Erythrocyte fatty acids and desaturase indices in early pregnancy are associated with risk of preeclampsia.

Preeclampsia (PE) is a pregnancy disorder that may be associated with inadequate maternal nutrition. Fatty acids are vital for placental and fetal growth. Fatty acid desaturases, key enzymes influencing the metabolism of polyunsaturated fatty acids, are reported to be associated with cardiometabolic risk. Any imbalance in the levels of omega-3 and omega-6 fatty acids can result in increased inflammatory response. The current study reports the levels of erythrocyte fatty acids and desaturase index across gestation in women who develop PE (n = 108) and compares them with non-PE women (n = 216). Maternal erythrocyte fatty acids were measured at 4 time points during pregnancy (i.e., 11-14, 18-22, 26-28 weeks and at delivery) using gas chromatography. Maternal total erythrocyte saturated fatty acids and omega-6/omega-3 fatty acid ratio was higher in the PE group as compared to the non-PE group at 11-14 weeks and 18-22 weeks respectively. Maternal Δ5 desaturase index was lower while Δ6 desaturase index was higher in the PE group at 11-14 and 18-22 weeks. Maternal stearoyl CoA desaturase-18 (SCD-18) index was lower at 11-14 weeks and at delivery. These changes were mainly observed in the early onset PE (EOP) group. Δ6 desaturase index at 11-14 weeks predicted the risk of EOP. Imbalance in fatty acid levels and desaturase indices predate the clinical diagnosis of PE, indicating their role in its pathophysiology. Measurement of fatty acids and desaturase indices in early pregnancy merits evaluation as predictors of risk of PE.

Study of the distribution of Malassezia species in patients with pityriasis versicolor and healthy individuals in Tertiary Care Hospital, Punjab.

PURPOSE: Pityriasis versicolor (PV) is a chronic superficial fungal disease caused by Malassezia species. Our aim was to identify Malassezia species from PV patients and healthy individuals in Punjab. MATERIALS AND METHODS: Modified Dixon agar was used as isolation culture medium. Identification was based on morphological observation and biochemical evaluation. The biochemical evaluation consisted of culture onto Sabouraud dextrose agar, catalase reaction, Tween assimilation, Cremophor EL assimilation, splitting of esculin and growth at 38 0 C. RESULTS: Out of 58 microscopically diagnosed cases of PV, growth was obtained from 54 (93.10%) cases. The most frequently isolated species were M. globosa, M. sympodialis and M. furfur which made up 51.79%, 31.42% and 18.51% of the isolated etiological agents respectively. However, the major isolate from the back of healthy individuals was M. sympodialis (47.61%), followed by M. obtusa (19.04%), M. globosa (14.20%), M. furfur (9.52%), M. pachydermatis (4.76%) and M. slooffiae (4.76%). CONCLUSIONS: M. globosa in its mycelial phase was the main etiological agent, but as normal flora from the back of healthy subjects, it was found in significantly less number (P = 0.01), suggesting that the higher pathogenicity of M. globosa in terms of enzymatic endowment, might be the cause of its predominance in PV lesions.

Type II diabetes increases mitochondrial DNA mutations in the left ventricle of the Goto-Kakizaki diabetic rat.

Mitochondrial dysfunction has a significant role in the development of diabetic cardiomyopathy. Mitochondrial oxidant stress has been accepted as the singular cause of mitochondrial DNA (mtDNA) damage as an underlying cause of mitochondrial dysfunction. However, separate from a direct effect on mtDNA integrity, diabetic-induced increases in oxidant stress alter mitochondrial topoisomerase function to propagate mtDNA mutations as a contributor to mitochondrial dysfunction. Both glucose-challenged neonatal cardiomyocytes and the diabetic Goto-Kakizaki (GK) rat were studied. In both the GK left ventricle (LV) and in cardiomyocytes, chronically elevated glucose presentation induced a significant increase in mtDNA damage that was accompanied by decreased mitochondrial function. TTGE analysis revealed a number of base pair substitutions in the 3' end of COX3 from GK LV mtDNA that significantly altered the protein sequence. Mitochondrial topoisomerase DNA cleavage activity in isolated mitochondria was significantly increased in the GK LV compared with Wistar controls. Both hydroxycamptothecin, a topoisomerase type 1 inhibitor, and doxorubicin, a topoisomerase type 2 inhibitor, significantly exacerbated the DNA cleavage activity of isolated mitochondrial extracts indicating the presence of multiple functional topoisomerases in the mitochondria. Mitochondrial topoisomerase function was significantly altered in the presence of H2O2 suggesting that separate from a direct effect on mtDNA, oxidant stress mediated type II diabetes-induced alterations of mitochondrial topoisomerase function. These findings are significant in that the activation/inhibition state of the mitochondrial topoisomerases will have important consequences for mtDNA integrity and the well being of the diabetic myocardium.

