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Study of Serum Homocysteine and Vitamin B12 levels in Eclampsia, Pre-Eclampsia, and the effectiveness of treatment with Inj vitamin B12 on the outcome of these patients.
Study of Serum Homocysteine and Vitamin B12 levels in Eclampsia, Pre-Eclampsia, and the effectiveness of treatment with Inj vitamin B12 on the outcome of these patients.
Study of Serum Homocysteine and Vitamin B12 levels in Eclampsia, Pre-Eclampsia, and the effectiveness of treatment with Inj vitamin B12 on the outcome of these patients.
Study of Serum Homocysteine and Vitamin B12 levels in Eclampsia, Pre-Eclampsia, and the effectiveness of treatment with Inj vitamin B12 on the outcome of these patients.
Study of Serum Homocysteine and Vitamin B12 levels in Eclampsia, Pre-Eclampsia, and the effectiveness of treatment with Inj vitamin B12 on the outcome of these patients.
Study of Serum Homocysteine and Vitamin B12 levels in Eclampsia, Pre-Eclampsia, and the effectiveness of treatment with Inj vitamin B12 on the outcome of these patients.
Study of Serum Homocysteine and Vitamin B12 levels in Eclampsia, Pre-Eclampsia, and the effectiveness of treatment with Inj vitamin B12 on the outcome of these patients.
Study of Serum Homocysteine and Vitamin B12 levels in Eclampsia, Pre-Eclampsia, and the effectiveness of treatment with Inj vitamin B12 on the outcome of these patients.
Study of Serum Homocysteine and Vitamin B12 levels in Eclampsia, Pre-Eclampsia, and the effectiveness of treatment with Inj vitamin B12 on the outcome of these patients.
Study of Serum Homocysteine and Vitamin B12 levels in Eclampsia, Pre-Eclampsia, and the effectiveness of treatment with Inj vitamin B12 on the outcome of these patients.
Study of Serum Homocysteine and Vitamin B12 levels in Eclampsia, Pre-Eclampsia, and the effectiveness of treatment with Inj vitamin B12 on the outcome of these patients.
Study of Serum Homocysteine and Vitamin B12 levels in Eclampsia, Pre-Eclampsia, and the effectiveness of treatment with Inj vitamin B12 on the outcome of these patients.
Study of Serum Homocysteine and Vitamin B12 levels in Eclampsia, Pre-Eclampsia, and the effectiveness of treatment with Inj vitamin B12 on the outcome of these patients.
Study of Serum Homocysteine and Vitamin B12 levels in Eclampsia, Pre-Eclampsia, and the effectiveness of treatment with Inj vitamin B12 on the outcome of these patients.
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Study of Serum Homocysteine and Vitamin B12 levels in Eclampsia, Pre-Eclampsia, and the effectiveness of treatment with Inj vitamin B12 on the outcome of these patients.

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The present study was carried out to evaluate the occurrence of association between homocysteine, and vitamin B12 in patients with Preeclampsia , Eclampsia and those with history of previous PIH. 30 …

The present study was carried out to evaluate the occurrence of association between homocysteine, and vitamin B12 in patients with Preeclampsia , Eclampsia and those with history of previous PIH. 30 such patients from obstetric ward were studied for estimation of serum homocysteine, and vitamin B12 over a period of Jan10 to Jun 2011. Serum homocysteine and vitamin B12 were determined by means of Immulite 1000 analyzer. The statistical analysis of study group of preeclampsia compared with normotensive control group, showed significant alterations in serum homocysteine, and vitamin B12 concentrations in preeclampsia and eclampsia group. Inverse association between serum homocysteine and vitamin B12 levels were observed in preeclampsia and eclampsia. The present study found hyperhomocysteinemia and deficiency of vitamin B12 along with increased blood pressure as a risk factor in preeclampsia. Final outcome of these patients after Inj Vitamin B12 therapy has improved at par with control group without any neonatal or maternal mortality in all four groups

