Study of genetic polymorphism in solvent exposed population and its correlation to in vitro effect of trichloroethylene on lymphocytes.

Trichloroethylene (TCE) is major industrial pollutant that contaminate environment. Its exposure may lead to hepato-renal toxicity along with the cancer progression. Although extensive research is done on its toxicity, still not much is known about its genotoxic potential on humans in relation to genetic polymorphism. Cytochrome P450 (CYP P-450) and glutathione-S-transferases (GSTs) are important in cellular detoxification of TCE. Variations in gene sequences result in population specific regional genetic variations (polymorphism). Genotyping of CYP1A1, GSTM1, GSTT1 and GSTP1 polymorphism was performed in 220 normal and 97 solvent-exposed individuals from northern part of India using real time PCR, PCR and restriction digestion techniques. The parameters examined to study genotoxicity were chromosomal aberration (CA) and cytokinesis block micronucleus assay (CBMN) in lymphocyte culture in vitro. The observed average frequencies for GSTM1 (null) and GSTT1 (null) were 41, 22 and 12.7%, respectively in normal subjects whereas frequencies of CYP1A1/GSTP1 with (ile/ile) or (ile/val) or (val/val) were found to be 76.2/52, 21.4/42.1 and 2.4/5.9% respectively. It was further observed that the frequencies of above genes were found to be similar in solvent exposed groups. The distribution frequencies of GST genes, when compared with other reports from various regions of India show variations. In vitro TCE exposure (2, 4 and or 6 mM) did not show any significant genotoxic effect. TCE may be toxic due to its metabolite.

Ventilator-associated pneumonia caused by carbapenem-resistant Enterobacteriaceae carrying multiple metallo-beta-lactamase genes.

Context: Ventilator-associated pneumonia (VAP) is a leading nosocomial infection in the intensive care unit (ICU). Members of Enterobacteriaceae are the most common causative agents and carbapenems are the most commonly used antibiotics. Metallo-beta-lactamase (MBL) production leading to treatment failure may go unnoticed by routine disc diffusion susceptibility testing. Moreover, there is not much information on association of MBL-producing Enterobacteriaceae with ICU-acquired VAP. Therefore, a study was undertaken to find out the association of MBL-producing Enterobacteriaceae with VAP. Settings: This study was conducted in a large tertiary care hospital of North India with an eight-bed critical care unit. Materials and Methods: The respiratory samples (bronchoalveolar lavage, protected brush catheter specimens and endotracheal or transtracheal aspirates) obtained from VAP patients (during January 2005-December 2006) were processed, isolated bacteria identified and their antibiotic susceptibilities tested as per standard protocols. The isolates of Enterobacteriaceae resistant to carbapenem were subjected to phenotypic and genotypic tests for the detection of MBLs. Results: Twelve of 64 isolates of Enterobacteriaceae were detected as MBL producers, bla IMP being the most prevalent gene. Additionally, in three strains, simultaneous coexistence of multiple MBL genes was detected. Conclusion: The coexistence of multiple MBL genes in Enterobacteriaceae is an alarming situation. As MBL genes are associated with integrons that can be embedded in transposons, which in turn can be accommodated on plasmids thereby resulting in a highly mobile genetic apparatus, the further spread of these genes in different pathogens is likely to occur.

Aeromonas caviae septicemia in immunocompetent gastrointestinal carriers

Aeromonas caviae strains have been isolated from blood and stool cultures of three immunocompetent patients, residents of Northern India, who presented with community acquired septicemia without any recent history of diarrhea. Cell culture infectivity test performed on Hep-2 cells have shown substantial degree of invasiveness in the isolated strains. This case unleashes a possibility of asymptomatic gastrointestinal carriage of such strains of A. caviae in a very large population of India, as several areas of India have very high rates of Aeromonas induced acute diarrhea/gastroenteritis (up to 13 percent). It needs to be appraised further in India as well as other countries having high rates of Aeromonas induced acute diarrhea/gastroenteritis.

Dorsal onlay buccal mucosal graft urethroplasty in long anterior urethral stricture

OBJECTIVE: To assess the success of buccal mucosal graft (BMG) urethroplasty by the dorsal onlay technique in long anterior urethral stricture (> 2 cm long) through the midline perineal incision. MATERIALS AND METHODS: From January 1998 to December 2003, 43 patients with long anterior urethral strictures were managed by dorsal onlay BMG urethroplasty. After voiding trial, they were followed up at 3 months with uroflowmetry, retrograde urethrogram (RGU) and American Urological Association symptoms score (AUA symptoms scores). Successful outcome was defined as normal voiding with a maximum one attempt of VIU after catheter removal. Patients were further followed-up with uroflowmetry at 3 months interval and RGU every 6 months interval. RESULTS: Mean stricture length was 4.8 cm (range 3 to 9 cm) and mean follow up was 48 months (range 12 to 84 months). Only five patients were found to develop stricture at anastomotic site, during follow-up. Two of them voided normally after single attempt of VIU. Other three patients (6.9 percent) required further open surgery or repeat VIU during follow up and were considered as failure. CONCLUSION: Dorsal onlay BMG urethroplasty is a simple technique with good surgical outcome.

