrat Sadeghzadeh University Department of Nursing, Zanjan Branch, Islamic Azad University, Zanjan, Iran E-mail: [email protected] Retrovirology 2012, 9(Suppl 1):P127 Objective(s): The objectives of the study were: 1) To assess the General knowledge and awareness among secondary school students in Winam Division, Kisumu District on: a) Modes of transmission of HIV and AIDS b) Signs and symptoms c) Prevention methods against HIV and AIDS. Design: The study design was descriptive and cross-sectional and was carried out in the study area between September and November 2007. Setting It covered a sample of student population of 405 drawn from 9 out of 30 schools in Winam Division. Simple random and Probability proportional to size sampling methods were used to sample the schools and the actual participants from each class. Subjects or participants Students attending secondary schools in the study area. Results: About (99%) indicated that they had heard about AIDS compared to only 4 (1%) who had not. Knowledge had no statistically significant relationship with risk of HIV and AIDS. About 53.3% of the respondents reported to have had sex with males being more likely to have an early sexual debut. Sexual activity was higher among peri-urban respondents (37%) who also had more than 3 sexual partners. About 71.4% of the respondents were willing to change their behaviour to avoid contracting HIV. On bivariate analysis, exposure to risk factors was dependent on gender (p < 0.05), perceived risk and condom used were related (p < 0.05). Conclusion: This study concluded that despite their high knowledge and awareness on HIV and AIDS, not all students who were exposed to risk perceived themselves to be at risk. Introduction: According to UNAIDS estimates, there are now 33.3 million people living with HIV, including 2.5 million children. During 2009 some 2.6 million people became newly infected with the virus and an estimated 1.8 million people died from AIDS. By mid 2011, 23125 people were infected with HIV, from which, 4311 people died in Iran. The vast majority of people with HIV and AIDS live in lower- and middle-income countries. But HIV today is a threat to men, women and children on all continents around the world. Since there is no vaccine for AIDS and there is no certain cure for AIDS, curing, prevention and secondary infection is essential. This study was conducted to determine the level of knowledge on AIDS and the way of preventing it among faculty members of Zanjan branch, Islamic Azad University. Materials and methods: This descriptive study involves 90 people. Data was collected by a questionnaire in five parts (Demographic questions, factors related to transmission, pathology, complications, and finally prevention of AIDS). Analysing of data was conducted by SPSS software. Results: Findings revealed that awareness of faculty members of Islamic Azad University of Zanjan about transmission of disease was 72.2, regarding pathology of disease was 67.7, related to complication of disease was 73.3, and about prevention of disease was 74.4.Finally, the level of knowledge of faculty members of Islamic Azad University of Zanjan was higher than moderate (average %70). Conclusion: It can be concluded that the rate of awareness is almost satisfactory but it is not enough and it is necessary to perform educational programs. Key Words: AIDS, Faculty member, Knowledge, Prevention. P126 The effect of teaching on awareness and attitude of the students of Zanjan Branch, Islamic Azad University, about AIDS SeyedehSusan Raoufikelachayeh University Department of Nursing, Zanjan Branch, Islamic Azad University, Zanjan, Iran E-mail: [email protected] Retrovirology 2012, 9(Suppl 1):P126 P128 HIV AIDS and substance abuse primary prevention in minority adolescents John Wodarski*, Sam MacMaster University of Tennessee, Knoxville, USA E-mail: [email protected] Retrovirology 2012, 9(Suppl 1):P128 Retrovirology 2012, Volume 9 Suppl 1 http://www.retrovirology.com/supplements/9/S1 Introduction: The research prevention project entitled “HIV/AIDS and Substance Abuse Primary Prevention in Minority Adolescents”, funded by the Substance Abuse and Mental Health Services Administration (SAMHSA), targets minority male and female adolescents 12-17 years old in the implementation and evaluation of a program designed to prevent adolescents from engaging in substance abuse and sexual activities which place them as risk for contracting the HIV/AIDS virus. The five-year intervention will serve 750 adolescents and 750 parents. Materials and methods: The program combines the effects of a program that consists of two evidence-based primary components: 1) refusal skills training and education for the adolescent focusing on issues relating to sexuality and substance abuse; and 2) a family prevention educational and skills component involving the parent(s) of the participating adolescents. The small group educational techniques employed with the adolescent component are based on the Teams-Games-Tournaments (TGT) Alcohol Prevention curriculum, cited as a Model Program in SAMHSA’s National Registry of Effective Programs and Practices (NREP) and as a Model Program by the Office of Juvenile Justice and Delinquency Prevention, and the Reducing the Risk (RTR) curriculum, both of which have been empirically evaluated as effective methods of teaching adolescent skills development in the areas of substance abuse and high-risk sexuality prevention. The parent prevention component is based on extensive research in which problemsolving skills and communication procedures have been used effectively with parents of adolescents. Results: The evaluation component of the project assesses participants (adolescents and parents) at baseline, post participation, and at 6-month follow-up periods. Dependent variables include the adolescents’ and parents’ knowledge of, attitudes toward, and behavior related to HIV AIDS and substance abuse. Additionally, adolescents’ self-efficacy and quality of peer and parental relationships are assessed. For all participating family members, conflict and communication measures are secured. Conclusion: Significant differences have been shown for adolescents and parents between baseline, post participation, and 6 month follow-up for all dependent variables. Training manuals provide program implementation requisites and materials to facilitate dissemination at local, state, and international levels. P129 Meeting the HIV prevention needs of substance using young adults in the United States Virgin Islands Samuel MacMaster*, John Wodarski University of Tennessee, Nashville Tn, USA E-mail: [email protected] Retrovirology 2012, 9(Suppl 1):P129 Introduction: The need for comprehensive HIV prevention in the United States Virgin Islands is tremendous. HIV rates in the Carribbean region of the world is second only to sub-Saharan Africa, and the USVI has one of the highest rates in the United States. The primary mode of transmission in the USVI is heterosexual contact fueled by substance using risks. The presenation will provide an overview of a model program designed to increase HIV testing access and reduce HIV