ABSTRACT
The present study was aimed to analyse the effect of acrylamide and Hybanthus enneaspermus leaf extract active principles on mice testis glutathione-s-transferases (GST; EC 2.5.1.18). These enzymes play a role in biotransformation of electrophilic compounds that cause damage to cells by conjugating with the substrate glutathione. Hybanthus enneaspermus, a spade flower, is an erect shrub of violaceae family, having free radical scavenging activity. Acrylamide is a known neurotoxicant that cause damage to almost all cells including liver, testis, brain and kidney. The GSTs purified from mice testis using glutathionyl linked agarose affinity chromatography were analyzed by using SDS-PAGE and were resolved into four sub units i.e. Yc, Yb, Yβ &Yδ. Also these subunits expression were confirmed by western blot analysis. During experimentation to analyze the effect of Hybanthus enneaspermus active principle (HE) mice were subjected to both acrylamide (AC) and also mixture of HE and AC. This exposure significantly altered the specific activity of mice GSTs in testis. Polyclonal antibodies produced against purified GSTs of mice testis on immunoblot analysis showed significant increase of μclass GSTs (Yb & Yβ) based on dose and time dependent manner. Therefore the present research of Hybanthus enneaspermus treatment on mice testis showed that, regulation of synthesis of μ-GSTs was depending on the dose of acrylamide concentration and also the active principles of HE. Hence it is proposed that μ-GSTs may be used as tumour markers for testis carcinoma, since their production is variable due to the increased dose concentration of synthetic chemical acrylamide and its regulation by plant product, HE.
ABSTRACT
PURPOSE: To evaluate the role of the radiometric BACTEC 460TB system and the conventional Lowenstein-Jensen (LJ) medium for isolation of M. tuberculosis from cerebrospinal fluid (CSF) samples of tuberculous meningitis (TBM) patients. METHODS: CSF specimens (n=2325) from suspected TBM patients were processed for isolation of mycobacteria by inoculating BACTEC 12B medium and the LJ medium. The isolation of mycobacteria in both media was confirmed by microscopy and biochemical identification. Drug sensitivity testing for the anti-TB drugs was carried out by BACTEC radiometric method. RESULTS: Among the total 2325 CSF specimens processed by both methods, M. tuberculosis was isolated from 256 specimens. The isolation rates were 93% and 39% for the BACTEC system and LJ medium respectively. Both the media supported growth in 32% of the culture-positive specimens. BACTEC system alone yielded growth in 61% and LJ alone in 7%, of the culture-positive specimens. Among 205 isolates tested for drug susceptibility 81% were sensitive to all the drugs tested and 19% were resistant. CONCLUSIONS: The BACTEC 460TB system provides a highly sensitive and rapid tool for the isolation and drug susceptibility testing of M. tuberculosis, from CSF of TBM patients. Use of a solid medium in conjunction with the BACTEC 12B medium is essential for optimal recovery for M. tuberculosis from CSF specimens.