Cord blood leptin levels in normal pregnancies, pregnancy induced hypertension and gestational diabetes mellitus.

OBJECTIVE: To compare cord blood leptin concentrations between normal pregnancy, pregnancy induced hypertension (PIH), and gestational diabetes mellitus (GDM). DESIGN: Cross-sectional study. SETTING: Academic institutes and a tertiary care maternal hospital. METHOD: 48 newborns of normal pregnancies (N=18), pregnancy induced hypertension (N=16), and gestational diabetes mellitus (N=14) were studied. Cord blood samples were collected and newborn anthropometric indices recorded at delivery. Leptin concentrations were measured using an enzyme immunoassay. RESULTS: Cord blood leptin levels were significantly different between the 3 groups (Kruskal-Wallis ANOVA; P=0.0064), and the difference resulted mainly from higher levels in GDM than in PIH [geometric mean (95% CI) for GDM: 10.89 (6.30, 18.84) vs PIH: 3.49 (2.14, 5.69) ng/ml (Dunn's multiple comparison: P<0.01). This pattern persisted even when leptin levels were normalized to the ponderal index (Kruskal-Wallis ANOVA P=0.0035; Dunn's multiple comparison: P<0.01). Leptin levels significantly and positively correlated with the ponderal index in normal pregnancy (Spearman r=0.506, p<0.05) and with birth weight in PIH (r=0.5463, p<0.05). CONCLUSION: In GDM cord blood leptin levels are significantly higher, and a source other than fetal adipocytes appears to contribute to this.

Maternal and cord blood levels of insulin-like growth factors--I and--II and insulin-like growth factor binding protein-1: correlation with birth weight and maternal anthropometric indices.

OBJECTIVES: To study the correlation of maternal and cord blood insulin like growth factor (IGF)-I and -II and IGF binding protein (IGFBP)-1 levels with birth weight and maternal anthropometric indices. DESIGN: Longitudinal prospective study. SETTING: Academic Institutions and a Tertiary Care Maternity Hospital. PARTICIPANTS: Women with uncomplicated singleton pregnancy (N = 35) and their newborns. MEASUREMENTS: Maternal weight, height, symphysiofundal height and serum levels of IGF-I, IGF-II, IGFBP-1 were measured thrice during the antenatal period, within 24 h of delivery and at 6 weeks and 6 months postpartum. Newborn anthropometric indices were recorded at birth, and at 6 weeks and 6 months of age. Cord blood levels of IGF-1, IGF-II, IGFBP-1, paternal height and weight, and placental weight measured. RESULTS: Maternal and cord blood IGF-I levels were lower than values reported for Caucasians. All newborns showed adequate growth at birth, and up to 6 months of age. Cord blood IGF-1 positively correlated with chest circumference (r = 0.4532, P = 0.0262), IGFBP-1, negatively with birth weight (r = -0.4024, P = 0.0461) and IGF-II had no effect. Cord blood IGF-I positively correlated with maternal levels at 28 +/- 2 (r = 0.4571, P = 0.0247) and 36 +/- 2 (r = 0.4291, P = 0.0364) weeks of amenorrhoea, whereas IGF-II and IGFBP-1 did not correlate with maternal values. Maternal IGF-I, IGF-II and IGFBP-1 did not correlate with newborn or maternal anthropometric indices. Placental weight correlated significantly with birth weight (r = 0.5299, P = 0.0348) and head circumference (r = 0.5031, P = 0.0470). CONCLUSIONS: Cord blood IGFBP-1 and placental weight appear to be determinants of birth weight variation even among appropriately grown for gestational age newborns.

Insulin-like growth factors-I and -II and insulin-like growth factor binding protein-1 during normal pregnancy: pattern of secretion and correlation with other placental hormones.

OBJECTIVES: To describe pattern of secretion of insulin-like growth factor (IGF)-I, IGF-II, IGF binding protein (IGFBP)-1 and their correlation with each other and major placental hormones during normal pregnancy. DESIGN: Longitudinal study. SETTING: Academic Institutions and a Tertiary Care Maternity Hospital. PARTICIPANTS: Healthy women with singleton uncomplicated pregnancies (N = 35). MEASUREMENTS: Serum levels of IGF-I, IGF-II, IGFBP-1, chorionic gonadotrophin (HCG), placental lactogen (HPL), prolactin, oestradiol and progesterone were studied thrice during the antenatal period and within 24 h of delivery. RESULTS: IGF-I, IGFBP-1, HPL, prolactin, oestradiol and progesterone increased and HCG decreased significantly with advancing gestation (Repeated measures ANOVA: P < 0.01 to 0.0001). IGF-II levels were not significantly affected by period of gestation. Significant negative correlations (multiple regression analysis) were seen between IGFBP-1 and prolactin at 28 +/- 2 (P = 0.0226) and 36 +/- 2 (P = 0.0417) weeks of amenorrhoea (WOA) and between oestradiol and IGF-II at 36 +/- 2 WOA (P = 0.037). Prolactin and IGF-I at 14 +/- 2 WOA (P = 0.0225) and progesterone and IGFBP-1 at 28 +/- 2 WOA (P = 0.0216) correlated positively. CONCLUSIONS: Maternal IGF-I and IGFBP-1 but not IGF-II significantly increase as pregnancy advances. Components of the IGF system regulate or are affected by some of the placental hormones and the effects vary with the period of gestation.

Wuchereria bancrofti: detection of microfilariae in asymptomatic microfilaremic individuals with Setaria digitata antigens.