infections. Materials and methods: The project is a collaboration between local entities and the Univeristy of Tennessee and Norfolk State University. The program seeks to provide culturally appropriate early intervention, HIV testing, and substance use services to young adults at high risk for contracting HIV. Results: Over the first two years of the project over 160 individuals have participated in the project and have experienced statistically significant improvements in HIV risk behaviors and levels of substance use. Conclusions: The project serves as an example of culturally appropriate interventions for high risk young adult populations. P130 Behavioral factors associated with HCV and HIV co-infection in residents of São Paulo, Brazil Norma Farias*, Umbeliana Barbosa de Oliveira, Iára de Souza, Débora Moraes Coelho, Claudia Afonso Binelli State Secretary of Health of São Paulo, São Paulo, Brazil E-mail: [email protected] Retrovirology 2012, 9(Suppl 1):P130 Page 48 of 54 Introduction: To date, there are no surveys in Brazil on the occurrence of co-infection hepatitis and HIV in the general population. The aim of the present study was to investigate factors associated with HIV/HCV coinfection among residents in the State of Sao Paulo, Brazil, notified at the National Databank of Major Causes of Morbidity. Material and methods: We reviewed 3,032 cases of HIV/HCV coinfection among 46,969 bank records of viral hepatitis from January 2007 to March 2010. The hepatitis C cases were confirmed by the presence of HCV RNA using reverse transcription-polymerase chain reaction (RT-PCR) in anti-HCV-positive samples. The diagnosis of HIV/ AIDS and data on demographic and behavioral aspects were collected through epidemiologic investigation forms. Variables associated with HCV/HIV co-infection were identified with Poisson regression model and confidence intervals of 95%. Results: The majority were male (73%), white (65%) and had less than 50 years (65%). In a adjusted analysis, the prevalence ratio was 1.27 (CI 95%:1.04-1.55) for sexual contact with patients with HVB or HCV, 1.48(CI 95%: 1.27-1,73) for 3 or more sexual partners, 1.73 (CI 95%: 1.38-2.16) for STD, 2.95 (95% CI:2.42-3.59) among IDU and 1.78(95% CI:1.49-2.17) among inhalable drug users or crack. Conclusions: These data show that illicit drug use is the main factor associated with co-infection HCV / HIV in São Paulo. Sexual transmission suggested to play a role in co-infected HCV / HIV. P131 Effective HAART reduces the incidence of high grade cervical neoplasia in HIV positive women Deborah Morris-Harris1*, Charmaine Miller-Spencer1, Clara Jones2, Song Zhang3, James Luby3 1 Parkland Health and Hospital System, HIV Services, Dallas, Texas, USA; 2Tufts University School of Medicine, Boston, Massachusetts, USA; 3University of Texas Southwestern Medical School, Dallas, Texas, USA E-mail: [email protected] Retrovirology 2012, 9(Suppl 1):P131 Introduction: Highly active antiretroviral therapy (HAART) has been shown to restore immunity and reduce the burden of Human Immunodeficiency Virus (HIV) in patients with HIV infections. Co-infection with HIV and the Human Papilloma Virus has been estimated to be as high as 51% in HIV positive women. Recently effective HAART has been shown to reduce the prevalence of HPV and intraepithelial lesions on Papanicolaou (pap) smear. It is unclear if HIV viral suppression can reduce the incidence of high grade cervical neoplasia . Materials and methods: A retrospective cohort of 1090 women, in care from 2005 through 2008, had 389 referrals to colposcopy clinic. 142 women mean age of 31.2 (range 17-54); 66.2% Non -Hispanic black,9.9% NonHispanic white, and 23.9% Hispanic with HIV, median baseline CD4 283(IQR 87.75-508) had a colposcopy and a second procedure, either colposcopy or excisional biopsy after a baseline abnormal Pap smear. Follow-up biopsies were performed with a median of 11.9 months (IQR 4.4-22.5) There were no significant baseline differences in clinical or demographic parameters between patients who were suppressed with VL. Results: Of 125 cases in which HAART was started before the first colpsocopy; 26 had a normal colposcopy and 48 had CIN1. From the normal and low grade group, 19 women developed high grade CIN (7, suppressed and 12, not suppressed.) Effective HAART reduced the risk of CIN2-CIS on a second biopsy by 83.2% (AHR 0.168; C.I. = 0 .057 -.498; p< 0.001) adjusted for age at diagnosis, race, smoking and baseline CD4. Higher baseline CD4 was associated with a reduced risk of high grade neoplasia (AHR=0.996; C.I. 0.993-0.999; p Conclusions: Reducing HIV viral load and preserving CD4 cells early in the course of HIV and HPV co-infection decreases the incidence of high grade cervical neoplasia. P132 Subclinical atherosclerosis in young persons horizontally infected with HIV-1 during infancy Augustin Cupsa, Florentina Dumitrescu*, Dina Maria Cupsa, Andreea Cristina Stoian, L Giubelan, Irina Niculescu, Cristina Iocu Infectious Diseases at University of Medicine and Pharmacy From Craiova, Craiova, Romania E-mail: [email protected] Retrovirology 2012, 9(Suppl 1):P132 Retrovirology 2012, Volume 9 Suppl 1 http://www.retrovirology.com/supplements/9/S1 Objectives: To evaluate subclinical atherosclerosis and to identify the cardiovascular risk (CVR) profile in young adults horizontally infected with HIV-1 during infancy. Methods: Retrospective randomized study carried out between 31.12.2009 and 30.06.2010 on 56 HIV infected persons(HIP) parenterally infected with HIV-1 during 1988-1990, following ART, under surveillance of HIV/AIDS Regional Center – Craiova. Variables followed: history and clinical data, traditional and additional CVR factors, metabolic, immunological and virusological parameters, inflammation markers (hs-CRP), ultrasound data regarding carotid intima-media thickness (IMT). Twenty-six HIVseronegative young adults were assigned as control group (CG) for metabolic parameters, hs-CRP and IMT. Results: General characteristics of the group: average age = 20.82 ± 1.1 years, equal distribution by gender, 47 HIP (83.93%) classified as clinical and/or immunological AIDS, 26 HIP (46.43%) with CD4>500/mm3, 40 HIP (71.43%) with undetectable RNA-HIV when evaluated, average ART duration = 9.09 ± 3.2 years, average number of ART regimens = 3.2 ± 1.63, 40 HIP (71.43%) experienced to protease inhibitors (PIs). In HIP – hsCRP = 2.17 mg/l, equivalent with a moderate CVR, statistically different compared with CG (p<0.0001); IMT = 0.76 ± 0.12 mm in HIP vs 0.6 ± 0.11 mm in CG. From the traditional CVR factors dyslipidemia levels were higher in the HIP group vs. CG (p<0.0001). In HIP, linear analysis of the evaluated parameters identified direct correlations between hs-CRPerytrocites sedimentation rate (ESR) (p=0.04), number of ART regimens and PIs exposure (p=0.007), IMT (p=0.000) and HIV-RNA (p=0.000) and also between IMT – triglycerides (p=0.004), PIs exposure (p=0.004), CD8+ (p=0.0000) and HIV-RNA (p=0.001). Conclusions: Young HIP have had an average value of hs-CRP equivalent with a moderate CVR; the CVR profile in young seropositive infected with HIV-1 during infancy