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  • 1. Title of the article: Study of Serum Homocysteine and Vitamin B12 levels inEclampsia, Pre-Eclampsia, and the effectiveness of treatment with Inj vitamin B12 onthe outcome of these patients.Type of article: Original articleName of the Author:Dr.Radha Yegnanarayan MD (Pharmacology)* (Corresponding & Principal Author)Dr G S Shekhawat MD (Obst & Gyn) +Dr Hemant S Damle MD(Obst & Gyn )≠Place of Research work: Dept of Pharmacology and Dept of Obstetrics &Gynecology, Smt Kashibai Navale Medical College, Narhe, Pune-411041,Maharashtra.Email of Principal & co worker: gsshekhawata@yahoo.co.in (M) 09372897090 anddrradha @sknmcgh.org, Tel :( O) (020-24106155)Address of the corresponding Author:Dr.Radha Yegnanarayan (Prof & HODPharmacology)Smt Kashibai Navale Medical College, Narhe, Pune-411041, Maharashtra.*Professor &HOD (Pharmacology), Associate Professor (Obstetrics & Gynecology), ≠ +Professor (Obstetrics & Gynecology), S mt Kashibai Navale Medical College, Narhe,Pune-411041, Maharashtra.Word Count: 2464
  • 2. AbstractThe present study was carried out to evaluate the occurrence of association betweenhomocysteine, and vitamin B12 in patients with Preeclampsia , Eclampsia and thosewith history of previous PIH. 30 such patients from obstetric ward were studied forestimation of serum homocysteine, and vitamin B12 over a period of Jan10 to Jun 2011.Serum homocysteine and vitamin B12 were determined by means of Immulite 1000analyzer. The statistical analysis of study group of preeclampsia compared withnormotensive control group, showed significant alterations in serum homocysteine, andvitamin B12 concentrations in preeclampsia and eclampsia group. Inverse associationbetween serum homocysteine and vitamin B12 levels were observed in preeclampsiaand eclampsia. The present study found hyperhomocysteinemia and deficiency ofvitamin B12 along with increased blood pressure as a risk factor in preeclampsia. Finaloutcome of these patients after Inj Vitamin B12 therapy has improved at par with controlgroup without any neonatal or maternal mortality in all four groupsKeywords : Homocysteine ,Vitamin B12 , Pre Eclampsia, EclampsiaIntroductionPregnancy induced hypertension may occur in about 3–10% of all pregnancies [1]. Itremains a major cause of perinatal and maternal morbidity and mortality world-wide,because of complications such as eclampsia, fetal growth retardation, premature birthor abruptio placentae[1][2]. An increased concentration of total circulating homocysteinein serum is recognized as an independent risk factor for Pre eclampsia[3][4]. Moreover,
  • 3. determinants of hyperhomocysteinemia, such as low concentrations of folic acid andvitamin B12 involved in homocysteine metabolism are also associated with increasedrisk of vascular damage & Pre eclampsia [5]. It is uncertain whetherhyperhomocysteinemia per se or low concentrations of vitamin B12 and folic acid areatherogenic factors that trigger Pre eclampsia[6]. The present study was undertaken todetermine the levels of serum homocysteine, and vitamin B12 and their correlation shipin patients with preeclampsia. We also studied the effectiveness of treatment withInjection B12 in patients who showed low levels of vitamin B12.Material & Methods This study was carried out at Department of Pharmacology and Department ofObstetrics & Gynecology, Smt Kashibai Navale Medical College and General HospitalPune after obtaining Institutional Ethics Committee approval. All participants completeda medical history form and provided informed consent. 40 patients in the age group of18–35 years were studied for estimation of serum total homocysteine, and vitamin B12over a period of 18 months. Detailed dietary history with reference to vegetarian or nonvegetarian status and consumption of folate rich foods were recorded in all cases.Peripheral blood smears were examined in all 40 cases for presence of megaloblasts.In all those cases where homocysteine concentrations were high and vitamin B12 levelswere low, we administered Injection B12, 1500 µgm I/M in three divided doses.Inclusion CriteriaThis prospective study was conducted among 40 patients, who were divided in 04groups. 10 patients with Eclampsia, 10 patients with pre-eclampsia , 10 patients withpast history of pre- eclampsia/eclampsia and another 10 normotensive patients ascontrol without any sign, symptoms, lab test suggestive of pre-eclampsia were included.
