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SEVENTH
INTERNATIONAL
CONFERENCE
ON
BIOTHERAPY
JUNE 21 - 24, 2007
The Convention Hall,
The KimKoo Museum
SEOUL, KOREA
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Organized by
International Biotherapy Society
CAM, Pochon CHA University
SEVENTH INTERNATIONAL
CONFERENCE ON BIOTHERAPY
JUNE 21 ~ 24, 2007
The Convention Hall, The KimKoo Museum
SEOUL, KOREA
ORGANIZATION
 International Biotherapy Society
 Complementary & Alternative Medicine, Pochon CHA University
SPONSOR
 Korean Oriental Medical Practitioners Association
 World Foundation of Apitherapy (Apimeds, Inc.)
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CONTENTS
Page
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2
3
6
12
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78
112
115
IBS, Organizing Committee
Welcome
General Information
Scientific Program
Poster Presentation
Abstracts – Speakers
Abstracts – Contributing Papers
Notes
Subway Map
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INTERNATIONAL BIOTHERAPY SOCIETY
President: Kosta Y. Mumcuoglu, Ph.D. (Israel)
Vice-President: Terence J. Ryan, M.D. (UK)
Secretary: Andrew Jarvis, M.D. (UK)
Treasurer: John C. T. Church, M.D. (UK)
Executive Directors:
 Theodore Chebuliez, M.D. (USA)
 Wim Fleischmann, M.D. (Germany)
 Olga S. Gileva, D.D.S. (Russia)
 Carita Hansson, M.D. (Sweden)
 Christopher MH Kim, M.D. (Korea)
 Theo Rufli, M.D. (Switzerland)
 Kathryn Vowden, R.N. (UK)
Home Page ☞ http://biotherapy.md.huji.ac.il
LOCAL ORGANIZING COMMITTEE
Honorary President: Sae I. Chun, M.D.
President: Christopher MoonHo Kim, M.D.
Members:
 Bank S. Choi, O.M.D.
 Duck H. Kim, V.M.D.
 Hyun S. Kim, O.M.D.
 Kyung H. Kim, O.M.D.
 Young J. Lee, M.D.
 Dae W. Shin, M.D.
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WELCOME
On behalf of the International Biotherapy Society (IBS) and the
Organizing Committee, we would like to welcome and the Seventh
International Conference on Biotherapy. Our warm and sincere greeting
goes out to every participant, particularly oversea guests and to those who
are visiting Korea for the first time.
We hope this Conference will give you a good opportunity to renew
existing links with friends and colleagues around the world, and to fulfill
new collaborations and friendships. The program of conference is going to
take three days and comprise about 17 keynote lectures, 15 contributed
papers, and 12 posters will be presented. We hope you enjoy the scientific
program to your full extent.
The Organizing Committee will assist you in any way during the
Conference.
We hope you have a pleasant time in Seoul.
The Conference Organizing Committee
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GENERAL INFORMATION
The Convention Hall, The KimKoo Museum
255 Hyochang-dong, Yongsan-ku, Seoul, Korea
Phone: xxx822-719-1311 Fax: xxx-822-718-1311
☞ www.kimkoomuseum.org
June 21-24, 2007
The official language of the Conference will be English.
The weather in Seoul in June is mainly sunny with temperatures ranging
from 15ºC/60ºF to 28ºC/82ºF.
Clothing is informal for all occasions.
All participants, accompanying persons and exhibitors are kindly requested
to wear their badges throughout the meeting in order to be admitted to the
lecture halls and other scheduled activities.
The local currency is the Korean Won (1 US Dollar = 940-960 Korean
Won). Cash and traveler's checks can be exchanged at the airport, hotels, or
at local banks. Visa, Mastercard, American Express, and other credit cards
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are accepted at most restaurants, department stores, and for travel.
Certificate of attendance will be available for all participants at the
registration desk from June 23, 2007.
Upon request, the Conference will send a personal invitation to participate.
This invitation is not a commitment on the part of the organizers to provide
any financial support. Applications for invitation letters must be received
by May 30, 2007.
Please check with your travel agent if you require an entry visa to Korea. It
is the responsibility of the participant to obtain a visa, if required.
The IBS and ICB do not accept responsibility for individual, medical,
travel or personal insurance. Participants are recommended to take out their
own travel policies.
- The registration fee will be $350 for regular participants, $175 for
accompanying persons and students, and $150 for one day registration.
- The breakfast is included in the hotel, and the lunch will be provided all
participants at the conference.
- On Friday (June 22) for a fee of $80, the participants will have the
opportunity to participate to one full day tour to the Korea's most famous
tourist sides of the Capital city of Seoul and Korean Folk Village. The
price included lunch and dinner.
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East Side No. 4B, West Side No. 11A
→ Koreana Hotel
→ Seoul Plaza Hotel
→ Lotte Hotel Get OFF!
- President Hotel : 1 minutes on foot
→ KAL Building
The conference will provide it.
SEVENTH INTERNATIONAL
CONFERENCE ON BIOTHERAPY
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____________________________________________________________
SCIENTIFIC PROGRAM
JUNE 21, 2007 THURSDAY
07:30 - 17:00 Registration
09:00 - 10:00 Opening session, welcome and overview
Byung S. Kim, M.D. (Korea), Chairman,
College of Medicine, Pochun Cha University, Korea
Christopher MoonHo Kim, M.D. (Korea),
President of the Local Organizing Committee
Kosta Mumcuoglu, Ph.D. (Israel),
President of the International Biotherapy Society
09:40 - 12:30 SESSION: MAGGOT THERAPY (Part 1)
Keynote Lectures
Chair: John Church, M.D. (U.K.) and Ronald Sherman,
M.D. (USA)
09:40 - 10:25 Clinical aspects of modern maggot therapy
Wim Fleischmann, M.D. (Germany)
10:25 - 11:00 Coffee Break
11:00 - 11:45 Antibacterial substances in maggots of Lucilia sericata
Kosta Y. Mumcuoglu, Ph.D. (Israel)
11:45 - 12:30 Integral Medicine & Biotherapy in the 21st Century
Sae I. Chun, M.D. (Korea)
12:30 - 14:00 Poster Presentation
12:30 - 14:00 Lunch
14:00 - 15:30
14:00 - 14:45
SESSION: MAGGOT THERAPY (Part 2)
Keynote Lectures
Chair: Wim Fleischmann, M.D. (Germany) and Andrew
Jarvis, M.D. (U.K.)
Maggot Therapy - reflections from the past, foretelling the
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14:45 - 15:30
15:30 - 16:00
16:00 - 17:45
16:00 - 16:15
16:15 - 16:30
16:30 - 16:45
16:45 - 17:00
17:00 - 17:15
17:15 - 17:30
17:30 - 17:45
17:45 - 18:00
18:00 - 18:30
future. Ronald Sherman, M.D. (USA)
Maggot therapy in clinical practice and mode of actions
Kristin Hasenoehrl, M.D. (Germany)
Coffee Break
Contributed Papers
Aletha W. Tippett, M.D. (USA)
Maggots to the Rescue: Using maggot therapy for wounds
in hospice patients
Sergey Chernysh (Russia)
Maggot immunity and drug development
Peter H. Nibberin, Ph.D. (The Netherlands)
Maggot Excretions/Secretions Inhibit Multiple Neutrophil
Pro-Inflammatory Responses
Kang-Jun Yoon, M.D. (Korea)
Effectiveness of Maggot Therapy in Chronic Infected
Wound with MRSA
Peter Takac, Ph.D. (Slovakia)
Maggot Debridement Therapy (MDT) in Slovakia
Shouyu Wang, M.D. (China)
The healing and antibacterial function research of the
maggot secretion on the ulcer area
M.J.A. van der Plas (The Netherlands)
Maggot Excretions/Secretions are effective against
biofilms of Staphylococcus aureus and Pseudomonas
aeruginosa
Anders Anderson (Denmark)
Maggot debridement therapy (MDT) in diabetic foot
wounds – Quorum sensing dependent bacterial niches may
promote infection or MDT failure
Round-table Discussions (Keynote Speakers)
JUNE 22, 2007 FRIDAY
City Tour
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JUNE 23, 2007 SATURDAY
08:30 - 17:00 Registration
09:00 - 12:30 SESSION: HIDUROTHERAPY (Part 1)
Keynote Lectures
Chair: Kosta Mumcuoglu, Ph.D. (Israel) and Vladmir
Savino, M.D. (Russia)
09:00 - 09:45 Hidurotherapy in modern medicine
Olga S. Gileva, D.D.S. (Russia)
09:45 - 10:30 Veterinary hidurotherapy
Sagiv Ben-Yakir, D.V.M. (Israel)
10:30 - 11:00 Coffee Break
11:00 - 12:30 SESSION: HIDUROTHERAPY (Part 2)
Keynote Lectures
Chair: Olga S. Gileva, D.D.S. (Russia) and Sagiv BenYakir, D.V.M. (Israel)
11:00 - 11:45 Symbiotic bacterium from medicinal leech as the
acting agent of hirudoautohaemotherapy, autoblood from
medicinal leech as a means of active nonspecific
immunotherapy in oncology.
Vladimir Savino, M.D. (Russia)
11:45 - 12:30 1) The use of medical leeches in plastic surgery
2) Elimination of symbiotic bacteria from the
gastro-intestinal track of the medicinal leech, Hirudo
medicinalis.
Kosta Y. Mumcuoglu, Ph.D. (Israel)
12:00 - 14:00 Poster Presentation
12:30 - 14:00 Lunch
14:00 - 16:00 SESSION: HELMINTHOTHERAPY,
ICHTIOTHERAPY AND BIODIAGNOSTICS
Chair: David Pritchard, Ph.D. (U.K.) and Martin
Grassberger, M.D. (Austria), Edwin Cooper, Ph.D. (USA)
14:00 - 14:50 1. Understanding Mechanism of Maggot & Clinical Datas
2. Hookworms in clinical trials for the treatment of
seasonal rhinitis and Crohn's disease
David Pritchard, Ph.D. (U.K.)
14:50 - 15:20 Ichtiotherapy for patients with psoriasis
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15:20 - 16:00
16:00 - 16:30
16:30 - 16:45
16:45 - 17:45
16:45 - 17:00
17:00 - 17:15
17:15 - 17:30
17:30 - 18:00
Martin Grassberger, M.D. (Austria)
Evidence Based Complementary and Alternative Medicine
Edwin Cooper, Ph.D. (USA)
The evolution of biotherapy
John C. T. Church, M.D. (U.K.)
Coffee Break
Contributed Papers
Galina Korobeinikova (Russia)
The Use of Medicinal Leeches and Medicinal Preparation
from them in the Complex Treatment of Skin and Mucosal
Diseases in UST Health Resort
17:00 - 17:15
Yavuz Dinler, Ph.D. (Turkey)
New development about medical leech therapy
Jiangning Wang, M.D. (China)
Application of maggot therapeutics for repairing serious
infective wound
Round-table Discussions (Keynote Speakers)
JUNE 24, 2007 SUNDAY
08:30 - 12:00 Registration
09:00 - 12:30 SESSION: APITHERAPY (Part 1)
Keynote Lectures
Chair: Christopher M-H Kim, M.D. (Korea) and
Anatoly A. Gribkov, M.D. (Russia)
09:00 - 09:45 Honeybee Venom Clinical and Discovery Research in The
United States and Canada in the 20th Century
Robert Brooks, Ph.D. (USA)
09:45 - 10:30 Use of Apitherapy in Clinical Medicine
Stefan Stangaciu, M.D. (Romania)
10:30 - 11:00 Coffee Break
11:00 - 12:30 SESSION: APITHERAPY (Part 2)
Keynote Lectures
Chair: Robert Brooks, Ph.D. (USA) and Stefan Stangaciu,
M.D. (Romania)
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11:00 - 11:45
11:45 - 12:30
12:30 - 14:00
14:00 - 15:30
14:00 - 14:15
14:15 - 14:30
14:30 - 14:45
14:45 – 15:00
15:00 – 15:15
15:15 - 15:30
15:30 - 15:45
15:45 - 16:15
16:15 - 17:15
1. Prerequisites and methods for intervertebral hernia
treatment by live bee stings and other bees' products
2. Video presentation
Anatoly A. Gribkov, M.D. (Russia)
Bee Venom Therapy in the modern medicine (1985-2005)
C. MoonHo. Kim, M.D. (Korea)
Lunch
Contributed Papers
Duck-Hwan Kim, V.M.D. (Korea)
Treatment by Injection-Acupuncture with Bee-Venom
(Apitoxin) and Apitoxin Combined by Chinese Herbal
Medicine in Patients with Canine Hind Limb Paralysis
Duck-Hwan Kim, VMD (Korea)
Bee venom (Apitoxin) therapy on canine facial nerve
paralysis
Byung-Soo Koo, O.M.D. (Korea)
Effects of Bee Venom on Glioma Cells
Ai-Young Lee, M.D. (Korea)
Effects of bee venom on human melanocyte proliferation,
melanogenesis and dendricity
Nasser M. Shahri, Ph.D. (Iran)
A new in vivomodel for testing effects of topically
biotherapy wound healing
Edwin L. Cooper, Ph.D. (USA)
Hepatoprotective potential of earthworm extract (Lampito
mauritii, Kinberg) against paracetamol induced liver
damage in rats
Edwin L. Cooper, Ph.D. (USA)
Therapeutic Potential of Tunicates
Coffee Break
Panel Discussion: Biotherapy
Moderator : C.T. Church, M.D.
Panel : Wim Fleischmann, M.D.
Olga S. Gileva, D.D.S.
C. MoonHo. Kim, M.D.
Kosta Y. Mumcuoglu, Ph.D.
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Ronald Sherman, M.D.
Stefan Stangaciu, M.D.
17:15
Closing
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POSTER PRESENTATIONS
 Anders Andersen (Denmark)
Maggot debridement therapy (MDT) in a diabetic foot wound patient
suffering from severe gout
 Takuya Kawabata, Ph.D. (Japan)
Induction of antibacterial activity in medicinal maggots by infected
environment
 Myung-Sang Kwon, Ph.D. (Korea)
Water extracted propolis inhibits TNF- α release following LPS injection in
the rat
 Myung-Sang Kwon, Ph.D. (Korea)
Water extracted propolis attenuates kainite-induced neurotoxicity via
adenosine A1 receptor in the rat
 Hideya Mitsui, M.D. (Japan)
Maggot Therapy for severe diabetic foot ulcer
 Alicia Fonseca Muñoz (Mexico)
Efficiency Maggot Debridement Therapy in Fournier’s Gangrene
 Alicia Fonseca Muñoz (Mexico)
Study of Maggot Debridement Therapy in necrotic pressure ulcers, Case
Series
 Nasser M. Shahri, Ph.D. (Iran)
Effect of a natural polymer "rabbit vitreous" to wounds therapy on an
experimental model
 Nasser M. Shahri, Ph.D. (Iran)
Macroscopic and microscopic surve of the effectiveness degree of some of
the biostimulators used in traditional medicine in the process of healing of
sheep skin wounds
 Zohrabyan A. Sureni (Armenia)
Maggot therapy in special wound treatment
 Hitoshi Takase M.D. (Japan)
The optimal conditions for medicinal maggots before and during shipment
ABSTRACTS
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for
Keynote Speakers
Veterinary Hirudotherapy
Dr. Sagiv Ben-Yakir BSc(in Biology), DVM(in honor), MRCVS,
CVA(IVAS), CVHomotox’(Baden-Baden,Germany)
“ORSHINA” – The Israeli Veterinary Institute for Holistic Medicine
Medicinal leeches were used for bloodletting, and have been applied to
congested or inflamed parts of the animal body for over 3,500 years. They
are used for engorged blood vessels as in acute equine laminitis, canine
aural hematoma, swollen testicles, laryngitis, canine prolapsed rectum or
bovine prolapsed uterus and inflamed vulva. Leeches are used also in
reconstructive surgery on swollen faces, limbs and digits following
successful arterial re-vascularization but limited venous repair. Leeches are
also used in the treatment of such disorders as inflammation and peripheral
venous and arterial diseases as in cat’s saddle thrombus. We can use
leeches to treat inflammatory and traumatic processes such as keratitis,
chorioretinitis, periorbital hematoma, subretinal hemorrhage, glaucoma and
cataract.
Hirudotherapy is being used to treat inflammatory diseases of the ear, nose
and throat such as acute and chronic otitis, sinusitis, aural hematoma, and
laryngitis, and to treat clinical forms of dermatitis and dermatosis (e.g
eczema, paronychia, acral lick dermatitis etc). Leeches can be used to treat
acute and chronic inflammatory immune--associated oral mucosa and
peridontium lesions as in Feline Immunodeficiency Virus(F.I.V) positive
cats with severe stomatitis and gingivitis, diseases of the salivary glands as
sialadenitis and sialoadenosis.
Furthermore, leeches are used to treat inflammatory conditions of the
urogenital tract as endometritis, dysfunction and injuries of the genital
organs in postoperative rehabilitation etc.
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Leeches are used to rehabilitate animals with postpartum pyo-septic
complications. Hirudotherapy was also found to be effective in healing
intractable and non-granulating wound or ulcers, especially in elderly and
immunocompromized animals.
Leeching has been used as well for treating cats with polycythemia vera, or
to reduce scrotal edema post-castration procedure in adult dogs. In
osteoarthritis leeches do lessen the pain and restore lost mobility as in
canine hip dysplasia (H.D) cases or knee osteoarthrits. Keep in mind that
osteoarthritis is more than a disease of joint’s wear and tear, it is also a
vascular disorder. Small clots can occur in the blood vessels supplying
bone & joint, and starve the tissue for oxygen. Chemicals that thin the
blood and break clots apart could ease animal’s misery as well as assisting
in regeneration.
Before applying the leeches the area to be treated should be cleansed
with clean non-chlorinated water. Remove any traces of povidone-iodine
(Betadine) or any other skin preps, as well as flea spray, dip or trans dermal
preparations. These traces of solutions on the skin may discourage the leech
from attaching. Leeches should be applied in adequate numbers to the
general area of maximal congestion or other treated area/tissue. One or two
leeches may be sufficient to treat an area of 5 square cm. A larger area may
require 3-4 leeches depending on initial clinical response.
The History of Honeybee venom in North America
Robert Brooks, Ph.D.
President, PVA Pharmaceutical Consultant
Honeybee venom in North America emerged as a topic of discussion and
interest in 1935 in New York City with the publishing of the book Bee
Venom Therapy by a Hungarian/American physician Dr. Bodog Beck. Dr.
Beck maintained an active bee hive in his office to provide a supply of live
bees for his patients. In 1941 Dr. Joseph Hollander, then a young
Rheumatologist at the University of Pennsylvania reported the
disappointing results of his well designed double blind clinical trial. Dr.
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Hollander’s trial drug did not utilize stings or the whole venom but rather
the filtrate of ground up whole bee parts containing little if any whole bee
venom. From 1938 through 1964 several attempts were made to
demonstrate the effectiveness of bee venom in the treatment of arthritis. In
1966 a group of scientists gathered at Walter Reed Army Institute of
Research including Biochemist William Shipman from San Francisco,
California. Shipman began to use a then new technique to separate the
components of bee venom, column chromatograph. Using G75-120 and
G25-80 sephadex columns Shipman was able to separate the major
components of bee venom. Shipman named two components mellitin and
apamine identified by their molecular weights and concentrations in the
whole venom. Quickly following him other scientists in England and
Bulgaria published confirmation of his findings with technical additions
and observations. Because of the available technology Shipman was
unable to move beyond the 6-7 fractions he was able to separate but he
always thought that their were a number more whose molecular weights
were closely aligned. He would be surprised to learn how many more
fractions were actually found in the whole venom. Shipman also
characterized the stability of not only the whole venom but all of the
fractions he identified. Working in the pharmacology laboratory at Walter
Reed Army Institute of Research Dr. James Vick and I defined the LD50s
for the whole venom in mice, dogs and monkeys and published our first
work on bee venom. We confirmed the results of Artemov’s work in
Russia. Artemov was the first to publish his in vivo results of ascorbic
acid depletion from which he theorized indirectly the significant increase in
endogenous circulating plasma cortisol. Artemov speculated that this
amount of endogenous cortisol had a therapeutic effect similar to the
injection of cortisone without the unwanted side effects that caused the
abandonment of cortisone as a standard treatment. Dr. Vick and I
confirmed this theory through a new blood study that measured circulating
plasma cortisol directly in the blood. We studied and evaluated this in a
number of ways including a dramatic study showing injected whole bee
venom increased cage activity in older dogs. We interpreted our data as a
demonstration of the therapeutic effectiveness of whole bee venom in
arthritic conditions. We similarly studied the fractions of bee venom with
special attention to their cardiovascular effect. We believed that the
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therapeutic activity of bee venom was much more complicated than the
production of endogenous cortisol but venom fractionation produced only
small amounts of the fractions which limited out ability to explore our
ideas and theory. From our beginning studies which I will provide in
detail in this presentation many hundreds of papers have followed with
increasing depth of knowledge that the improving technology has
allowed. I can specifically offer you some interesting anecdotal details of
the beginnings of a few of these works. As our knowledge of medicine
has also evolved so have the medical specialties of our time like geriatric
medicine and more recently genomic medicine. Only recently have
physicians questioned the need of patients to be free of pain and the
development of pain management as a medical specialty has led to a better
use of whole bee venom for what it has been known for centuries, not the
treatment of arthritis, but the often dramatic and sustained relief of pain.
The Evolution of Biotherapy.
Leeches, Maggots, Worms, Rats & Dogs; parasites or colleagues?
John C T Church, M.D.
In the evolution of scientific discovery, and its exploitation, over the past
two centuries, physics and chemistry can be said to have dominated the
scene. By contrast, the twenty-first century is already being identified as
the Biological Century (1).
The scientific emphasis throughout has been reductionist, but with no end
in sight as the complexities of quantum physics and the genome unfold.
By contrast again, natural biological evolution is holistic. A multiplicity
of unicellular organisms interact, giving rise to a staggering diversity of
multi-cellular organisms, all in their turn altering the environment and
creating ecosystems which are ‘fuzzy’.
‘Biotherapy’ can be defined as ‘the use of living organisms in medicine’.
Most of the natural biological ‘research and development’ in this arena has
occurred over formidably long periods. Thus, for instance insects and
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mammals have been interacting for over 200 million years. Host/parasite
interactions, whether symbiotic or hostile, are highly sophisticated and
complex, perhaps nowhere more so than in the interplay of their respective
immune systems. Dogs and men have lived together since the earliest
days of their respective evolution. The relationships that can develop
between sentient individuals of difference species, such as dog and man,
can be quite as complex as those between individuals of the same species.