Urethrorectal fistula by bull horn injury.

Bull horn injuries are common in rural India. Here one such case causing urethrorectal fistula has been reported. The patient was presented with history of bull horn injury 6 hours back. He was examined under general anaesthesia and found to have lacerations in the anus and anterior wall of rectum. Urgent retrograde urethrography and cystography showed partial rupture of bulbar urethra and urethrorectal fistula. Initially sigmoid colostomy and suprapubic cystostomy was done. Later optical internal urethrotomy was done. The patient was catheterised for 3 weeks and the fistula healed completely.

Alteration in the acceptor side of photosystem II of chloroplast by high light.

The effect of high light on the acceptor side of photosystem II of chloroplasts and core particles of spinach was studied. Both Vmax and apparent Km for DCIP were altered in photoinhibited photosystem II core particles. The double reciprocal plot analysis as a function of actinic light showed increased slope in chloroplasts photoinhibited in the presence of DCMU. Exposure of chloroplasts to high light in the presence of DCMU did not protect the chloroplast against high light induced decrease in Fm, level. Further the high light stress induced decrease in Fm level was not restored by the addition of DCMU. These results suggest that the high light stress induced damage to chloroplast involves alteration in the binding site for QB on the DI protein on the acceptor side of photosystem II.

Cytochrome b6-f complex : The carburettor of exciton distribution in oxygenic photosynthesis.

Efficient oxygenic photosynthesis not only requires synchronous turover and operation of photosystem I (PS I) and photosystem II (PS II) but also the preferential turnover of PS I for cyclic photophosphorylation to maintain required ATP and NADPH ratio during carbon dioxide reduction. Ohe initial higher rate of turnover of PS II in viva is accounted by the fact that (i) PS I contains only about one-third of total chlorophylls, (ii) about 90% of light harvesting a/b protein (LAC) which accounts for about 50% of the total chlorophylls, remains associated with PS II as PS II-LHC II complexes (PS IIa) and (iii) the ratio of PS II/PS I is always greater than unity, in the range of 1-2 : 1 under different environmental regimes. Ohe initial preferential feeding of PS II, due to its larger antenna, is bound to result in faster rate of turn over of PS II than PS I, leading to higher rate of reduction of an intersystem carrier than the rate of its oxidation by PS I. Ohe light dependent phosphorylation of a 'mobile' and small pool (– 20%) of LHC II of PS IIα (possibly located at the edge of appressed regions of the membranes) increases the repulsive forces of LHC II resulting in its migration to non-appressed region associating itself with PS 1. Ohe phosphorylation itself is controlled by the redox state of an intermediate of electron transport. Several experimental approaches have provided evidence which suggest that (i) phosphorylation of LAC II involves interaction of cyt b5-f complex with LAC II kinase and the interaction of QA with cyt b5-f complex and (ii) different kinases may be involved in phosphorylation of LHC II versus PS II polypeptides. Ohe major purpose of light dependent LAC II phosphorylation and its consequent migration close to PS I appears to balance the rate of cyclic versus non-cyclic photophosphorylation. Ohe mechanism by which cyt b5-f complex controls the activation of LAC II is not known. Ohe role of membrane bound ealmodulin, electron transfer through cyt b6-f complex in activation of LAC II kinase should be explored.

Mechanism of reductive photoactivation of enzymes of C4 pathway.

Light, besides initiating primary photochemical processes, alters the redox state of soluble components in chloroplast. The present review attempts to cover the mechanism of reductive photoactivation of enzymes of photosynthetic carbon reduction cycle using key enzymes as examples. The reduced soluble components — ferredoxin, thioredoxin and NADPH, in turn, cause the reduction of disulphides to dithiols of chloroplastic enzymes. NADP-malate dehydrogenase is subject to activation by light through changes in NADPH/NADP. The key enzyme of C4 photosynthesis-PEP carboxylase, though cytosolic, has been shown to be activated by disulphide/sulphhydryl interconversion by reductants generated in light through chloroplast electron transport flow. Pyruvate Pi dikinase activity is controlled by the adenylate energy charge. It remains unclear how light controls the activation of cytosolic enzymes.

Site-site heterogeneity in isocitrate lyase of castor seed endosperm: evidence from kinetics of inactivation by tetranitromethane.

Inactivation of isocitrate lyase (native and EDTA-dialysed) by excess tetranitromethane (TNM) exhibits, biphasic kinetics, in which half of the initial activity is lost in a fast and the remaining half in a slow phase each following the pseudo-first order kinetics. Rate constants of the two phases are proportional to the TNM concentration. High succinate concentration protects the enzyme against TNM inactivation only in the slow phase without any effect on the fast phase. With the EDTA-dialysed enzyme, no such protection (against inactivation by TNM) is observed in the presence of succinate or Mg2+ ions. Addition of both these ligands together brings about protection against the slow phase (as with the native enzyme). It has been proposed that the site-site heterogeneity of isocitrate lyase is a consequence of its quaternary structure constraints.