  • 4. Besides routine base line ANC investigations, all patients were subjected to specialinvestigation including renal, liver and coagulation function tests for pre- eclampsia /eclampsia patients.Exclusion CriteriaPatients having use of medications (therapy involving S-adenosyl-methionine,carbamazepine, phenytoin, 6-azauridine, xanthopterin, antifolic acids, anticonvulsantagents, tamoxifen, and theophylline), cancer, severe anemia, systemic illness and thosewith major illness were excluded from study [7].Blood Sample CollectionVenous blood samples were collected in test tube with aseptic precautions. After 2 h ofcollections sample was centrifuged at 3000 rpm for 5 min. Serum was separated andcollected in polythene tube with cork. The sera with no sign of haemolysis were codedand used for the analysis of total circulating homocysteine and vitamin B12. Theinvestigator carrying the estimation was unaware of the clinical history and treatmentstatus of the patients.Biochemical AnalysisSerum homocysteine concentration was measured by competitive chemiluminescentenzyme immunoassay method [6]. Serum vitamin B12 concentration was evaluated bysolid phase, competitive chemiluminescent assay method. We used fully automatedenzyme amplified chemiluminescent immuno assay based Immulite 1000 analyzer.Hyperhomocysteinemia was defined as a serum homocysteine concentration greaterthan 15 µmoles/l. Vitamin B12 deficiency was defined as Vitamin B12 level lowerthan223 pg/ml.Statistical Analysis
  • 5. Numerical variables were reported in terms of mean and standard deviation or standarderror of mean. Statistical analysis of results was done by Student t test with correction&Yates corrected chi square test wherever applicable. In this analysis, variablesshowing P-value less than 0.05 and 0.001 were considered to be statistically significantand highly significant, respectively. Pearson correlation test was used to test correlation.ResultsDemographic data of pre eclamptic patients such as mean age of patients showedsignificant fall (P<0.05). Systolic blood pressure (SBP) and diastolic blood pressure(DBP) were significantly increased (P<0.05) in pre eclamptic/ eclamptic group ascompared with control group (Table 1). Table 2 depicts changes in serum profile whencontrol group was compared with study group of preeclampsia. As can be seen,significant increase (P<0.05) were observed in serum homocysteine whereas, vitaminB12 levels showed significant decrease (P<0.05). A negative and significant correlationwas observed between serum homocysteine when compared with vitamin B12 (Table2and 3). All patients of Eclampsia and pre eclampsia were treated with vitamin B12whereas only 02 patients with past history of pre eclampsia needed Vitamin B12treatment as their homocysteine levels were above normal ( Table 3).When pre and post vitamin B12 levels were compared, decrease & normalization ofhomocysteine levels and increase & normalization in vitamin B12 levels were seen afterInj B12 treatment in both eclampsia and pre eclampsia patients.(Table 4) Thisimprovement was statistically highly significant (p<0.001). A negative and statisticallysignificant correlation (r = -0.335 and P<0.05) was found between serum homocysteineand vitamin B12 in preeclampsia(Table 5). Final outcome of these patients after InjVitamin B12 therapy has improved at par with control group without any neonatal or
  • 6. maternal mortality in all four groups, however maternal and perinatal morbidity wasmuch higher among preeclamptic and eclamptic group because of pre existingpathology (Table 6).DiscussionOur findings suggest that levels of serum homocysteine, and vitamin B12 are altered inpreeclampsia and eclampsia patients as compared in age-matched normotensivepregnant control subjects.The present study shows that there was significant hyperhomocystinemia & deficiency of Vitamin B12 in patients with preeclampsia andeclampsia. Several prospective studies with rather small cohorts of patients withpreeclampsia have shown an independent association between elevated serumhomocysteine level and untoward obstetric outcome [7] [8]. Several factors mayincrease homocysteine levels in women with preeclampsia [9]. Metabolism in the kidneyis the major route by which homocysteine is cleared from plasma and this route ofelimination may be affected by preeclamptic changes in the kidney [10].Hyperhomocysteinemia in preeclamptic patients found in our study might be due tomodulation in homocysteine metabolism, which corroborates with the work of Walker etal, Hogg et al , Vollset et al. Several studies have demonstrated serum concentrationsof elevated homocysteine in preeclampsia [11]. These studies support our results. Inour study, the levels of vitamin B12 were also significantly lower in the preeclamptic andeclamptic group as compared to control groups ,suggesting raised homocysteine wasdue to vitamin B12 deficiency. Carmel R found differences in folic acid concentrationsbetween preeclamptic and normal pregnant women. Similarly, in a systematic review by
