Subclinical hypothyroidism in eastern Nepal: a hospital based study.

BACKGROUND: Subclinical hypothyroidism itself is associated with serious complications and also there is a known risk of subclinical hypothyroidism patients getting converted into overt disease. OBJECTIVES: The objective of the present study was to fi nd out the prevalence of subclinical hypothyroidism in the suspected cases i.e. amongst the cases attending the thyroid laboratory at B.P. Koirala Institute of Health Sciences, Dharan, Nepal. MATERIALS AND METHODS: It was a retrospective cross sectional study. Data of the free T3, free T4 and TSH estimations of the year 2007 of the Thyroid lab at BPKIHS, Dharan, Nepal was analyzed. ELISA based free T3, free T4 and TSH tests in the serum had been performed in all the cases. RESULTS: Total cases were 1714 including 24.446% males and 75.554% females. Cases with raised TSH levels were 26.021%, cases with normal TSH levels were 54.66% and cases with low TSH levels were 19.316%. Total 350 cases (20.42 %) had subclinical hypothyroid dysfunction which includes 84 (4.901 %) males and 266 (15.519%) females. And the maximum percentage of cases in either gender was between the age groups 20 -59 years. CONCLUSION: The prevalence of subclinical thyroid hypothyroidism amongst the suspected cases was 20.42 % which is much higher compared to the other parts of the world. The highest percentage was found in the female age group 20 - 59 years. The routine screening of the whole population is not cost effective and on the basis of the present study it is suggested that there may be routine screening of the selected populations, especially women between 20 to 59 years of age in Nepal region. The preferred screening method advised is a sensitive ELISA based TSH test.

Acetoxy-4-methylcoumarins confer differential protection from aflatoxin B(1)-induced micronuclei and apoptosis in lung and bone marrow cells.

The ability of various acetoxy derivatives of 4-methylcoumarins to inhibit the genotoxic changes due to aflatoxin B(1) (AFB(1)) is reported here. Several 4-methylcoumarins (test compounds), such as 7,8-diacetoxy-4-methylcoumarin (DAMC), monoacetoxy-4-methylcoumarin (MAC), 5-N-acetyl-6-acetoxy-4-methylcoumarin (NAMC) and 7,8-dihydroxy-4-methylcoumarin (DHMC) were separately administered intraperitoneally (i.p.) to male wistar rats followed by AFB(1) administration i.p. or intratracheally (i.t.) (2-8 mg/kg b.wt.) and another dose of the test compound. The animals were sacrificed 26h after AFB(1) administration. From animals receiving AFB(1) i.p., bone marrow (BM) cells were isolated and stained with Mayer's haematoxylin and eosin. Micronuclei (MN) in BM were scored by light microscopy. From animals receiving AFB(1) i.t., bronchoalveolar lavage (BAL) was obtained, lung cells (LG) were isolated and stained with fluorochrome 6-diamidino-2-phenylindole (DAPI) for the analysis of MN, apoptotic bodies (AP) and cell cycle variations. Rats were separately treated with the vehicle DMSO to serve as the proper control. AFB(1) caused significant dose-dependent induction of MN in BM as well as LG. AP were observed in LG of rats receiving AFB(1) and was found to correlate with MN induction. DAMC injection caused significant decrease in AP due to AFB(1) in LG and MN in both BM and LG. The effectiveness of MAC was approximately half that of DAMC, thereby indicating that number of acetoxy groups on the coumarin molecule determine the efficacy. The fact that NAMC had no effect either on MN or AP indicate that neither acetoxy group at C-6 nor the N-acetyl group at C-5 facilitate the transfer of acetyl group to P-450 required for inhibition of AFB(1)-epoxidation. DHMC, the deacetylated product of DAMC had no normalizing effect on the induction of MN and AP. These findings confirm our earlier hypothesis that DAMC-mediated acetylation of microsomal P-450 (catalysing epoxidation of AFB(1)) through the action of microsomal transacetylase is responsible for the protective action of DAMC. The relative number and position of acetoxy groups on the coumarin nucleus determine the specificity to the transacetylase necessary for the chemopreventive action.

Distribution and ethnic variation of â-thalassemia mutations in Nepal.

This is the first study characterizing spectrum of beta-thalassemia mutations in Nepalese population. Mutations were analyzed in 22 patients using 10 sets of allele-specific primers. Five of the mutations, namely F.S 41/42 (--TCTT), IVS1 nt5 (G-->C), IVS1 nt1 (G-->T), 619 bp deletion and F.S 8/9 (+G), were found to constitute 87.82% of total alleles studied. F.S 41/42 (--TCTT) was the commonest mutation. -88 (C-->T), Codon 16 (--C) and Codon 15 (G-->A), had a combined frequency of 12.18%. Distribution of mutations causing beta-thalassemia in different ethnic Nepalese groups was analyzed. The mutational profile in Nepal share several similarities with that from the two neighboring countries, India and China. Detection of more than one mutation in three cases of thalassemia trait raises the likelihood of existence of multiple mutations in cis in Nepalese thalassemic carriers. Such possibility has to be carefully considered while developing prenatal screening program for Nepalese population.