Maternal nutrition and birth size among urban affluent and rural women in India.

BACKGROUND: Varying results of worldwide intervention programs to pregnant mothers necessitate the need to understand the relationship between maternal nutrition and birth size among well nourished and undernourished mothers. OBJECTIVE: To examine this relationship among urban affluent mothers and to compare the findings with those on rural Indian mothers. SUBJECTS: Data collected on urban affluent mothers (n = 236) was compared with rural mothers (n = 633). DESIGN: Mothers were contacted at 18 +/- 2 and 28 +/- 2 wk of gestation for anthropometry, dietary intakes [24-hr recall, Food Frequency Questionnaire (FFQ)] and after delivery for neonatal anthropometry. RESULTS: Despite large differences in nutritional status of urban and rural mothers ( pre-pregnant weight 55.9 +/- 9.2 Vs 41.5 +/- 5.2 kg, respectively) maternal fat intakes at 18 wk were associated with birth weight (p < 0.05), length (p < 0.01) and triceps skin fold thickness (p < 0.05) of the newborn in urban and rural mothers. Consumption of fruits was associated with birth length (p < 0.05) in urban (18wk) and with birth weight (p < 0.01) and length (p < 0.01) in rural (28wk) mothers, when their energy intakes were low. Maternal consumption of milk too, was associated with newborn's triceps (p < 0.01) in urban (28wk) while with birth weight (p < 0.05) and length (p < 0.05) in rural (18wk) mothers. The findings mainly underscore the importance of consumption of micronutrient rich foods, when energy intakes are limiting during pregnancy, for improving birth size. CONCLUSIONS: Creating nutritional awareness and motivating rural mothers for consuming micronutrient rich foods like green leafy vegetables and seasonal fruits that are easily available in rural areas, will be a much affordable solution for combating the problem of low birth weight rather than waiting for improvement in the existing nationwide programs for pregnant women.

Frequency of partial D in Western India.

Partial D is of clinical importance as the partial D-positive individuals who lack some epitopes of D antigen can develop anti-D if exposed to normal D antigen. The frequency of partial D varies in different populations. The majority of molecular studies on D variants have been reported in European, African and some East Asian populations, but no study has been reported in the Indian population so far. The aim of the study was to screen Indian population for detection of partial D by serology and classify them by multiplex polymerase chain reaction (M-PCR). The study population, consisting of 10,000 RhD-positive individuals from West India, was screened for detection of partial D using the partial D kit. In addition to these, blood samples referred because of serological RhD discrepant results from blood banks of West India were also investigated. The samples identified as partial D from these two groups were further characterized by M-PCR. Fifteen partial D cases were identified by population screening and 45 were identified from referred samples. Population screening revealed that one third of partial D was DFR when tested by partial D kit. We were able to classify 63.4 and 76.6% of partial D by partial D kit and M-PCR, respectively. The incidence of partial D in West India was found to be at least 0.15% when tested with partial D kit. DFR partial D was found to be predominant in the present study.

Varied distribution of RhD epitopes in the Indian population.