None of these interactions lend themselves readily to scientific
measurement.
For the greater part, ‘Biotherapy’ involves the use of normal healthy
organisms, so managed that natural aspects of their behaviour are
appropriately utilised.
Leeches (hirudotherapy), maggots (maggot
débridement therapy, MDT), and scavenger fish (ichthyotherapy), all have
had a place in traditional medicine. In the ‘evolution’ of medical practice,
all these branches of ‘Biotherapy’ are now being incorporated into modern
medical practice, in a variety of ways, albeit mostly in a somewhat
unstructured environment. To this list we can now add helminths
(helminotherapy), and, in the recognition of disease, ‘sniffer’ rats and
‘cancer sniffer’ dogs (biodiagnostics).
Hirudotherapy
The use of leeches in medicine, for bloodletting, dates back to the ancient
civilizations of India and Egypt. Bloodletting in ancient Greece was used
to restore the balance of the four humors, blood, lymph, black and yellow
bile. Throughout history the use of leeches this way has waxed and
waned. In medieval Europe, religious and superstitious ideas caused a
surge in the popularity of bloodletting. In 1833 alone France had to
import ‘over 40 million leeches’!(2).
With the development of microsurgical techniques leeches now have a vital
place in plastic and reconstructive surgery, particularly where venous
congestion hinders healing [Illustration slide 1]. An excellent example of
this is the following case report: A patient was admitted to a plastic surgical
unit having had his ear bitten off by a dog. The dog was caught and the
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ear retrieved. The ear was then taken to hospital with the patient. Its
surgical reattachment included finding a small divided arteriole and
suturing it to restore arterial blood flow into the ear. But no veins of
sufficient size could be found for such suturing. This left a precarious
situation which, if left that way, would have led to engorgement of the reattached ear and failure. But leeches were applied to the rim of the ear for
a sufficiently long period to allow capillary re-growth across the wound,
with ultimate restoration of normal circulation [Illustration slide 2]. So
this patient regained his ear, by dint of prompt action at the roadside,
meticulous micro-vascular surgery, and the judicious application of hungry
leeches (3).
Modern hirudotherapy includes the use of leeches over arthritic joints, in
venous ulcers, in a variety of ophthalmic and neurological conditions (4).
A number of physiologically active biochemical agents in the leech saliva
have been studied in the search for mechanisms for the beneficial effects of
this treatment in these conditions. A word of caution must however be
added. In spite of meticulous culture in dedicated laboratories it is
impossible to fully eradicate bacteria from leech secretions, and appropriate
antibiotic cover is advised during hirudotherapy.
Maggot Débridement Therapy (MDT)
‘Harvested’ maggots have been used for centuries as cleansing agents in
open wounds. They were first cultured in dedicated laboratories, for
clinical use, in the 1930s (5). With the advent of antibiotics maggot
therapy almost disappeared. But with the renaissance of this treatment by
Sherman in the 1980s (6), and in the UK in 1995 (7), Maggot Débridement
Therapy (MDT) has now not only been accepted internationally but has in
many centres become an integral part of modern wound care [Illustration
slide 3-6, or 7]. This development however is by no means uniform, due in
part to negative or cautious attitudes of health care professionals and
managers. It is also due to the difficulties of adequately monitoring and
regulating the production and use of maggots this way. After extended
deliberation FDA approval in the USA was granted in January 2004, and
maggots have been available on prescription in the UK from February 2004.
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Maggot feeding activity in wounds is characterised by at least the following
factors:
 Enzymic digestion of decomposing organic material.
 Antibacterial action, in the wound and in the maggot gut. This
includes the destruction of bacterial biofilm, and suppression of its
formation.
 Enhancement of granulation tissue, presumably by the action of
cytokines.
 Mobility. Maggots are dynamic and respond actively and
appropriately to changes in the wound environment.
 pH. Maggot alkaline exo-secretions reverse acid wound pH.
 Wound warming. Maggot metabolism is exothermic, particularly
so when they are closely packed, in group feeding.
 Enzyme ‘shelf life’. Maggot exo-secretions have a zero ‘shelf life’,
being secreted directly from the salivary glands into the wound.
A number of prospective studies have been undertaken or are under way,
comparing MDT with other wound care products. However, this whole
exercise, though essential for proper scientific assessment, presents
considerable intrinsic challenges. There is no single manufactured
product available than can simultaneously deliver all the factors listed
above, and thus be compared with maggot activity. Equally, it is
impossible to separate or isolate these various factors when healthy
maggots are allowed to feed normally in open wounds.
Ichthyotherapy
This can be defined as the use of fresh water or marine organisms as agents
of wound cleansing. The history of such treatment in traditional medicine
is sparsely documented. In a museum near the River Kwai, recording the
privations of prison camps, a sketch drawn by a prisoner showed him up to
his waist in water, but with small fish attending to his leg ulcers (8).
There is widespread use of such fish in India, particularly in rural areas. In
Kerala State Professor Padmanabham PhD studied this phenomenon in
detail (9) During the sixth International Biotherapy Conference in Turkey,
in June 2003, the Kangal Balikli spa was visited. Warm water pools are
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stocked with three species of small fish that scavenge lesions of the skin,
particularly psoriasis [Illustration slide 8 ] (10).
Helminthotherapy
A number of diseases are associated with auto-immunity dysregulation.
Typical of these are Crohn’s disease and ulcerative colitis. Acting on the
hypothesis that the auto-immune response in these patients is due to lack of
helminth antigenic stimulus in the alimentary tract, studies have been
conducted giving such patients doses of helminth eggs orally.
The
chosen helminth, porcine whipworm, Trichuris suis, is not pathological to
man, but has a positive effect on the host immunity, with reversal of
symptoms (11).
This work opens up a further horizon on the potential to use living
organisms that interact with each other, particularly through immunological
reactions, thereby maintaining healthy co-existance.
Biodiagnostics
Biodiagnostics can be defined as the use of living organisms in the
detection of disease. Though modern medical screening systems are
thorough, they are not universally available, and they are often still far
short of requisite levels of sensitivity and specificity. There are still no
reliable tests for the early detection of most cancers. National screening
in the UK is currently only available for breast and cervical cancer.
Trained sniffer-dogs are now used extensively in the detection of drugs and
explosives, in forensic work, and in a variety of other ways such as the
detection of dry rot and termites in houses. Anecdotal stories have been
published of pet dogs that have apparently caused their owners to seek
advice, and have an early cancer diagnosed and treated, with good result
(12, 13) A controlled pilot study has shown that dogs can be trained to
recognise cancer of the urinary bladder [Illustration slide 9] (14). A certain
species of rat in Tanzania has been trained to recognise tuberculosis in
samples from patients afflicted with this disease (15)
23
This presents us with a extensive field for similar studies, using any species
with a well developed olfactory system that can be trained to recognise
disease at high levels of specificity and sensitivity.
The future of ‘Biomedicine’
Nature provides us with an almost limitless array of species that respond to
chemical or other signals in their environment. There is thus a vast
potential to harness or control such behaviour to give us models for the
early recognition of disease, perhaps most readily when it is characterised
by volatiles expressed by the patient.
Working well with living organisms is exacting.
It demands an
understanding of their biology, and enough control of their ‘working’
environment to result in ‘optimal’ behaviour. Their use in the clinical arena
is thus an art. With skilful and appropriate use, excellent clinical results
can be obtained.
It must be expected that all this ‘fuzzy-edged’ activity will become
progressively more integrated into modern clinical practice. This already
presents its own array of challenges for its requisite monitoring and
evaluation, and the assessment of clinical outcomes, but guidelines for all
this activity are being established.
References:
1. Benford G. The Biological Century
http://reason.com/9511/BENFORDfeat.shtml
2. El-Awady A. Maggots and leeches make a comeback. Science in
Africa July-Aug 2003
http://www.scienceinafrica.co.za/2003/july/leech.htm
3. Teo TC. Personal case.
Sherman RA. Maggot Therapy – The Last Five Years. ETRS Bulletin
Vol 7 Issue 3. 2000
4. Gilyova O. Modern Hirudotherapy – a Review.
http://www.scienceinafrica.co.za/2003/july/leech.htm
5. Lederle - Surgical Maggots. Council of Pharmacy and Chemistry.
New and non-official remedies.
JAMA 1932;98:401
24
6. Pechter EA, Sherman RA. Maggot therapy: the surgical
metamorphosis. Plast Reconstr Surg 1983;72(4):567-570
7. Church JCT. ETRS working party consensus paper on wound
debridement. Surgery 1995;13(10):228c-d
8. JCTC. Personal observations April 1998
9. Cohen J. Feeding the fish. BMJ 2000;320:181
10. Church JCT. 2003 Report on the Sixth International Biotherapy
Conference, Turkey
http://biotherapy.md.huji.ac.il/newsletter07.htm
11. Weinstock J, Summer WR. Will Helminths Become the Future
Treatment for Inflammatory Bowel Disease?
http://www.uihealthcare.com/news/currents/vol2issue1/1helminths.html
12. Williams H, Pembroke A. Sniffer dogs in the melanoma clinic?
Lancet. 1989;1:734
13. Church J, Williams H. Another sniffer dog for the clinic? Lancet
2001;358:930
14. Willis CM, Church SM, Guest CM, Cook WA, McCarthy N,
Bransbury A, Church MRT, Church JCT. Olfactory detection of human
bladder cancer by dogs: proof of principle study. BMJ 2004 329: 712.
15. Balile D. Tanzania trains rats to detect tuberculosis. Science and
Development Network December 28 2003
http://www.scidev.net/News/index.cfm?fuseaction=readNews&itemid=116
9&language=1
Acknowledgments.
The author is grateful to Mr .T C Teo for illustrations nos. 1 and 2, Mr. A
Jarvis for illustrations nos. 3-8, and to the Medical Illustration Department,
Wycombe General Hospital for illustration no. 9.
25
Evidence Based Complementary and Alternative Medicine
Edwin L. Cooper, Ph.D., Sc.D.
Professor, Department of Neurobiology
David Geffen School Of Medicine at UCLA, USA
University of California, Los Angeles
Time and contributions have moved rapidly since organizing the first
International Congress, Complementary and Alternative Medicine,
Kanazawa Japan in 2003. There we established the international peer
reviewed journal: Evidence Based Complementary and Alternative
Medicine (eCAM) published quarterly by Oxford University Press. Our
journal, truly international with an impressive list of Editorial Board
Members, seeks to create dialogue between and across disciplines in order
to fully understand CAM and its numerous practices worldwide that
include for example acupuncture and moxibustion. Kanazawa Japan was
also the site for organizing and editing the Proceedings with Prof. N.
Yamaguchi (Biomedical Approaches to Complementary and Alternative
Medicine published by Plenum). We strongly believe that eCAM will
flourish through imagination, scientific rigor and cooperative
enthusiasm. The first objective in a serious approach to complementary
and alternative medicine (CAM) should be to obtain a broad understanding,
with a minimum of detail, of how CAM fits into the pattern of biology—of
the way in which the neuroendocrineimmune system evolved, its function
and coordination with other body systems, and its development from the
embryo onwards. At the same time, such an outline should provide an
adequate background for easy application of CAM ideas to the detail of
practical CAM work in public health, clinical and medical practice, and yet
not stray far away from the very biology that under girds it. CAM is
organismic, inclusive and not reductionist and exclusive. Quality control is
important for the safe use of natural products. Our eCAM is launched in a
desire to ameliorate this situation, by encouraging the publication of
original scientific papers based on sound scientific guidelines, but without
prejudice against the possible efficacy of these new and ancient treatments.
Turning to products from animals, particularly those from the sea, marine
natural product bioprospecting has yielded a considerable number of drug
26
candidates. Most of these molecules are still in preclinical or early clinical
development. For terrestrial animals, Lombrokinase is a product isolated
from earthworms and marketed as a fibrinolytic agent. Their use dates to
ancient Ayurvedic practices in India and Traditional Chinese Medicine
(TCM) in China. Propolis from bees and antimicrobial peptides from
maggots (fly larvae) are all effective CAM therapies. We must remember
that such products are often associated with the immune systems of these
creatures and that they evolved millions of years ago—thus their immune
systems have been an effective survival strategy. And if it has worked for
them, then humans should harness these as new-wave antibiotics or
anticancer molecules, just to offer two biomedical (CAM) applications.
Clinical Aspects of Modern Maggot Therapy
Wim Fleischmann, MD
Department of Trauma and Reconstructive Surgery
Academic Hospital of the University of Heidelberg, Germany
1. Indications:
When the orthopaedic surgeon William S. Baer presented his method and
results of maggot therapy in 1931 his focus of interest was acute soft tissue
and chronic bone infection. Later, it was mainly the work of Ronald
Sherman which enlarged the scope of indications to chronic wounds, like
stasis ulcers and bed sores. Presently, clinical emphasis is put to the
problem of impaired wound healing as seen in diabetic wounds.
2. Mode of action:
Maggots release digestive juices into the wound which contain growth
factors, antiseptic substances and proteases. The high proteolytic activity
liquefies necrotic tissue, which is the main food source of maggots.
Clinically, there is an effective, atraumatic debridement of the wound and
together with dissolved dead tissue its contaminating bacteria are removed
as well.
3. Application of maggots:
Maggots may either be applied directly to the wound surface or enclosed in
27
porous pouches which might be compared with tea bags. Using the latter
the maggots are physically separated from the wound and the fluids
streaming through the pores of the pouch convey active substances and
nutrients. As it is superior to "free range maggots" in terms of patients`
comfort, pain reduction, ease of maggot application, hygiene and aesthetics
maggot pouch dressings could be regarded as an important innovative step
in the logical progression of maggot therapy. In Germany pouch dressings
account for more than 85% of sold maggot products.
4. Surgical technique:
In superficial wounds like some bed sores, stasis and diabetic ulcers as a
first measure mechanical “macro-debridement” (scalpel etc.) should be
performed to remove dead tissue, mainly to accelerate healing and reduce
costs of treatment. Maggots are then used for biological “microdebridement” and as an antiseptic agent. In deep soft tissue and bone
infections the septic focus has either to be completely removed
by surgery or at least extensively exposed by tissue incisions or excisions to
give maggots and their secretions enough space to do their work properly.
To prevent suffocation of maggots when put into deep wounds either textile
spacer materials or polyvinyl-alcohol sponges are used to keep the wound
edges apart. Profuse wound secretions need to be drawn off using drainage
systems. To enhance the survival rate of maggots in particularly deep
wounds special drains for ventilation purposes may be necessary.
5. Secondary measures:
Having achieved a clean, well healing wound vacuum dressings (VAC)
may be applied for acceleration of wound closure or external tissue
augmentation procedures (EasApprox) are used for rapid closure of
remaining wound defects (Biological-Mechanical Closure of Wounds,
BMW).
6. Conclusion:
Maggot Debridement Therapy is an excellent low-risk procedure for the
treatment of wound infections and a variety of chronic wounds, given that
this therapy is embedded in an effective concept of interdisciplinary wound
management.
28
Hirudotherapy in Modern Medicine
Olga Gileva, DDS
Perm State Academy of Medicine, Russia
In spite of the serious evolution of our scientific findings of the
HTH
performance, we haven’t totally covered its mechanism even now.
From the modern biochemistry position we can speak about polyfactoral
effects of leech saliva substances with anticoagulant, antithrombotic,
antiaggregative, antiflogistic, vasodilatative activities from leech saliva,
secreted to blood and tissues during the blood drainage. HTH has the
positive influence on posttraumatic inflammation and regeneration,
promoting antiexudative effect, improving the microcirculation and oxygen
regimen, sells balance and etc.
In the medical practice physicians have usually used Hirudo
Medicinalis Oficinalis (HM), reared in special biofactories. All the
biomaterial used in clinics has conformity certificate of regional centers of
certification and quality control of drugs, and that permits doctors to use
Hirudo Medicinalis in Curative purposes.
Leech therapy may be used alone, as monotherapy or in addition to
the main treatment, can be combined with pharmacotherapy.
The usual treatment involves leeches applying on dermal or mucosal
surfaces, with limited or unlimited time of hemoextraction, in non
aspirative regimen - without hemoextraction and in aspirative regimen –
with limited time of hemoextraction.
The treatment course depends on the character of the disease and
lasts from 3 days up to 5 weeks.
HTH has low number of contraindications. The absolute ones are
hemophilia, hemorrhagic diathesis, expressed and firm hypotension,
expressed and firm anemia, sepsis. Clinicians also must remember about
the cases of individual intolerance to leeches and should be very careful to
prescribe leeching in pregnancy and during anticoagulant treatment.
The lecture will be focused on a spectrum of clinical indications
for HTH and it’s efficacy:
- cardiovascular diseases (myocardial ischemia, postinfarction
cardioscleroses, hypertension and it’s complications)
29
-
-
phlebopathology (chronic tromboflebitis and varicose dilatation of
veins)
leg ulcers, post- thrombotic symdrome
dermatological diseases (psoriases, eczema, sclerodesmia,
neurodesmite, ruber lichen planus)
different forms of ophthalmology (inflammatory diseases of eyes
– keratitis, uveitis, traumatic jnjuries of organ of vision, cataract,
glaucoma)
gynecology diseases (parametritis, mastitis, genital endometriosis)
neurological pathology (radiculitis, neuritis, plexitis, chronic
myofacial syndrome, infantile cerebral paralysis)
lesions of oral cavity (glossalgia, facialgia, aphtose stomatitis,
periodontitis, gingivitis)
Practitioners emphasize the value of leeches in microvascular and
reconstructive surgery).
Side effects of HTH (local allergic skin reaction,
hyperpimentation of the skin, regional lymphadenitis, subfebrile condition,
headache, the formation post procedural cixatrix) will be analyzed in
lecture.
The majority of these side effects are transient, can be cupping
rapidly and easily by means of pharmacotherapy.
The wide array of bioactive products, combined with its minimal
side effects led many clinicians and researches to apply leeches to a variety
of illnesses with high efficacy.
Ichthyotherapy for patients with psoriasis: first results and future
aspects
Martin Grassberger, MD, PhD
University Medical Center Hamburg-Eppendorf, Germany
Ichthyotherapy (therapy with the so-called “Doctorfish of Kangal”, Garra
rufa) has been shown to be effective in patients with psoriasis and other
30
skin diseases in the Kangal hot springs in Sivas, Turkey.
In a pilot study we set out to evaluate the efficacy and safety of
ichthyotherapy in combination with short-term UVA sunbed radiation in the
treatment of psoriasis under controlled conditions. Sixty-seven patients
diagnosed with psoriasis who underwent 3 weeks of ichthyotherapy at an
outpatient treatment facility in Lower Austria between 2002 and 2004 were
analyzed retrospectively.
Main Outcome Measures were (1) overall relative reduction in Psoriasis
Area Severity Index score; (2) proportion of patients with an improvement
in their PASI score of ≥75% (PASI-75) and ≥50% (PASI-50); (3) patient
reported outcomes assessed with a custom questionnaire; (4) patient
follow-up with a questionnaire sent out in March 2005. Safety was
evaluated by reviewing adverse events and vital signs.
Overall there was a 71.71% reduction in PASI score compared to baseline
(p<0.0001). Of the 67 patients studied, 31 (46.3%) achieved PASI-75 and
61 patients (91%) achieved at least PASI-50. Patients reported substantial
satisfaction with the treatment. The reported mean remission period was
8.58 months (95% CI, 6.05 to 11.11). 87.5% of patients reported a more
favourable outcome with ichthyotherapy, when asked to compare
ichthyotherapy to other previously tried therapies. Sixty-five percent stated
that after the relapse their symptoms were less severe than before treatment.
There were no significant adverse events.
The benefit demonstrated in this study along with the favourable safety
profile suggests that ichthyotherapy combined with UV-phototherapy could
provide a viable treatment option for patients with psoriasis.
Additionally we conducted a morphological study on some features of the
fish’s anatomy, shedding more light onto the mechanism of the observed
therapeutic effect. A prospective controlled trial to further validate efficacy
of this unusual treatment modality is currently in the planning phase.
Keywords: Garra rufa, ichthyotherapy, Kangal fish, psoriasis treatment,
ultraviolet A.
31
Introduction
Psoriasis is a common skin disorder with a worldwide distribution; the
average prevalence in Europe and the USA has been estimated at 2%.1
Whilst considerable advances have been made in the management of this
disease in recent years, there is no cure, and no simple, safe and invariably
effective treatment.2 The disease carries a substantial burden even when not
extensive, and is associated with widespread treatment dissatisfaction. 3 A
wide range of treatments is offered for psoriasis. Some patients rely on
conventional pharmacological methods; others try alternative and
complementary treatment modalities.
Surely one of the most unusual alternative treatments is the so-called
“Doctor Fish of Kangal” in the Central Anatolia region of Turkey. This
treatment was first mentioned in The Lancet in 19894 but the details of the
treatment were published only recently by Özcelik et al.5 According to the
authors two different types of fish live in the pools of the Kangal hotspring:
Cyprinion macrostomus and Garra rufa. Both fish are members of the carp
and minnow family (Cyprinidae). Garra rufa is regarded as the main
therapeutic. Garra rufa is normally a bottom dweller, where it adheres by
suction to rocks with its ventral crescent-shaped mouth to feed on phytoand zooplankton.6 However, in the hot pools of Kangal, where phyto- and
zooplankton are scarce, these fish feed on the skin scales of bathers,
reportedly reducing illnesses such as psoriasis and atopic dermatitis.5
Whether this remarkable treatment is also effective outside of the Kangal
hotspring in Turkey is unknown. Since there have been many unscientific
and misleading names for this kind of therapy, we suggest the term
“ichthyotherapy”, in accordance with other so-called biotherapy concepts
such as maggottherapy (use of sterile fly larvae), hirudotherapy (use of
leeches) and apitherapy (use of bee venom). In this retrospective study, we
set out to evaluate the efficacy and safety of ichthyotherapy when used in
combination with short term UVA radiation in patients with psoriasis
vulgaris in an outpatient treatment facility.
32
Patients and Methods
Patients
We retrospectively analysed 67 patients diagnosed with psoriasis who
underwent 3 weeks of ichthyotherapy combined with a short course of
UVA sunbed treatment at an outpatient treatment facility in Lower Austria
between 2002 and 2004. All patients had moderate to severe chronic plaque
psoriasis. Patients referred themselves to the treatment facility to find relief
from their symptoms. The costs of this treatment were not covered by the
patients’ health care insurance. All patients had signed informed consent
and were under constant medical supervision throughout the course of
treatment.