includes elevated triglycerides, ESR, CD8+, HIV-RNA values and long time of PIs exposure. IMT in young HIP experienced to ART suggests a premature “aging” of the vessel by about two decades. P133 Positive predictive value of Interferon-gamma release assay for incident active tuberculosis in HIV-infected persons Susan Shin-Jung Lee*, Hsi-Hsun Lin, Hung-Chin Tsai, Yen-Yun Ni, Yao-Shen Chen, Chi-Tai Fang Kaohsiung Veterans General Hospital, Kaohsiung, Taiwan, Province of China E-mail: [email protected] Retrovirology 2012, 9(Suppl 1):P133 Introduction: Tuberculosis (TB) is the leading cause of death in HIV/AIDS. The incidence of active TB is 30 times higher in HIV-infected persons. Treatment of latent TB infection (LTBI) is pivotal to the control of TB. We aim to determine the positive predictive value of an interferon-gamma release assay, QuantiFERON-TB GOLD test (QFT), for incident active TB in HIV-infected persons. Materials and methods: This prospective, 5-year, cohort study enrolled HIV-infected adults without active TB. Demographic data, past exposure to TB and previous TB disease, HIV risk factors, CD4 counts, and HIV viral loads were recorded. QFT tests were done at entry. Cases of incident TB disease were ascertained by linking to our national TB database registry. Results: We recruited 774 HIV-infected adults, with a mean age of 36.9 years, mostly men (96.8%). HIV risk factors included intravenous drug user (67.2%), men-who-have-sex-with-men (24.2%), and heterosexual (7.9%). QFT was positive in 90 (11.6%,95% CI: 9.5-14.1%) and indeterminate in 31 (4.0%). On multivariate logistic regression analysis, significant risk factors for QFT positivity included older age, females, past TB disease and exposure to TB. Fifteen incident active TB cases (rate: 5.19/1000 person-years 95%CI:3.138.61) occurred during a mean follow up time of 3.73 person-years, with the majority (91.3%) followed up for over 2 years. Incident active TB disease occurred in 5.6% (5/90) of those with a positive QFT result, 3.2% (1/31) indeterminate results, and 1.4% with a negative QFT result (p=0.03). Hazard ratio for developing active TB was 3.10 (95%CI:1.03-9.30, p=0.04) for a positive QFT and 1.72 (p=0.60) for an indeterminate QFT result. Conclusions: Our study demonstrated that QFT test predicted incident active TB disease in HIV-infected persons, with a hazard ratio of 3.10. QFT can be used for diagnosis of LTBI in HIV-infected persons, to allow targeted treatment in this high risk group. Page 49 of 54 P134 Nocardiosis - an emerging complication in the clinical management of HIV infected patients Chioma Onyinye Nwuba*, Gabriel Kogo, Ngozi Ogbu, Oluwafemi Abolarin, Robert Okonkwo Pro-Act, Msh, Ilorin, Nigeria E-mail: [email protected] Retrovirology 2012, 9(Suppl 1):P134 Introduction: In Nigeria as well as other parts of Africa, little is known regarding the prevalence of nocardiosis among HIV positive patients. Nocardiosis is usually not considered in the differential diagnosis for tuberculosis (TB) since it is not regarded as an AIDS defining illness. The aim of this study is to determine the prevalence of nocardiosis in HIV positive patients suspected of having tuberculosis. Materials and methods: In this prospective study, sputum samples of 234 HIV positive patients with suspected cases of pulmonary tuberculosis were analyzed for Mycobacterium TB and Nocardia specie. Each sample was processed by conventional Ziehl Neelsen stain and examined for the presence of acid fast bacilli (AFB). AFB negative samples were streaked on slopes of Sabouraud’s dextrose agar and paraffin coated glass rods which acted as bait for Nocardia was introduced into each of these inoculated media and incubated at 37°C. Cultures were examined after two weeks for the presence of cream/orange tufts around the rods suggestive of nocardia. The isolates were scraped and further identified using biochemical tests. The CD4 cell count of each patient was estimated using Becton Dickenson FACS count system. Results: Of the 234 patient samples examined, 8 had positive culture for Nocardia. The prevalence of TB was 10.3% while that of Nocardia spp was 3.4%. All cases of nocardiosis detected was found in patients with CD4 count of <200cells/ul with 75% of these cases having CD4 count below 100cells/ul. Out of the 8 patients diagnosed with nocardiosis in this study, 7 (7.4%) were not receiving antiretroviral therapy (ART) while 1(0.7%) with a CD4 count of 109 cells/ul had already commenced ART. We also observed that all 8 patients diagnosed with nocardiosis all had a negative AFB result after producing three sputum samples for TB analysis. Conclusions: In Nigeria where HIV-related tuberculosis occurs frequently, some patients diagnosed as having sputum smear-negative pulmonary tuberculosis actually have nocardiosis. It is pertinent that TB laboratories include gram staining during routine investigations of sputum samples especially for patients who present with typical features of active tuberculosis but whose smears are repeatedly negative for AFB. P135 Marginal zone lymphomas in HIV patients Philippe Genet*, Christine Fourcade, Virginie Masse, Bouchra Wifaq, Laurent Sutton, Dris Chaoui, Ahmad Al Jijakli, Nina Arakelian Hématologie at Centre Hospitalier Victor Dupouy, Argenteuil, France E-mail: [email protected] Retrovirology 2012, 9(Suppl 1):P135 Incidence of Non-Hodgkin’s Lymphoma (NHL) remains high in the context of HIV infection. High-grade NHL are the sub-types commonly seen. Indolent lymphomas have been rarely described although several cases of follicular or marginal zone lymphoma (MZL) have been published. Among MZL, MALT lymphomas are predominant. We describe 3 cases of MZL with leukemic presentation. In our on-going cohort of 580 HIV-patients, 3 cases of MZL were identified. The following characteristic were recorded: clinical exam, thoracoabdominal CT-scan, cytological aspect of the peripheral blood smear, immunologic and cytogenetic analysis. There were 2 male and 1 female. Duration of HIV infection was 8, 13 and 18 when MZL was diagnosed. HCV serology was positive in 2 cases with pcr negativity in one case. In all cases, MZL was diagnosed during the occurrence of a mild hyperlymphocytosis: 5000, 6000 and 9000/mm3 respectively while hemoglobin and platelets remained normal. All patients were received HAART for more than 8 years and all have CD4 above 500/mm3 and viral load under 50 copies/ml. All patients were asymptomatic without peripheral adenopathy neither hepatosplenomegaly. On thoracoabdominal CT scan, only infracentrimetric adenopathy were detected. On peripheral smears, several lymphocytes with a villous aspect were detected for each patient. Retrovirology 2012, Volume 9 Suppl 1 http://www.retrovirology.com/supplements/9/S1 In all cases, immunologic phenotype by FACS was consistent with the diagnosis of MZL. Karyotype of the peripheral lymphocytes was normal in one case, revealed an isochromosome 3 in the second case and showed 46, XX, der(1)t(1;?), t(8;14;18)(q24;q32;q21) in the last case. After a follow-up of 2, 2 and 5 years, all patients are alive with a stable disease without chemotherapy. These data