BACKGROUND: Inhabited by more than 4000 caste and tribal groups, India has an extremely heterogenous population. For thousands of years many tribal groups have practised endogamy and are practically genetically isolated. Traditionally, polyclonal anti-D reagent has been used for RhD typing; though monoclonal antibodies are increasingly being used. As a result, blood banks find it difficult to assign the RhD status to an increasing number of people. As monoclonal anti-D typing reagents may not detect all RhD antigen epitopes, we studied the RhD antigen epitope heterogeneity in different population groups in India. METHODS: Red cells of 5315 RhD-positive individuals belonging to different castes and tribes of India were tested with 30 different epitope-specific monoclonal anti-D antibodies. RESULTS: No single monoclonal antibody could detect all RhD-positive red cells detected by polyclonal antisera. The highest proportion of D antigen was detected by LHM 76/55 and BRAD-8 (98%) monoclonal antibodies. CONCLUSION: We need to determine the correct mix of monoclonal antibodies that will detect nearly all RhD antigens detected by polyclonal anti-D sera. Similarly, before accepting monoclonal anti-D for therapeutic use, it would be necessary to determine the appropriate ones for use in the Indian population.

Iron status of Hindu brahmin, Jain and Muslim communities in Surat, Gujarat.

AIM OF THE STUDY: To determine iron status of healthy, unrelated Brahmin, Jain and Muslim participants having different dietary habits. METHODOLOGY: Control participants other than above three communities, consumed vegetarian or non-vegetarian diet. Brahmin and Jain were strictly vegetarian but Jain did not consume roots or tubers. Muslims consumed non-vegetarian food. Standard techniques were used to measure hematological parameters, serum iron, total iron bindings capacity (TIBC), serum ferritin, transferrin and transferrin saturation. For statistical evaluation mean, standard deviation, pair t test, χ2 and ANOVA (F test) were employed. RESULTS: Study includes 565 male and 198 female children and adults. Among them 205 were children and remaining adults. All four categories i.e. control, Brahmin, Jain and Muslims showed higher incidence of anemia and iron deficiency in females compared to males. Mean values of hematological parameters did not vary significantly in four groups. Serum iron, TIBC, transferrin and ferritin levels indicated iron deficiency anemia more frequently in Jains and less frequently in Muslims (p<0.05). Iron status of Brahmin was comparable with controls (p<0.01). Majority of the participants had serum ferritin concentration >15 ng/mL. Except one male Jain child none of the participants had serum ferritin concentration <12 ng/mL. Jain subjects more frequently had serum iron concentration <60 µg/dL. CONCLUSION: Jain participants had higher incidence of iron deficiency anemia. Vegetarian diet consumed by Gujarati Hindu Brahmin community provided them with a sufficient iron to maintain their iron profile like Muslims consuming non-vegetarian diet.

Hb Q(India) and its interaction with beta-thalassaemia: a study of 64 cases from India.

Haemoglobin Q (Hb Q), a relatively uncommon alpha-chain structural Hb variant, has been reported either in the heterozygous state or interacting with beta-thalassaemia. Individuals inheriting Hb Q generally are asymptomatic and are diagnosed by chance during population screening or as a part of a family study. This paper represents the first large study from India of 64 cases of Hb Q, documenting the haematological and molecular findings on 36 cases of Hb Q trait, 22 of Hb Q beta-thalassaemia trait and three of Hb Q beta-thalassaemia major, as well as a family of Hb Q homozygous cases. Hb Q is detected by Hb electrophoresis and chromatography. Hb Q levels in homozygous cases ranged from 32% to 35%, while in Hb Q heterozygotes the level was 20%. When there was an interaction of beta-thalassaemia heterozygotes the level was 14%, and in interacting beta-thalassaemia homozygotes the levels ranged from 7% to 9%. beta-thalassaemia mutations were characterised in cases showing elevated Hb A2 levels, which were markedly reduced in the majority of cases in which beta-thalassaemia was absent. Hb Q is rare and not a single homozygous case has been reported. However, Hb Q disease showed wide variation in clinical and haematological presentation in the same family.

Flow cytometric quantification of antigen D sites on red blood cells of partial D and weak D variants in India.