Treatment regimen
Treatment lasted three weeks. A daily two hour “fish bath” (Fig. 1) was
taken in a bath tub at a comfortably warm temperature (36-37°C). Patients
with no contraindication to UV exposure (reported history of UV-related
skin malignancies) used a commercially available stand-up rapid-tan
facility (Cyclone 60 with 60 Cosmolux VHR160 lamps, CMC Sun Capsule
USA) for 3-5 minutes after each bath session according to skin type. The
spectral emission of the lamps is shown in figure 2. After UV exposure, the
patients applied a generic skin lotion (Pharmacy Neunkirchen, Austria)
containing glycerine, Butyrospermum parkii (Shea butter) and Aloe vera
extract. If psoriasis lesions severely involved the scalp, the patient’s head
was shaved before treatment.
Treatment tubs
The treatment tubs, made from food-safe plastic (Chemo®, Weinstadt,
Germany) had a capacity of 1100 litres, filled to approximately 80%
capacity. Between 250 to 400 fish were used, depending on the size and
severity of the skin lesions. The bath tubs were equipped with an elaborate
fresh water system, the technical details of which will be published
elsewhere. The water in the tubs was constantly filtered and sterilized (700
litres per hour) by a filter pump and a UVC water sterilisation device and
was simultaneously enriched with oxygen. The water was exchanged
completely 3 to 4 times a day. A thermostat was used to maintain a
comfortable water temperature between 36 and 37°C.
33
The fish, reared in an adjacent breeding facility, ranged from 5 to 10 cm in
length (approx. 1.5 years old) when used for treatment. In contrast to the
Kangal hotspring, where they lack nutrients due to the high temperatures of
the spring, the fish were fed commercially available fish food (TetraMin®,
Tetra GmbH, Melle, Germany) daily after the treatment sessions.
Water samples were tested monthly for Legionella species and
Pseudomonas aeruginosa. The fish were examined for Aeromonas
hydrophila, A. sobria, A. caviae, Mycobacterium marinum and M. piscium
to rule out any risk for potential zoonotic infections. Since the fish were
given only commercially available fish food, the risk of Diphyllobothrium
latum (fish tapeworm) infestation could be ruled out based on the absence
of an intermediate host.
Each patient was allocated to a single bathing tub for the duration of the
three-week treatment, and no two patients shared the same tub at any time
during that period. After the three-week treatment, each tub along with the
technical equipment was disinfected (2% solution Dodarcana® Rapid,
Schülke & Mayr, Austria) for 1 hour.
Assessment
Clinical assessment. The primary efficacy outcome measure was the overall
total reduction in Psoriasis area severity index (PASI) score and the
proportion of patients with 50% and 75% improvement in PASI score
(PASI-50 and PASI-75) at week 3, relative to baseline. The PASI is a
physician-accessed score, recognized by the US Food and Drug
Administration to assess efficacy of psoriasis therapies in clinical trials.
The PASI score takes into account the extent of involved skin surface and
the severity of erythema, desquamation, and plaque induration. The
composite score ranges from 0 to 72, with higher numbers indicating more
severe disease and a reduction in score representing improvement. The
PASI-75 is the currently recognized benchmark of end-points used in
psoriasis clinical trials. PASI-50 is also regarded as a clinically significant
endpoint in the assessment of psoriasis.7 PASI was assessed on highresolution digital colour photographs taken at baseline and at the end of the
34
three-week treatment period. Baseline measurements were those made
closest but prior to the beginning of treatment.
Additionally, response to treatment was defined according to the rate of
improvement in PASI score, as follows: complete response, more than 95%
improvement; good or marked, 75% to 94%; moderate, 50% to 74%; slight,
25% to 49%, and none, less than 25%.
Patient-reported outcomes were evaluated by a short custom questionnaire
immediately after the three-week course of treatment (Table 1) and by a
follow-up questionnaire sent to all patients after treatment in March 2005 to
assess the duration of remission, the number of different treatment
regimens prior to ichthyotherapy, the severity of a possible relapse and the
personal satisfaction with ichthyotherapy when compared to previous
treatments (Tables 2&3). At this point the time elapsed since the end of
therapy was between 3 months to almost 3 years.
Safety evaluation. The safety and tolerability of the therapy were evaluated
by reviewing adverse events (assessed weekly), vital signs and a weekly
physical examination.
Statistical analyses
The primary endpoint (efficacy of treatment) was evaluated by comparing
the mean PASI score before and after three weeks of treatment. The PASI
scores were analysed using a Wilcoxon signed rank test for paired
comparison. P values ≤ 0.05 were considered statistically significant.
Analysis was carried out using Prism 4 for Macintosh (GraphPad Software,
Inc. San Diego, California).
35
Results
Patients’ Characteristics
Sixty-seven patients, 39 male and 28 female, were included in this
retrospective study. All 67 patients completed the three-week treatment.
Ages ranged between 10 and 75 years (mean 41.01 years, 95% confidence
interval, CI 37.51, 44.52). The mean duration of psoriasis at baseline was
13.9 years (range 1 to 35 years; 95% CI 11.6, 16.21).
Treatment Efficacy
Physician-Assessed outcomes. At the end of the three-week treatment
course, 31 of the 67 patients (46.3%) achieved PASI-75 and 30 other
patients (44.8%) achieved PASI-50. The mean PASI score at baseline for
the entire study cohort was 18.9 ± 12.37 (95% CI 15.89, 21.9). PASI score
at the end of the treatment period was 5.34 (95% CI 4.27, 6.42). Overall
there was a 71,71% reduction in PASI score compared to baseline
(p<0.0001) (Fig. 3).
Response to treatment was complete in 3 patients (4.5%), marked in 29
(43.3%), moderate in 29 (43.3%), and slight in 6 (8.9%). No patient
failed to respond at all.
Patient-Reported outcomes. In the short questionnaire given immediately
after the three weeks of treatment, patients reported substantial satisfaction
with the treatment (Table 1).
Of the 67 follow-up questionnaires sent to the patients in March 2005, 64
were deliverable. Forty questionnaires were returned, giving a response rate
of 60%. The mean time since the end of treatment was 21.8 (95% CI 18.99,
24.68). The reported mean remission period was 8.58 months (range: 1 to
30 months; 95% CI 6.05, 11.11) with two patients (5.1%) still in remission
at the time the questionnaire was received (time elapsed since the end of
therapy was 12 and 26 months, respectively). The mean number of
previously used other therapies was 5 ± 3 (± SD) (range: 0 to 12; 95% CI
4.02, 5.93) (Table 2). When asked to compare their treatment results to all
previously used therapies, 87.5% of patients reported a more favourable
outcome with ichthyotherapy. 65.1% stated that after the relapse their
symptoms were less severe than compared to baseline (Table 3).
36
Safety evaluation, Side effects
No severe side effects were recorded during the treatment period. Mild,
transient bleeding from open crusted lesions was reported by one patient
with eczema, and UV-radiation-related erythema by two others. No
clinically significant pattern of changes in vital signs were observed during
the study period. Ichthyotherapy in combination with UVA-treatment was
generally very well tolerated.
Discussion
This retrospective study indicates that ichthyotherapy used in combination
with short term UVA radiation is an effective and safe treatment for
psoriasis vulgaris. 46.3% of the 67 patients achieved PASI-75 and an
additional 44.8% patients achieved at least PASI-50 after a three-week
course of treatment with the “doctorfish of Kangal” Garra rufa (Fig. 4). It
is well established that a 75% improvement in PASI score is a clinically
meaningful endpoint for clinical trials, and there is strong evidence
demonstrating that a 50% improvement in PASI score is also associated
with a significant improvement in the patients quality of life. 7 These
findings are further supported by the results of the questionnaire completed
by the patients immediately after treatment (Table 1).
The results of the follow-up survey indicate that most of the patients were
more satisfied with ichthyotherapy than with any other previously tried
treatment. This satisfaction might be explained, at least in part, by the
rather long reported mean remission period of 8.6 months. The response
rate of the survey is comparable to those of other mail-based surveys.8
However, we are aware that this approach leaves room for potential nonresponse bias; patients with unfavourable results (i.e., early relapse) may
have been less likely than others to respond to our survey.
In this study it was also shown, that ichthyotherapy combined with UVA
radiation was generally well tolerated with only three patients experiencing
mild side effects. Two of these three patients showed UVA treatment related
erythema. Whereas a prolonged UVA treatment course, or frequently
repeated courses, may have an, as yet, undefined risk with regard to
melanoma and ageing changes, a recent study suggests that UVB but not
UVA radiation initiates melanoma.9 However, we are aware of the risks as
identified by the British Photodermatology Group Workshop 10, and
37
therefore suggest to further evaluate ichthyotherapy in combination with
broadband UVB and narrowband UVB treatment.
Several mechanisms have been suggested regarding the observed efficacy
of ichthyotherapy in Turkey.5 One obvious mechanism is the physical
contact with the fish, which feed on the desquamating skin, thus leading to
a rapid reduction of the scales. This phenomenon was also consistently
observed in our patients, who reported a pleasing micro-massage like
feeling while the fish nibbled at their skin. The fish seem to prefer affected
to healthy skin possibly it is easier to nibble at this surface. Another
suggested mechanism is the direct effect of natural ultraviolet radiation due
to the high altitude (1650m) of the Kangal spa.5 Phototherapy is a well
recognised option for patients with widespread psoriasis lesions with
climatotherapy being the simplest form.2 However, a simultaneous removal
of scales by the fish probably facilitates the penetration of UV rays to the
dermis. This exposure of the lesions may explain the better outcome of
combined ichtyotherapy/UVA-treatment when compared to the poor results
of UVA sunbed treatment alone.11 A third suggested mechanism of
ichthyotherapy in Kangal is the presence of a high level of selenium (1.3
mg/L) in the hot spring water.5 While Özcelik et al.5 argue, that selenium
constitutes an important factor to treatment success in Kangal, an analysis
of the water used in our study revealed a selenium concentration of less
than 5 µg/L. Therefore selenium is not likely to have contributed to the
observed efficacy of ichthyotherapy reported in the present study. A fourth
mechanism suggested by Özcelik et al.5 is the reverse Koebner
phenomenon. The reverse Koebner phenomenon is seen when an area of
psoriasis clears following injury.12 However, it had been shown that
destruction of the dermal papillae within the injured tissue plays the key
role in preventing epidermal hyperplasia of psoriasis.13 Therefore it seems
unlikely that the reverse Koebner phenomenon contributes to the observed
efficacy of ichthyotherapy. Finally, psychological factors like stress are
regarded a causal or exacerbating factor in psoriasis.1 Thus, the two hour
daily fish bath, which most patients referred to as relaxing and pleasing,
might have contributed to the observed treatment effect by reducing
patients’ stress and enhancing psychological well-being.
There are two important key differences between this study and the study
by Özcelik et al.5 concerning treatment protocol. First, in our study the
38
patients were required to stay in the treatment tubs for only two hours per
day, whereas the mean stay in the pools in Kangal was 7.4 ± 1.1 hours a
day.5 This shortened treatment time makes ichtyotherapy more acceptable
for the patients and considerably improves compliance. Second, at the
treatment facility described herein, each patient was allocated a personal
bathing tub, and the fish only came into contact with a single patient.
Conversely, in the pools of the Kangal hot spring patients are required to
take their bath with 10 to 20 patients simultaneously (personal observation
MG). This approach might be unacceptable for some patients and a
possible hygienic risk cannot be entirely excluded.
The present study is limited by the relatively small number of patients
treated and by lack of a control group. Randomized studies would be
needed to compare the ichthyotherapy treatment with controls (e.g. water
with the UV-therapy alone) and to assess treatment with standardized
health related quality of life questionnaires. Therefore it is evident that
many questions remain unanswered concerning the optimal protocols for
ichthyotherapy. In the absence of studies to address these issues, we
recommend that the described protocol may be used for guidance.
In summary, the benefit demonstrated in this study, along with the
favourable safety profile, suggests that ichthyotherapy combined with a
short course UVA treatment could provide a viable treatment option for
patients with psoriasis vulgaris.
Maggot therapy - mode of actions and clinical practice
Hasenöhrl K. M.D., Wollina U. Prof.
Department of Dermatology, Dresden-Friedrichstadt Hospital, Germany
Biosurgery has received increasing interest for the debridement of chronic
wounds. Maggot therapy employs the use of freshly emerged, sterile larvae of
the common green bottle fly, Lucilia sericata and is a form of artificially
induced myasis in a controlled clinical situation.
How maggots remove necrotic tissue from the wound is currently investigated.
39
There are several proposed mechanisms: removing of necrotic tissue by their
secretion and mechanical effects by attacking the tissue with their
mouthhooks.
Maggots secretions contain a rich soup of digestive and proteolytic enzymes
while feeding. They produce for instance carboxypeptidase A and B, leucine
aminopeptidase, collagenase and trypsin-like and chymotrypsin-like serine
proteases. Serine proteinases exhibited degradation of extracellular matrix in
its components laminin, fibronectin and collagen types I and III. Both effects,
mechanical action and the secretion of digestive enzymes seem to be the clue
in efficient wound debridement.
Despite great success in clinical use and technology of breeding and
application the knowledge about larval changes in the wounds is limited.
Therefore our clinic investigated larval morphology before and after feeding
in the wound by histopathology. These results demonstrate the changes in
larval morphology during biosurgery.
After removing necrotic tissue larvae encouraged wound healing and
formation of granulation tissue. The enhanced tissue oxygenation was
measured by remittance spectroscopy. Our results showed an improvement of
tissue oxygenation as revealed by the characteristic oxygen doublet peak.
Beside direct promotion of granulation tissue maggots has been observed to
combat infection. The majority of wounds hosts a variety of both anaerobic
and aerobic bacteria. Bacteria are directly killed in the alimentary tract of
maggots and they produce several antibacterial factors. Also maggots are able
to kill clinical isolates of MRSA.
Our lecture gives further a brief review of medical indications of biosurgery
and the practical handling of maggots. It is used in wounds which have
previously failed to respond to conventional treatment like chronic venous or
mixed leg ulcers, traumatic wounds, pressure sores, diabetic foot ulcers and
malignant tumours of skin. We provide clinical data from literature and our
own experience.
40
Studies of the Apitox (Apitoxin)
Christopher MoonHo Kim, M.D.
Visiting Professor, Biomedical Center, CHA University, Korea
President, International Pain Institute, USA
(1)
Pharmacology Written Summary of the Apitox
1. Brief Summary
The pharmacology profile of bee venom has been determined in in vitro
systems and in various animal models. The results from some of these
animal studies are summarized in the following sections and show that
Apitoxin has biological activity in animal models that are suggestive of
potential efficacy for the treatment of refractory osteoarthritic pain and
inflammation in humans.
2. Primary Pharmacodynamics
Adjuvant-induced polyarthritis in the rat has been established as an
experimental model for human rheumatoid arthritis.i Arthritic symptoms
are induced by administering a single SC injection of 1 mg Mycobacterium
butyricum (Freund’s adjuvant or CFA) suspended in mineral oil into a hind
paw of the rat. Other studies, including those in rabbits, dogs, and monkeys,
are also described.
The ability of bee venom (dose range: 0.01 to 1.0 mg/kg/day) to suppress
adjuvant-induced arthritis was studied in rats following daily SC
administration over a period of 17 days. ii Bee venom suppressed the
development of adjuvant arthritis in a dose-related manner. A single SC
administration of bee venom also suppressed the development of
carrageenan-induced paw edema and, when administered SC the day before
or on the day of injection of adjuvant, effectively suppressed the
development of polyarthritis. This suppressive effect decreased
progressively as dosing was delayed. Bee venom was found to be most
effective when mixed and injected together with CFA, the disease-inducing
agent. Similarly, antigens such as egg albumin, when incorporated into
CFA and injected into the hind paw, prevented the development of arthritis.
These results suggest that at least two mechanisms are involved in the antiarthritic action of bee venom: (1) alteration of the immune response, most
likely via antigen competition, and (2) an anti-inflammatory action.
41
One of the earliest studies was conducted to evaluate the use of bee venom
prophylactically (beginning 2 weeks prior to adjuvant injection) and
therapeutically (beginning 1 week after adjuvant injection) to reverse
adjuvant-induced arthritis in the rat.iii Bee venom (1 and 4 mg/kg injected
SC three times a week for 4 weeks) was shown to prevent the arthritic
syndromes (foot volume, secondary lesions, and reduction in inflammation
units) both as a therapeutic and as a prophylactic (more pronounced effect).
The molecular mechanisms of the anti-inflammatory effects of bee venom
were investigated in the rat model of carrageenan-induced acute edema in
the paw and the rat model of chronic adjuvant-induced arthritis.iv Bee
venom at 0.8 and 1.6 µg/kg administered daily for 14 days into the plantar
surface of the right hind paw reduced hind paw edema. These animal
results were consistent with in vitro data that showed an inhibitory effect of
bee venom at 0.5, 1, and 5 µg/mL and melittin (a major component in bee
venom) at 5 and 10 µg/mL on lipopolysaccharide-induced expression of
cyclooxygenase 2, cytosolic phospholipase A2, inducible nitric oxide (NO)
synthase, generation of prostaglandin E2 and NO, and the intracellular
calcium level, providing information on the mechanism of action of
anti-arthritic effects of bee venom.
The effect of chromatographic fractions (designated as Oa, Op, and the
protease inhibitor) and pure proteins and peptides (melittin, apamin, and
phospholipase A2) from bee venom were tested in the rat models of arthritis
and inflammatory edema. v In rats with adjuvant-induced arthritis, the
components of venom (100 µg/kg once daily for 20 days) caused a 15-25%
inhibition of arthritic symptoms whereas phospholipase stimulated arthritic
symptoms by 30%. Rats with hind paw edema induced by CFA were
injected SC with various fractionated bee venom components (10 µg/kg)
24 hours and/or 0.5 hour (n = 6/group) prior to administration of the
inflammatory agent. The inhibition of the carrageenan and prostaglandin
E1 inflammation under the effects of Oa and Op fractions and the protease
inhibitor were the strongest.
Rats (n = 4/group) with induced adjuvant disease (which includes a severe
and persistent polyarthritis) were treated for 21 days with SC administered
bee venom (300 or 500 µg or 1.2 and 2 mg/kg for a 250 g rat) twice a day;
saline (control); bee venom intraperitoneal (IP); or melittin, apamin, or
42
phospholipase A2 SC (at the amount contained in 1 mg of bee venom).vi
Rats treated SC with bee venom developed little or no arthritis with the
higher dose being more effective than the lower dose while rats injected IP
with bee venom or with SC melittin, apamin, or phospholipase A 2 had no
apparent benefit on the polyarthritis. When SC bee venom injections
were initiated after moderate to severe polyarthritis had developed, rats did
not have the course of the disease altered.
The ability of a daily administration of bee venom (2 mg/kg/day) to
suppress adjuvant-induced arthritis in rats was studied over a period of
24 days. The bee venom treatment suppressed but did not abolish the
primary and secondary inflammatory responses to the adjuvant as
monitored by decreases in the swelling of the left (where the adjuvant was
injected) and right hind paws, respectively.vii The response was delayed,
with statistically significant suppressive effects being noted 2-3 weeks
following the administration of the adjuvant. A sex difference in the
suppressive effect of bee venom was noted, with female rats demonstrating
a more pronounced beneficial effect than male rats. The dosage used in this
study (2 mg/kg) was rather high, as compared to 50-70 µg of solids in the
average honeybee sting.viii,ix
In a study performed to assess the clinicotherapeutic effect of bee venom,
90 rats were administered bee venom (1 bee [about 0.1 mg bee venom or
0.4 mg/kg for a 250 g rat], SC), prednisolone (10 mg/kg, orally), or saline
control (0.1 mL, SC) every other day over a period of 14 days.x Clinical
findings of lameness score, edema volume, hematological values, and
histopathology (interphalangeal joint of the right hind paw) were observed
during the treatment period. In the treatment groups, the development of
inflammatory edema and polyarthritis was suppressed. No significant
differences of hind paw edema volume and lameness score between the
prednisolone and bee venom groups were observed during treatment. No
differences in red blood cell count, hematocrit, or hemoglobin
concentration were noted between the groups although significant
leucocytosis was observed in the control group (p < 0.01). Erosions of
articular cartilage and inflammatory cell infiltrations into the
interphalangeal joint were effectively suppressed in treated groups. Thus,
whole honeybee venom was found to suppress arthritic inflammation in the
43
rat.
A study was conducted to determine whether some of the in vivo effects of
bee venom may be mediated by alterations in lymphokine production and
to observe the in vitro effect of lymphokines on reduced mitogenic
responses of bee venom-treated rats.xi When 0.5 mg/kg/day bee venom
was administered intramuscularly (IM) to rats over a period of 17 days, a
reduction in interleukin (IL) production by splenocytes was observed. In
vitro addition of IL-1 or IL-2 to the cultures resulted in an increase in
responses to normal levels, suggesting that bee venom affects the
production of IL-1 by macrophages. This finding suggests that the
modifying effects by bee venom on inflammatory responses in local
therapy may be due to interference with certain functions of inflammatory
cells.
In an attempt to explain the mechanism of bee venom’s anti-inflammatory
action, the effect of bee venom on neutrophil O2- production was
investigated. xii Neutrophil production of toxic oxygen radicals and
metabolites are thought to play a role in chronic inflammation and tissue
destruction in a wide variety of diseases. Using human peripheral blood
leukocytes, the polymorphonuclear fraction was isolated and used in in
vitro assessments on the ability of melittin and other bee venom peptides to
affect the production of O2-. The results showed that melittin, but not other
bee venom fractions, inhibited O2- production both pre- and poststimulation, suggesting the melittin may have a role in the in vivo
regulation of radical production and suggesting a possible mechanism of
action for this major bee venom component.
The effect of bee venom on inflammatory cell function was studied
following injection of immune complexes into rabbit knee-joints. xiii
Treatment with single SC injections of bee venom (1.2 to 20 µg/kg)
significantly reduced the leukocyte counts in the immune complex
challenged joints as compared to untreated rabbits (p < 0.02). Bee venom at
a dose of 12 µg/kg decreased leukocyte counts 3 and 6 hours after
challenge with immune complexes but this effect was not noted at 9 hours.
Significant effects on leukocyte random migration, chemotactic
responsiveness, or phagocytosis were not observed, indicating that bee
venom did not interfere with normal phagocyte motility and ingestion. The
44
modifying effects by bee venom on the inflammatory response to immune
complexes in vivo was, therefore, thought to be most likely due to
interference with other components of the inflammatory response.