  • 7. Mignini et al., folic acid and vitamin B12 concentrations were lower in preeclampticwomen when compared with those of normotensive women[12] . In another study, therewas no difference in folic acid and vitamin B12 levels between pooled normal andpreeclamptic groups but these levels were significantly lower in patients with the 677 CTmutation of MTHFR[13]. The serum homocysteine was found to have negative andinsignificant correlation with serum folic acid in preeclamptic patients. In our studynegative and statistically significant correlation (r = -0.335 and P<0.05) was foundbetween serum homocysteine and vitamin B12 in preeclampsia. There are twopathways by which homocysteine is metabolized:- remethylation and transsulfuration.Folic acid and vitamin B12 are required for the remethylation of homocysteine tomethionine; vitamin B6 is required for the transsulfuration of homocysteine to cysteine.A good correlation between serum homocysteine, and vitamin B12 levels observed inour study support this view. It is justifiable to administer Inj vitamin B12, 1500µgm to allpatients developing pre eclampsia as prophylactic dose to prevent further complicationsof PIH.From the above discussion we can assume that biochemical screening such ashomocysteine, vitamins B12 are of paramount importance in preeclampsia. The inverserelation between homocysteine, and vitamin B12 indicates that severity associated withmetabolic disturbances in preeclampsia can contribute to obstetric complications[14].On the other hand, there is an absolute need for large studies designed to answer thequestion as to whether hyper homocysteinemia and vitamin B12 deficiency areassociated with increased risk for pre eclampsia and whether therapy of these disordersmight influence maternal mortality & morbidity [15]. Further studies should help define
  • 8. the role of genetic polymorphism in enzymes of homocysteine, folic acid, vitamin B12metabolism and their role in pre eclampsia [16].References 1. Hogg BB, Tamura T, Johnston KE, DuBard MB, Goldenberg MA, Goldenberg RL. Second-trimester plasma homocysteine levels and pregnancy-induced hypertension, preeclampsia, and intrauterine growth restriction. Am J Obstet Gynecol. 2000; 183:805–9. 2. Vollset SE, Refsum H, Irgens LM, Emblem BM, Tverdal A, Gjessing HK, et al. Plasma total homocysteine, pregnancy complications, and adverse pregnancy outcomes: the Hordaland homocysteine study. Am J Clin Nutr. 2000;71:962–8. 3. Gambhir D S, Gambhir J K (2000) Homocysteine metabolism in health & disease, Indian Heart Journal 52(suppl) 59-515 . 4. Hankey G L, Eikelboom J W(2000) Homocysteine and vascular disease. Indian Heart Journal, 52 (suppl) 518-526. 5. Barron WM, Murphy MB, Lindheimer MD. Management of hypertension during pregnancy. In: Laragh GH, Brenner BM, editors. Hypertension: pathophysiology, diagnosis and management. Raven: New York; 1990. p. 1809–27. 6. Desouza C, Keebler M, McNamara D M & Fonseca V(2002)- Drugs affecting homocysteine metabolism; Drugs 62(4) 605-606. 7. Clarke R, Daly L, Robinson K, Naughten E Cabalane S, et al (1991) Hyperchromocystenemia; an independent risk factor in vascular disease, New England Journal of Medicine 324 (17) 1149-1155.
  • 9. 8. Wald DS, Law M, Morris JK. Homocysteine and cardiovascular disease: evidence on causality from a meta-analysis. Br Med J. 2002; 325:1202–6.9. Voutilainen S, Rissanen TH, Virtanen J, Lakka TA, Salonen JT. Low dietary folic acid intake is associated with an excess incidence of acute coronary events: the Kuopio ischemic heart disease risk factor study. Circulation. 2001; 103:2674–80.10. Klerk M, Verhoef P, Clarke R. MTHFR studies collaboration group. MTHFR 677C T polymorphism and risk of coronary heart disease: a meta-analysis. J Am Med Assoc. 2002; 288:2023–31.11. Ueland PM, Refsum H, Stabler SP, Malinow MR, Andersson A, Allen RH. Total homocysteine in plasma or serum: methods and clinical applications. Clin Chem. 1993; 39:1764–79.12. Walker MC, Smith GN, Perkins SL, Keely EJ, Garner PR. Changes in homocysteine levels during normal pregnancy. Am J Obstet Gynecol. 1999; 180:660–4.13. Bostom AG, Lathrop L. Homocysteinemia in end-stage renal disease: prevalence, etiology, and potential relationship to arteriosclerotic outcomes. Kidney Int. 1997;52:10–20.14. Mignini L, Latthe P, Villar J, Kilby M, Carroli G, Khan K. Mapping the theories of preeclampsia: the role of homocysteine. Obstet Gynecol. 2005;105: 411–25.15. Lachmeijer AM, Arnigrimsson R, Bastiaans EJ, Pals G, ten Kate LP, de Vries JIP. Mutation in the gene for methylenetetrahydrofolate reductase, homocysteine levels and vitamin status in women with history of preeclampsia. Am J Obstet Gynecol. 2001;184:394–402.