The D antigenic sites are diminished in weak D and partial D variants. The aim of this study was to estimate D antigen on RBC in weak D and partial D variants in Indian population by using flow cytometric method. Blood samples of 42 cases of partial D, eight cases of weak D and 123 normal Rh phenotypes identified by serological methods were used in the study. An indirect immunofluorescence method was employed using 29 monoclonal anti-D as primary antibody. The D antigenic sites were calculated using control RBC as internal standard. The D antigenic sites in weak D and partial D variants are less than that found in normal Rh phenotypes. In weak D, the D antigenic sites were between 1500 and 7000 D antigens per cell. Among partial D variants, DVI had minimum and DVa had maximum number of D sites. Flow cytometry is a very good tool for demonstrating minor variation in D antigen when serological methods are inconclusive. The D antigenic sites per RBC in partial D variants identified in Indian population are reported for the first time.

Production of murine monoclonal anti-B.

BACKGROUND & OBJECTIVE: Monoclonal antibodies against red blood cell antigens used in research and as diagnostics in India are commercially procured from western countries. Indigenously generated potent clones are not available in India. Hence, the objective of the present study was to raise potent murine monoclonal antibodies against A, B and H blood group antigens indigenously and establish a stable clone of anti-B secreting cells. METHODS: Spleen cells of female BALB/c mice immunized with B group red blood cells were fused in presence of polyethylene glycol (PEG) 1500 with a mouse myeloma cell line Sp 2/0 Ag. 14 in hypoxanthine aminopterine thymidine (HAT) selective medium and incubated at 37 degrees C, 5 per cent CO(2) and 95 per cent humidity for a week. RESULTS: The culture supernatant of the wells showing anti-B activity, were further subcloned and a clone 2C4D5F10 was generated which showed a good potency, avidity and specificity. INTERPRETATION & CONCLUSION: The anti-B clones thus produced indigenously provided a useful reagent in blood group typing. The unlimited availability unlike polyclonal antisera makes this reagent more cost-effective. It also ensures a regular supply with the similar specificity.

The burden and determinants of reproductive tract infections in India: a population based study of women in Goa, India.

BACKGROUND/OBJECTIVE: Reproductive tract infections (RTI) present major health, social, and economic problems in developing countries. Our objective was to describe the prevalence and risk factors of RTIs in a population based sample of women aged 18-45 years. METHOD: 2494 women of 3000 randomly selected from the population defined by a primary health centre catchment area consented to participate. Participants were interviewed regarding complaints and risk factors. Laboratory specimens were collected for the diagnosis of RTIs. Analyses of risk factors were carried out separately for the outcomes of sexually transmitted infections: chlamydia, gonorrhoea, trichomoniasis; and endogenous infections: bacterial vaginosis (BV) and candida. RESULTS: Endogenous infections were relatively common (BV 17.8%; candida 8.5%), and sexually transmitted infections (STI) were infrequent (4.2%). Factors indicative of poverty and marginalisation were associated with STIs and BV. Gender disadvantage, particularly spousal violence, was associated with BV, while concern about a husband's extramarital relationships, an indicator of sexual risk, was associated with STI. Husband's discharge was strongly associated with STI, and a non-white vaginal discharge was associated with both STI and BV. Condom use and oral contraceptive use were associated with a reduced risk of BV. CONCLUSIONS: Most of the population burden of RTIs is attributed to endogenous infections. Socioeconomic deprivation and gender disadvantage are associated with raised risk for BV, while the risk factors for STIs indicated that disadvantaged women were likely to be infected by their husbands.

The burden and determinants of dysmenorrhoea: a population-based survey of 2262 women in Goa, India.