Aflatoxin B1-induced micronuclei and cell cycle alterations in lung and bone marrow cells and their modulation by Piper argyrophyllum extract.

Aflatoxin B, (AFB1) is a clastogen that causes cellular damage by covalent modification of nucleic acids. In this investigation, male rats were injected i.p. with AFB1 (8 mg/kg b.w.) in DMSO and the same dose of AFB1 was also administered intratracheally (i.t.) to the animals separately. The animals were killed after 26 h of the carcinogen treatment, femur bone was removed, and bone marrow cells were isolated and stained with Mayer's hematoxylin and eosin. Micronuclei (Mn) were scored by using light microscopy. Bronchoalveolar lavage (BAL) was prepared from rats administered AFB1 i.t. A part of BAL was fixed with 70% ethanol, stained with the fluorochrome DAPI, and analysed for cell cycle variations; the other part of the lavage was used for making slides to record Mn with a fluorescent microscope. A significantly greater proportion of lung cells were found to enter cell cycle with extended S-phase due to AFB1 treatment. Mn were induced in polychromatic erythrocytes (PCE) as compared to normochromatic erythrocytes (NCE) in the bone marrow of AFB1-treated rats, where there was nearly a three-fold increase in the number of Mn of bone marrow cells. The administration of AFB1 resulted in a two-fold rise in the Mn in the lung cells. The effect of BSO, DEM, and PB, the modulators of AFB1 metabolism, was studied on AFB1-induced Mn formation. A significant increase in the Mn score in PCEs of BSO- and DEM-treated rats was noted, while a slight reduction in the Mn score was noted in the case of PB-treated rats. The administration of the methanol extract of the leaves of Piper argyrophyllum (taken up in DMSO) to rats for a week exhibited normalising effect on AFB1-induced Mn in bone marrow cells. These observations record the induction of Mn in lung cells due to AFB1 for the first time. We propose the utility of AFB1-induced Mn as a model for screening plant extracts as inhibitors of genotoxicity. Prevention of genotoxic changes described above by phytochemicals is being pursued in our Laboratories.

Chemoprevention of benzene-induced bone marrow and pulmonary genotoxicity.

Our earlier studies documented the ability of 7,8-diacetoxy-4-methylcoumarin (DAMC) to cause irreversible inhibition of cytochrome P-450 linked mixed function oxidases (MFO) mediated by membrane bound DAMC: protein transacetylase. Since P-450 catalyzed oxidation of benzene is crucial to its toxic effects, the action of DAMC and related analogues were considered promising in preventing the genotoxicity due to benzene. For this purpose rats were pretreated with various acetoxy-4-methylcoumarins (test compounds), which was followed by the administration of benzene either intratracheally (IT) or intraperitoneally (IP), and sacrificed 26 h after the injection of benzene. The incidence of micronuclei (MN) in bone marrow (BM) and lung (LG) were assessed by light and fluorescent microscopy, respectively. A dose-dependent induction of MN in BM and LG cells was observed in rats administered with benzene. A significant reduction in benzene-induced MN in BM and LG was observed as a result of DAMC administration to rats; a higher dose of DAMC resulted in greater inhibition of clastogenic action of benzene as revealed by MN incidence. 7,8-dihydroxy-4-methylcoumarin (DHMC), the deacetylated product of DAMC, demonstrated relatively lesser potency to inhibit the clastogenic action of benzene. This observation is consistent with the ability of DAMC to inhibit the formation of benzene oxide as well as to scavenge the oxygen radicals formed during the course of benzene metabolism. The fact that DHMC can only scavenge the oxygen radicals and is ineffective in inhibiting benzene oxidation in vivo explains the reduced capability of dihydroxy coumarin to prevent MN due to benzene. 7-Acetoxy-4-methylcoumarin (MAC) inhibits the MN due to benzene being roughly 50% of that produced by DAMC. DAMC is also effective in normalizing the cell cycle alterations produced by benzene in BM and LG. These observations further substantiate our hypothesis that the biological effects of acetoxy coumarins are mediated by the action of membrane bound transacetylase that catalyzes the acetylation of concerned proteins. Teratogenesis Carcinog. Mutagen. 21:181-187, 2001.

Mechanism of biochemical action of substituted 4-methylbenzopyran-2-ones. Part II: Mechanism-based inhibition of rat liver microsome-mediated aflatoxin B1-DNA binding by the candidate antimutagen 7,8-diacetoxy-4-methylcoumarin.

7,8-Diacetoxy-4-methylcoumarin (DAMC), with no prerequisite for oxidative biotransformation has been reported to produce suicide inactivation of microsomal cytochrome P-450-catalysed formation of aflatoxin B1-8,9-oxide that binds to DNA. Parenteral administration of DAMC to rats caused significant inhibition of AFB1 binding to hepatic DNA in vivo as well as AFB1-induced micronuclei formation in bone marrow cells. These results highlight the antimutagenic potential of DAMC.