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(2006) The Indian Journal of Nutrition and Dietetics, Vol.43, No.6, Pp. 229-237. http://www.banglapedia.org/httpdocs/HT/F0142.htm http://111.parentree.in/parentree-editors/journal-161/pregnant-indianwomen-heatlhy. http://cbhi-hsprod.nic.in/listdetails.asp?roid = 213. http://en.wikipedia.org/wiki/adolescentpregnancy http://india-reports.in http://www.groundreport.com http://www.indiadiets.com/diets/normal_diets/diets_in_pregnancy.htm. http://www.whoindia.org/LinkFiles/Adolescent-Heath-and-Development(AHD)-Mapping Adolescent-program-and measurement-overview: Pdf. www.betterhealth.vic.gov.au www.cpcip.fsu.edu, 2005. www.whoindia.org ANNEXURE I INTERVIEW SCHEDULE TO ELICIT INFORMATION REGARDING THE SOCIO-ECONOMIC CONDITIONS OF THE FAMILIES OF THE PREGNANT ADOLESCENTS Block : Village : I. II. S. No. III. Identification particulars 1. Name of the Investigator : 2. House No. / Address : 3. Type of Family : 4. Name of the Interviewee : 5. Size of the Family : 6. Religion : Particulars of household members Name of the family members Relation to Sex Age the head of the family Educational level Other sources of income, if any (specify) Occupation Income per month IV. Monthly expenditure pattern S.No. 1 Food 2 Clothing 3 Shelter-Rent / Repaying house loan 4 Education 5 Medicine / Hospital 6 7 Cooking fuel and electricity Utensils and furniture 8 9 10 Transportation Savings Recreation (Movies, Park, TV) 11 Others (Social events, donations, miscellaneous) V. Rs. Spent / Month Items % of total Income Monthly food expenditure pattern S.No. Items 1 2 Cereals Pulses 3 4 5 Green leafy vegetables Roots and tubers Other vegetables 6 7 8 9 10 11 12 Fruits Milk and milk products Meat, fish and poultry Fats and oils Sugar and jaggery Beverages Processed / readymade foods Rs. Spent/ % of Food Month Expenditure VI. Dietary pattern of the expectant woman 1. Meal pattern of pregnant adolescents a) b) Number of meals / day Number of snacks / day 2. Frequency of intake of various food by the pregnant adolescence S.No. 1 Cereals Rice Rice Products Ragi Wheat / Wheat products Others 2 Pulses Horse gram Green gram Red gram Bengal gram Red gram Bengal gram dhal Black gram Cowpea Peas Soyabean Others 3 Green leafy vegetables Chekkurmanis Amaranth Drumstick leaves Pumpkin leaves Cabbage Colacasia leaves Coriander leaves Others 4 Roots and tubers Tapioca Yam Colacasia Carrot Beetroot Potato Onion Others 5 Other vegetables Papaya green Brinjal Beans Pumpkin Bottle gourd Ash gourd Bitter gourd Snake gourd Banana Tender jackfruit Cauliflower Plantain green Ladies finger Drumstick D TRW TW OW OFN OM R 6 7 8 9 10 11 12 13 Kovaikkai Cucumber Others Fruits Banana Guava Papaya Amla Jackfruit Mango Dates Grapes Watemelon Orange Pineapple Raisins Tomato ripe Apple Others Milk and milk products Milk Curd Buttermilk Fleshy foods Fish Mutton Chicken Beef Egg Others Nut and oil seeds Groundnuts Cashewnut Coconut Others Fats and oil Coconut oil Palm oil Sunflower oil Gingelly oil Dalda Ghee Others Sugar and Jaggery Sugar Jaggery Beverages Coffee Tea Others Prepared / processed food D-Daily, TRW – Thrice in a Week, TW – Twice in a week, OW – Once in a week, OFN – Once in fortnight, OM – Once in a month, R - Rarely VII. Risk scoring schedule (Paul and Vijayalakshmi, 1994) Risk factors Scores Age less than 18 1 Maternal height less than 145cm 1 Weight less than 45 kg more than 90 kg 1 Primi / multi 5+ 1 Previous obstetric losses 2 Rh –ve 2 Previous low birth weight infant 2 Antepartum haemorrhage 2 Anaemia (Hb less than 8g/dl) 1 Pregnancy induced hypertension 2 Febrile ailment during pregnancy 1 Previous premature rupture of membranes 1 Foetal distress 2 Previous prolonged labour > 20 2 ANNEXURE II HEALTH ASSESSMENT CARD Block : Height : Name : Pre-pregnant weight : Age : Date of last menstrual cycle : Address : Para of pregnancy Religion : Income of family 1. : Anthropometric measurement of the expectant women Month of Weight Mid arm Complications, pregnancy in kg circumference if any 4 5 6 7 8 9 2. Foods included during pregnancy S.No. Special foods included Reasons : 3. Foods avoided during pregnancy S.No. 4. Foods avoided Reasons Are you taking any nutrient supplementation as prescribed by the physician ? Yes No If yes, details S.No. Nutrient composition 1 Iron and folate 2 Iron and calcium 3 Iron, folate and calcium 4 Iron, folate and zinc 5 Iron, folate, calcium with vitamin B3 6 Iron, folate and B Complex 7 Iron, vitamins with amino acids and minerals 5. Interval between previous and present pregnancy 6. Details regarding present delivery a. Complications, if any Frequency/day i) Obstetric loss ii) Foetal distress iii) Rupture of amniotic membrane before delivery iv) Antepartum haemorrhage v) Change in position of the foetus b. 7. 