A dot-ELISA for detection of microfilariae of Wuchereria bancrofti in an endemic area was developed. This test can differentiate the endemic normals from the microfilaraemic asymptomatic individuals. Antigens of molecular weight 130 and 52 kDa of the cattle filaria worm Setaria digitata were used for this test. It was observed that these two antigens were also present in the serum of asymptomatic microfilaraemic individuals.

A preliminary analysis of Wuchereria bancrofti microfilarial antigens for potential use in diagnosis.

Several antigens from the microfilarial stage of Wuchereria bancrofti have been identified using immunoblots of microfilarial antigens and screening with immune sera and tropical pulmonary eosinophilia (TPE) sera. This analysis revealed an array of antigens with apparent molecular weights of 14kDa, 35kDa, 42kDa, 63kDa, 88kDa, 97kDa and 200kDa. Among these only the 14kDa and 42kDa antigens were consistently recognized by most of the immune sera. A 132kDa antigen was recognized only by TPE sera. Analysis of rabbit immune sera revealed that the 42kDa antigen was shared by two developmental stages of W. bancrofti, namely L3 and mF. This antigen could become a potential vaccine candidate. The 14kDa antigen seems specific for the microfilarial stage and therefore could be a diagnostic marker for active infection. The 132kDa antigen could aid in the diagnosis of TPE.

A simple and rapid non-radioactive oligonucleotide based hybridization assay for the detection of Wuchereria bancrofti.

Five biotin labeled oligonucleotides was designed based on a previously cloned and characterized repetitive DNA sequence specific for Wuchereria bancrofti. The oligonucleotide mix (containing five probes) when used in a hybridization assay, detected as little as 100 pg of purified W. bancrofti, microfilarial DNA, a single infective stage larva and a single microfilaria in 50 microl blood sample. A simple protocol was followed for the hybridization assay. Blood samples lysed with sterile distilled water and digested with proteinase K in the presence of a detergent were directly applied on to nylon membranes for dot blot assays. The DNA extract of mosquitos carrying infective stage larvae was eluted through sephadex G-50 minicolumns prior to blotting. The assay was also able to detect DNA extracted from microfilariae infected samples stored over five days at room temperature (28 degrees C). This simple and rapid oligonucleotide hybridization protocol with the highly sensitive chemiluminescent-based detection has significant potential for the development of a field kit to detect W. bancrofti infection.

PCR based detection of Mycobacterium tuberculosis: effect of sample preparation.

Tests based on the polymerase chain reaction (PCR) for the detection of the Mycobacterium tuberculosis complex in clinical samples have a lower sensitivity when compared to culture. This has been attributed to the presence of inhibitors to Taq polymerase and/or suboptimal DNA extraction procedures. We tested different methods of processing smear negative culture positive sputum (n = 52) using different detergents, including nonidet P-40 (NP-40), sodium dodecyl sulphate (SDS), tween 20, triton X 100 and N-lauryl sarcosine. The detergents were used in combination with lysozyme and proteinase K enzymes. NP-40 was significantly better than SDS, tween 20 and N lauryl sarcosine (p < 0.05). When NP-40 was used as the detergent, 42 out of 52 specimens gave positive results with the standard amplification protocol which amplifies a 245 bp sequence of the insertion element IS 986. The 10 specimens that were negative were further diluted ten fold and/or eluted in sephadex G-50 columns before standard DNA amplification. A further 8 specimens then became positive. Elution in sephadex G-50 was better than ten fold dilution in processing of samples. The two negative samples had very low colony counts (n < 5). The study demonstrates that the sensitivity of the PCR is dependent on the sample preparation technique and the amount of target sequence available for amplification.

Effect of age on serum prolactin concentration in presumably fertile men in Sri Lanka.

OBJECTIVE: To examine possible age related variation in serum prolactin in men. DESIGN: A cross sectional study. SUBJECTS: Healthy married men aged 21 to 85 years with one or more children. MEASUREMENTS: Serum prolactin concentrations measured by immunoradiometric assay. RESULTS: Prolactin concentrations were significantly higher (p < 0.001) from 61 years of age onwards when compared with 31 to 60 years. Prolactin concentrations between 21 to 30 years were significantly higher than between 31 to 50 years (p < 0.05), but lower than between 61 to 70 (p < 0.05) and 75 to 85 (p < 0.01) years. CONCLUSIONS: Serum prolactin concentrations show age related variations in presumably fertile men.

Serum protein binding of antiepileptic drugs in Sri Lankans.

The percentage protein binding of antiepileptic drugs was investigated in epileptic patients (n = 90) undergoing treatment with phenobarbitone, phenytoin and carbamazepine either as a single drug therapy or in different combinations. When administered individually, the percentage (mean +/- SEM) protein binding of phenobarbitone, phenytoin and carbamazepine were 50.84 +/- 7.03, 87.23 +/- 2.98 and 76.80 +/- 6.30 respectively. Combination of phenobarbitone and phenytoin resulted in percentage (mean +/- SEM) protein binding of 51.94 +/- 6.09 for phenobarbitone and 83.54 +/- 7.01 for phenytoin, while the combination of phenobarbitone and carbamazepine resulted in percentage (mean +/- SEM) protein binding of 49.60 +/- 2.59 for phenobarbitone and 79.10 +/- 3.31 for carbamazepine. When phenytoin was given with carbamazepine percentage (mean +/- SEM) protein binding was 87.22 +/- 4.48 for phenytoin and 72.50 +/- 5.92 for carbamazepine.