OBJECTIVE: To describe the prevalence and determinants of dysmenorrhoea, the most common menstrual complaint, in a community in India. DESIGN: Cross-sectional survey. SETTING: Catchment area of primary health centre in Goa, India. POPULATION: Three thousand women aged 18-45 years randomly selected. A total of 2494 women consented to participate (83.1%). METHODS: Eligible participants were asked standardised questions regarding menstrual complaints over the past 12 months, and socio-demographic, psychosocial and reproductive risk factors. Vaginal or urine specimens were collected for the diagnosis of reproductive tract infections. MAIN OUTCOME MEASURES: Dysmenorrhoea of moderate to severe intensity. RESULTS: A total of 2262 women were eligible. More than half reported dysmenorrhoea; moderate to severe dysmenorrhoea was reported by 755 participants (33.4%, 95% CI 31.4-35.4). There was a linear association between severity of pain and impact (medication and taking rest) and the onset of pain (premenstrual onset associated with more severe pain). On multivariate analyses, the risk of moderate-severe dysmenorrhoea was associated with the experience of violence (OR 2.23, 95% CI 1.5-34); other somatic complaints (OR 3.67, 95% CI 2.7-4.9 for highest somatoform symptom score category compared with the lowest); gynaecological complaints (non-menstrual lower abdominal pain: OR 1.78, 95% CI 1.3-2.3; dysuria: OR 1.98, 1.4-2.7); menorrhagia (OR 1.92, 95% CI 1.4-2.6); and illiteracy (OR 1.32, 95% CI 1.0-1.7). Having had a pregnancy (OR 0.53, 95% CI 0.4-0.7), older age of menarche (OR 0.70, 95% CI 0.5-0.9, for age >14 compared with <13 years) and older age (OR 0.43, 0.3-0.6 for age 40-50, compared with 18-24 years) were protective. CONCLUSIONS: The burden of dysmenorrhoea is greater than any other gynaecological complaint, and is associated with significant impact. Social disadvantage, co-morbidity with other somatic syndromes and reproductive factors are determinants of this complaint.

Heme oxygenase-1 gene expression increases vascular relaxation and decreases inducible nitric oxide synthase in diabetic rats.

Hyperglycemia represents the main cause of complication of diabetes mellitus and oxidative stress, resulting from increased generation of reactive oxygen species (ROS), and plays a crucial role in their pathogenesis. Impairment of vascular responses in diabetic rats, as a result of an increase in superoxide (O2-), formation is a major complication in diabetes. Since heme oxygenase (HO) expression regulates the level of ROS by increasing antioxidant, such as glutathione and bilirubin, we investigated whether upregulation of HO-1 modulates the levels of iNOS and eNOS and altered vascular responses to phenylephrine (PE) and acetylcholine (Ach) in aorta and femoral arteries of diabetic (streptozotocin (STZ)-induced) rats. Our results showed that iNOS expression was increased, but HO activity was reduced, in diabetic compared to nondiabetic rats (p<0.05). Upregulation of HO-1 expression by cobalt protoporphyrin (CoPP), an inducer of HO-1 protein and activity, conferred an increase in eNOS and differentially decreased iNOS protein levels (p<0.05). Isolated aortic and femoral arteries obtained from diabetic rats exhibited contraction to PE and relaxation to Ach, which were markedly increased and decreased, respectively. However, HO-1 induction in diabetic rats normalized relaxation compared to controls. Therefore, overexpression of HO-1 may mediate an increase in eNOS and a decrease in iNOS, potentially contributing to restoration of vascular responses in diabetic rats.

Activity of the new quinolone WCK 771 against pneumococci.

The activity of WCK 771, a new experimental quinolone being developed to overcome quinolone resistance in staphylococci, against quinolone-susceptible and -resistant pneumococci was determined. Comparative activities of ciprofloxacin, levofloxacin, gatifloxacin, moxifloxacin, clinafloxacin, vancomycin, linezolid, amoxycillin, cefuroxime, azithromycin and clarithromycin were determined with MIC and time-kill experiments. Animal experiments were also performed to test the in-vivo anti-pneumococcal activity of WCK 771 compared to levofloxacin. WCK 771 MIC50/90 values for 300 quinolone-susceptible Streptococcus pneumoniae isolates (108 penicillin-susceptible, 92 penicillin-intermediate and 100 penicillin-resistant) were 0.5/0.5 mg/L; the MICs of beta-lactams and macrolides rose with those of penicillin G, and all isolates were susceptible to vancomycin and linezolid. WCK 771 MIC50/90 values for 25 quinolone-resistant pneumococcal isolates were 4/8 mg/L, compared to 0.5/1 mg/L for clinafloxacin, 2/4 mg/L for gatifloxacin and moxifloxacin, 8/16 mg/L for levofloxacin, and 16/>32 mg/L for ciprofloxacin. Time-kill studies showed that WCK 771 was bactericidal against pneumococci after 24 h at 4 x MIC, as were the other quinolones tested. Animal model studies showed that WCK 771 had efficacy comparable to that of levofloxacin, by both the oral and subcutaneous routes, for systemic infection caused by three quinolone-susceptible isolates of pneumococci. Overall, WCK 771 was potent both in vivo and in vitro against quinolone-susceptible, but not quinolone-resistant, S. pneumoniae, regardless of penicillin susceptibility.