suggest that, as for non-HIV infected patients, MZL have an indolent course in HIV patients. P136 HIV-associated multicentric castelman disease, a report of 5 cases Sylvie Jonckheere*, Jean-Cyr Yombi, Anne Vincent, Leila Belkhir, Dunja Wilmes, Bernard Vandercam Medecine Interne Infectiologie at Centre Refrence St Luc Ucl, Bruxelles, Belgium E-mail: [email protected] Retrovirology 2012, 9(Suppl 1):P136 Introduction: Multicentric Castleman’s disease (MCD) is a rare, non-clonal lymphoproliferative disorder characterized by constitutional symptoms, anaemia and generalised lymphadenopathy. Aim: The present study intends to compare demographic features, clinical presentation, laboratory studies, imaging results as well as treatment regimens and out- come in our MCD patients to those of larger reported series. Method: We reviewed the files of 930 HIV-1-infected patients from our AIDS Reference Centre. Data was collected from the operating software for the patients’ medical records of our institution. Results: We report a series of five cases of MCD among our HIV/AIDS patients’ cohort. Three were of African origin. They were diagnosed after 2003, after a mean duration of 85 months of HIV-seropositivity. All presented with characteristic clinical features and laboratory findings (table 1), and all but one patient were started on HAART only a few months before or upon MCD diagnosis. Four patients were treated with chemotherapy, and one with HAART only. One patient who was given Adriamycin/Bleomycin/ Vinblastin is in continuous remission after 6 years of follow-up. Two are alive, with good symptom control, regardless of the treatment they received. One recently relapsed, and one unfortunately died before completing the intended 6-courses chemotherapy regimen. Conclusion: MCD is a rare, but rising issue among HIV-infected patients. The clinical and paraclinical features of our series of five patients are in keeping with those of larger reported series. Currently, treatment is mainly chemotherapy-based, but a wide variety of protocols have been used, mainly because of the lack of available evidence. New approaches such as anti-CD 20 antibodies seem highly effective, and the role of HHV-8 needs to be further investigated, as it might be an important target for future treatment. P137 Study of depression and its associated factors among women living with HIV AIDS in coastal South India B Unnikrishnan*, V Jagannath, JT Ramapuram, S Hegde Kasturba Medical College (Manipal University), Mangalore, India E-mail: [email protected] Retrovirology 2012, 9(Suppl 1):P137 Backgound: Depression is one of the most prevalent psychiatric diagnoses seen in HIV-positive individuals. Women with HIV are about seven times more likely to be depressed than those who are not infected, Depressive symptoms in women are associated with impaired adherence to antiretroviral therapy (ART), higher HIV plasma viral loads, higher mortality, less social support and a worse quality of life.The study was carried out with the objective to assess the sociodemographic and clinical correlates of Depression among Women living with HIV/AIDS. Method: The crossectional study was carried out in one Public and one Private hospital in Mangalore, Coastal South India. The study subjects included 137 HIV Positive women enrolled after obtaining written informed consent. The data was collected using a pretested semi structured proforma which elicited information about socio demographic variables, HIV related Page 50 of 54 clinical and laboratory data .Depression was assessed using BDI (Becks Depression Inventory), Lubben Social Network Scale was used to assess social support. The data was analyzed using SPSS Version 11.5, for univariate analysis chisquare test and t test were used and for multivariate analysis step wise logistic regression model was used. The study was approved by the Institutional Ethics Committee of Kasturba Medical College, Mangalore. Results: Among 137 HIV positive women, 51% were depressed. The mean (SD) age of the subjects was 35 (7) years. The mean (SD) CD4 count among those with Depression was 328 (125). Majority (63%) of the subjects were not on ART and 16% were having moderate to high risk for isolation. In multivariate regression model, living in rural area, Widowed, Lower socio economic status were significantly associated with depression. Conclusion: Depression is highly prevalent among women living with HIV and there is a need to incorporate mental health services as an integral component of HIV care. P138 Adverse pregnancy outcomes in HIV positive women. A study from a District General Hospital in the UK Priya Thayaparan*, Mohanarathi Kawsar, Thambiah Balachandran Luton and Dunstable Hospital, Luton, UK E-mail: [email protected] Retrovirology 2012, 9(Suppl 1):P138 Background: Increasing number of women with HIV are choosing to become pregnant as there is reduction in vertical transmission. However, management of HIV in pregnancy still poses a variety of challenges and adverse pregnancy outcomes are still common. We aimed to explore the factors associated with adverse outcomes of pregnancy in our HIV cohort. Methods: It is a retrospective case notes review of all the women attended to our unit and had the HIV care from 2008-2011. A total of 87 women were followed up. Three women had two pregnancies during the study period. Data collected from Genitourinary Medicine and maternity records were analysed by using SPSS program. Results: Mean age was 34 yrs ranging from 20-43 yrs. Majority (91%) were of African origin; 67% had HIV subtype C; 26% resistant to one or more class of HIV drugs; 55% had a nadir CD4 fewer than 350; 44% diagnosed at an antenatal setting and 62% were planned pregnancies. Prior to the current pregnancy, these women had 121 children: 5% of the children have HIV and 33% not tested for HIV. None of the children born during the study period were infected with HIV; there were 3 sets of twins; one still birth and one child died soon after birth. Around 46% were on anti retroviral therapy (ART) during conception, 6% had miscarriage and 16% had emergency caesarean section. 