Meena Desai

Effect of alcohol consumption on bone mineral density and hormonal parameters in physically active male soldiers

BACKGROUND Previous studies on the influence of alcohol intake and smoking on bone mineral density (BMD) in men are inconsistent and the effect of these variables on BMD in physically active men is yet to be explored. OBJECTIVE To investigate the influence of alcohol intake and smoking on BMD in a cohort of males with well-defined lifestyle conditions. DESIGN Men from the armed forces (n=400) having uniform and defined routines were enrolled. BMD was measured by DXA and participants were grouped according to lifestyle variables. Hormonal parameters were measured by immunoassays. RESULTS Participants with intake of >24 g/wk of alcohol had significantly higher BMD at femur compared to non-alcohol consumers (p=0.0001) and a linear increase in mean femoral BMD over increasing categories of alcohol intake (p(trend)<0.0001) was observed. Smoking was negatively associated with femoral BMD. In multiple regression analysis, age, BMI, alcohol consumption and smoking were independent predictors of femoral BMD, explaining 10.6% variance. At lumbar spine, age, height and BMI were independent predictors, explaining 9.4% variance in BMD. The concentrations of total testosterone, free testosterone, bioavailable testosterone and PTH were low (p<0.0001) whereas estradiol (p=0.02), free and bioavailable estradiol (p<0.001) were high in alcohol consumers compared to non-consumers. In multiple regression analysis alcohol intake and height explained 5.5% variance in estradiol(.) CONCLUSIONS In physically active men with well-defined lifestyle conditions, alcohol consumption was associated with higher femoral BMD, the effect of alcohol is complex and is probably partly mediated by influencing the sex steroid levels.

Neonatal screening for congenital hypothyroidism in a developing country: problems and strategies

Neonatal screening in India poses more organisational and socio-economic rather than medical challenges. Based on the pilot study of 450 cord sera, the plan for screening considered cord TSH<30 μU/ml as normal, 30 to 80 as borderline with recall by letters and >80 as indicative of hypothyroid state, with recall by home visits. Of the 17,240 live births only 12,407 cord sera were collected. Envisaging follow-up difficulties, T4 was assayed in cord sera when TSH was>30 μ U/ml. 2·81% (350) babies needed recall. Only 30% of 302 (2·43%) babies with cord TSG 30 to 80 responded, to recall letters and were normal; availability of both cord TSH and T4 helped in excluding hypothyroidism in majority of non-respondents. Forty-eight (0·38%) newborns had TSH>90 μU/ml; 80% of this group and 100% with TSH> 100 μU/ml were traced by home visits. Hypothyroidism was confirmed in 5/48, biochemically and by thyroid scan. All five hypothyroids had cord TSH>300 μU/ml. The incidence in this nonendemic region of India was 1∶2481. Thus false elevation of cord TSH 30 to 300 μU/ml was noted in 0·34% with a chance of detecting a hypothyroid 1 in 10 when TSH>80 μU/ml. Screening strategies in a developing country must ensure meticulous clerical assistance, co-operation and education of nurses and parents, precise and cost effective technics and facilities for continued surveilance of detected hypothyroids.

A comparison of performances of four enzymes used in ELISA with special reference to beta-lactamase.

Horse radish peroxidase, alkaline phaosphatase and beta-D-galactosidase are widely used as labels in the development of enzyme immunoassays (EIAs). Enzyme beta-lactamase, though introduced as a label in late seventies has not yet become very popular inspite of having the necessary features of an enzyme to be used in EIAs. The present article reviews assays developed with this enzyme, highlights its salient features and brings out an argument in favour of its wide spread use in EIAs.

A sensitive ELISA for 6β-hydroxycortisol in urine using enzyme penicillinase (β-lactamase)

Abstract A sensitive and specific, enzyme labelled immunosorbent assay (ELISA) for 6β-hydroxycortisol in diluted urine using penicillinase was developed. 6β-Hydroxycortisol-21-hemisuccinate was conjugated with enzyme penicillinase. Antibody immobilized on a polyvinylchloride ELISA plate (Dynatech) was used for separation of bound from free ligand. The sensitivity of the assay was between 2.0–3.0 pg per well and recovery of 6β-hydroxycortisol from urine ranged between 85.0–108.0%. The assay is simple, rapid and precise.

Monocyte Proteomics Reveals Involvement of Phosphorylated HSP27 in the Pathogenesis of Osteoporosis

Peripheral monocytes, precursors of osteoclasts, have emerged as important candidates for identifying proteins relevant to osteoporosis, a condition characterized by low Bone Mineral Density (BMD) and increased susceptibility for fractures. We employed 4-plex iTRAQ (isobaric tags for relative and absolute quantification) coupled with LC-MS/MS (liquid chromatography coupled with tandem mass spectrometry) to identify differentially expressed monocyte proteins from premenopausal and postmenopausal women with low versus high BMD. Of 1801 proteins identified, 45 were differentially abundant in low versus high BMD, with heat shock protein 27 (HSP27) distinctly upregulated in low BMD condition in both premenopausal and postmenopausal categories. Validation in individual samples (n = 80) using intracellular ELISA confirmed that total HSP27 (tHSP27) as well as phosphorylated HSP27 (pHSP27) was elevated in low BMD condition in both categories (P < 0.05). Further, using transwell assays, pHSP27, when placed in the upper chamber, could increase monocyte migration (P < 0.0001) and this was additive in combination with RANKL (receptor activator of NFkB ligand) placed in the lower chamber (P = 0.05). Effect of pHSP27 in monocyte migration towards bone milieu can result in increased osteoclast formation and thus contribute to pathogenesis of osteoporosis. Overall, this study reveals for the first time a novel link between monocyte HSP27 and BMD.