Role of HLA antigens in Rh (D) alloimmunized pregnant women from Mumbai, Maharashtra, India.

Immunogenetic studies in various diseases provide potential genetic markers. We have studied the incidence of HLA A, B, C, DR and DQ loci antigen in Rh (D) antigen isoimmunized mothers compared to those nonimmunized isoimmunized Rh negative mothers. Seventy six mothers who were immunized to Rh (D) antigen due to pregnancy (responders) and fifty four mothers who did not develop Rh (D) isoimmunization despite positive pregnancies (nonresponders) were selected for the study. Standard methods of serological HLA typing, ABO and Rh (D) groups, and screening for Rh D antibodies were used. 392 unrelated individuals from the population were compared as controls. In addition 45 unrelated individuals from the same population were typed for HLA DRB and DQB gene using PCR-SSP kits. The genotype frequencies of HLA A2, A3, A28, B13, B17, B35, B52, B60, Cw2, Cw6, DR4, and DQ3 were significantly increased, while the frequencies of the HLA A11, A29, A31, B7, B37, B51, Cw1 and DR9 were decreased in the responder women when compared to the non-responder women. HLA A30 (19) split antigen was not identified in immunized women while HLA A23 (9) split antigen was not identified in non immunized women. HLA A3, B17, Cw2 and DR4 showed a significant relative risk among the immunized responder women. When compared with Rh immunized women (responders) reported from USA, England and Hungary the phenotype frequencies of HLA A11, A24, A28, B5, B17, B40, DR2 and DR5 were increased while HLA A23, B8, B18, and DR6 were decreased in the Indian Rh immunized women. Two locus haplotype frequency analysis observed among the responders women revealed that among the significant haplotypes expressed A2-B5, B7-Cw1, DR2-DQ1 were highly significant haplotypes in positive linkage, while A1-B5, and A1-B7 were in significant negative linkage disequilibrium. The haplotype frequencies were

Prostaglandins and nitric oxide mediate superoxide-induced myocardial contractile dysfunction in isolated rat hearts.

Oxygen-derived free radicals have been implicated in the pathogenesis of myocardial injury. We therefore investigated the pathophysiology of myocardial injury induced in isolated rat hearts by perfusion with superoxide radical generated by reacting 2.5 mmol/l purine, 0.03 U/ml xanthine oxidase and 300 U/ml catalase. Perfusion with superoxide significantly (P<0.05) increased left ventricular end-diastolic pressure within 15 to 20 min. During the same time period, heart rate and left-ventricular developed pressure significantly declined to 44.6+/-8.2% and 31.0+/-4.9% of control, respectively. Superoxide perfusion also significantly increased production of prostaglandins, nitric oxide (detected as nitrites) and peroxynitrite (detected immunohistochemically as nitrotyrosine). N(G)-nitro-l-arginine (100 micromol/l), a nitric oxide synthase inhibitor, attenuated superoxide-induced generation of peroxynitrite, increased synthesis of prostacyclin, and partially blocked myocardial dysfunction, as did 1H-[1,2,4]oxadiazolo[4,3-a]quinoxalin-1-one (30 micromol/l), a selective inhibitor of soluble guanylate cyclase, and ONO-3708 (10 micromol/l), a selective thromboxane A(2)receptor antagonist. In contrast, nitroglycerin (4 micromol/l) and sodium nitroprusside (1 micromol/l) each exacerbated the superoxide-induced myocardial dysfunction. These results suggest that nitric oxide and related reactive species contribute to myocardial injury induced by superoxide. Moreover, they suggest that oxidative stress can be delayed or inhibited by reducing levels of nitric oxide, by inhibiting soluble guanylate cyclase, and by blocking thromboxane/prostaglandin receptors.