  • 10. 16. Finkelstein JD. Methionine metabolism in mammals. J Nutr Biochem. 1990;1:228–37.Table 1 Demographic data in controlsParameters Eclampsia Pre- Past History Control group P-value group eclampsi of PET/ without any (n=10) a Eclampsia past history or Group group (n=10) PET/ (n = 10) Eclampsia in present pregnancy (n = 10)Age (years) 21.6±2.3 22.1±3.1 25.5±3.3 25.1±2.4 <0.05Gestation age 32 36 36 36(weeks)Parity P1=02 P1=06 Eclampsia=02 P1=04 P2=06 P2=02 Pre P2=4 P3=02 P3=02 eclampsia=06 P3=02SBP (mm of Hg) 158.7±7.1 144.3±4.6 118.1±8.2 120.3±6.1 <0.05DBP (mm of Hg) 108.1±5.2 96.3±3.1 90.6±2.9 84.0±3.9 <0.05Proteinuria Significant Significant Not Significant Absent <0.05mg/24hrThe results were compared between preeclampsia/eclampsia groups & control group.The values are presented as mean ± S.D.
  • 11. Table2–Baseline Serum total homocysteine and vitamin B12 levels in four groups.Parameters Eclampsia Pre- Past History Control P-value group eclampsia of PET/ group (n=10) Group Eclampsia (n = 10) (n = 10) group (n=10)Homocysteine 34.1±6.2 22.3±4.8 10.97±1.61 8.8±3.2 <0.01(μmol/l)Vitamin B12 131.9±20.5 157.4±44.8 411.4±8.3 624.6±11.9 <0.05(pg/ml)Hb% 9.28±2.5 9.39±3.2 9.22±3.8 9.65±1.9 >0.05The results were compared between preeclampsia/eclampsia groups & control.The values are presented as mean ± SEM.Table 3:- No of patients treated with Inj B12 in each group. Drug Eclampsia Pre-eclampsia Past History P-value group Group of PET/ (n=10) (n = 10) Eclampsia group (n=02) Vitamin B12 10 10 02 <0.05 (pg/ml)Table 4:- Serum homocysteine & Vit B12 levels after Inj B12 treatment in eachgroup. The values are presented as mean ± SEM.
  • 12. Drug Eclampsia Pre-eclampsia P-value group Group (n=10) (n = 10) Homocystein(µmol/l 14.51±7.14 11.03±2.34 <0.001 ) Vitamin B12 (pg/ml) 1239.1±755.7 1412.4±615.1 <0.01Table 5 -Correlation of total homocysteine and vitamin B12 in preeclampsiapatients Parameters ‘r’ Value P-value Homocysteine -0.71 <0.0001 Vitamin B12 (pg/ml) -0.52 <0.01r = Correlation coefficientTable 6- Obstetric and perinatal outcome among all 04 groups
  • 13. Parameters Eclampsia Pre- Past Control P-value group eclampsia History group (n=10) Group of PET/ (n = 10) (n = 10) Eclampsia group (n=10)Perinatal outcomeLow Birth weight 100% 40% 20% 10% <0.001IUGR 60% 30% 20% 10% <0.001IUD 10% 0% 0% 0% -Abruptio Placentae 10% 0% 0% 0% -Need for NICU 80% 60% 20% 20% <0.001Need for 60% 40% 10% 10% <0.001Resuscitation1 Min APGAR<7 40% 30% 20% 10% <0.001Maternal outcomeNormal delivery 40% 60% 70% 80% <0.001Cesarean Section 40% 40% 30% 10% <0.001Instrumental 20% 0% 0% 10% -Delivery( Forceps/Vacuum)DIC 2% 0% 0% 0%

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