Association of interleukin‑28B rs12979860 and rs8099917 polymorphisms with sustained viral response in hepatitis C virus genotype 1 and 3 infected patients from the Indian subcontinent.

Background: Polymorphisms of the IL28B gene (rs12979860 and rs8099917) have been shown to impact treatment responses in hepatitis C virus (HCV) infected patients. The association of these polymorphisms with sustained viral response (SVR) has been studied in HCV genotype 3 infected patients in India, but not in genotype 1. Objectives: This study aimed to determine the association of IL28B gene polymorphisms and other host and viral factors with treatment response in patients with HCV genotype 1 and 3 infection. Materials and Methods: DNA from 42 HCV‑infected patients on antiviral therapy was analysed for the IL28B polymorphisms using polymerase chain reaction‑restriction fragment length polymorphism (PCR‑RFLP). Bidirectional sequencing was performed on a subset of samples for verification of PCR‑RFLP results. Information on age, weight, height, diabetic status, pre‑treatment viral load and alanine aminotransferase (ALT) levels was obtained from clinical records. The IL28B genotypes and the other factors were analysed for their association with SVR. Results: The frequency distribution of rs12979860 CC/CT/TT genotypes was found to be 66.7%, 26.2% and 7.1%, respectively. For rs8099917 genotype, the TT/GT/GG distribution was 73.8%, 21.4% and 4.8%, respectively. SVR was seen in 61.9% of cases (55.6% in genotype 1 and 62.5% in genotype 3). CC genotype at rs12979860 and TT genotype at rs8099917 were significantly higher in responders (P = 0.013 and 0.042, respectively). Lower baseline ALT and rapid viral response were also found to be associated with SVR. On logistic regression analysis, CC genotype at rs12979860 emerged as the most powerful predictor of treatment response. Conclusion: IL28B polymorphisms are strong predictors of SVR in patients from the Indian subcontinent infected with HCV genotype 3 and genotype 1.

Mother to child transmission of hepatitis B virus: A cause for concern.

Mother to child transmission of hepatitis B virus (HBV) is an important public health issue. India introduced HBV vaccine in 10 states as part of its Universal Immunization Program (UIP). Here we show evidence of mother-to-child transmission of HBV in three families from Jharkhand and Bihar states where HBV vaccination is not yet included in the UIP. This report illustrates the need for active screening of HBV in pregnant women and implementation of HBV vaccine across all states in India to reduce the burden of disease.

In-vitro antibacterial activity of coral reef associated bacteria and optimization of bioactive metabolite production.

This study reports the antibacterial activity of coral reef associated bacteria against bacterial pathogens and optimization of metabolite production. Twelve morphologically different bacterial were isolated from stony coral reef collected from Rameshwaram coastal area, Tamil Nadu. Cell free supernatant of all the bacterial isolates were tested against Staphylococcus aureus, Bacillus cereus, Salmonella typhi, Escherichia coli, and Klebsiella pneumoniae by agar well diffusion method. Of the 12 isolates tested, seven isolates showed antibacterial activity again at least one of the test organisms with the zone of inhibition in the range of 9-14 mm. The antibacterial compounds were extracted from the cell free supernatant using ethyl acetate and tested for antibacterial activity. In diffusion method, crude extract from strain RC12 showed 11-17 mm inhibition against all the test pathogens and the same was selected for optimization studies. Effect of culture conditions and medium components such as different carbon, nitrogen, mineral, pH, temperature on antibacterial metabolite production was studied by adopting one-factor-at-a- time method. The metabolite production was influenced by lactose, ammonium sulphate, K2HPO4 while the optimal cultivation conditions were pH 7 and 37°C. The potent strain RC12 was identified as Pseudomonas sp based on their phenotypic and biochemical characteristics. The finding of the present study concludes that coral reef bacteria found to be the best source for bioactive metabolites with broad spectrum activity.

Bioactive potential of actinobacteria against drug resistant pathogens.