38% of the women experienced an obstetric complication, premature labour 9%; premature rupture of membranes and gestational diabetes both accounted to 4% whilst 3% had post partum haemorrhage. On ART during conception and late HIV diagnosis, nadir CD4, less than 350 cells were significantly associated (P< 0.05) with having a foetal complication such as prematurity 8%, low birth weight 7% or having a foetal abnormality 2.3%. More analysis is awaited as to drug exposure and adverse outcomes. Conclusion: Late diagnosis of HIV and ART during conception is significantly associated with adverse outcomes of pregnancy. Widespread HIV testing is essential and has to be extended to non traditional settings. P139 Gastric adenocarcinoma is not an HIV related malignancy Violet Kayamba1*, Akwi Asombang2, Mpala Mwanza1, Edford Sinkala1 1 Tropical Gastroenterology and Nutrition Group, University of Zambia School of Medicine, Zambia; 2Washington University School of Medicine in St Louis, Missouri, USA E-mail: [email protected] Retrovirology 2012, 9(Suppl 1):P139 Introduction: HIV infection has been shown to increase the risk of developing some malignancies. We evaluated the possibility of an association between gastric cancer and HIV infection in patients seen at Retrovirology 2012, Volume 9 Suppl 1 http://www.retrovirology.com/supplements/9/S1 the University Teaching Hospital, Lusaka, Zambia. Other known risk factors such as infection with Helicobacter pylori (H. Pylori), presence of CagA, serum pepsinogen 1 to 2 ratios, smoking and alcohol intake were also evaluated. Methods: This was a prospective case-control study with cases being patients with gastric adenocarcinoma confirmed by histopathology while controls were patients without visible mucosal abnormality. Two controls were enrolled for each case after matching for age and sex. The presence of HIV and H.pylori antibodies, the virulence factor CagA and serum pepsinogen 1 and 2 levels were determined using ELISA. Odds ratios were calculated to determine the presence of any association. Results were analysed using STATA 10. Results: A total of 38 cases and 76 controls were enrolled. There was no association between gastric cancer and HIV infection (Odds Ratio 1.41, 95% CI 0.3-6.4; P=0.73). Smoking and alcohol were found to increase the odds of developing gastric cancer, with P values after multivariate logistic regression of 0.04 and 0.02 and odds ratios of 3.5 and 3.1 respectively. Overall, 81% of the patients were found to be positive for H. pylori infection, with no significant difference between the cases and the controls (P=0.24). The presence of antibodies to CagA was also not different between the two groups (P=0.79). Serum levels of pepsinogen 1 were not significantly different between the two groups (P=0.45). However, the presence of a low pepsinogen 1 to 2 ratio was more discriminating, with a higher proportion among the cases (P=0.009). Conclusions: No association was found between gastric cancer and HIV infection. Alcohol and smoking increase the odds of developing gastric cancer. Patients with gastric cancer have lower pepsinogen 1 to 2 ratios, although there is no difference in the levels of pepsinogen 1, H.pylori infection and CagA between gastric cancer patients and healthy controls. P140 Treatment outcome and survival for HIV and AIDS patients coinfected with Mycobacterium Tuberculosis in Nava Kiran Super Speciality Center for AIDS care in Nepal Ujjwal Baral*, Prakash Yogi, Anita Pradhan, Ramesh Pandey National Association of People Living With Hiv and Aids in Nepal, Kathmandu, Nepal E-mail: [email protected] Retrovirology 2012, 9(Suppl 1):P140 Topic: Treatment outcome and survival for HIV and AIDS patients coinfected with Mycobacterium Tuberculosis in Nava Kiran Super Speciality Center for AIDS care in Nepal. Introduction: Tuberculosis (TB) is a frequent disease among People Living with HIV/ AIDS (PLHAs) in Nepal.Prevalence is more than 70% and TB is still a major killer in PLHAs and first Oppertunistic Infection (OI) among PLHAs. We intend to assess prevalence, clinical features, treatment outcome, resistant to first line antitubercular drugs and survival of PLHAs with TB. Method: A longitudinal retrospective-prospective study of clinical records from co-infected before and after Antiretroviral Treatment (ART) with confirmed TB diagnosis or clinical suspect was done. Sensitivity of drugs performed in cases of Treatment failure or epidemiological issues. Treatment success was defined as cure; poor outcome included incomplete treatment, failure or death. Results: 8913 patients since 2007 to 2009 admitted at Nava Kiran Super Speciality Center, 680 clients were co-infected. TB incidence was 68.9%. Age ranged from 16 to 61 years, 75% were under 40 years and 520 clients that is 76.47% were male. HIV infection route; intravenous drug users 59.26%, heterosexual 28.32%, MSM 8.15%, and others were 28.32%. Mean CD4 was 150 cells/cml. TB was first incident in 59.6%. Pulmonary TB account 590 (86.76%) with acid fast smear positive in 87% and extrapulmonary TB counted 90 (13.23%). Multiple Drug Resistant (MDR) TB account 52 (8.8%) of pulmonary TB. In pulmonary TB complete cure was obtained in 64.03% and 22.97% in MDR TB inspite of adequate treatment. Success was high in Pulmonary TB. Conclusion: TB is devastating situation in PLHAs of Nepal. A high suspect, early diagnosis and documented sensitivity improve the outcome and diminished sequel and mortality. Page 51 of 54 P141 Depression, testosterone concentration, sexual dysfunction and methadone use among men with hypogonadism and HIV infection Mahmood Amini Lari*, Hosain Faramarz, Mesbah Shams, Maryam MarzbanA, Nader Parsa Shiraz Hiv Aids Research Center, Shiraz, Iran E-mail: [email protected] Retrovirology 2012, 9(Suppl 1):P141 Background: Hypogonadism is known to be prevalent among patients with HIV infection. Low testosterone levels are associated with depression and impaired sexual performance. The purpose of this study was to determine the impact of hypogonadism on sexual function and depression among men with HIV infection in Shiraz, Iran. Material and methods: A total of 237 patients referred to voluntary counseling centers were recruited based on convenience sampling. All patients provided informed consent and completed the Beck Depression Inventory and Brief Male Sexual Function Inventory (BMSFI) from May to October 2009. Early morning fasting blood samples were collected to measure free testosterone (FT) concentration. Results: According to FT level, 67.8% of the participants had hypogonadism. About 68% had moderate and severe depression. According to the t-test, serum FT levels were significantly lower in patients with depression (t=1.97 & P=0.046). By univariate regression analysis the odds ratio of having a higher depression score was 1.96 times higher in men with hypogonadism than in eugonadal one (CI: 1.09-3.58). Methadone use was significantly associated with FT (OR = 1.8, 95% CI: 1.01-3.21). We found a significant inverse relationship between sexual drive, erectile and ejaculatory function domains of BMFSI with hypogonadism. In an analysis of four subgroups we investigated the effect of methadone and depression on BMSFI domains. An inverse association was found in methadone non-user and non-depressed patients were seen between hypogonadal and eugonadal men in three domains of BMSFI. However, there was no significant association between methadone user status and depression. Conclusion: Depression and hypogonadism had a reciprocal effect. Depression and methadone use were associated with hypogonadism and had significant effects on sexual function. P142 Intellectual quotient score comparison between HIV-infected and HIV exposed children at the Peruvian National Institute of Child Health, Lima Peru Peggy C Martinez1*, Silvia M Montano2, Julio Flores3, Viviana Granados3, Jose Rodriguez1 1 Instituto Nacional de Ciencias Neurologicas, Lima, Peru; 2U.S. Naval Medical Research Unit-6 Lenka Kolevick; 3Instituto Nacional de Salud del Nino, Lima, Peru E-mail: [email protected] Retrovirology 2012, 9(Suppl 1):P142 Introduction: Pediatric HIV encephalopathy includes motor