In a study designed to investigate the possible role of the soluble fraction of
bee venom in producing the anti-arthritic actions of bee venom acupuncture,
whole bee venom was extracted into two fractions according to solubility: a
water soluble fraction (BVA) and an ethylacetate soluble fraction (BVE). xiv
Subcutaneous BVA injection (0.9 mg/kg/day) into the Zusanli acupoint was
found to dramatically inhibit paw edema and radiological change (i.e. new
bone proliferation and soft tissue swelling) caused by CFA injection. The
BVA treatment also reduced the serum IL-6 increase caused by rheumatoid
arthritis induction to levels observed in the non-arthritic animals. In
addition, BVA therapy significantly reduced arthritis-induced nociceptive
behaviors (i.e. nociceptive scores for mechanical hyperalgesia and thermal
hyperalgesia). Finally, BVA treatment significantly suppressed adjuvantinduced Fos expression in the lumbar spinal cord at 3 weeks post-adjuvant
injection. In contrast, BVE treatment (0.05 mg/kg/day) failed to show any
anti-inflammatory or antinociceptive effects on rheumatoid arthritis. These
results demonstrate that BVA is the effective fraction of whole bee venom
responsible for the antinociception and anti-inflammatory effects of bee
venom acupuncture treatment. Further study is necessary to clarify which
constituents of the BVA fraction are directly responsible for these antiarthritis effects.
Previous studies in experimental animals suggested that the therapeutic
effect of bee venom on arthritis is dependent on the site of administration.
In particular, local injection of bee venom near the site of inflammation,
such as the hind limb in the rat model, is more effective in inhibiting the
development of adjuvant-induced arthritis than injections into a more
distant site (e.g. on the back).vii,ix Because of this potential site specificity, a
study was designed to evaluate the anti-nociceptive effect of bee venom
injections into a specific acupoint (Zusanli) compared to a non-acupoint in
the rat model of chronic arthritis.xv Subcutaneous bee venom treatment (1
mg/kg per day) was found to dramatically inhibit paw edema caused by
CFA injection and significantly reduced arthritis-induced nociceptive
behaviors (i.e. the nociceptive scores for mechanical hyperalgesia and
45
thermal hyperalgesia). These anti-nociceptive/anti-inflammatory effects of
bee venom were observed from 12 days through 21 days post-bee venom
treatment. In addition, bee venom treatment significantly suppressed
adjuvant-induced Fos expression in the lumbar spinal cord at 3 weeks postadjuvant injection. Finally, injection of bee venom into the Zusanli
acupoint resulted in a significantly greater analgesic effect on arthritic pain
as compared to bee venom injection in to a more distant non-acupoint. The
study demonstrated that bee venom injection into the Zusanli acupoint has
both anti-inflammatory and anti-nociceptive effects on CFA-induced
arthritis in rats. These findings suggest that bee venom acupuncture may be
an effective therapy for the treatment of rheumatoid arthritis.
The antinociceptive and anti-inflammatory effects of bee venom
pretreatment on carrageenan-induced inflammation in the rat was studied in
order to determine if bee venom, which is nociceptive under normal
conditions, serves as an anti-inflammatory and/or antinociceptive agent
under conditions of inflammation.xvi Rats were injected SC with bee
venom at 0.8 mg/kg (n = 8) or 0.08 mg/kg (n = 6) 30 minutes prior to
carrageenan-induced acute paw and thermal hyperalgesia. At 0.8 mg/kg,
bee venom significantly suppressed carrageenan-induced edematous paw
volume (p < 0.001) and thermal hyperalgesia (p < 0.01) and induced
expression of Fos positive neurons in the spinal cord (p < 0.01). These
results suggested that bee venom may be useful in the treatment of pain and
edema associated with chronic inflammatory diseases.
The effect of bee venom on cortisol levels and increase in activity in dogs
with hip dysplasia was investigated.xvii The 24 dogs, 8 of which had hip
dysplasia and 16 of which were normal, were divided into 4 treatment
groups. Groups I and II included 8 normal dogs each and Groups III and IV
each had 4 dogs with hip dysplasia. Groups II and IV received 1 mg (about
0.067 mg/kg for a 15 kg dog) of bee venom SC on Days 30, 37, 50, and 60
and then were crossed over to received saline control treatment on Days 90,
97, 110, and 120. Groups I and III began with saline control treatment and
then were crossed over to the bee venom treatment. Dogs receiving bee
venom injections had increased plasma cortisol levels and arthritic dogs
had increased daily cage activity. On day 90, the treatments were crossedover and again dogs receiving bee venom had increased plasma cortisol
46
levels and arthritic dogs had increased daily cage activity. The results
suggest that bee venom stimulates the production of cortisol and enhances
the cage activity of arthritic dogs.
The effect of whole bee venom on plasma cortisol levels was investigated
in the unanesthetized monkey following single SC injections of bee venom
(1.0 to 100 mg) or melittin (1.0 to 10 mg).xviii Both bee venom and melittin
injections produced marked and sustained elevations in plasma cortisol
levels. These increases occurred at approximately 1 hour following SC
injection and lasted for 2-4 days. The effect appeared to be dose-related
with the higher doses of bee venom or melittin producing the earliest and
most pronounced plasma cortisol elevations. When a second dose of bee
venom (1 mg) and melittin (0.1 mg) was administered to 1 monkey each at
72 or 96 hours, respectively, there was an immediate and sustained rise in
cortisol, which lasted for 20 to 30 days. Melittin appeared to be 10 times
more potent than bee venom. Necropsy results from 4 monkeys receiving
the highest doses of bee venom or melittin indicated no significant gross or
microscopic tissue changes. Surgical removal of the pituitary gland from
4 monkeys prevented the effect, indicating that bee venom and melittin
may be stimulating the production of cortisol from the adrenal gland, which
may provide an explanation for the beneficial effects of bee venom therapy
in a variety of disease conditions that respond to adrenal steroid therapy.
Using HTB-94 human chondrosarcoma cells, gene expression profiles were
determined following treatment with bee venom, lipopolysaccharide (LPS),
or both.xix Of 344 genes profiled, 35 were down regulated by bee venom,
16 were up regulated and 7 down regulated by LPS, and 32 were down
regulated by bee venom and LPS combined. Bee venom reversed the
upregulation caused by LPS for some genes, such as the IL-6 receptor,
matrix metalloproteinase-15 (MMP-15), tumor necrosis factor,
superfamily-10, caspase-6, and tissue inhibitor of metalloproteinase-1
(TIMP-1). These results provided information for understanding the
pharmacologic activity of bee venom for the treatment of arthritis.
3. Secondary Pharmacodynamics
1) General Pain
In a study designed to evaluate the potential antinociceptive effect of bee
venom pretreatment on formalin-induced pain behavior and its associated
47
spinal cord Fos expression, rats were administered a single SC injection of
bee venom (dose range: 0.0016 mg/kg to 0.08 mg/kg) or saline control into
the Zusanli acupoint (5 mm lower and lateral to the anterior tubercle of the
tibia).xx Pretreatment with bee venom significantly decreased paw-licking
time in the late phase of the formalin test. In contrast, bee venom injected
into a non-acupoint in the back region did not suppress the paw-licking
time. Bee venom pretreatment into the Zusanli acupoint markedly inhibited
spinal cord Fos expression induced by formalin injection. These findings
indicate that bee venom pretreatment into the Zusanli acupoint has an
antinociceptive effect on formalin-induced pain behavior.
In a follow-up study to the one describe above, an abdominal stretch assay
in mice and rats and formalin test in rats were used to further investigate
bee venom’s antinociception effect.xxi Bee venom was administered SC to
the mice at a 1 to 100 or 1 to 1000 dilution and to rats at a 1 to 1000
dilution in 20 µL of saline. Bee venom at the 1 to 100 dilution into an
acupoint or non-acupoint produced antinociceptive effects while bee venom
at the 1 to 1000 dilution was effective only when dosed into an acupoint,
indicating that bee venom may be a promising method for the relief of pain.
2) X-Irradiation Protection
The response of animals to whole-body X-irradiation in the lethal range can
be modified by certain changes in their physiological state if induced prior
to exposure. The ability of bee venom to produce a degree of physiological
stress in animals, thereby eliciting a neuroendocrine response (pituitaryadrenal stimulation) that would increase radiation resistance, was studied in
mice.xxii In a series of experiments, pre-treatment with bee venom (IP dose
range: 1.1 to 1.24 µg; SC dose range: 4.3 to 5.6 µg) resulted in greater 30day survival rates as compared to a saline control. Bee venom administered
SC afforded the greatest protection (70-80% 30-day survival rate). When
melittin, a major component of bee venom, was separated and injected SC
at 5.4 µg, the 30-day survival rate was 7%. Based on these results it was
proposed that at least 3 mechanisms of action may account for the
radioprotective effect of bee venom in mice: 1) it has a stressor-like action
that elicits an “adaptation syndrome,” 2) it produces changes in the
hematopoietic system, or 3) it has antibacterial properties.
In a separate study, melittin provided statistically significant x-irradiation
48
protection to mice that received an SC injection doses up to 60 mg/kg
24 hours prior to the irradiation. xxiii Whole bee venom did not show
conclusive evidence for the same protection.
Pearson CM, Wood FD. Studies of arthritis and other lesions induced in rats by
the injection of mycobacterial adjuvant. VII. Pathologic details of the arthritis
and spondylitis. Am J Pathol. 1963 Jan;42:73-95.
Chang YH, Bliven ML. Anti-arthritic effect of bee venom. Agents Actions.
1979 Jun; 9(2):205-211.
Lorenzetti OJ, Fortenberry B, Busby E. Influence of bee venom in the
adjuvant-induced arthritic rat model. Res Commun Chem Pathol Pharmacol.
1972 Sep;4(2):339-52.
Park HJ, Lee SH, Son DJ, Oh KW, Kim KH, Song HS, Kim GJ, Oh GT, Yoon
do Y, Hong JT. Antiarthritic effect of bee venom: inhibition of inflammation
mediator generation by suppression of NF-kappaB through interaction with the
p50 subunit. Arthritis Rheum. 2004 Nov;50(11):3504-15.
Shkenderov S. New pharmacobiochemical data on the anti-inflammatory
effect of bee venom. Animal, Plant, and Microbial Toxins. 1976;2:319-336.
Zurier RB, Mitnick H, Bloomgarden D, Weissmann G. Effect of bee venom on
experimental arthritis. Ann Rheum Dis. 1973;32:466-70.
Eiseman JL, von Bredow J, Alvares AP. Effect of honeybee (Apis mellifera)
venom on the course of adjuvant-induced arthritis and depression of drug
metabolism in the rat. Biochem Pharmacol. 1982 Mar 15;31(6):1139-46.
Lichtenstein LM, Valentine MD, Sobotka AK. A case for venom treatment in
anaphylactic sensitivity to hymenoptera sting. N Engl J Med. 1974 May
30;290(22):1223-7.
Hadjipetrou-Kourounakis, Yiangou M. Bee venom and adjuvant induced
disease. J Rheumatol. 1984 Oct;11(5):720.
Kang SS, Pak SC, Choi SH. The effect of whole bee venom on arthritis. Am J
Chin Med. 2002;30(1):73-80.
Hadjipetrou-Kourounakis, Yiangou M. Bee venom, adjuvant induced disease.
J Rheumatol. 1988 Jul;15(7):1126-8.
Somerfield SD, Stach JL, Mraz C, Gervais F, Skamene E. Bee venom melittin
blocks neutrophil O2- production. Inflammation. 1986;10(2):175-80.
Thomsen P, Bjurtsten LM, Ahlstedt S, Bagge U, Bjorksten B. Inhibitory effect
of bee venom on immune complex mediated leukocyte migration into rabbit
knee-joints. Agents and Actions. 1984; (14(5/6):662-66.
49
Kwon YB, Lee HJ, Han HJ, Mar WC, Kang SK, Yoon OB, Beitz AJ, Lee JH.
The water-soluble fraction of bee venom produces antinociceptive and
anti-inflammatory effects on rheumatoid arthritis in rats. Life Sci. 2002 May
31; 71(2):191-204.
Kwon YB, Lee JD, Lee HJ, Han HJ, Mar WC, Kang SK, Beitz AJ, Lee JH. Bee
venom injection into an acupuncture point reduces arthritis associated edema
and nociceptive responses. Pain. 2001 Feb 15;90(3):271-80.
Lee JH, Kwon YB, Han HJ, Mar WC, Lee HJ, Yang IS, Beitz AJ, Kang SK.
Bee venom pretreatment has both an antinociceptive and anti-inflammatory
effect on carrageenan-induced inflammation. J Vet Med Sci.
2001;63(3):251-59.
Vick JA, Warren GB, Brooks RB. The effect of treatment with whole bee
venom on daily cage activity and plasma cortisol levels in the arthritic dog.
Inflammation. 1975-1976;1(2):167-74.
Vick JA, Mehlman B, Brooks R, Phillips SJ, Shipman W. Effect of bee venom
and melittin on plasma cortisol in the unanesthetized monkey. Toxicon.
1972;10:581-6.
Yin CS, Lee HJ, Hong SJ, Chung JH, Koh HG. Microarray analysis of gene
expression in chondrosarcoma cells treated with bee venom. Toxicon.
2005;45:81-91.
Kim HW, Kwon YB, Ham TW, Roh DH, Yoon SY, Lee HJ, Han HJ, Yang IS,
Beitz AJ, Lee JH. Acupoint stimulation using bee venom attenuates
formalin-induced pain behavior and spinal cord expression in rats. J Vet Med
Sci. 2003;65(3):349-55.
Kwon YB, Kang MS, Kim HW, Ham TW, Yim YK, Jeong SH, Park DS, Choi
DY, Han HJ, Beitz AJ, Lee JH. Antinociceptive effects of bee venom
acupuncture (apipuncture) in rodent animal models: a comparative study of
acupoint versus non-acupoint stimulation. Acupunct Electrother Res.
2001;26:59-68.
Shipman WH, Cole JL. Increased resistance of mice to X-irradiation after the
injection of bee venom. Nature. 1967;215:311-12.
Ginsberg NJ, Dauer M, Slotta KH. Melittin used as a protective agent against
X-irradiation. Nature. 1968 Dec;220:1334.
50
(2) PREVIOUS HUMAN EXPERIENCES OF APITOX
Several studies have been conducted with Apitox(Apitoxin), including a
Phase 3 clinical trial that formed the basis of the marketing approval of
Apitox in Korea. These studies are summarized below.
1.
Studies in Healthy Normal Human Volunteers
Fourteen healthy male (10) and female (4) subjects between the ages of 26
and 52 years of age were enrolled in and completed this study. Among
the male subjects, 3 were beekeepers who were included in order to obtain
data from the maximum end of the spectrum without having to
administered venom in high dosages (they received 10-350 stings per week
during the course of their normal work). Injections were administered
subcutaneously twice weekly, with each dose being dependent upon
tolerance to the previous dose. All subjects were administered a maximum
dose of 0.07 mg of Apitox and maintained on that dose for a week before
continuing with an arthritic injection schedule. Each subject received
13 doses of Apitox according to the following schedule:
Concentration
per Injection
Injection #
1
2
3
4
5
6
7
8
9
10
11
12
13
1×
1×
2×
2×
3×
3×
4×
4×
5×
5×
5×
5×
5×
0.07
0.07
0.07
0.07
0.07
0.07
0.07
0.07
0.07
0.07
0.07
0.07
0.07
+ 0.05
+ 0.05
+ 0.05
+ 0.05
Total Dose
(mg)
0.07
0.12
0.14
0.19
0.21
0.26
0.28
0.33
0.35
0.35
0.35
0.35
0.35
Skin tests, vital sign measurements, and blood and urine laboratory
evaluations were performed. One subject who became ill and was
51
subsequently hospitalized was shown to have contracted an adenovirus
infection. A female subject had a delayed local reaction: 18 hours after
injection, she developed a 4+ wheal with a very large flare. A repeat
injection of the same concentration and all subsequent injections were
normal. None of the subjects had a systemic reaction and there were no
changes noted in vital sign measurements. None of the subjects reported
significant pain.
In a second study, 20 healthy (10 male, 10 female) subjects between the
ages of 23 and 45 years of age were enrolled and 15 completed the study.
Five subjects dropped out of study because of failure to keep study
appointments, not following study directions, or due to consumption of
alcohol during the study. Each subject received an initial test dose of
0.05 mL and then 12 doses of Apitox, starting with 0.1 mL with the first
intradermal injection and increasing to 0.2 mL (second injection), 0.25 mL
(third injection), 0.3 to 0.7 mL (fourth through twelfth injections).
Injections were administered 2 to 3 times per week over a period of 4 to
6 weeks. Physical examination, blood and urine laboratory evaluations,
and vital sign measurements were performed.
There were no significant changes from baseline noted. Localized itching
was the most common adverse experience (11/15). Edema (5/15), pain at
injection site (2/15), and blister at injection site (1/15) were also reported,
but no serious adverse experiences were reported. Thus, it was concluded
that Apitox can be safely administered to humans when applied in
therapeutic doses.
2.
Studies in Patient Populations
Studies evaluating the safety and efficacy of Apitox have been conducted in
patients with chronic pain and inflammation (due to rheumatoid arthritis,
osteoarthritis, fibromyositis, or peripheral neuritis) and those with
osteoarthritis.
1) Chronic Pain and Inflammation Study (United States)
In the chronic pain and inflammation study, 180 subjects were randomized
to receive, over a period of 6 weeks, twice weekly injections of either
Apitox (1 mg/mL) or histamine phosphate (0.275 mg/mL). The number
of injections increased with each subsequent visit (e.g. 3, 6, 9, 12, 15, 18,
52
20, 20, 20, 20, 20, and 20 injections). The injections were administered to
the area of pain first and then beginning with the fifth session, as the
number of injections increased, to the corresponding dermatomal area of
the spine.
The visual analogue scale (VAS) and McGill Pain
Questionnaire (MPQ) were used to assess the level of pain.
Thermographic evaluations and physical examinations (to evaluate
swelling, tenderness, and range of motion limitations) also were performed.
Both groups had reductions in pain scores following treatment, with the
Apitoxin treatment group demonstrating a greater improvement (pain
scores: control 57 vs. Apitox 18). In addition, there were significant
differences between control and the Apitox treatment group at the 6-month
follow-up visit (pain scores: control 83 vs. Apitox 29). The Apitoxin
treatment group also demonstrated greater improvement in the physical
examination and thermographic findings.
2) Pain and Inflammation of the Osteoarthritis Study (Korea)
This was a randomized, active-controlled (nabumetone) study in which
101 subjects with osteoarthritis of the knee or spine were given twice
weekly injections of Apitox (maximum doses of 0.7 mg [Group A], 1.5 mg
[Group B], or 2.0 mg [Group C]) or an oral once daily dose of the control
(1000 mg nabumetone, Group D) over a period of 6 weeks. A 4-point
Likert-like symptom severity rating scale was used to assess pain, disability,
and physical signs. A 5-point scale was used for subject self-evaluation.
Safety was assessed through observation of adverse experiences and blood
and urine laboratory measurements.
A total of 81 subjects completed the study. Those subjects assigned to an
Apitox treatment group demonstrated a statistically significant greater
improvement than those in the nabumetone group (p < 0.01). Within the
Apitox groups, Groups B and C demonstrated greater improvements than
Group A (p < 0.01). The most common adverse experiences reported
were injection site itching and generalized body aches.
3) Other Experience (Korea)
Following approval of Apitox in Korea, a post-marketing survey was
conducted in November 2005. Included in this survey were 1,596 patients
who received Apitox therapy.
53
Patients, without any compensation, voluntarily filled out the survey forms.
The information collected included personal information, present illness,
past history, and present medications. The physicians recorded the
treatment records, including the diagnosis, treatment dates for 12+ sessions,
doses, and adverse experiences (including a detailed description). A
complete blood count was performed before treatment and after the last
treatment. The physician immediately notified the pharmaceutical company
and Korean FDA in the event of a serious adverse experience.
According to the survey, no major adverse experiences were reported.
In addition to this survey, The Pain Center, PC University Medical Center
located in Korea has documented the use of Apitox in 3, 679 intractable
medical condition and autoimmune disease patients between
September 2003 and November 2005. No major adverse experiences were
reported. Minor adverse experiences included itching (injection site),
swelling (injection site), pain, low-grade fever, flushing, headache, and
diarrhea.
Antibacterial Properties of Whole Body Extracts and Haemolymph of
Maggots of Lucilia sericata
Kosta Y. Mumcuoglu1, Lea Huberman2, Nathan Gollop2, Colin Bloch3 &
Rachel Galun1
1
Department of Parasitology, Hebrew University-Hadassah Medical School,
Jerusalem, Israel; 2Department of Food Sciences, The Volcani Center, ARO,
Beit Dagan, Israel; 3Clinical Microbiology Unit, Hadassah-Hebrew
University Medical Centre, Jerusalem, Israel
The aim of this study was to partially characterize maggot-secreted
antibacterial substances and determine their range of activity against
different bacteria. Sterile and non-sterile maggots maintained in the
laboratory and taken from wounds of treated patients were used. Whole
body extracts and haemolymph was fractionated and their range of activity
54
against bacteria was tested by the zone inhibition assay. The mode of action
of bacterial destruction was examined by viable counts, influx of K+,
changes in the membrane potential by scanning electron microscope
(SEM). Extracts of sterile and maggots non-sterile showed an activity of
200 arbitrary units (AU)/ml and 400AU/ml respectively. Maggots removed
from chronic wounds had an activity of 1200AU/ml. Injuring sterile
maggots with a sterile needle doubled the antibacterial activity within 24
hours, while the antibacterial activity of haemolymph increased fourfold
after injuring with a sterile needle and sixteenfold with an infected needle.
The fractions with a molecular weight of <1kDa and 3–10kDa showed
antibacterial activity against Gram-positive and Gram-negative bacteria
including Pseudomonas aeruginosa, Klebsiella pneumoniae and
methicillin-resistant Staphylococcus aureus (MRSA) isolated from wounds.
The fraction with a MW <1kDa lysed over 90% of the bacteria within 15
minutes by causing an influx of K+ and changing the membrane potential
of bacteria. The nature of the antibacterial materials extracted from
maggots indicates their potential significance in wound healing in addition
to the actual ingestion of the necrotic tissue on the wound.