8. Labour pain i) 1st delivery ii) 2nd delivery iii) 3rd delivery Type of delivery a. Normal b. Caesarean c. Vacuum d. Forceps Anthropometric measurements of the new born of the present pregnancy a. Weight b. Crown heel length c. Head circumference d. Chest circumference e. Mid upper arm circumference ANNEXURE III PROFORMA TO ASSESS THE NUTRITIONAL KNOWLEDGE Block Name : Village : Age : Name of the interviewee : House No. / Address : Religion : : 1. What is the desirable age of pregnancy ? 2. What is the desirable weight gain during pregnancy ? 3. Mention the importance of nutrient supplementation 4. What is the importance of health checkups in pregnancy ? 5. What are the foods to be avoided during nausea ? 6. List out the foods to be included in liberal amounts during pregnancy. 7. Should the requirements of iron and calcium be increased during pregnancy? 8. Mention the foods to be included to overcome anaemia 9. Should you increase the intake of green leafy vegetables during pregnancy ? 10. What is the effect of malnutrition during pregnancy ? 11. What is the best exercise during pregnancy ? 12. What is the average birth weight of the new born baby ? ANNEXURE V PROCEDURE FOR BIOCHEMICAL ANALYSIS DETERMINATION OF BLOOD HAEMOGLOBIN (Cyanmeth haemoglobin method) (Raghuramulu et al., 2003) PRINCIPLE In solution the ferrous ions (Fe2+) of the hemoglobins (Hb) are oxidized to the ferric state (Fe3+) by potassium ferric cyanide to form methemoglobin. In turn, methemoglobin reacts with the cyanide ions (CN-) provided by potassium cyanide to form cyanmethemoglobin, which has the absorbance at 540nm. REAGENTS a. Cyanmethemoglobin solution (Drabkin’s solution) : Dissolve 0.05g potassium cyanide, 0.200g potassium ferric cyanide and 0,140g dihydrogen potassium phosphate in 1L of distilled water . Add 1ml of Triton X-100 and mix. Stable for atleat six months. b. Haemoglobin standard : Lyophilized human methemoglobin (supplied by Sigma USA). Each vial is equivalent to haemoglobin concentration of 18g/dl whole blood when reconstituted in 50ml of Drabkin’s solution. Stable for six months when refrigerated at 2-6 C. PROCEDURE Transfer 0.02ml of blood using a calibrated haemoglobin pipette, into a tube containing 0.5ml of Drabkin’s reagent. Rinse the pipette several times with the reagent. Allow diluted heamoglobin solution to stand for atleast five minutes to achieve full colour development. Measure the absorbance at 530550nm of the unknown sample (Aunk) and that of a standard of known heamoglobin content (Astd) against a reagent blank. CALCULATION Haemoglobin unknown (g/dl)= Aunk x Con. of Hb S tan dard (g / dl) Astd ANNEXURE VI DETERMINATION OF SERUM TOTAL PROTEIN (BIURET METHOD) (Raghuramulu et al., 2003) PRINCIPLE Protein forms a purple colour complex with copper ions in alkaline solution. The biuret reaction takes its name from compound, which react in the same way. The purple color developed is read colorimetrically at 500 m . REAGENTS 1. Sodium chloride diluent – Dissolved 9g of sodium chloride in one litre of water. 2. Stock biuret reagent – Dissolved 45g of potassium sodium titrate (Rochelle’s salt) in 400ml of 0.2N sodium hydroxide. Added 15g of cupric sulphate. Stirred until it is dissolves. Added five grams of potassium iodide and diluted it to one litre with 0.2N sodium hydroxide. 3. 0.2N sodium hydroxide : 50g of sodium hydroxide dissolved in 1000ml of water. 4. Dilute biuret reagent - .Diluted 250ml of stock biuret and reagent to one litre with 0.2N sodium hydroxide and added four grams of potassium iodide. 5. Standard protein solution – Weighed 400mg of albumin and dissolved in 0.9 per cent saline solution. Made up the volume to 100ml with saline. 6. 