and cognitive deficits and can result in poor school performance, borderline intelligence or mental retardation. We compared intellectual quotient (IQ) between HIV-infected children and a group of HIV-negative children at the National Institute of Child Health in Peru. In addition, we studied the relationship between verbal and executive function and the clinical stage of HIV infection (CDC classification), HIV viral load, CD4 count and other potential risk factors. Materials and methods: We evaluated 28 HIV-infected children and 28 HIV-uninfected children matched by age and sex. The second group was exposed to HIV during pregnancy. Neuropsychologic testing to determine the intellectual quotient (IQ) included the Wechsler-Revised (WPPSI-R) for children between 3 - 7 years and 3 months, and the Wechsler Third Edition (WISC-III) for children older than 7 years and 4 months. Clinical records were reviewed to gather medical history, clinical and demographic data. T-test and Pearson’s correlation (R) were used to compare groups. IQ score was categorized as average or above average, low average, and borderline intellectual function or below for IQ scores > 90, 80-89, < 80, respectively. Retrovirology 2012, Volume 9 Suppl 1 http://www.retrovirology.com/supplements/9/S1 Results: The average IQ among HIV positive children was lower than the control Group (84.6 vs. 91.7, p =0.05). This difference was driven by the verbal sub test (81.2 vs. 90.3, p =0.05). The percentage of children with average IQ or higher was higher among the control group (57.1%) in comparison with the cases (32.1%); more HIV-infected children scored in the borderline intellectual function or below range (35.7% vs. 17.9%, respectively), but these differences were not statistically significant. Children with advanced HIV infection had lower IQ scores and verbal performance than children with less-advanced HIV infection, but these differences were not statistically significant. Verbal test scores were negatively correlated with HIV viral load (R=- 0.424, p=0.024). Conclusions: HIV positive children show lower IQ scores in comparison with HIV negative children. Further studies are needed to confirm our findings. P143 Adrenal insufficiency in patients with acquired immunodeficiency syndrome-an underestimated problem Deepak R Madi*, Shashidhar Khanapure, John Ramapuram, Basavaprabhu Achappa, Sathish Rao Manipal University, Mangalore, India E-mail: [email protected] Retrovirology 2012, 9(Suppl 1):P143 Introduction: Adrenal insufficiency is a serious complication of AIDS. Usually the integrity of HPA (Hypothalamo pituitary) axis is assessed by measuring cortisol response to 250μg of ACTH. Recent studies have shown that lower ACTH dose increases the sensitivity of the procedure. In the present study we have tried to estimate prevelance of adrenal insufficiency using low dose ACTH test (1μg). Primary objective: To estimate the prevelance of adrenal insufficiency in AIDS patients using low dose ACTH test. Research design and methods: 50 patients with confirmed diagnosis of HIV were included in the study. History and physical examination were recorded. Lab investigations included Cd4 count, serum cortisol and low dose acth stimulation test. Data of patients with adrenal insufficiency (GROUP 1) was compared with those without adrenal dysfunction (GROUP 2). Stastical analysis was done using appropriate tests. Results: 37/50 (74%) of study subjects had adrenal insufficiency. Basal cortisol in (GROUP 1) and (GROUP 2) was 10.09μg/dl and 21.95μg/dl (P < .05). Cortisol post Acth stimulation test in (GROUP 1) and (GROUP 2) was 9.49 μg/ dl and 19.93μg/dl (P < .05). Mean Cd4 count in (GROUP 1) and (GROUP 2) was 138.7±56.17 cells/μl and 171.8.7±25.41cells/μl (P < .05). Blood glucose, serum sodium was low and serum potassium, eosinophil counts were high in (GROUP 1) when compared to (GROUP2). Conclusion: Adrenal insufficiency in patients with acquired immunodeficiency syndrome is a common problem in clinical practice. P144 Causes of death in vertically infected paediatric HIV-seropositivesKarnataka experience Suresh Shastri*, Bharat Rewari, Pavithra Boregowda Karnataka State Aids Prevention Society, Bangalore, India E-mail: [email protected] Retrovirology 2012, 9(Suppl 1):P144 Background: Children represent a population at higher risk of HIV-1 infection and AIDS-related death. Tuberculosis is a common cause of severe lung disease and death in children infected with HIV, particularly those living in areas of high tuberculosis prevalence. We investigated the causes of death in HIV-infected paediatric patients. Methodology: A retrospective survey conducted in 29 ART centres in Karnataka, India. Medical records of all deaths that occurred between January and September 2011 amongst paediatric patients were reviewed. Immediate and underlying causes of death were described. Results: Ninety-seven deaths occurred between January-September 2011. 55% of the deceased were males. The mean duration of survival on antiretroviral therapy was 36 weeks. Median age at time of death was 10 years (range 1-19) median CD4 count was 210 cells/μl (7-2500); 57% had CD4 Page 52 of 54 cell count <250 cells/μl. In all, 64 causes of death were reported. In 44% (28/ 64), the causes were WHO clinical stage IV AIDS-defining illness, 36% (23/64) and 2% (1/64) were stage III and stage II conditions, respectively. Adverse effects to anti retrovirals were noted in 6% (4/64) of patients. Multiple causes were also reported in 6% (4/64). Other immediate causes of death were cardio respiratory arrest (2), suicide (1) and intra-cerebral haemorrhage (1). Infections were noted in 22% (14/64) patients. 43% (23/64) patients had tuberculosis at the time of death. Patients dying from AIDS-related events were more often men (17 out of 31). Conclusion: Although antiretroviral therapy has substantially and dramatically decreased AIDS-related opportunistic infections (OIs) and deaths, prevention and management of OIs remain critical components of care for HIV-infected children. P145 Clinical, brain imaging and therapeutic evaluation of toxoplasma encephalitis in HIV-infected patients in Yaounde Annick Mélanie Magnerou*, V Sini, P Ongolo-zogo, JY Fonsah, AK Njamnshi, L Kaptue Université des Montagnes, Douala, Cameroon E-mail: [email protected] Retrovirology 2012, 9(Suppl 1):P145 Introduction: Toxoplasma encephalitis is very common in patients with AIDS. The diagnosis is based mainly on the response to medical treatment. Objectives: To evaluate clinical diagnosis, CT-scan findings, therapy and evolution of AIDS patients with toxoplasmic encephalitis. Material and methods: It was a descriptive cohort study with a prospective and a retrospective phase. Neurologic deficit was scored by using the NIHSS (National Institute of Health Stroke Scale). Results: Sixty consenting patients were recruited, the mean age was 38.7 ± 9.7 years. The F/M sex ratio was 1.6. The mean CD4 cell was 53.5 ± 42.6 /μl. The main presenting complaints were motor deficits (65 %), seizures (40 %), headaches (31.7 %), language and the speech disturbances (35 %). Signs of meningeal irritation and raised