The Use of the Medicinal Leech, Hirudo medicinalis, in the
Reconstructive Plastic Surgery
Kosta Y. Mumcuoglu1, Rivka Cohen1, Andre Ofek2, Howard Lipton2 and
Carole Pidhorz2
1
Department of Parasitology, Hebrew University-Hadassah Medical School,
Jerusalem, Israel
2
Department of Plastic Surgery, Hadassah Medical Center, Jerusalem, Israel
The medicinal leech, Hirudo medicinalis, is being used to salvage
compromised microvascular free-tissue transfers, replanted digits, ears, lips
and nasal tips due to venous congestion. Twenty-three patients (14 males
and 9 females), 8-79 years old average: 35.9 years) with devascularized or
amputated fingers, open wounds after accidents or surgical wounds after
scar revision were included in the study. Of the 15 fingers treated by leech
55
therapy, 10 fingers were saved (4 out of 9 replanted fingers and 6 out of 6
revascularized fingers), while out of 18 flaps treated by this treatment
modality, 17 were salvaged (3 out of 4 free flaps and all 14 island and
random flaps). Fifteen patients received 1-13 units of packed blood cells
(average 2.9). The patients with revascularized or replanted fingers were
treated in average of 2.5 days and each finger was treated with an average
of 5.7 leeches. The 15 patients with flaps were treated in average of 3.4
days and each flap was treated with an average of 9.2 leeches. In
conclusion, leech therapy should be considered as an integral part of the
armamentarium used in reconstructive surgery. It improves greatly the
success rate of the surgery in cases of post-operative venous congestions,
allowing blood drainage until angiogenesis is established.
Elimination of Symbiotic Bacteria from the Intestinal Tract of the
Medicinal Leech, Hirudo medicinalis
Kosta Y. Mumcuoglu1, Colin Block2, Lea Huberman1, Rivka Cohen1,
Violetta Temper2 & Rachel Galun1
1
Department of Parasitology, Hebrew University-Hadassah Medical School,
Jerusalem, Israel and 2Department of Clinical Microbiology and Infectious
Diseases, Hadassah-Hebrew University Medical Centre, Jerusalem, Israel
In recent years, recognition of a number of antithrombotic substances, a
vasodilator and other factors in the saliva of the medicinal leech, Hirudo
medicinalis, has generated renewed interest in its use in promoting venous
drainage in tissues whose vitality is threatened by venous congestion and
obstruction, especially in plastic and reconstructive surgery. Hirudotherapy
has been complicated by infections, occasionally of extreme severity,
caused by Aeromonas spp., which are considered endosymbionts of the
leech. Therefore, antibiotics such as cephalosporins or fluoroquinolones are
given prophylactically during treatment. Up till now, methods of
eliminating these bacteria prior to treatment have not proved successful.
The purpose of this study was to use a unique method of inducing leeches
to ingest a blood-free antibiotic solution in an attempt to eliminate the
56
aeromonads, thus possibly eliminating the need for chemoprophylaxis.
Eighty control leeches were examined bacteriologically for colonization by
aeromonads at 1-2 week intervals before the definitive experiment.
Cultures were taken from the external surface and then by dissection from
the crop and intestine. Test animals were fed in an artificial feeding
chamber using an arginine solution in physiological saline as a
phagostimulant: 63 received the solution with ciprofloxacin 100μg/mL; 54
of these were given blood meals at intervals after treatment, to evaluate
whether the drug influenced their ability to take blood; 7 controls were not
given blood. After treatment, leeches were held in sterilized chlorine-free
tap water containing 20μg/mL ciprofloxacin. Aeromonas spp. were grown
from 57/80 control leeches (71.25%). Readiness to take a blood meal was
unaffected in treated animals. All 54 treated animals that were subsequently
given blood meals were free of detectable aeromonads when cultured at
intervals from 1 week to 2 months. All 7 treated leeches that were not fed
blood meals were still alive after 2 months. Other environmental bacteria
and some filamentous fungi were isolated from 28.6% of treated animals
that had taken blood meals, but were not fully characterized in this phase of
the study. These preliminary results indicate that our strategy reduced the
population of leech-associated aeromonads to undetectable levels for
extended periods, and that treated leeches are likely to take a blood meal
from patients undergoing hirudotherapy. This might permit the avoidance
of antimicrobial prophylaxis in these patients. Outstanding questions to be
resolved are, among others: 1) the maximum time leeches can survive with
undetectable quantities of their aeromonad symbionts, 2) the amount of
time that will be needed between treating the leeches and their becoming
ready to take a blood meal, 3) the nature and potential clinical significance
of the microbial population remaining after the elimination of the
aeromonads and 4) the true role of these organisms in leech biology.
57
58
59
60
61
Autoblood from medicinal leeches as a means of active nonspecific
62
immunotherapy in oncology
V.A. Savinov and O.A. Kursakova
Moscow Alliance of Hirudotherapeutists, Russia
Autoblood from medicinal leech contains symbiotic bacterium
Aeromonas hydrophila which is a nonanamnestic antigen for man and is
therefore used with the purpose of active nonspecific/adjuvant
immunotherapy. Autoblood diluted 1:10 with normal saline is administered
intradermally in the amount of 0.1 mL, or 0.5 mL of the native autoblood –
subcutaneously, or 1.0 mL of autoblood diluted in 100.0 mL of normal
saline – intravenously in droplets. Intracutaneous administration is
accompanied by development of the delayed-type hypersensitivity reaction
with the formation of a macrophagal-lymphocytic infiltration, while
subcutaneous administration is followed by fever and hyperdermia up to
41°C during 5 to 6 hours with no subsequent negative aftermath, which is
related to invasion of the human body with the symbiotic bacterium.
From 1992 to 2002, in the complex with the standard therapeutic
methods autoblood from medicinal leech was used for treatment of 60
patients with morphologically verified diagnosis: 23 – prostatic cancer, 16
– urinary bladder cancer, 14 – breast cancer, 5 – lymphgranulomatosis, 2 –
colorectal cancer. Improved quality of life and elevated level of the
immune status were noted.
Maggot Therapy - Reflections from the Past, Foretelling the Future
Ronald A. Sherman, M.D.
University of California, U.S.A
Purpose
To explore the history and current status of maggot therapy world-wide, in
order to predict and better guide its future.
Methods
63
Review of published and unpublished records of maggot therapy use,
research, and regulatory intervention.
Results
Several trends were identified: Within the past 12 years, the use of maggot
therapy has spread throughout the world. Some communities have adopted
maggot therapy more readily than others. In most countries where it is used,
it is now accepted, if not embraced, by the medical establishment. Factors
inhibiting more widespread use include concerns about cost recovery, ready
access, hassles associated with dressing application, emotional distress of
patients, and lack of scientific studies. Some of these concerns are
legitimate impediments; others are groundless. All must be addressed and
overcome.
Conclusions
Some communities have adopted maggot therapy more quickly and
completely than others. Understanding the reasons for such trends may help
us to overcome the impediments to access.
Frequently asked Questions in Apitherapy
Dr. Stefan Stangaciu
President of “Apitherapy Consulting & Trading International” President of
the German Apitherapy Society
• What are the best bee products and the best Apitherapy protocols to
improve health, beauty and strength?
The best bee products for us are the ones that clearly can improve our
health, strength or beauty. To know how to choose the best bee products we
need to follow the following guidelines:
* the best bee products are the fresh ones, collected and processed in the
areas where we live; these products can usually ensure the maximum of
nutrients and active compounds that our body needs; if the local bee
products are not available in enough amount, we can order other ones,
but ideally also from our region; the reason is that the bee products are
collected by the bees from plants and trees that are perfectly adapted to
64
the climatic conditions where we also live; the plants that lives in the
mountains secretes more bio-flavonoids than the plants that are at the
sea level, due to the higher radiation level in the mountains; as a result,
the honey and propolis made in the mountains protects better the people
that lives also in the mountains area, than the products made/collected
at the sea level; same kind of example is valid also for the people
suffering from pollen allergies (hay fever); if the bee pollen is collected
from the same area where the patient lives, and administered in very
small doses at the beginning, with slow increase of the administered
amount, after meals, the chance to desensitize the patient of that allergy
is higher; the explanation is in the fact that the pollen allergens that are
present in the atmosphere of that city/area will be present for sure, in
very small amounts, in the bee pollen collected by the bees from the
same area too, but with greater probability will be not present in same
amount in the bee pollen collected more than 30 Km. away
* once we decided to use the local bee products and we can get them in
enough amount, we need to ask our local medical doctors and pharmacists
what are the best administration forms to use. Here the doctors will need
to ask the local pharmacists to prepare specifically for each patient (if
necessary) the exact preparation/product that can reach faster the affected
area. If we have conjunctivitis (eye inflammation) for example, we should
use mainly eye drops and not suppositories…
Here below we insert a table that can orient you in asking your local
doctors/pharmacists for a more specific administration method:
System/Organ/Tissue to
be treated
Nervous system
Best pharmacological
forms
Administration method
Oral forms: fresh collected Per oral (products that can
bee products, or at least
fresh frozen, in any form
that can be swallowed
(honey-pollen
mixtures,
tablets, capsules, powder,
granules, tincture, syrups,
in form of liquids, juices,
65
be swallowed).
Injectable
bee
venom
solution and/or live bee
stings applied on the
painful spots and/or on the
acupuncture
points/meridians.
etc.); bee venom in
solution form for injection
on acupuncture points;
ointments and/or creams to
stimulate the blood flow in
the head and vertebral
column area
Endocrine system
Same as above. The Same as above, but the
preparations that can be acupuncture points to be
kept under the tongue for stimulated
will
differ
minimum 4-5 minutes according with the location
should be preferred
of the endocrine gland that
is currently treated
Immune system
Face
Same as above + Propolis Same as above, but
based suppositories
acupuncture points to
stimulated
will
specifically elected by
specialist in apipuncture
Creams, ointments and/or Local application following
masks with all bee
products + oral forms +
bee venom solutions for
micro-apipuncture
Eyes
the
be
be
the
the cosmetic related rules,
ideally on the facial related
acupuncture
points;
swallowing the oral forms
after
chewing
and/or
sucking of the freshly
collected (frozen) raw bee
products
Eye drops + ointments and Eye
drops
creams
+
oral conjunctival
in
sac
the
+
pharmacological forms that ointments and creams that
can be swallowed
can be applied on the eye
lids and on the surrounding
acupuncture points + oral
forms
that
can
be
swallowed
Nose
Nose drops + solutions for Dripping the nose drops
66
inhalations with honey,
propolis
extract
and
essential oils + sunflower
or olive oil based propolishoney
mixtures
+
ointments + oral forms +
bee venom solution
solution in small amount +
inhaling the vapours from
the solution specially made
for inhalation + superficial
inserting the creamy-oily
mixtures in the nostrils (not
too deep) + stimulation of
the local acupuncture points
with
ointments/creams
and/or micro-injectable bee
venom
for
microapipuncture + swallowing
the oral forms (ideally,
except the capsules, after
sucking them for minimum
3 minutes)
Ears
Sunflower or olive oil inserting gently the creamybased
propolis-honey oily mixtures in the external
mixtures + solutions for ears entrance, in very small
inhalations with honey,
propolis
extract
and
essential
oils
+
ointments/creams + oral
forms + solutions bee
venom solution + ear
candles
amount + stimulation of the
local acupuncture points
with
ointments/creams
and/or micro-injectable bee
venom
for
microapipuncture + inhaling the
vapours from the solution
specially
made
for
inhalation + swallowing the
oral forms (ideally, except
the capsules, after sucking
them for minimum 3
minutes)
+
specific
application of beeswax ear
candles
Mouth
Various
honey-pollen- Local application inside the
propolis-royal
jelly mouth + swallowing of the
67
mixtures + water extract of
propolis + propolis tincture
+ chewing-gum + tooth
paste with propolis + oral
forms + bee venom
solution
Teeth
oral forms + acupressure
and/or micro-apipuncture
on the local bioactive points
+ injectable bee venom on
distant acupuncture points
that
are
related
energetically
with
the
mouth tissues
Liquid forms + injectable Ask your dentist to apply
soft propolis extract +
crystallized honey (small
amount)
mixed
with
propolis extracts, royal
on and under the gums
surrounding the affected
tooth/teeth mixtures of bee
products, according to the
jelly and fresh bee pollen + local condition + swallow
oral forms
the oral forms + stimulate
the
teeth
related
acupuncture points
Head
Propolis and/or Bee venom stimulate the head and neck
ointments + oral forms
acupuncture points with the
ointments + through microapipuncture + swallow the
oral forms (after sucking
them for minimum 3
minutes, except for the
capsules)
Neck
Same as above
Back
Bee venom and Propolis Stimulation of the painful
Creams and Ointments +
raw honey + beeswax,
propolis cataplasms + bee
venom solutions + bee
stings + oral forms
Chest
Same as above, but use the
neck related acupuncture
points
spots + the acupuncture
points + honey massage +
local application of warm
beeswax-propolis
cataplasms + eating raw,
fresh (frozen) bee products
Bee venom and Propolis Same as above, but no
68
Abdomen
Upper and lower limbs
Creams and Ointments +
raw honey + beeswax,
propolis cataplasms + oral
forms
injections of bee venom
solutions, due to the higher
sensitivity of the skin
(exception:
microapipuncture) + eating raw,
fresh (frozen) bee products
Same as above
Same as above
Bee venom and Propolis Same as above; the best
Creams and Ointments + acupuncture points are
beeswax,
propolis located on the “Yang”,
cataplasms + injectable bee more hairy areas.
venom solutions + oral
forms
Foot soles
Bee venom and Propolis
Ointments + beeswax,
propolis cataplasms + oral
forms
Use of creams and/or
ointments
on
the
reflexologic
areas
(reflexotherapy
=
Fußsohlenreflexmassage)
Cardiovascular apparatus All oral forms + ointments Swallowing of the oral
+ injectable solutions + bee forms after minimum 4-5
+ Blood
stings + suppositories
minutes of chewing and/or
sucking; stimulation of the
acupuncture points with
ointments, injections and/or
(micro) bee stings +/- use of
suppositories or ointments
in cases of hemorrhoids
Digestive Apparatus
All oral forms, used Liquids or feshly made
according with the distance “cocktails” for treatment of
to the distance to the mouth, esophagus, stomach
“target” + injectable bee and
small
intestine;
venom
solutions
+ capsules, solid propolis
suppositories
“beans” and suppositories
for the large intestine,
rectum and anus areas +
stimulation
of
the
69
acupuncture
points
according to the location of
the affected organs
Respiratory Apparatus
Solutions for inhalations +
creams/ointments
+
cataplasms + injectable
solutions + chewing-gum
with propolis + oral forms,
especially butter-propolis
extract
in
tuberculosis
Uro-genital Apparatus
cases
Inhalation through the nose;
stimulation of the nose,
chest and back areas
(between
shoulder
bladders)
with
the
ointments and/or injectable
of bee venom solutions +
swallowing the oral forms
For women: Suppositories
+ vaginal suppositories +
creams and ointments +
vaginal solutions with
propolis and/or royal jelly
+ injectable solutions +
oral forms, especially
propolis extracts + fresh
Local use of suppositories
(intra-rectal)
+
local
stimulation
of
the
acupuncture points with the
creams and/or ointments +
solutions to be used by
women intra-vaginally.
frozen royal jelly.
Ingesting of the oral forms
For men:
Suppositories + creams and
ointments + injectable
solutions + oral forms,
especially propolis extracts
+ pollen preparations and
products
Osteo-articular and
muscular system
Bee venom and Propolis Local stimulation of the
ointments + injectable bee affected
tissues
and
venom + bee stings + oral acupuncture
points
forms + propolis-beeswax including through microwarm/cold cataplasms
apipuncture
+
gently
“inject” the bee venom and
propolis ointments through
methods
specific
to
physiotherapy (like electro-
70
and
phonophoresis)
+
application
of
the
cataplasms according to the
local signs and symptoms +
swallowing of the oral
forms, especially propolis
extracts
Skin
Raw,
fresh
collected Local application according
Honey, Propolis, Royal to the symptoms and signs
jelly and Pollen in various
extracts and cocktails +
Ointments and Creams +
oral forms + (micro) bee
of the
Medicinal
wounds
Tincture
stings + shampoos, soaps dermic
and lotions with bee lesions
products according to your
skin type and/or disease
affected area.
honey for deep
and
Propolis
for superficial
and
epidermic
• What are usually the best doses?
The ones that bring relief, which improves immediately or on a long
term our health, strength and beauty. It is always recommended (except for
the emergencies like burns or scalds) to use the bee products initially in
very small doses (to test for a possible allergy or intolerance), than to
increase gradually until a clear improvement occurs (see the time/quantity
relationship)
• What are the best pharmacological preparations and/or products for me?
The ones that reach faster and in enough amount the affected tissues,
organs or systems in your body. Ask always your local health practitioner
and your pharmacist what could be the best forms. Use also, as guidelines,
the above table.
• What is the best way to administer the bee products for my health,
strength and beauty?
Ideally is to administer as many as possible pharmacological forms (see
71
above), to be sure that you will reach the affected area with enough
nutrients, oxygen and active healing compounds.
• What are the best doses and administration methods for children and old
people?
For children under 3 years ¼ of a dose of an adult. For children
between 3-8 is better to use 1/3 of the adult dose and for the children
between 8-12 use half (1/2) of the adult dose.
• What is the best time/quantity relationship for me when I use bee
products for health? How much time and in what amount shall I take the
bee products?
For those diseases that have less symptoms than usually (like blood
diseases or cancers in initial stages) the level of “smile” can be obtained
through specific laboratory analysis.
After complete healing/improvement pauses can be taken, according to
the medical advice.
72
• Are there any adverse reactions when I take the bee products together
with other remedies or practice simultaneously Apitherapy with other
healing methods?
Usually not, but in some cases the use of bee venom therapy is
restricted for those taking heart related drugs. Also, rheumatic and multiple
sclerosis patients are usually advised to refrain from taking steroidal and
non-steroidal anti-inflammatory drugs during bee venom therapy. In cases
when your medical doctor prescribed you mandatory some drugs, you can
use though the “soft” bee products like honey, bee pollen, royal jelly and
propolis, and once the situation is improved, with the agreement of your
doctor, you can start also bee venom therapy.
• What persons can practice bee venom therapy?
Only the licensed medical doctors (MD) and the Naturopathic Doctors
(HP) can inject various bee products extracts and/or bee venom in solution
form.
Any person can though use propolis and bee venom ointments but
respecting the leaflet recommendations.
• How can we improve the efficacy of bee products through other remedies
and/or healing methods? Is my diet and lifestyle important in Apitherapy?
Any other method or remedy that can improve the absorption of the bee
products related nutrients and/or active compounds should be used.
Also, any other healing method that detoxifies the body, mind and spirit
should be used.
Once the necessary nutrients, the water and the oxygen have reached the
affected area in perfect quantity and quality, the regeneration/improvement
of that area should start. In this phase, a very good local or general
systemic relaxation (sleep) can make wonders. Learn and use any technique
of relaxation, Yoga, Taiji Quan recommended by your local psychotherapist
in order to get faster the results you wish.
All our living cells from our body needs besides nutrients, water and
oxygen, also a warm enough environment; so learn to use daily massage
and calisthenics (gymnastic) techniques.
Our health depends 50% of our diet, 40% of our lifestyle and only 10% of
73
the treatments we make, so be sure to have also a very good diet, specific to
your body needs and to your constitution. Here, the specific advises of your
nutritionist (ideally specialized in Ayurveda too) can help you also
tremendously.
• What are the major counter-indications and limits of bee products?
The allergies (especially the bee venom allergy), the intolerances
(especially to honey and pollen) and the excessive (non-controlled) use of
honey in cases of diabetes type 1.
If the dose is correctly selected and the administration method is proper,
even the people having normally counter-indications to the use of bee
products can use them, but of course ONLY under strict medical control.
• Where do I find more specific information?
Ask for help your local health and Apitherapy practitioners registered
in the nearest to your place Apitherapy Society and you will receive it!
Morphologic Changes of Larvae during Biosurgery
U. Wollina1, N. Kunath1, G. Haroske2
1
Department of Dermatology, Dresden-Friedrschstadt Hospital, Germany
2
Institute of Pathology, Dresden-Friedrschstadt Hospital, Germany
Maggot therapy or biosurgery is discussed with increasing interest not only
by scientists but also by the public. Maggot therapy is based upon the use
of living larvae of necrophagous flies to treat chronic non-healing wounds
and leg ulcers. While there is great success in clinical use and the
technology of breeding and applying larvae, the knowledge about the
changes of larvae within the wounds is still quite limited. For this reason
we investigated the larval morphology before and after the use thereof in
chronic ulcers by light microscopy. The results demonstrate dramatic
changes in larval morphology during the biosurgical process.
Introduction
74
The therapy of chronic wounds and ulcers with maggot (larvae of Lucilia
sericata) was widely used during 1920s and 1930s, as no antibiotics for
their treatment existed. [1,7,9] This therapy has experienced a renaissance
in the last 10~15 years. Clinical studies have shown that the larvae were
remarkably successful and efficient in the case of debridement of chronic
wounds such as leg ulcer, diabetic gangrene, or decubitus. [ 10,12,13,17 ]
Based on the level of present knowledge, the mechanism of healing
supported by larvae is to be described as follows :
* Larvae separate the proteolytic enzyme, which dissolves the necrotized
tissues.
* The mixture of tissue and fluid is soaked up by larvae.
* In the presence of larvae the amount of the produced exudate of wounds
increases, and as a result the bacteria will be washed out.
* Through the secretion of larvae the pH-value of the wounds changes.
* Effective antibacterial substances in the digestive tract of the larvae
reduce the number of germs.
* The secretion of larvae promote the healing process. [ See Table 1. ]
* The movement of larvae inside the wounds promotes granulation.
Thus the digestive tract plays the central role for the clinical activities of
larvae. During antibacterial activities the separation of enzymes and
Zytokinen appears to be important. [ 3,14,15 ]
In one particular study, haemolymph, digestive fluid, and skinning
(sloughing ??) hormone ( 20-Hydroxyecdyson ) of Lucilia sericata were
brought together, and their effect on human Fibroblasten was tested. And
it was confirmed that each of these extracts stimulated the growth of
Fibroblasten cultivation, albeit merely by 12 %. The epidermal growth
factor alone can achieve that much stimulation. A significant increase in the
build-up of Fibroblasten occured, when the extracts were used together
with epidermal growth factor. The digestive fluid had significant effects on
the Fibroblasten, when they were combined with Zytokin Interleukin 6.