22.5 per cent sodium sulphate. PROCEDURE Into a series of test tubes 0.5, 1.0, 1.5, 2.0, 2.5ml of standard protein solution was pipetted out and then the volume was made upto 3ml with distilled water. Into another test tube, pipetted out 0.2ml of serum and diluted to 0.9 per cent saline upto five ml. From this, 2ml of the solution was taken, made upto 3ml with distilled water and treated as unknown. Now added 3ml dilute biuret reagent to all the test tubes. Along with this a blank was prepared. The colour developed was read colorimetrically at 500nm after 30 minutes. The amount of protein in the serum was calculated. This gives the value of total protein. PRECIPITATION OF GLOBULIN Mixing 0.21ml of serum with 4.8ml of 22.5 per cent sodium sulphate solution precipitates globulin. Stoppered the tube, inverted several times and kept in the incubator at 40 C overnight. Filtered the solution the next day using Whatman No. 42 filter paper. Took 2ml of the filtrate and carried out the experiment as for total protein. The concentration in gram per cent of albumin present in the globulin free filtrate is determined from the standard graph. Globulin value can be obtained by deducting albumin value from total protein. CALCULATION Total protein g/1000ml = Amount of globulin O.D of test x concentration of standard O.D of s tan dard = Total protein - Serum albumin ANNEXURE VII DETERMINATION OF VITAMIN A (RETINOL) (HPLC METHOD) (Raghuramulu et al., 2003) PRINIPLE The vitamin A is extracted with a suitable organic solvent and an aliquot of the organic phase is injected onto a normal or reversed phase HPLC column, followed by an eluting solvent of suitable polarity. Retinol, which is eluted as a sharp peak within 1-6 min is detected by a sensitive UV detector set at 325328nm. Retinol is quantitated by use of peak height rations or peak area ratios relative to an internal standard (retinyl acetate or other appropriate analogues). REAGENTS Solvents of HPLC graph must be degassed and be free of particles. Standards : A stock standard solution of retinyl acetate in ethanol (50 g retinol/ml) is prepared by dissolving about one mg of retinyl acetate in 10ml of ethanol, determining the concentration in a 1/30 dilution of an aliquot in % ethanol by use of the E11cm at 325nm as 1795 and then diluting the stock standard appropriately with ethanol. Then 10ml of this solution is diluted to 19.4ml to yield the stock standard with 50 g retinol/ml. PROCEDURE 1. Normal phase Transfer 100 l of serum (or plasma), 15 l of internal standard solution (4 g/ml of retinyl acetate or propionate) and 100 l of methanol to a conical centrifuge tube. Mix the contents of the tube with a vortex mixer. Add 200 l of extraction solution (petroleum ether 80, dichloromethane 19.3, isoporpanol 0.7 by volume) and cap the tube. Extract by interrupted mixing on the vertex mixer for 60s. After centrifugation (3000 rpm, 2 min) inject 100 l of the supernatant into the column by the use of a Hamilton syringe. Elute with the same solvent as used for extraction. 2. Chromatographic conditions Procedure 1 Column 15 x 0.2c i.. Micro Pak Si-10 Mobile phase Petroleum ether : Dichloromethane : isopropanol (80:19.3:0.7) Flow rate 0.5 ml/min Pressure 10 kg/cm2 Detector wavelength 328 nm Detection sensitivity 0.04 AUFS* on recorder Temperature Ambient Recorder Speed Not specified Retention time (min) Retinol 5.0 Retinyl acetate 6.2 CALCULATION By use of an internal standard, losses due to incomplete extraction, inaccurate aliquots, oxidation, etc. are automatically corrected. The internal standard should have physical and chemical properties sufficiently similar to retinol, is suitably separated from retinol on HPLC, should not coincide with other 325nm absorbing materials in serum and is not converted to retinol under the assay conditions. A precisely known amount of the internal standard is added to the aliquot of plasma to be analyzed. By determining the relative extraction efficiency and detector response of retinol and the internal standard, a standard curve is fashioned in which the ratio of peak heights (or areas) is plotted against the retinol concentration in plasma. In experimental samples, the peak height (or area) ratio is determined and the appropriate plasma retinol concentration determined from the standard curve. A standard curve is prepared by adding varying amounts of retinol (i.e., 10-120ng) to a fixed amount (i.e. 50ng) of internal standard in a final volume of 100 l of eluting solvent, injecting the solution of HPLC under assay conditions, measuring the peak heights and calculating the peak height ratio. The peak height ratio is then plotted as the abscissa with the standard retinol concentration (for a 100 l plasma aliquot) as the ordinate.