Development of an ELISA for sPSP94 and utility of the sPSP94/sPSA ratio as a diagnostic indicator to differentiate between benign prostatic hyperplasia and prostate cancer.

BACKGROUND The serum PSA (sPSA) test has low specificity for prostate cancer (PCa), since sPSA also rises in benign prostatic hyperplasia (BPH). Serum PSP94 (sPSP94), a major secreted prostate protein, is indicated as a PCa marker. The potential of sPSP94 and sPSA in conjunction with each other to improve specificity of diagnostic test for PCa needs to be evaluated. METHODS PCa patients (n=33), BPH patients (n=44) and healthy controls (n=50) were recruited. A serum-based sandwich ELISA was developed to measure sPSP94 concentrations. Utility of sPSP94 in improving specificity of sPSA test was evaluated by studying sPSP94/sPSA ratios of study participants. RESULTS Considerable decrease in overlap among sPSP94/sPSA ratio values of BPH and PCa patients was observed, as compared to sPSP94 or sPSA alone. For differentiating between BPH and PCa patients, this ratio had a maximum area under the curve (AUC) of 0.859 (P=0.0132) and had a comparable sensitivity (90.91%) to sPSA with an increased specificity of 70.45%. Further, decision curve analysis (DCA) showed that sPSP94/sPSA ratio had a superior net benefit in identifying PCa, in patients opting for biopsy. CONCLUSION The sPSP94/sPSA ratio can be a better differentiating marker between BPH and PCa, than sPSP94 or sPSA alone.

Hypothalamic hamartomas and precocious puberty

Ten children, five boys and five girls with true precocious puberty at an early age were found to have hypothalamic hamartomas on brain imaging. Very early onset of puberty, varying from a few weeks to three years of age, and rapid progression were characteristic. Accelerated growth velocity and markedly advanced bone age were evident in all. Gonadotropin and gonadal hormone levels were elevated above the prepubertal range. Six children had associated developmental delay or hyperactivity.

Screening infertile women for the assessment of corpus luteal function and their response to therapy by ELISA of pregnanediol glucuronide

Corpus luteal function was assessed by estimating pregnanediol 3-alpha-glucuronide (PdG) in three midluteal-phase urine samples collected from 85 women attending the infertility clinic. The previously established cut off limits based on PdG estimations were useful in detecting anovulation in 23 cases, corpus luteal adequacy in 42 cases and corpus luteum deficiency (CLD) in 20 cases. In 8 women CLD could be corrected with 50 mg of clomiphene citrate (CC) therapy whereas 6 women required 100 mg of CC and 3 pregnancies were achieved. This rapid screening method is thus useful in segregating a large number of women according to their ovulatory status and in the subsequent treatment of CLD.

Metachlopromide-induced hyperprolactinemia fails to affect ovarian cyclicity in the common marmosets (Callithrix jacchus)

Intramuscular administration of metachlopromide (2.5, 5, and 10 mg) induced a dose-dependent increase in plasma prolactin levels. The magnitude and duration of metachlopromide-induced hyperprolactinemia were also dose related. However, metachlopromide treatment (5 mg/day) for 60 days failed to affect ovarian function in the common marmoset as evidenced by ovulatory plasma estradiol and progesterone profiles. During the pretreatment cycle, there was no consistent pattern in plasma prolactin levels depending on the stage of cycle. During lactation, higher levels of plasma prolactin were observed.

Serum levels of phosphorylated heat shock protein 27 (pHSP27) are associated with bone mineral density in pre- & postmenopausal women: A pilot study

Background & objectives: Phosphorylated heat shock protein 27 (pHSP27) has been implicated in the pathogenesis of osteoporosis. Oxidative stress and proinflammatory cytokines, which are known to be involved in aetiology of osteoporosis, can trigger HSP27 phosphorylation. Since pHSP27 is present in circulation, it was hypothesized that serum pHSP27 would be elevated in low bone mineral density (BMD) condition and might serve as an indicator of osteoporosis/osteopenia. Hence, the aim of this study was to examine serum levels of pHSP27 in relation with BMD in pre- and postmenopausal women. Methods: Premenopausal (30 to 40 yr) and postmenopausal (50 to 60 yr) women having either low BMD (osteopenia/osteoporosis) or high BMD were selected (n=80) from a prospective cohort (n=200). Serum levels of pHSP27; along with levels of oestradiol, malondialdehyde, total antioxidant capacity, interleukin (IL)-1, IL-6, tumour necrosis factor - alpha, (TNF-α), c-telopeptide fragments of collagen type I (CTX-1) and osteocalcin were estimated. Results: The serum levels of pHSP27 were significantly elevated in low BMD groups in premenopausal and postmenopausal categories (P<0.05). It also exhibited a significant odds ratio (OR) to differentiate between low and high BMD in both premenopausal (OR=1.734, P=0.013) and postmenopausal (OR=1.463, P=0.042) categories. Additionally, area under the curve to predict low BMD was non-significantly higher for pHSP27 than CTX-1 in premenopausal and postmenopausal categories. Interpretation & conclusions: This study highlights a novel relation between serum pHSP27 and BMD in Indian women however, these findings need to be confirmed in larger studies.