[11]
In the following investigation, we analyzed the morphological changes
of larvae under the light microscope during the clinical biosurgical process.
Material and Method
75
It was the larvae of Lucilia sericata species that were utilized by Biomonde,
a Hamburg-based firm. A few of them were conserved in formalin, colored
with Hematoxylin-Eosin (HE) and were fixed on the microscope slides for
investigation. The remaining larvae were used for biosurgery on chronic
leg ulcers. After 5 days of activities on the ulcer, these larvae were
collected, colored likewise with HE. They were placed on the slides, and
examined under the microscope. The morphology was then compared with
the unused larvae.
Outcome
The larvae freshly delivered from the company are small and agile. Their
average length amounted to about 6.2mm, with maximum diameter
0.34mm (Fig. 1 a). They were covered with finely folded skins
(“Kutikula”). Around the head area, there were mouth-like tools with a
hook visible. (Fig. 1 a & b ). The saliva-glands were well developed. At
the rear end of larvae “Vakuolen” (= cellular cavity) were to be observed.
The connective tissue existed only in narrow areas, although the “Malphigi
vessel” were enlarged (Fig. 1 d). The intestines of the larvae were empty
(Fig. 1 e).
After the biosurgical usage, that is, after 5 days of application to
chronic wounds, during which the larvae feed themselves with fluid
necrotic materials, they grew up (Fig. 2b). The length was now about 1415 mm, and its diameter was grown up to 0.77 mm. “Kutikula” was now
thicker, and the fat tissue was distinctly increased. (Fig 2b). The hooks of
the mouth-like tools were not recognizable any more, and the saliva-glands
had also disappeared. The connective tissues revealed themselves more
strongly, and the Lipid- and Saccharid-filling were now visible (Fig 2c & d).
Diameter of Malphigi-vessel had become smaller(Fig 2d).
Discussion
Lucilia sericata larvae form, in their natural living environment,
amorphous mass of maggots, which protects each individual larva and
offers the optimal conditions for growth. The larvae are necrophilia,
which dissolve and take in the necrotized tissues through secretion of
Lipasen and Proteasen. In the middle section of these larvae’s intestines,
the materials thus taken experience a further corrosion of enzyme.
Around the end of larvae stages the size of each individual maggot gets
increased by up to 100 times. The length of the larva’s digestive tract
76
amounts to something like 5 times its body length, and is longer than the
fully-grown flies. The digestive tract is divided into three parts. In the
first (fore intestine) and the second (middle intestine) segments Proteasen –
and perhaps Lipasen, too – are secreted. The pH-value in these areas is
sour, standing at 2.4 up to 4.8. The most active segment is the middle one.
Following this is the hinter segment, i.e., the end of intestine, which leads
to the rectum. [3, 14, 15] The larvae finish their taking up of nutrition and
digestion in 5~6 days, and prepare themselves for “metamorphosis” or
“pupation”.
Malphigi-vessels are an absorptive organ at the end of the intestine,
which helps principally the excretion. They contain calcium carbonate in
the form of small stones. The calcium is required in order to ensure the
mechanical strength of the cocoons during the pupation process. [6] The
reason Malphigi-vessel becomes smaller seems to be related to the fact that,
around the end of the nutrition take-up phase, the end of intestine is empty.
On the other hand, the fat deposits become notably bigger (Fig. 2 b), in
order to satisfy the demand of energy during the pupation process.
In the case of biosurgery for chronic wounds, the larvae achieve
principally debridement and decontamination, and demand the formation of
granulation tissues. The bacterial strains of chronic wounds are clearly
diminished through the intra-intestinal anti-bacterial activities. The larvae
are effective against a wide spectrum of “pathogenen” germs such as, for
example, MRSA. [ 2, 3, 6, 11, 14, 15 ]
The volume of bacteria thus taken will reduce itself very clearly, as they
pass through middle- and end-intestines. [10] In a clinic study on the
maggot therapy for the Osteomye patients, a much higher effectiveness in
the case of the wounds infected with Gram-positive bacteria was confirmed,
corresponding to the in-vitro results. [14] However, the Gram-negative
bacteria are cultivated out of the wounds after maggot therapy more often
than before. In the case of wounds infected with Gram-positive bacteria
the opposite effect was to be noted. In “Vivo” the maggots appear to be
less effective against the wounds which are infected with gram-negative
bacteria. The authors of the study are of the opinion that, not only for the
larger wounds but also for the wounds with larger scab on the surface, more
maggots are needed, and also for the wounds infected with gram-negative
bacteria. [14]
77
The Sezenierung of the substances looking like growth factors (?)
stimulates the healing of the wounds through the larvae (Interleukin-6 and
Interleukin-10, substances which are similar to the epidermal growth
structure and insulin-like growth factor, as well as Interferon-y). [11] The
excretive-secretive products (ESP) of the larvae clearly reduce the in-vitroadhesion of Fibroblasten to Fibronektin, and in closer environment, to
collages.
The expansion of Fibroblasten cells was equally reduced, although the
cells were capable of surviving. As far as the expansion as well as the
adhesion was concerned, the heat-treated ESP was essentially less active
than ESP not so treated. Compared with ESP-Blank, the activity was far
more considerable. ESP seems to indirectly change the adhesion of
Fibroblasten, and in fact, on the way over a proteolytic fragmentation of the
surface of the Fibronektin.
Observed altogether, the secretion of Lucilia sericata larvae changes
the adhesion of the Fibroblasten and the expansion of extra-cellular matrix
over the protein surface, through which the cells nevertheless remain
capable of surviving. Here the proteolytic activity of ESP plays an
important role. Transferred to the situation of a wound, the amended
interactions of Fibroblasten with the extra-cellular matrix can require the
formation of new tissues. [8] As was displayed in a remissionspectroscopic way, the micro-circulation gets improved after the biosurgery,
but the exact reasons for this are not yet known.
It follows, from our own results as well as from the literature, that
complex morphological changes take place with the maggots – either on a
skin damage or in natural field – which is necessary, in order for the larvae
to be able to pupate. The physiological properties of Lucilia sericata
support the healing of chronic wounds. Their ESP and digestive tracts are
essential factors of these medically desirable effects of biosurgery.
78
ABSTRACTS
for
Contributed Papers
Maggot debridement therapy (MDT) in a diabetic foot wound patient
suffering from severe gout
A. Andersen1, 2, B. Joergensen2, T. Karlsmark2 ,K. Kirketerp-Moeller2, K. A.
Krogfelt1.
1.
Statens Serum Institut, Copenhagen Dk, 2. Copenhagen Wound Healing
Center Dk.
Purpose/Background: 57 year old male diabetic, through 25 years
suffering from severe gout (arthritis urica AU), was confined to a
wheelchair due to wounds on both feet. The severity of the AU in this
patient makes him a unique case in Denmark. With the occurrence of large
sloughy and necrotic diabetic wounds on both feet, distending from the big
toes covering approximately 35 cm2 down both forefeet. The excessive
deposition and release of monosodium-urate crystals, mm to cm in size
made wound debridement difficult. The removal of the crystals was
accompanied by excruciating pain for the patient and required lots of brute
force from the nurses. Furthermore, bacterial colonization and the crystals
in combination caused the wounds to be extremely malodorous.
Methods: The patient was treated with MDT on both feet during two
consecutive periods of two times 3 days with three weeks pause between
79
the two periods. 3 Biobags was applied to each wound. In the periods
between the treatments the patient was treated with Biatain Ibu and
standard conservative treatment by the home nurse. Following MDT the
patient was admitted for Versajet, debridement of the deeper cavities and
split thickness skingrafts on both feet.
Results: MDT facilitated debridement of the wounds, odour and bacterial
burden control and softening of the crystals, which could subsequently
easily be rinsed from the wound. Versajet debridement finished the
preparation of the wounds for skin grafts, which took nicely.
Conclusion: Through the enzymatic action of the maggot secretions, the
monosodium-urate crystals were softened and easily mechanically
removed from the wounds. The wound balance was shifted as to allow skin
grafting and the patient who had been confined to a wheel chair for almost
one year walked out the Copenhagen Wound Healing Center supported by
cane and prescription footwear.
Maggot debridement therapy (MDT) in diabetic foot wounds –
Quorum sensing dependent bacterial niches may promote infection or
MDT failure
A. Andersen1, 2, B. Joergensen2, T. Karlsmark2, T. B. Rasmussen3, T
Bjarnsholt3, M. Givskov3, K. A. Krogfelt1.
1.
Statens Serum Institut, Copenhagen Dk, 2. Copenhagen Wound Healing
Center Dk. 3.CBM, DTU Lyngby Dk.
Background: Many diabetic patients with foot wounds are ideal candidates
for MDT. The maggots gently and thoroughly remove necrotic tissue by
mechanical action and by proteolytic digestion. The maggots secrete
antimicrobial peptides into the wound, kill ingested bacteria in their gut and
alkalinize the wound. However a few reports show that MDT in some cases
subsequently facilitates an infection, when specific bacterial species are
present in the wound prior to treatment. MDT has also been observed to fail
in some cases due to the maggot’s death in the wound environment i.e.
Their bio surgical properties were not applicable.
Wounds and particularly diabetic wounds are susceptible to infection due to
the development of microbial communities within and around the wound
environment. It is now realized that chemically based cell-cell
80
communication (denoted quorum sensing (QS)) and biofilm formation play
important roles in a multitude of human infections. QS enables the bacteria
to keep track of their numbers and pool their efforts in order to successfully
cause disease. Bacterial biofilms show an inherent tolerance to a variety of
antimicrobial treatments including the action of the immune system. This
makes biofilm infections impossible to completely eradicate. Several
pathogenic bacteria show QS mediated organisation and speculation
regarding the role of QS and biofilm formation in wound healing is
appealing. In this preliminary study we wish to investigate the possible role
of QS in relation to infections related to MDT.
Methods: In an in vitro setup using blood agar plates, we challenged L.
sericata maggots with different concentrations of green fluorescent protein
(GFP) tagged Pseudomonas aeruginosa wild type (PAO1 WT), a GFP
tagged QS deficient mutant (PAO1 RR) and a Staphylococcus aureus. The
survival of the maggots was determined and the clearing efficiency
assessed during 3 day periods. The uptake of P. aeruginosa was determined
using fluorescence microscopy.
Results: The maggot survival and clearing efficiency was unaffected by S.
aureus and PAO1 RR compared to negative controls, but maggot survival
was significantly impaired by the PAO1 WT. Furthermore the florescence
microscopy showed reduced uptake of PAO1 WT compared to the PAO1
RR.
Conclusion: The results indicate that infections following maggot
debridement therapy may originate from preferential feeding by the
maggots creating micro niches in which bacteria survive and subsequently
unchallenged cause infections and that this preference is QS dependent.
Furthermore QS controlled virulence factors are toxic for the maggots.
Therefore the presence of specific bacterial species may be counter
indications for MDT. This suggests that QS inhibitors may be a useful
future adjuvant for maggot debridement therapy in wounds harbouring
specific bacterial species or as supplement to standard antibiotic therapy.
MAGGOT IMMUNITY AND DRUG DEVELOPMENT
81
Sergey Chernysh1, Natalia Gordja1, Natalia Chernysh1, Dmitry Tulin1,
Vadim Anikin1, Valery Pleskach1, Myung Jeom Ryu,2 Cheolju Lee2.
1
Laboratory of Insect Biopharmacology and Immunology, St. Petersburg
University, St. Petersburg, Russia
2
Life Sciences Division, Korea Institute of Science and Technology, Seoul,
Korea
Purpose: To analyze mechanisms of Calliphora vicina maggots’ immunity
with respect to drug development.
Methods: Range of experimental immunology, peptide chemistry and
microscopy methods has been used.
Results: Septically injured maggots synthesize about dozen antibacterial
and antifungal peptides of insect defensin, cecropin, diptericin and 3kDa
peptide families. The peptides natural composition demonstrates
antibacterial activity dramatically exceeding the activity of synthetic
antibiotic, chloramphenicol.
The maggot hemolymph contains cytotoxic hemocytes able to kill human
tumor cells and soluble peptides stimulating cytotoxic activity of
mammalian natural killer cells. The peptides referred to as alloferons
were structurally characterized and subjected to full scale preclinical and
clinical studies. Nowadays alloferon-1 is registered as antiviral drug to treat
genital herpes and hepatitis B. Alloferon mode of action is mediated by NFκB signalling pathway and directed to boost recognition of viral and tumor
antigens.
Conclusion: Blow flies developed surprisingly effective immune system to
survive in environments abundant in conventionally pathogenic
microorganisms. Effector and regulatory molecules of maggot immunity
demonstrate biological activities prospective for medical use. Alloferons
are currently used as antiviral medicines and may have broader applications
to treat viral, oncological and immunocompromized conditions in the
future. The maggot antibacterial and antifungal peptides are other
prospective drug candidates.
Further research of maggot immunobiology may contribute new insight to
the biotherapy and drug development.
82
Hepatoprotective potential of earthworm extract (Lampito mauritii,
Kinberg) against paracetamol induced liver damage in rats
Edwin L. Cooper, Ph.D., Sc.D.
Professor, Department of Neurobiology
David Geffen School Of Medicine at UCLA, USA
Our pre-historic ancestors have been exploring natural compounds to cure
their ills, improve and enrich their own lives. Most of these compounds
were derived from plants and animals. The science of curing of various
ailments by using therapeutics obtained from animals is known as
Zootherapy. Animal-based medicines have been elaborated from parts of
the animal body, from their products (secretions and excrements), or from
non-animal materials (nests and cocoons). Research on traditional
medicines has given a vital tool to the upcoming art of Bioprospecting for
pharmaceuticals compounds. Totally 252 essential compounds have been
selected by the World Health Organization, 11.1% derived from plants and
8.7% from animals. For thousand of years, the earthworm and its products,
has been used for therapeutic benefits. The traditional medical knowledge
of indigenous people internationally, more particularly in Asia: India,
Myanmar, China, Korea and Vietnam has played a vital role in identifying,
extracting
and using biologically active
compounds
from
earthworms. Earthworms have a dense nutritional content because they
live in the soil. Various studies of the earthworm reveal several therapeutic
properties: antipyretic, antispasmodic, detoxic, diuretic, antihypertensive,
antiallergic, antiasthmatic, spermatocidal, antioxidative, antimicrobial,
anticancer, antiulceral and anti-inflammatory (Cooper et al., 2004;
Balamurugan, et al. 2007). Earthworm L.mauritii, found throughout India
has been reported in Siddha medicine and is used mainly for antioxidative,
antimicrobial, anticancer, antiulceral and anti-inflammatory. In our present
study, the extract of L.mauritii was investigated for its hepatoprotective and
antioxidant effects on paracetamol (2 mg/kg) induced acute liver damage in
Wistar albino rats. Hepatoprotective activity was determined by using
biochemical parameters such as serum alkaline phosphatase (ALP), serum
83
glutamate oxalate transaminase (SGOT), serum glutamate pyruvate
transaminase (SGPT), bilirubin and total protein. Extract at doses of 100,
200, 300 mg/kg produced significant hepatoprotective effect by decreasing
the activity of serum enzymes, bilirubin and lipid peroxidation whereas it
significantly increased the levels of glutathione (GSH) and super oxide
dismutase (SOD) in a dose dependant manner. The effects of extracts were
comparable to those of standard drug silymarin. These results suggest that
earthworm extracts may have potential therapeutic value in treating certasin
liver disorders, perhaps by antioxidative effect on hepatocytes.
Balamurugan M., Parthasarathi K., Cooper, E.L. Ranganathan
L.S. Earthworm paste (Lampito mauritii, Kinberg) alters inflammatory,
oxidative, haematological and serum biochemical indices of inflamed
rat. European Review for Medical and Pharmacological Sciences 2007
(11) 79-89..
Cooper, E.L., Hrzenjak, T.M. and Grdisa, M.. Alternative sources of
fibrinolytic, anticoagulative, antimicrobial and anticancer molecules. Inter.
J. of Immunopathol. And Pharmacol. (2004) 7(3): 237-244.
Therapeutic Potential of Tunicates
Edwin L. Cooper, Ph.D., Sc.D.
Professor, Department of Neurobiology
David Geffen School Of Medicine at UCLA, USA
Biotherapy may be entering a new era with the recognition of the utility of
looking to other sources of therapies especially by turning to products from
animals, particularly those from the sea. In recent years bioprospecting
using marine natural product has yielded a considerable number of drug
candidates. Most of these molecules are still in preclinical or early clinical
development but some are already on the market. Research into the
ecology of marine natural products has shown that many of these
compounds function as chemical weapons and have evolved into highly
potent inhibitors of physiological processes in the prey, predators or
competitors of the marine organisms that use them. Some of the natural
84
products isolated from marine invertebrates (sponges, mollusks, tunicates)
have been shown, or are suspected to be of microbial origin and this is now
thought to be the case for the majority of such molecules. Marine
microorganisms, whose immense genetic and biochemical diversity is only
beginning to be appreciated, may also become a rich source of novel
chemical entities for the discovery of more effective drugs. This
recognition offers a word of caution for maintaining the purity of the
isolated products. With respect to products derived from complex
multicellular organisms, we must remember that these products are often
associated with the immune systems of these creatures and that they
evolved millions of years ago—thus their immune systems have been an
effective survival strategy. And if it has worked for them, then humans
should harness these products as new-wave antibiotics or anticancer
molecules, as offerings for biomedical applications, particularly those that
may apply to complementary and alternative medicine (CAM). Many
marine invertebrates are immobile, attached to the ocean floor and use
highly evolved chemical compounds to attract food, block the growth of
intruding neighbors or repel predators. Biologists, especially comparative
immunologists in general and invertebrate immunologists in particular,
believe that these survival demands triggered the evolution of a particularly
abundant mixture of bioactive compounds. For example tunicates are one
of the richest sources of interesting chemicals from the marine
environment. They have an elaborate innate immune system and these
various compounds some of them are being assayed as sources of anticancer molecules: lung, breast and ovarian cancer melanoma. Recently
new work reveals the antiviral treatment of hepatitis B virus-transgenic
mice using the sessile tunicate Styela plicata. Clearly the sea offers
enormous potential for discovery of compounds that can be utilized as
therapeutic agents.
Cooper, E.L., Wright, R.K., Stein, E.A., Roch, P.J. and Mansour,
M.H. 1987. Immunity in earthworms and tunicates with special reference
to receptor origins. In Invertebrate Models, Cell Receptors and Cell
Communication, ed. A.H. Greenberg, 79-103, New York, Karger.
Cooper, E.L.1995. Immunology: A look toward the sea and what we have
learned from tunicates. Aquaculture 132: 1-15.
Cooper, E.L. 2004 Drug disvcovery, CAM and natural products. ECAM 1:
85
215-217.
86
The Use of Medicinal Leeches and Medicinal Preparation from them in
the Complex Treatment of Skin and Mucosal Diseases in UST Health
Resort
1
Sidorov B.B., 1Korobeinikova G.A, 2Gileva O.S.
1
UST Health resort, Russia
2
Perm State Academy of Medicine, Russia
Hirudotherapy (HTH) – the use of the leeches for treatment – is one
of the oldest practices in medicine. The past 3 decades have demonstrated a
resurgence of interest in leeches among physicians and patients as a form of
effective treatment. In spite of the serious evolution of our scientific
findings of the hirudotherapy performance, we haven’t totally covered it’s
mechanism even now. Among the important biological properties of
medicinal leeches are blood – letting action and the production of a variety
of active substances that suppress inflammation and pain, improve tissues
microcirculation, metabolism and regeneration.
Various skin and mucosal disorders and diseases still remain a
serious medical problem. Leeches are now applied to treat a wide array of
dermatological problems. Russian special literature of the last years
contains isolated references on the dermatological aspects of leeching
(Smirnoff A. V., 1994; Dyomina T.A. et al, 2003); the spectrum of
indications for HTH of the skin pathology is wide enough: from the severe
forms of dermatoses (psoriases, eczema, neurodermite, ruber lichen planus,
lupus erithrematodes, sclerodermia) to traumatic and infectious lesions of
the skin and mucosae (furuncule, carbuncle, purulent skin wound).
Sometimes the leech can not be a comfortable medicinal preparation:
in some clinical cases it is simply impossible to apply the leech to the focus
of lesion, to standardize the curative effects of leeching, to avoid some side
effects of HTH. That is why today the progress of hirudology is connected
with creation and clinical aprobation of medicinal preparation with
biological active components of leeches. Among them Pijavit –
ecologically pure preparation with a complex of biological active
substances produced by Hirudo Medicinalis. In the forms for external use it
hasn’t shown any irritative, toxic and allergenic influence, standarts
microcirculation in skin and mucosae, provide anti-inflammatory and
87
hydrotative effects.
The purpose of the study was to assess the effectiveness of native
HTH (with medicinal leeches) and hirudopharmacotherapy (HPTH)
(medicinal preparation with biological active substances from Hirudo
Medicinalis) in the complex treatment of dermal and mucosal diseases.
Materials and methods: our own clinical experience in the
treatment of different dermal disorders with the help of leeches (HTH and
HPTH) we have cumulated in the department of Hiridotherapy of UST –
Katchka Health Resort – one of the biggest balneological health resorts of
Russia.
The effect of the complex treatment with Piyavit use in the form of
cream for external use was investigated in 36 children (6-13 years) with
manifestations of neurodermite and atopic dermatitus, 17 patients (17-32
years) with herpetic chilitis and stomatitis. The skin, oral and lip mucosae
were irrigated with antiseptics, dried and covered with thin layer of the
cream twice a day. The duration of the treatment – 7-9 days. The native
HTH (aspirative regimen) was used in 23 patients with ruber lichen
planus (with dermal and mucosae lesions). The whole course consisted of 7
sittings over day.
Results: in 63 % of clinical cases of lichen planus we saw
positive results after 1-2 courses of HTH: the decrease of papul’s number,
the decrease of their capacity, the healing of erosive arrears of the skin and
mucosa, the diminish of edema, erythema and perifocal inflammation.
Positive clinical dynamic was supported by means of polarographic
method (increase of initial and maximal oxygen saturation, speed of
oxygen saturation and etc.) . Results demonstrated the objective and
subjective improvement just on the 1-2 days after the beginning of the
treatment with Pijavit cream: diminish of the strain and pain, edema,
induration and eczematisation of the derma, disappearance of the pruritus
and xeroderma, recovery of salivation and etc. The results after the first
course of HTH in patients with neurodermite and dermatitis were the
following: completely healing of affected derma in 45% of cases,
considerable improvement – in 37%, improvement – in 11% and condition
without visible improvement - in 7% of the patients. The efficiency of the
HPTH raised significally after the second course of treatment.