intracranial pressure were found in 21 % and 10 % of the cases respectively. The brain lesions were all heterogeneous with contrast enhancement in 80.8 % of cases. Most lesions were supratentorial in 92.3 % of cases, and multilobar in 69.2 %of cases. Abscesses were multiple in 51.7 % of cases, and associated with brain herniation in 61.5 % and hydrocephalus in 30.8 %. With adequate treatment, 61.7% had complete resolution and 13.3 % of the patients, had persistence of neurologic signs at the end of the intensive treatment period. The neurologic deficits improved by 50 % by the 7th day of treatment and even more by the 14th day. The most encountered treatment options with comparable outcomes were sulfadiazinepyrimethamine in 61.7 % of the cases, followed by Trimethoprimesulfamethoxazole in 31.1 %. Conclusion: Focal neurologic deficits of progressive onset, with or without headache and seizures, are the signs and symptoms that alert the suspicion of toxoplasma encephalitis in HIV-infected patients. Adequate treatment leads to improvement in neurologic deficits from the first week of treatment, measured with the NIHSS. P146 Profiles of opportunistic infections in people living with HIV followed at the Military Hospital of Kinshasa Reference (Camp Kokolo), DRC NE Kamangu*, NH Situakibanza, LG Mvumbi, IL Kakudj, TD Tshienda, TG MESIA Université de Kinshasa, Kinshasa, Congo E-mail: [email protected] Retrovirology 2012, 9(Suppl 1):P146 Introduction: In the Democratic Republic of Congo (DRC), Opportunistic Infections (OI) are still a major problem in the care of People Living with HIV (PLHIV). Through the military medical program, few data are available regarding the treatment and prevention of OI. Because of their high mobility and the environment in which they live, the security forces may be considered a population at risk. This study aims to determine the profile of opportunistic infections encountered in PLHIV supported the Military Hospital of Kinshasa Reference (Camp Kokolo / DRC). Retrovirology 2012, Volume 9 Suppl 1 http://www.retrovirology.com/supplements/9/S1 Methodology: This study was conducted at the Military Hospital of Kinshasa Reference (HMRK). It is a literature review that focused on issues of HIV adult patients 18 years followed in the course of January 1st to December 31st 2010. The elements of interest were: age, sex, infections diagnosed. Records that did not contain all of these data were not included. Results: Tuberculosis (43.2%), candidacies (oral (16.7%), vaginal (2.3%) and esophagus (1.5%)) prurigo (15.9%), pneumonia (11, 4%), malaria (10.6%), herpes zoster (9.8%), isosporiasis (5.3%), condyloma (3.8%) and salmonella (3.8%) infections were found in patients with HIV attending HMRK. Conclusion and recommendations: Tuberculosis, oral candidacies, prurigo, pneumonia and malaria are the opportunistic infections found in these patients. We believe it is necessary to systematize the search for co-association of tuberculosis in any PLHIV. P147 Disparities in the prevalence of AIDS related opportunistic infections in Nigeria- implications for initiating prophylaxis based on absolute CD4 Count Chioma Onyinye Nwuba*, Robert Okonkwo, Oluwafemi Abolarin, Ngozi Ogbu, Pauline Modebelu Pro-Act Msh, Ilorin, Nigeria E-mail: [email protected] Retrovirology 2012, 9(Suppl 1):P147 Page 53 of 54 Introduction: Coinfection with viral hepatitis (VH) and some opportunistic infections (OI) can decrease HIV-infected patients’ survival. The purpose of this study was evaluation of humoral immunity factors in the commonest concomitant and OI. Materials and methods: 110 HIV-infected patients were involved in the study. Among them 76% had candidiasis, 74% VH, 23% HSV-infection. Determination of IgG, IgM, and IgA was performed by immune turbidimetry method by using Human test kits, Germany. Statistic analyses were performed by parametric and non-parametric procedures for paired samples T-test. Results: There was no significant difference in serum IgG levels between overall study population and individual patient groups. The most significant difference in IgM concentrations was found in the group with HSV. Thus, IgM level in patients with HSV was significantly lower in comparison with group without HSV (p=0.014) as well as with patients with both HSV and VH (p=0.029). IgA level was higher in patients without candidiasis (p=0.022) in comparison with general study population. With purpose to clarifying the degree of intra-group heterogeneity in Ig parameters patient were additionally divided into subgroups. It was revealed that IgG concentration in patients with oral candidiasis (1663±59 mg/dl, n=32) was different from that in the patients with not only oral candidiasis, but other localizations (1945±89 mg/dl, n=48, p=0.05). It is important to mention that higher IgM и IgG were enrolled in VH with moderate and high activity of ALT and AST. Figure 1 Conclusions: Difference in humoral immunity disorders in HIV-infection depend on certain associated infections as well as on their severity. Introduction: The differences that exist between the CD4 count strata at which certain opportunistic infections occur in sub Saharan Africa suggest that targeting prophylaxis at HIV patients in this part of the world based on data generated from developed countries may be one of the reasons why there is high HIV-related morbidity in Africa. The aim of this study was to investigate the changing incidence of some AIDS defining illnesses in Northern and Southern parts of Nigeria, relating them to different CD4 count strata. Materials and methods: In this study, sputum samples of 234 HIV positive patients were analyzed for Mycobacterium tuberculosis and Pneumocystis jiroveci while 202 stool samples of HIV patients were analyzed for the presence of opportunistic intestinal parasites. We considered five CD4 strata (0 to 99, 100 to 199, 200 to 399, 400 to 499 and > 500 cells/μL). Incidence of the various opportunistic infections and their occurrence within each CD4 strata were estimated using simple statistical method. CD4 count of each patient was estimated using the Becton Dickenson FACSCount system. Results: The prevalence of Mycobacterium tuberculosis, Pneumocystis jirovecii, Cryptosporidium parvum, Isospora belli and Cyclospora spp were 10.3%, 41.9%, 30.8%, 24.2% and 4.4% respectively. All the opportunistic infections occurred at higher rates in CD4 counts less than 200cells/ul (p < 0.0001) except for tuberculosis which occurred highest at CD4 counts >200cells/ul (16 out of the 24 positive sputum smears were recovered in patients with CD4 counts >200 cells/ul). Despite pronounced immunosuppression, P. jiroveci was not detected in sputum samples of 8% patients with CD4 count <200 cells/ul. Opportunistic parasites occurred almost exclusively at CD4 count <200 cells/ul. However, 6.4% of these parasites were isolated in patients with CD4 >200 cells/ul. Cryptosporidium parvum (30.8%) was the most frequently encountered opportunistic parasite, followed by Isospora belli (24.2%) and Cyclospora specie (4.4%). Conclusions: Although HIV related opportunistic infections are often reported to occur exclusively