LH/FSH ratio: a better marker of secondary amenorrhea in patients from eastern Nepal.

FSH, LH and prolactin (PRL) levels were assessed by ELISA in 50 cases with secondary amenorrhea and 52 age and sex-matched healthy controls from eastern Nepal. Cases were diagnosed by differential diagnosis, and data were analyzed using standard statistical tools. Early stage (3-6 months) and long standing (> 6 months) secondary amenorrhea had no effect (p > 0.05) in hormonal parameters studied. Pulse, SBP, DBP, weight, height, age of menarche, cycle interval and duration of flows were homogenous (p > 0.05) in patients and controls. Median age of menarche, median cycle interval and median duration of flows in healthy subjects were 14 years, 30 and 4 days respectively. FSH in cases (15.38 +/- 7.24 mU/ml) was significantly elevated (p < 0.01) as compared to controls (9.38 +/- 6.34 mU/ml). LH in cases (35.44 +/- 24.35 mU/ml, median 36.5 mU/ml) was significantly (p < 0.01) elevated by almost 5 times of its mean value and 9 times of its median value as compared to that of controls (7.58 +/- 6.604 mU/ml, median 4.2 mU/ml). LH/ FSH ratio in cases (2.44 +/- 1.73, median 2.00) was significantly higher (p < 0.01) as compared to controls (0.82 +/- 0.42, median 0.76). FSH e"12 mU/ml, LH e"10 mU/ml and LH/FSH ratio e"1 cut offs were significantly associated (p = 0.000 in each) with the cases as revealed by chi-square analysis, and LH/FSH ratio e"1 (Sensitivity = 84.0%, specificity = 77.0%) was found to be a stronger marker of secondary amenorrhea. As the elevation of LH was more pronounced than that of FSH, this study hints towards possible LH receptor mutation, which is generally found in premature ovarian failure (POF). Diagnosis of cases in this region may need a new cut off level for POF, as the elevation of FSH itself was not as pronounced as reported by other workers.

Age related changes in cholesterol level among the patients with subclinical hypothyroidism in eastern Nepal.

This study was undertaken at Biochemistry Department (BPKIHS) from 2001 to 2002. The aim of this study was to assess whether subclinical hypothyroidism is associated with abnormal cholesterol level in different age groups. Of the cases referred to the department, 398 euthyroid controls, 189 hypothyroid cases, 179 hyperthyroid cases and 181 subclinical hypothyroid cases were enrolled for further analyses. Both the sexes showed increased total cholesterol levels in hypothyroidism, and were not significantly changed in hyperthyroidism than in euthyroid controls. Female subclinical hypothyroid cases of age group 45-59 years and both the male and female cases of age group > or = 60 years had significantly (p < 0.01) elevated total cholesterol levels (196 +/- 37.36 mg/dl vs.169.37 +/- 29.12 mg/dl, 211.5 +/- 30.48 mg/dl vs. 151.54 +/- 55.84 mg/dl and 225 +/- 25.05 mg/dl vs. 181.73 +/- 32.95 mg/dl respectively) as compared to euthyroid controls. When data were analyzed at 33 and 45 years of age cut offs, the cases of > or = 33 years age in both hypothyroid male and female showed significantly (p < 0.01) elevated cholesterol levels (222.66 +/- 29.26 mg/dl vs. 156 +/- 37.09 mg/dl and 231.66 +/- 46.17 mg/dl vs 198.1 +/- 48.72 mg/dl respectively) where as subclinical hypothyroid female showed increased total cholesterol level (211 +/- 31.2 mg/dl vs. 157.95 +/- 45.92 mg/dl) at > or = 45 years age cut off. It is concluded that hypothyroidism not the hyperthyroidism is associated with increased total cholesterol level in the cases of this region, and we advise routine screening for cholesterol level in both the sexes of hypothyroid (for > or = 33 years) and female subclinical hypothyroid (for > or = 45 years) cases in this area, as they have high risk for higher cholesterol levels and developing related disorders.

Easy approach to DNA shuffling: its potential implication in health sciences.

Directed evolution experiments rely on the cyclical application of mutagenesis, screening and amplifications in a test tube. During the laboratory evolution of biological molecules, recombination has been used to generate novel sequences in a process known as DNA shuffling. DNA shuffling is a recently developed technique that allows accelerated and directed protein evolution for desired properties in vitro, which recombines and evolves genes to rapidly obtain molecules with improved biological activity and fitness. DNA shuffling is generally achieved by DNaseI treatment and by PCR. This has led to the creation of novel proteins for a wide range of applications. The use of improved enzymes for medical, industrial and environmental purposes is prevalent today, and will be expanding. New applications in vaccine development and disease diagnosis are among the key features of DNA shuffling. However, directed evolution currently requires an uncertain, typically large number of labor-intensive and expensive experimental cycles before proteins with improved function are identified. A simplified and low-cost DNA shuffling protocol for random recombination of homologous genes in vitro is described here.