Native HTH and the use of medicinal preparations from Hirudo
88
Medicinalis can be useful adjunct to the complex treatment of some types
of skin and mucosal diseases.
Induction of antibacterial activity in medicinal maggots by infected
environment
Takuya Kawabata
Okayama university Graduate Sc, Japan
Purpose: Maggot debridement therapy (MDT) has received increasing
interest for the treatment of infected foot ulcers. It is reported that maggot
has three effects on infected wounds; to debride necrotic tissue, to kill
micro-organisms and to stimulate the wound healing. Previous studies on
larval antibacterial activities in vitro have been performed using larvae
grown up under sterile condition. But no attempt was made to investigate
the antibacterial activity of larvae exposed to infected environment.
The purpose of this study was to investigate the antibacterial activity of
larvae exposed to infected environment.
Methods: Larvae of Lucilia sericata were used. Pretreatment with bacteria
was obtained by exposing sterile larvae to Staphylococcus aureus or
Pseudomonas aeruginosa for 24 hours. Larvae were homogenized in sterile
phosphate-buffered saline (PBS) and centrifuged. The supernatant was
incubated with suspension of S. aureus of P. aeruginosa for 30 seconds, 3
hours or 6 hours at 37℃. Aliquots of the mixture were plated on nutrient
agar and the number of Colony Forming Units (CFU) was counted after an
overnight culture at 37℃.
Results: Effect on growth of S. aureus
The number of CFU in S. aureus and P. aeruginosa pretreated group was
significantly lower than that of sterile and PBS-control group (n=8, p).
89
Treatment by Injection-Acupuncture with Bee-Venom(Apitoxin) and
Apitoxin Combined by Chinese Herbal Medicine in Patients with
Canine Hind Limb Paralysis
Hyung-kyou Jun*, Se-kun Park*, Duck-hwan Kim*,1,
Christopher Moon-ho Kim**, Chin-yuan Hsu***, Chin-ling Hsu***,
Jim-cai Liao#, Hao-jen Chueh## and Han-wen Cheng###
* College of Veterinary medicine, Chungnam National University,
Daejeon, Korea
** Anapunsesang Clinic, Seoul, Korea
*** Yeon Chang Veterinary Clinic, Miaoli City, Miaoli Province, Taiwan
# Shan Qun Animal Hospital, Shang Cheng City, Taipei Province, Taiwan
## Zoo Animal Hospital, Taipei City, Taiwan
### Sino Union Animal Hospital, Young He, Taipei, Taiwan
Purpose: The therapy by injection-acupuncture(AP) with bee venom
(Apitoxin) and injection-AP with Apitoxin combined by administration of
Chinese herbal medicine was applied in 2 cases with canine intervertebral
disc disease(IVDD).
Method: Case 1 was diagnosed as thoraco-lumbar IVDD (T11-T12, T12T13, L3-L4 and L4-L5) and case 2 was diagnosed as IVDD at T 10- T11
and T 12- T 13, respectively. Injection-AP with Apitoxin(total 200 ㎍ of
Apitoxin, 0.1 ml /acupoint) plus physical exercise(walking with gocart,
TID/day) and aquatherapy(swimming treatment, BID/week) were given to
each patient. The used acupoints were GV20(Bai Hui), GB30(Huan Tiao),
ST36(Zu San Li), GB34(Yang Ling Quan), ST40(Feng Long), ST41(Jie
Xi) and BL40(Wei Zhong), the lesions, and trigger points. In addition,
Chinese herbal medicine(Koda Pharmaceutical Co., Taiwan) including
Zheng Gu Zi Jin Dan(正骨紫金丹: 1 g), Shiuh Duann(續斷: 0.2 g), Du
Zhong(杜仲: 0.2 g), Mo Yao(沒藥: 0.2 g), Ru Xiang(乳香: 0.2 g) and
Pyrite(自然銅: 0.2 g) were orally medicated BID for 9 days in case 2.
Result: Walking was possible after session 11 for 4 weeks in case 1 and
after session 6 for 2 weeks in case 2, respectively.
Conclusion: The present cases were canine IVDD which showed favorable
therapeutic responses to injection-AP with Apitoxin only and injection-AP
with Apitoxin combined by Chinese herbal medicine.
90
Bee venom(Apitoxin) therapy on canine facial nerve paralysis
Hyung-Kyou Jun*, Hyun-Uk Oh*, Hyun-Hwa Lee*, Ji-Won Han*,
Cristopher Moon-Ho Kim** and Duck-Hwan Kim*
* College of Veterinary Medicine, Chungnam National University, YusungGu, Daejeon 305-764, Korea
** Anapunsesang Clinic, Seoul, Korea
Purpose: To elucidate the therapeutic effect by Apitoxin therapy for canine
facial nerve paralysis(FNP), the present study was performed.
Method: The 12 mongrel dogs (2-3 years old, 2.0-5.6 kg, BW) were used
in this experiment. The experimental dogs were divided intocontrol(4 dogs),
experimental group I(4 dogs) and experimental group II(4 dogs). FNP was
induced by clamping of facial nerve with hemostatic forceps for 20 minutes
under zoletil anesthesia. As for the treatment, control group was
intramuscularly injected with saline into head muscle, and experimental
group I was treated by injection-acupuncture(AP) with dexamethasone
(Huons Co., Korea, 1 mg/ml/head) and experimental group II was treated
with injection-AP with Apitoxin(Guju Pharmacological Co. and Apimeds,
Inc., Korea, 1mg/bottle, 200ug of Apitoxin: 2 % lodocaine =1:1) after
induction of FNP, twice per week for 2 weeks, respectively. The used
acupoints were LI04, LI20, ST02, ST07, GB03, SI18, TH17 and GB34.
The degrees of FNP improvement were evaluated as clinical scores(0:
normal, 1: mild, 2: moderate and 3: severe) and serum creatine kinase(CK)
activities were evaluated. Statistical significance was analyzed by student’s
t-test.
Result: The FNP symptoms were not improved at all until day 14 in control
group, however, the FNP symptoms were much improved until day 14 in
experimental group I and II, respectively. The significant differences of
clinical scores were detected on day 14 in experimental groups, compared
by those of control group (p<0.05).
Maggot Therapy for severe diabetic foot ulcer
H. Mitsui, T. Kawabata, S. Ugaki, S. Ohsawa, Y. Fujii, S. Sakurai, S.
Kasahara, S. Sano
91
Institution: Okayama University Graduate School of Medicine and
Dentistry, Department of Surgery, Division of Cardiovascular Surgery
Purpose: Although effectiveness of maggot therapy for diabetic intractable
ulcers has been advocated in Europe and United States, it has not been tried
so far in Japan.
Method: We started this therapy to treating diabetic foot ulcer for the first
time in Japan and had experienced 43 cases during these 26 months.
Result: The ulcers of 40 patients out of 43 patients were all healed after 3
months. During these treatments complications related to maggot therapy
were not experienced. Failed 3 cases had the same co-mobidity (chronic
renal failure and low cardiac output) and undergo major limb amputation
because of sepsis.
Conclusion: We concluded maggot therapy is effective to wound bed
preparation of diabetic ulcer. Maggot therapy has some benefits: 1. No
contraindication 2. Anesthesia is unnecessary 3. Cost-effective compared
to conventional therapy. 4. Long history and enough evidence of
effectiveness to diabetic ulcer treatment in western countries.
Perspectives: We are now trying to promote maggot therapy to save
diabetic foot which would have been amputated in Japan.
In this congress we want to present the outcome of those 43 cases and
discuss the indication and limitation of maggot therapy to diabetic foot
ulcer. Also we want to show our special dressing for maggot therapy to
make maggot to work effectively and comfortably on ulcer bed.
92
Effects of Bee Venom on Glioma Cells
Byung-Soo Koo, O.M.D.
Chief, Department of Neuropsychiatry, Dongkuk Oriental Medical School,
Korea
Objective: Bee venom (BV) has been used for the treatment of
inflammatory diseases such as rheumatoid arthritis and relief of pain in
Oriental medicine. The two main components of BV are melittin and
phospholipase A2 (PLA2). Of these, melittin, the major active ingredient of
BV, has been reported to induce apoptosis and to possess anti ‐tumor
effects. Several studies have established that the agents inducing apoptosis
in target organs suppress tumorigenesis. As the other component, PLA2 has
been reported to induce neurite outgrowth in PC12 cells. However, there
was no report about proliferative effect of BV in neuronal cells. In order to
examine the effect of BV on glioma cell, human glioma cell line, U87 was
used.
Methods: Analysis of proliferation was confirmed by MTT assay. BV
increased cell number through dose‐ and duration‐dependent manner
and these effects are apparent at a concentration of 10 ug/ml. To observe
which signaling molecules will be activated by BV, phosphorylation of Akt,
MAPK, PYK2 or CREB were examined by Western blot analysis. To study
the long term effect of BV in U87 cells, the image of cells treated with BV
for 4 days were obtained.
Results: The phosphorylation levels of PYK2 and Akt were increased at 5
min after addition of 10 ug/ml of BV and sustained to 2 hours. On the other
hand, phosphorylation of MAPK and CREB were increased at 5 min,
maximum at 10 min, and returned to 30 min. These imply that BV may
activate two different signaling pathways, PYK2/Akt and MAPK/CREB.
BV treated cells showed increased neurite number and length.
These results propose that BV may induce differentiation as well as
proliferation of U87 cells through the activation of PYK2/Akt and
MAPK/CREB.
Efficiency Maggot Debridement Therapy in Fournier’s Gangrene
93
Hugo E. Sarmiento Jiménez*, Jose Contreraz Ruiz**, Alicia Fonseca
Muñoz***
*Department of Surgery, Hospital “San Jose” Oaxaca Oaxaca,
** Department of Dermatology, Hospital General "Dr. Manuel Gea
Gonzalez," Mexico City, Mexico
*** Department of Biology, Hospital “San Jose” Oaxaca Oaxaca, Mexico
Maggot debridement therapy (MDT) has been shown to be a highly
effective method of debridement of necrotic tissue. The speed and efficacy
of this modality has made it a valuable aid in the treatment of chronic
wounds.
Purpose: To show the usefulness and safety of MDT in the treatment of a
serious case of Fournier’s Gangrene.
Method and Results: We present the case of a 59 year old male from
Oaxaca, Mexico. Relevant history for diabetes for 30 years, hypertension
for 5 years and renal insufficiency with peritoneal dialysis for the past 3
years. He was seen at the emergency room for necrosis of the scrotum and
gas that had begun 15 days earlier. The diagnosis of Fournier’s gangrene
was made and the patient underwent surgical debridement. After surgery
and due to recurrent areas of necrosis sessions of MDT were applied and
IV antibiotics continued. The patient had a favourable outcome with
increased granulation tissue, and decreased exudate and odour. that allowed
for treatment with honey-impregnated gauze and grafting.
Conclusions: MDT has proved to be an effective debridement method and
it can be used in Fournier’s gangrene after surgical debridement to increase
the healing rate and improve outcomes.
94
Study of Maggot Debridement Therapy in necrotic pressure ulcers,
Case Series.
Hugo E. Sarmiento Jiménez*, Jose Contreras Ruiz**, Alicia Fonseca
Muñoz***
*Department of Surgery, Hospital “San Jose” Oaxaca Oaxaca, México
** Department of Dermatology, Hospital General "Dr. Manuel Gea
Gonzalez," México City, México
*** Department of Biology, Hospital “San Jose” Oaxaca Oaxaca, México
Purpose: To show the efficacy of MDT in four diabetic patients with
necrotic pressure ulcers
Method: We present a clinical trial with four diabetic patients with necrotic
pressure ulcers in different areas: (2) ischiatic ulcers and (2) sacral
area .The time of developed of them cover a period of (1-11 months) before
MDT was applied. The cause of the development of wounds was
hemiplegia. We treated four patients two women and two men. The average
age was 60.83 years. The mean hospital stay was 18 days (range 7-45
days). Sterile maggots (100-1500) were administered to the wound two or
three times per week and replaced every 24-48 hours. The patients were
assessed daily by nurses in order to evaluate the maggots viability. Finally
the wounds were treated with honey honey-impregnated gauze and grafting.
Results: The complete debridement was achieved in three patients and
partial debridement in one case. After debridation, 2 ulcers were treated
with honey-impregnated gauze until they close and 2 ulcers required skin
graft.
Conclusions: In debridement of necrotic pressure ulcers MDT was
effective and safety method and proved to be a cost-effective method in
short time.
95
Water extracted propolis inhibits TNF- α release following LPS
injection in the rat
Myung-Sang Kwon, Ph.D.
Department of Immunopharmacology, Kangwon University, Korea
Purpose: Acute sepsis is a one of the most prevailing syndrome during
inflammatory response, and is a leading cause of death. Tumor necrosis
factor-α (TNF) is known to as a primary mediator of immune regulation
and the inflammatory response. Some kinds of drug application of blunting
the TNF response in sepsis have been attempted.
The purpose of this study is to know the antiinflammtory effect of propolis
on LPS induced acute inflammation.
Materials and Methods: In order to compare how much blunting TNF
between water extracted propolis (WEP) and pentoxifylline which inhibits
the production of TNF as a positive control. WEP contains lots of
flavonoids shown to inhibition of lipoxygenase. We infer this inhibition due
to WEP may be down regulation of the production of TNF. In this study,
we examined the effect of WEP in a rat model of lipopolysaccharide (LPS)
induced acute septic shock.
Results: Dose response was determined by two different doses injection of
WEP and pentoxifylline. Rats were injected intraperitoneally with
endotoxin. TNF was measured by ELISA.
We also investigated the chemiluminescence of PMA(phorbol-1,2myristate-1,3-acetate) stimulated peritoneal macrophage of rat. Serum TNF
highest elevation occurred after endotoxin injection with peak levels at 90
min. WEP and pentoxifylline treated rats had lower TNF levels and more
diminish chemiluminescence than saline treated control rats at 90 min.
We found that WEP is a potent inhibitor of LPS induced TNF levels and
diminished this chemiluminescence. Conclusions : Our data suggest that
propolis seems to contribute to alternative anti-inflammatory material for
treatment of some kinds of fever inducing bacterial infection.
96
Water extracted propolis attenuates kainite-induced neurotoxicity via
adenosine A1 receptor in the rat
Myung-Sang Kwon, Ph.D.
Department of Immunopharmacology, Kangwon University, Korea
Purpose: It is well recognized that kainic acid (KA)-induced neurotoxicity
is mediated, at least in part, by oxidative stresses. Because some
antioxidants block KA-induced neurotoxicity, The purpose of this study is
to know the antioxidant effects of propolis on seizure induced by kainic
acid.
Methods: we examined the effect of antioxidant propolis on KA-induced
neurotoxicity in rats. Sprague-Dawley rats received water-extracted
propolis (WEP) (50, 100, 200mg/kg, p.o.) five times at twelve times
intervals. KA (10mg/kg, i.p.) was injected 1 hour after last propolis
treatment.
Results: Convulsing behaviors were significantly decreased in the seizing
rat that pretreated with WEP in a dose-dependent manner. Malondialdehyde
(MDA), protein carbonyl (CO) and glutathione (GSH) levels of rat
hippocampus were examined as oxidative stress markers. MDA and CO
levels were significantly increased 4 hours after KA administration. GSH
levels were reduced by KA administration. Pretreatment with EEP blocked
these changes in a dose-dependent manner. These neuroprotective effects of
propolis were counteracted by adenosine A1 receptor antagonist, 8cyclopentyl-1,3-dimethylxanthine [CPT (1mg/kg, i.p.)] and adenosine A2
receptor antagonist, 3,7-dimethyl-1-propargylxanthine [DMPX (1mg/kg,
i.p.)]. However, CPT was more efficacious than DMPX in counteracting
protective effect of propolis.
Conclusions: Consequently, these results suggest that antioxidant
properties of propolis on KA-induced neurotoxicity are mainly via
adenosine A1 receptor.
97
Maggot Excretions/Secretions Inhibit Multiple Neutrophil ProInflammatory Responses
M.J.A. van der Plas1,2, A.M. van der Does1, M. Baldry1, H.C.M. DogteromBallering1, C van Gulpen1, J.T. van Dissel1, G.N. Jukema2 and P.H.
Nibbering1
1
Dept of Infectious Diseases, 2Dept of Surgery, LUMC, Leiden
Introduction:
Maggots of the blowfly Lucilia sericata are used
worldwide for the treatment of wounds that lack the tendency for
spontaneous healing like diabetic foot ulcers. In such wounds, activated
neutrophils and their products may precipitate tissue damage rather than
contribute to healing. Since the beneficial actions of maggots are contained
in their excretions/secretions (ES) we assessed the effects of maggot
excretions/secretions (ES) on pro-inflammatory responses of activated
human neutrophils.
Methods: Neutrophils were isolated from venous blood. g/ml of ES (one
maggotSubsequently, these cells were incubated with 0-100 g of ES/h)
and stimulated with fMLP (syntheticproduces approximately 2 bacterial
peptide) or PMA (activator of protein kinase C). The effects of ES on
neutrophils were analyzed measuring chemotaxis, the production of H2O2
and the release of elastase. Furthermore, phagocytosis and killing of C.
albicans by neutrophils was investigated. In an attempt to gain insight into
how ES affects neutrophils, we measured the changes in intracellular
concentrations of Ca2+ and cAMP.
Results: ES inhibited elastase release and H2O2 production by fMLPactivated neutrophils in a dose-dependent fashion with maximal inhibition
g of ES/ml inhibited neutrophilg of ES/ml. Already 0.5 seen with 5-50
g of ES/ml almost completely blocked it.chemotaxis towards fMLP and
100 Importantly, ES did not affect the ability of neutrophils to
phagocytose and intracellularly kill C. albicans. Interestingly, ES did not
affect the fMLP-induced rise in the [Ca++]i in neutrophils indicating that
ES acts down-stream of the phospholipase C-mediated activation of protein
kinase C. In agreement, ES inhibited the PMA-activated neutrophil
responses. Furthermore, ES induced a rise in the intracellular cAMP
concentration and pharmacological activators of cAMP-dependent
98
mechanisms mimicked its inhibitory effects.
Conclusions: The beneficial effects of maggots on chronic wounds may be
explained in part by inhibition of multiple pro-inflammatory responses of
activated neutrophils by ES without affecting their antimicrobial activities.
Although the mechanisms underlying these effects on neutrophils are not
fully elucidated, activation of cAMP-dependent mechanisms may be
involved.
Maggot Excretions/Secretions are effective against biofilms of
Staphylococcus aureus and Pseudomonas aeruginosa
M.J.A. van der Plas1,2, A.M. van der Does1, M. Baldry1, H.C.M. DogteromBallering1, C van Gulpen1, J.T. van Dissel1, G.N. Jukema2 and P.H.
Nibbering1
1
Dept of Infectious Diseases, 2Dept of Surgery, LUMC, Leiden, The
Netherlands
Introduction: Chronic wounds that lack a tendency of healing are common
in patients with chronic conditions like diabetes mellitus. In many of such
patients, the wound healing process is hampered by bacterial infection
especially when the bacteria form biofilms protecting them from the
actions of both antibiotics and the immune system. In this study we
assessed the effects of maggot excretions/secretions (ES) on biofilms of
Staphylococcus aureus and Pseudomonas aeruginosa, microorganisms that
are clinically most relevant in this respect.
Methods: g/ml ES (one maggotWe assessed the effect of 0-20 g of
ES/h) on modulation of biofilms using microtiterproduces approximately
2
biofilm assays. Furthermore, antibacterial activity of ES was
investigated using in vitro killing assays and radial diffusion assays. We
also used specific reporter bacteria to investigate whether ES affects
quorum sensing systems.
Results: g of ES prevented S. aureus biofilmAlready 0.2 g of ES
rapidly broke down established biofilms. In contrast,formation and 2 ES
initially promoted P. aeruginosa biofilm formation, but after 10 h the
99
biofilms collapsed. Break down of established P. aeruginosa biofilms
occurred after 10 h and required 10-fold higher amounts of ES than for S.
aureus biofilms. Interestingly, boiling of ES abrogated their effect on
biofilms of S. aureus, but not of P. aeruginosa, indicating that different
molecules within ES are responsible for the observed effects. Furthermore,
modulation of bacterial biofilms by ES did not involve bacterial killing or
effects on bacterial quorum sensing systems.
Conclusion: ES prevent S. aureus biofilm formation and degrade
established S. aureus biofilms, but appear less effective against P.
aeruginosa biofilms. Our results are consistent with clinical observations
that maggots are effective against chronic, infected wounds although
wounds infected with P. aeruginosa are more difficult to cure than wounds
infected with S. aureus.
Parasite role reversal
Worms on trial in rhinitis and asthma
University of Nottingham, NG7 2RD
FINDINGS
METHOD
INTRODUCTION
 The allergic phenotype has a controlling influence
over hookworm infection (IL5/eosinophilia/IgE)1-4
Group 1
ConA stimulation
 Therefore, hookworms may have a bystander
effect on the development of allergic disease6
through the combined production of antiantiinflammatory molecules, and (possibly) the
induction of regulatory T cell propulation5.
0
3
45000
40000
35000
30000
25000
20000
15000
10000
5000
0
IFNIFN-
6
0
Weeks
ILIL-5
pg/ml
400
300
300
200
200
100
100
0
0
0
3
0
3
6
Weeks
CONCLUSION
STUDY DESIGN
GROUP 1
CD4/CD25
0.5
0.4
0.4
X 109 per litre
X 109 per litre
To determine, in normal volunteers, an infective
dose of N. americanus which:
0.3
0.2
0.1
 gave the desired infection intensity, equating to
protection
against
respiratory
wheeze,
encountered in the study of Scrivener et al.
(2001)6.
 We can administer a dose of N. americanus to patients exhibiting
bronchial hyperhyper-responsiveness without adverse effects.
GROUP 2
CD4/CD25
0.5
CD4/CD25
0.3
0.2
0.1
0.0
0.0
0 1 2 3 4 5 6 7 8 9 10 11 12 13 14 15
0 1 2 3 4 5 6 7 8 9 10 11 12 13 14 15
Weeks
Weeks
GROUP 1
GROUP 2
17.5
WBC
Eosinophils
15.0
X 109 per litre
X 109 per litre
 was asymptomatic, yet induced a measurable
immunological response (as we will be looking in
the longer term to induce protective levels of
regulatory T cells).