at CD4 count <200 cells/ul in patients; the result of this study shows that disparities exist and so, the possibility of opportunistic pathogens must remain in the differential diagnosis of infections in HIV patients independent of absolute CD4 count. P148 Immunoglobulins concentration in HIV-infected patients’ with viral hepatitis, candidiasis and herpes simplex viral infection Narina Sargsyants*, Tigran Davtyan Armenicum Clinical Center, Yerevan, Armenia E-mail: [email protected] Retrovirology 2012, 9(Suppl 1):P148 Figure 1(abstract P148) Retrovirology 2012, Volume 9 Suppl 1 http://www.retrovirology.com/supplements/9/S1 P149 Clinical, diagnostic and Cryptococcosis neuroméningées evolutionary aspects in HIV infection Rais Mounira*, Amel Ouyahia, Abedelkader Gasmi, Wahiba Guenifi, Houda Boukhrissa, Abdelmadjid Lacheheb Maladies Infectieuses at Chu Setif Algerie, Setif, Algeria E-mail: [email protected] Retrovirology 2012, 9(Suppl 1):P149 Context: Cryptococcosis is a deep Mycosis of reserved in AIDS prognosis Goal. The objective of our study is to analyze the characteristics of clinical, diagnostic and scalable of the NJC in the PVIH. Materials and methods: Descriptive and retrospective study conducted from medical records of positive HIV patients hospitalized for neurological disorders of 2000-2009 in the service of infectious diseases CHU of Sétif. Results: In 12 patients hospitalized for neurological disorders, four cases of meningitis, cryptococcal (02 men and 02 women) have been diagnosed. The average age of our patients is 45 years; the HIV contamination was sexual in 100% cases. The NJC was the fact of discovery of the HIV infection in 01 case. All our patients had clinical events ranking stage C, CD4 performed in all patients were < 100/mm3. The clinical presentation was variable and the clinical signs most frequently encountered were headache, neck stiffness, the alteration of consciousness, fever and the seizures. Examination of CSF to the ink was contributing to the diagnosis in all cases. The culture of CSF made all our patients was positive. The total lymphocyte count showed lymphopenia with an average number of lymphocytes à 800 /mm 3 . Cases of co-infection were recorded (with Mycobacterium tuberculosis in 1 patient, and pneumocystis jirovecii in another) Amphotericin B (0, 7 mg/kg/day) monotherapy was used in all patients in first intention with relay by fluconazole, or in addition to treatment, or for drug intolerance. The evolution of the NJC has been marked by the death of 04 patients. P150 Toxoplasma encephalitis among AIDS patients in a tertiary care hospital in Mangalore, India Anand Venugopal*, Basavaprabhu Achappa Unnikrishnan, B Deepak Madi, Vishak Surendra, John T Ramapuram Kasturba Medical College, Mangalore, India E-mail: [email protected] Retrovirology 2012, 9(Suppl 1):P150 Background: Toxoplasmosis associated with HIV infection is caused by reactivation of a chronic infection and manifests primarily as toxoplasmic encephalitis. This study was done to determine clinical presentations and outcomes of CNS toxoplasmosis and to find out their association with CD4 counts at time of diagnosis and initiation of ART. Materials and methods: Retrospective study done by reviewing medical records of HIV-positiveS diagnosed with toxoplasmosis from Jan 2000 to Dec 2010. Diagnosis was based on clinical features, demonstration of elevated IgG by ELISA and associated CT findings. Data obtained was correlated with CD4 count and whether or not patient was on ART. Analysis was done using SPSS version 11.5. Results: 2826 HIV positives attended Infections Disease Cell from 2000 – 2010, of which 33 (1.12%) had CNS Toxoplasmosis. Among 33 cases, 29 were males (88%) and 4 females (12%). Mean age was 37.33 yrs. 10 cases (30.3%) had CNS toxoplasmosis as the initial manifestation of HIV. Most common clinical presentations were fever(58%) and headache (52%). Mean CD4 at diagnosis of toxoplasmosis was 160.6. Mean level of IgG Page 54 of 54 was 255.69. CT / MRI finding of ring enhancing lesion or cerebritis was seen in 79 % of the cases with 18% of lesions in both basal ganglia and parietal lobes. Cerebritis was most common lesion in CT/MRI, seen in 16 cases while ring enhancing lesions were seen in 10 cases. 82% improved with treatment and 18% expired. Conclusions: The possibility of cerebral toxoplasmosis should be considered in every HIV-positive patient with neurological symptoms. In our study, Toxoplasmosis occured at CD4 levels >150, which should warrant prophylaxis for Toxoplasmosis at higher CD4 count. Parietal lobe lesions were common in our study, contrary to other existing data which say toxoplasma lesions are usually midline lesions. P151 Prevalence of ulcer forming STIs among HIV-positive women clinic attendees in two Nigerian hospitals Victoria Awolade*, Fajuyi Cantonment, Odogbo Ibadan, Onogbogi Olarewaju, UCH Ibadan Health Initiatives for Safety and Stability in Africa, Ibadan, Nigeria E-mail: [email protected] Retrovirology 2012, 9(Suppl 1):P151 Background: The presence of an untreated STI also increases the risk of both acquisition and transmission of HIV by a factor of up to 10. Human Immunodeficiency Virus (HIV) is one of the most common and most dreaded Sexually Transmitted Infection (STI) worldwide. In order to reduce the prevalence of HIV/AIDS infection and subsequently reduce morbidity and mortality among adolescent women, there is need for proper exploration of the relationship between STIs and HIV infection. This study however sought to determine the association between ulcerforming locally endemic STIs and HIV infection. Methods: A seven year retrospective review of 300 case notes of female patients attending the STI clinics of two Nigerian hospitals was done. Those with ulcer-forming sexually transmitted infections were then reviewed to determine age at sexual debut, number of sexual partners and overall sexual behavioural pattern. The results of laboratory tests taken by the patients were also reviewed. Data was analyzed with the use of the SPSS data editor. Chi square tests (95% confidence) were used to determine whether the level of association observed was of statistical significance. Results: Fifty-two patients had been treated for ulcerative STIs (17.3%). The mean age at sexual debut was 16.7±1.3years. Thirty - one cases (59.6%) of those confirmed by laboratory tests to have ulcerative STIs were also found to be HIV+. Patients that had 3 sexual partners or more had the highest incidence of STIs and HIV (23.6%). Syphilis was the highest reported ulcerative STI 28.8% followed by Chancroid 21.1%. There is a significant association between ulcer-forming STIs and HIV infection (p=0.01). Conclusions: Ulcer producing STIs are associated with HIV infection. An aggressive management of STIs and a more effective contact tracing is needed to reduce new infections of HIV and transmission especially in women. Cite abstracts in this supplement using the relevant abstract number, e.g.: Awolade et al.: Prevalence of ulcer forming STIs among HIV-positive women clinic attendees in two Nigerian hospitals. Retrovirology 2012, 9 (Suppl 1):P151