14
13
12
11
10
9
8
7
6
5
4
3
2
1
0
12.5
10.0
7.5
WBC vs
Eosinophils
5.0
 did not cause adverse effects in the airways of
patients
demonstrating
bronchial
hyperhyperresponsiveness.
Weeks
 We have now embarked on Phase 3 of the trial in asthmatics.
0 1 2 3 4 5 6 7 8 9 10 11 12 13 14 15
REFERENCES
Weeks
1.5
1.0
0.5
Airway
responsiveness
0.0
-0.5
-1.0
-1.5
-2.0
 These patients responded with a measurable immune response to
the dose with data encouraging the belief that the immune system
is sufficiently engaged to follow up with an investigation into
regulatory T cell expansion.
2.5
0 1 2 3 4 5 6 7 8 9 10 11 12 13 14 15
Change in pd10AMP
(doubling dose)
To ensure that the safe dose selected in normal
volunteers:
 During the first four weeks bronchial reactivity was greater (not
significant) in the hookworm group compared with placebo.
Therefore, there appears to be no deleterious effect on bronchial
bronchial
reactivity during pulmonary larval migration.
0.0
 These data are published7.
PHASE 2 – SAFETY 2
 Whole blood was analysed for differential leucocyte counts.
 Changes in bronchial reactivity to AMP were measured.
6
Weeks
PHASE I - SAFETY I
 Human peripheral blood lymphocytes (PBL) were isolated and
stimulated with lectin (Concanavalin A) and compared with a
medium control. After 4 days in culture, supernatants were
assayed for cytokines (IFN
(IFN and ILIL-5) using a multiplex assay
system.
 PBL were labelled for CD4 and CD25 expression and analysed by
flow cytometry.
cytometry.
Group 2
ConA stimulation
500
*
400
pg/ml
6
600
*
500
3
Weeks
Group 1
ConA stimulation
600
 A double blind, controlled clinical trial, will allow
firm conclusions to be made.
Whole blood was taken over a period of 12 weeks post treatment,
treatment,
processed as follows.
Group 2
ConA stimulation
45000
40000
35000
30000
25000
20000
15000
10000
5000
0
pg/ml
pg/ml
 Hookworms modulate the allergic response to
survive.
Group 1
Group 2
1.
2.
3.
4.
5.
6.
7.
Pritchard and Walsh (1995) Parasite Immunol.
Immunol. 17, 605605-607.
Pritchard and Brown (2001) Trends Parasitol.
Parasitol. 17, 169169-172.
Culley et al.
al. (2002) Eur.
Eur. J. Immunol.
Immunol. 32, 13761376-1385.
Quinnell et al. (2004) J. Infect. Dis. 190, 430430-438.
Carvalho et al.
al. (2006) Parasite Immunol.
Immunol. 28, 525525-534.
Scrivener et al. (2001) Lancet 358, 14931493-1499
Mortimer et al. (2006) Amer. J. Trop. Med. Hyg.
Hyg. 75, 914914-920.
ACKNOWLEDGEMENTS
 induced a measurable immune response.
Clinical
Immunology
100
J. Britton, J. Feary,
Feary, A. Venn
D. Pritchard, D. Hooi,
Hooi, A. Brown
A New in-vivo model for testing effects of topically biotherapy wound
healing.
Nasser M. Shahri
Department of Biology, Azad University, Iran
Studying in texts, documents and sources of Islamic and Iranian traditional
Medicine suggests the point that taking care of wounds and injuries, is one
of the special clinical subjects in their rapid healing.
Most of the well-known ancient physicians of Iran have been aware of this
matter by experience. They knew that by using medicines the rapid healing
and recovery process can be made.
In doing this research it has been tried to study, from a scientific viewpoint
the efficacy of some of the traditional healing agents by conducting some
experiments so that views can be made concerning the efficacy of these
stuffs.
Therefore, after making 200 regular holes (circular incisions with a
diameter 8mm) on the ears of 30 rabbits, some of these holes were chosen
as the test wounds and some of them, with the same anatomical position,
were selected as evidence control wounds.
The evaluation standards for quick diagnosis of the healing and recovery
(reconstruction) procedures have been the measurement of the diameter of
the test and control wounds, comparing them and also comparing the
percentage of the recovered (reconstructed) levels in the test and evidence
samples, during 7th-13th days after formation of the scar thus, based on the
above-mentioned standards, the prepared charts and considering the
evaluation standards of the rapid diagnosis of the healing process in
following this research, in general it can be stated, with a scientific
standpoint, that some of the traditional healing agents of animal origin
(some of animals fats, in particular, cow butter oil and duck fat in some
cases mixed with the ash of human hair); provided that they are noninfectious can interfere effectively in rapid healing and
recovery/reconstruction of the damaged tissues of laboratory animals, in
other words they can shorten the time necessary for healing and recovering.
101
Macroscopic and microscopic survey of the effectiveness degree of
some of the biostimulators used in traditional medicine in the process
of healing of sheep skin wounds.
Dr. Nasser Mahdavi Shahri,
Colleges of science, Ferdowsi University, Iran
In view of the importance of the rapid healing of dermal (skin)
wounds in veterinary medicine and taking into consideration that some
of the stimulants of healing process, with animal origin in Iranian
traditional medicine have been introduced as healing medicines, in the
execution of this research, it has been tried to make studies on the
degree of effectiveness of these materials in the healing process and
recovery of sheep dermal (skin) wounds. For this purpose some
wounds were made in forms of splits or by taking epidermis, dermis
and hypodermis parts. Some wounds were selected as control and the
other group as test, and both groups were almost at the same
anatomical position. The wound of test group were locally treated
once a day by traditional healing agents. Those used in this test were
cow butter oil and duck fat. The principle of this research was to
compare and evaluate control and test wounds both macroscopically
and microscopically during 1-14 days. For this purpose , in
macroscopic evaluations of the wounds , rapid formation of the wound
crust , the wound extent (the distance between the wound edges) and
the clinical appearance of the wounds, were the diagnosis basis, and in
microscopic level has been possible. Cases: Rapid formation of
epithelium, quicker omission of blood clots and damaged tissues,
regarding the amount of the produced scar and the amount of the
inflammatory cells, were the comparison basis. The result of the
evaluations between the test and control samples showed that in all of
the mentioned cases, the test wounds heal more rapidly than the
control wounds. In making a general result, these cases in the test
samples can be related to using the traditional healing agents. Using
some of the synthetic healing ointment locally and by the comparison
of their functions with the afore-mentioned traditional healing agents,
the effectiveness of the traditional ones is proved in this research.
102
Evaluation of the efficacy of curcuma longa rizom ointment in healing
process of burning wounds
Nasser Mahdavi Shahri, H.Rakhshandeh, E.Nasrabadi
Department of Biology, Ferdowsi University, Iran
The application of curcuma longa has significant important in traditional
medicine of eastearn asia and IRAN.in this study we used ointment of
powder rizomCurcuma longa 0/010 on burned wound in pabbits skin. We
collected 14 male rabbits and shaved dorsal part of them and then we
burned wound part of them by chloidric for 50 seconds. We produced 6
burned wound on dorsal part of each rabbit and so we had 90 burned area
in adition. We collected 4 wounds as test and i wound as control asd i
wound as negative control. Every day and after bunting we used ointment
of powder of rizom on test wound vazeline on control wounds. This was
continued for 24 days. For historical studies we sampled from wounds in
days 3, 6, 8, 16 after burning. The specimens were examined for the
following elements of wound healing to determine a potential treatment
response. Percent of wound epithelized epithelizal thickness (cell layer
mm) and amound of scab formation every day, we measured tt of wound
recovery. Result of clinical studies show that healing process in wounds
that received ointment of powder rizom is faster than control wounds that
received vazeline.
Effect of a natural polymer "rabbit vitreous" to wounds therapy on an
experimental model
Nasser Mahdavi Shahri, S.samareh Mosavi
Department of Biology, Ferdowsi University, and
Department of Biology, Azad University, Iran
Today for production of natural polymer, used different cells culture or
isolation material from bioorganisms. There are much hyaluronic acid
as natural polymer in ultrastructure of rabbit vitreous. It can adhesion
103
to surface of cells and led to cell immigration, regeneration of rabbit
pinna punch hole.
The rabbits that we used in this study were the Newzealand white
rabbits. We used standard metal earpunched that marked for this
purpose. We created well-circum scribed circuler surgical wound of
8mm in diameter. One of the rabbit pinna day-to-day treated with
rabbit vitreaos and the other one was as control statistical analysis
showed that deviation pinna nonregeneration tissue between vitreous
treat and control was significant. However this deviation on day 15th
after ear punching was maximum level of significance (p<0.05)
vitreous treated wounds had an epithelial percentage of 76.16 and
control had an epithelial percentage of 63.9 (day 14). Vitreous treated
wounds and control had an epithelial percentage of 90.34, 79.1
respectively (day 30).
Vitreous treated wound had an epithelial thickness of 59.16 pm and control
had an epithelial thickness of 45 pm (day 14). Vitreous treated wounds and
control had an epithelial thickness of 107.66, 94.54 pm respectively (day
30).
MAGGOTS THERAPY IN SPECIAL WOUND TREATMENT
Zohrabyan A. Sureni
Surgical Department, Central Hospital, MOD,
Yerevan, Armenia
Purpose: To avoid from expanded surgical intervention on total removal
of prolene mesh implant after plastics of a ventral hernial gate and shor
tening of the treatment terms.
Methods: Using medical maggots of Lucilia Sericata for wound debrim
ent from necrotic tissue and infection.
Results: The patient (43 year old gentlemen) was presented to the surgical
department of Central Clinical Military Hospital with long-time
functioning purulent fistula on a place of postoperative scar .
In fight he was wounded in abdomen. After that case he was exposed to
104
numerous operations. Because of these numerous operations there was
formed a huge ventral hernia. For closing this ventral hernia was done
hernioplasty and the hernial gate was closed with mesh prolene implant of
30х30 cm size. Four years later purulent fistula was formed on the place
of postoperative scar. Conservative treatment and modern medicines had
not provided his recovery. Also he refused from total surgical removal
of the implant. For this reason was offered maggot therapy. Narrow
pass of a fistula was insufficient for the penetration of maggots, therefore
the part of skin with subcutaneous and fibrous tissue and part of mesh were
excised. Results from intraoperative cultures showed Pseudomonas
aeruginosa without sensitivities to antibiotics. When the wound formation
was completed two pots of maggots were applied. Three days later there
was noticed improvement in the wound condition. The maggots were appl
ied only two times. As a result the wound was cleared and implant was c
overed with granulations. A month later on the place of the wound was
generated scar.
Conclusions: This case shows new and unexpected opportunities of th
e maggot therapy. It was abundantly clear, that the maggots suppress
ed wound infection so, that it had epithelized without removal of a fore
ign body.
MAGGOT DEBRIDEMENT THERAPY IN SLOVAKIA
Takáč P, PhD.1 ;Čambal M, M.D.2; Krumpálová Z, PhD.1 Kozánek M,
PhD.1.
1
Institute of Zoology,Slovak Academy of Sciences, Bratislava, Slovakia
2
1st Depatment of Surgery, University Hospital and Faculty of Medicine,
Comenius University, Bratislava, Slovakia
The maggot debridement therapy is used in Slovakia since
August 2003. First sterile blowfly Lucillia sericata colony was established
in the Institute of Zoology, Slovak Academy of Sciences (SAS) in
Bratislava. First patients were treated on the 1st Department of Surgery,
105
University Hospital and Faculty of Medicine, Comenius University in
Bratislava.
The successful cooperation of scientific workers from SAS and
physicians resulted in establishing of non-profit organization MEDALT
(Medical alternative) settled in the building of the Technology Incubator
built up with the European Union support. In 2005, MEDALT won the
project supported by European Social Foundation. The main idea of the
project is to develop research and development centre, which will be
focused on the development and dissemination of the biomedical
therapeutic methods in Slovakia. On the example of maggot debridement
therapy, we would like to create some model applicable also for
development of further biotherapeutic methods (i.e. hirudotherapy,
apitherapy, helmintotherapy...) and introduce these methods to clinical
practice. Up to now MEDALT has successfuly spread maggot debridement
therapy in hospitals all around Slovakia. By straight application of Lucillia
sericata larvae in wounds two layered dressing is used to prevent larvae
escape.
The bottom layer (cage layer) is fixed to the intact skin surrounding
the wound. Its purpose is to keep maggots in the wound and to protect
surrounding skin from aggressive maggots juice side effects. We use
colostomy pads Coloplast. The second layer is chiffon, which is attached
by a special disperse glue (developed in the Institute of Polymers, SAS,
Bratislava) to the bottom layer. This layer allows maggots to breath oxygen
and to drain out liquefied necrotic tissue and wound secretion. One day old
sterile larvae are applied under plastered chiffon.
Top layer is a dry gauze which is usually placed over the cage layer to
absorb the liquefied necrotic tissue. Only this layer is regularly (every 4-6
hours) changed by the nursing staff.
“Biobag” is our challenge. Simuntaneously we are designing a new
“biobag” for maggot debridement therapy. By using biobags we didn´t
notice any larvae escape. The application, removal and disposal is simple
and practical. We can recommend this way of maggot’s application direct
into the wound.
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The optimal conditions for medicinal maggots before and during
shipment
Hitoshi Takase M.D.
Biotherapy Medical Co., Ltd, Japan
Purpose: To investigate the optimal conditions for medicinal maggots
before and during shipments, such as period, feed, temperature, and
ventilation.
Method: Newborn sterilized maggots are put on the various conditions for
certain periods, and then their medicinal effectiveness on wounds is
assessed by their size, survival rate, and amount of food ingested.
Result, conclusion: The experiment is now being conducted and the results
are reported at the congress.
Maggots to the Rescue: Using maggot therapy for wounds in hospice
patients
Aletha W. Tippett, M.D.
Wound Consultant; Hospice Medical Director, Ohio, USA
Purpose: Wounds are a problem that affect over 1/3 of hospice patients (1),
and often these wounds are severe, with various causes: ischemic, pressure,
diabetes, trauma and surgical. Many times debridement of the wounds is
indicated, due to infection, sepsis, pain, necrosis, or gangrene, but patients
usually are not candidates for surgical debridement, including amputation
of limb. Often other debridement forms such as wet to dry, or enzymatic,
are not appropriate due to issues of pain, cost, or time constraints (e.g.
infection with sepsis needs urgent debridement). Sometimes debridement
is not seen as consistent with palliative care. This paper presents a
selection of case studies of the use of Phaenicia sericata larval therapy
(maggots) for wound debridement in hospice patients.
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Methods: A selection of case studies from wound treatment of hospice
patients over a three year period to illustrate wound types, problems, and
outcomes are presented. The cases were selected to be representative of the
types of wounds encountered that could benefit from maggot therapy.
Results: In all cases goals of therapy were achieved. Results included:
resolution of sepsis, prevention of amputation, elimination of infection,
reduced pain and odor, and in some cases, wound healing. Therapy was
well tolerated in all cases.
Conclusions: Maggot therapy is a very viable option for treating complex
wounds in hospice patients when debridement is indicated. Maggot
therapy is rapid, inexpensive, nearly painless, simple, quite gentle, and
consistent with goals of palliative care. Standard protocols are published
and easy to follow (2). Families and patients are very involved, and
grateful for such innovative therapy, with newfound respect for one of
nature’s often maligned creatures.
Application of maggot therapeutics for repairing serious infective
wound
Jiangning Wang
Dalian University Medical Acad, Dalian, China
Objective: To research the application of maggot therapeutics for repairing
serious infective wound.
Methods: Two methods were used for getting clean maggot, one is flyblow
asepsis, the other is larva antiasepsis. No bacterium or virus exist in the
body of the maggots and the maggots are vivid. From 2005to 2007, we
took use of maggot therapeutics to repair 4 cases serious infective surfaces.
Fifty to one hundred aseptic maggots were put to the infective surfaces
every time, aseptic dressing overlied the surface, nylon net covered outside
the dressing. The maggots and dressings were changed every two days.
Results: Average cure period was 7 days, all necrotic tissues of the wound
were cleanup, fresh tissue developed. When bacterium culture of the wound
surface shows no bacterium, the wound was covered with skin graft, all
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results were satisfied.
Conclusion: Maggot therapeutics is an effective biological therapy for
repairing serious infective wound.
The postoperative wound infection is still one of major complications of
replantation. Failure to control infection can lead directly to vascular
thrombosis and in turn loss of replanted extremity. The use of maggots for
wound debridement has a long history and has been lately re-introduced for
treatment of intractable wounds. In this report, we present the experience of
successful debridement of a severe infected wound after forearm
replantation with the maggot therapy. The results and mechanism of the
maggot therapy are discussed. Today replantation has become clinical
routine for salvage of amputated extremities with great success. However,
since many cases of amputation are caused by agriculture and manufacture
injuries, gunshot, and traffic accident with relative wound contamination,
the postoperative wound infection is still one of major complications of
replantation. The presence of infection can lead directly to vascular
thrombosis, sepsis, and loss of replanted extremity. Treatment of infected
wound needs good debridement and coverage with vascularized tissue, so
we begin to choose maggots therapy for infected wound.
The healing and antibacterial function research of the maggot
secretion on the ulcer area
Shouyu Wang
Dalian Medical University No.1, Dalian, China
Objective: Investigate influence of treatment with maggot secretion on
wound healing of diabetes ulcer and to study the antibacterial function of
maggot therapeutics for decubitus ulcer after spinal cord injury.
Methods: (1) Some circle wounds were made in back of diabetes rats,
The injured area spreads the maggot secretion. The area size were detected
after injury with the transparent membrane method and carried on the
pathology examination. (2) Using Staphylococcus aureus and Pseudemonas
aeruginosa as the indicator organisms which from the infective wound of
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decubitus ulcer after spinal cord injury, the hemolymph secretion of the
larvae which was induced by the bacterium with pricking was collected, the
antibacterial activity was tested with disc method, recorded the
antibacterial effect, the hemolymph secretion of the larvae which was not
induced by the bacterium and Cefoperazone Sodium as the contrast groups.
5 cases suffered from decubitus ulcer after spinal cord injury was treated
with maggot therapeutics, observe the exchange of wound surface and
checked the bacterium.
Results: (1) The injured area achieved 100% recovery in latter 4 weeks,
at the same time, wounds of diabetes big rats did not recover, and expanded
unceasingly, in the pathology examination maggot secretion group
regenerated epidermis , the small blood vessels and the fibrinogen cells. (2)
The secretion of the larvae had antibacterial activity on Staphylococcus
aureus and Pseudemonas aeruginosa. The secretion of the larvae which was
induced by the bacterium had stronger antibacterial activity. Average cure
period of 5 cases was 9 days, fresh tissue developed, necrosis tissue
cleaned, no bacterium existed on the ulcer surface after bacterium culture.
Conclusions: (1) The maggot secretion has the obvious promotion
function to the ulcers. (2) Maggot therapeutics has effective antibacterial
function on decubitus ulcer after spinal cord injury.
Effectiveness of Maggot Therapy in Chronic Infected Wound with
MRSA
K.J. Yoon1, Y.B. Shim1, S.S. Ha1, H.T. Kim2, K.S. Kim, PhD2, N.H. LEE2
1
Department of neurosurgery, St. Peter’s Hospital;
2
Medilarvatech
Purpose: There have been reports of using maggots to treat the bacterial
infection that are resistant to antibiotics, especially, methicillin resistant
staphylococcus aureus(MRSA) or to clean and diminish the size of the
wounds for skin graft bed in burn or pressure sore patients. We produce the
sterilized maggots and report the result of clinical applications of maggot
therapy in the treatment of MRSA infection, pressure sore and tissue
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defects.
Material and Methods: From Aug 2005 to May 2007, 33 patients (7- 65
years old, mean 44.5, male 17, female 16) were treated: 7 MRSA infection
cases( 3 pressure sore; 4 postoperative wound), 15 Diabetic foot, 6 crushing
injuries, 5 acute burn wounds.
We used sterilized, 1 day old larvae (1-2mm in length) of the flies,
“Phaenicia sericata” (product by Medilarvatech). Maggot application was
confined to the wounded area with a specially designed bandage and the
larvae eat all the infected tissues. The bandage was removed after 3-4 days,
when the larvae are mature enough and cannot consume the infected tissues
anymore. The treatment was repeated 3 to 20 times (mean 8.3 times)
depending on the severity of each wounds and follow-up period was 3 to 24
months (mean 12.5 months). Each treatment used 100-800 maggots. No
antibiotics were used during the treatment.
Results: A patient with crushing injury in the ankle gave up the treatment
due to severe pain. In all the 7 MRSA cases, the discharge of the pus was
stopped and wound was spontaneously healed at the end of the treatment.
In all other patients with tissue defect, wound became clean and diminished
in size, and secondary skin graft was performed successfully. There were
no side effects.
Conclusion: Maggot therapy is very effective and could be the new
strategy in the treatment of MRSA infection. And also in cleansing and
diminishing tissue defect in burn, pressure sore and crushing injuries to
promote wound healing and get better outcome in skin graft
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New development about medical leech therapy
Yavuz Dinler, Ph.D. (Turkey)
Leech therapy has been used in Turkey for at least 400 years. Today leeches
are collected in large quantities from at least 600 lakes in Turkey. . More
than 20 hospitals and clinics in this country practice hirudotherapy. Since
1994 our group has been using medical leech therapy and we have
applied over 10.000 individual treatments. Dukin's method is used for the
treatments and we have achieved with very good results for more than 10
different clinical conditions. Since 2006, we have also treated patients with
glaucoma and other eye ailments. We intend to use this treatment modality
in the future for skin cancer and leukemia.
Effects of bee venom on
melanogenesis and dendricity
human
melanocyte
proliferation,
Ai-Young Lee, M.D., Professor, Department of Dermatology, Dongguk
University Hospital, Gyenggi-do, Korea
Bee venom (BV) has been clinically used to control rheumatoid arthritis. In
the present study, we examined the effect of BV on the human melanocyte
proliferation, melanogenesis and dendricity, and its signal transduction in
vitro. BV increased the number of melanocytes dose dependently through
PKA, Erk, and PI3K/Akt activation. Melanocyte dendricity and the level of
cAMP were also increased by BV treatment. Moreover, BV also induced
MITF and tyrosinase expression by phosphorylation of CREB through
activation of PKA and Erk. Overall, in this study, we demonstrate the effect
of BV on melanocyte proliferation, melanogenesis and dendricity through
complex signaling pathways in vitro, which suggest a possibility that BV
may be a useful substance in the treatment of re-pigmentation.
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