The present investigation was focused to study the antagonistic potential of actinobacterial strain TK2 isolated from Thirukurungudi Hills (Western Ghats, Tamil Nadu), against selected drug resistant bacterial pathogens. Of the 9 clinical bacterial strains, S. aureus was found to be resistant against methicillin and vancomycin while the remaining 8 gram negative pathogens were confirmed as extended spectrum beta lactamase (ESBL) producers. Strain TK2 showed good antagonistic activity against all the bacterial pathogens, except the three isolates of P. aeruginosa. Strain TK2 produced the antimicrobial activity on 4th day of incubation when growing on ISP2 agar medium whereas the same strain showed activity only on 8th day of incubation when it was grown on ISP2 broth. Of the various solvents tested for extraction, bioactive compound was extracted only in ethyl acetate. The crude extract showed 14-18 mm inhibition zone in disc diffusion method. The crude extract produced two spots in thin layer chromatography (TLC) in chloroform: methanol (30:60) solvent system. In bioautography, the second spot (Rf value 0.685) showed activity. The active compound was purified by preparative TLC, which showed maximum activity (15-20 mm inhibition) against test pathogens. Based on the results of chemical screening, the active compound was identified as sugar containing substance. Strain TK2 showed good growth on various growth media and culture conditions. Based on the studied phenotypic characteristics strain TK2 was identified as the species of the genus Streptomyces. Of the various growth parameters tested, ISP2 medium, glucose, pH 7, 1% NaCl and temperature 300C was influenced the antagonistic activity of strain TK2. Strain TK2 will be a potential for the isolation of bioactive compound(s) which will be a candidate for the development of antibiotic against drug resistant bacterial pathogens.

Exploitation of bacteria from forest ecosystem for antimicrobial compounds.

This study deals with bacterial prospecting from forest soil with special reference to antimicrobial substances. Total of 25 morphologically different bacterial colonies were isolated from soil samples collected from Anaimalai forest and Parambikulam tiger reserve forest, Western Ghats, India. About 12 (48%) out of 25 isolates showed antibacterial activity in which strain AF1 showed inhibitory activity against more number of test pathogens. Bioactive substance from strain AF1 was produced by adopting submerged fermentation and extracted using ethyl acetate and chloroform. In disc diffusion method, ethyl acetate extract showed good antibacterial activity (9-17 mm zone of inhibition). Active fraction present in the ethyl acetate extract was determined by thin layer chromatography based bioautography. Findings of this work supported that the forest ecosystems investigated in this study will be potential place for bacterial bioprospecting.

Biosynthesis of gold nanoparticles by actinomycete Streptomyces viridogens strain HM10.

Biosynthesis of gold nanoparticles by Streptomycetes from Himalayan Mountain was undertaken for the first time. Out of 10 actinomycete strains tested, four strains (D10, HM10, ANS2 and MSU) showed evidence for the intracellular biosynthesis of gold nanoparticles, among which the strain HM10 showed high potency. Presence of spherical and rod shaped gold nanoparticles in mycelium of the strain HM10 was determined by transmission electron microscopy (TEM) and X-ray diffraction analysis. The average particle size ranged from 18-20 nm. UV spectral analysis indicated that the reduction of chloroauric acid (HAuCl4) occurred within 24 h of reaction period. Further, the strain HM10 showed enhanced growth at 1 and 10 mM concentration of HAuCl4. The gold nanoparticles synthesized by the strain HM10 showed good antibacterial activity against S. aureus and E. coli in well-diffusion method. The potential actinomycete HM10 strain was phenotypically characterized and identified as Streptomyces viridogens (HM10). Thus, actinomycete strain HM10 reported in this study is a newly added source for the biosynthesis of gold nanoparticles.

Studies on extended beta lactamase producing, biofilm forming clinical bacterial pathogens and its invitro inhibition by Actinobacterial extracts.

At present scenario, the extended spectrum beta lactamase (ESBL) producing bacterial pathogen causes various life threatening infections especially by the members of the family Enterobacteriaceae in hospital settings. In order to study the prevalence of ESBLs in Kanchipuram hospital, the bacterial strains were isolated from patients having Urinary tract infections (UTI), diabetic foot ulcer, pregnancy women’s, surgical wound infections, deep wounds, and genitourinary tract problems. Totally 40 bacterial isolates were recovered from 30 samples and the isolates were identified as Escherichia coli (45%), Pseudomonas sp, (25%) and Klebsiella sp (30%). The ESBL production was confirmed with third generation cephalosporins (cetixime, cephoxitin, ceftazidime, cetepime, ceftriaxone, ceftazidime/clavulanic acid) using the Kirby- bauer disc diffusion method and also by double disc diffusion method. The highest ESBL production was found among E. coli (42%), followed by Pseudomonas sp. (25%) and Klebsiella sp (20%). All the ESBL producer were tested for biofilm formation by tube method in which E.coli (43%) was found to be the good biofilm producer followed by the Klebsiella sp (31%) and Pseudomonas sp (25%). An attempt was also made to study the in-vitro inhibition of biofilm forming ESBL pathogens by actinobacterial extracts by disc diffusion method. Of the five actinobacterial extracts tested, extracts produced from the strain MA7 inhibited (8-12 mm zone of inhibition) all the biofilm forming ESBL pathogens. Further purification and characterisation of active compound from actinobacterial strain MA7 is in progress.

Inadvertent insertion of nasogastric tube into the brain.

An unusual case of a nasogastric (Ryle's) tube inserted in the brain in a patient having severe head injury with skull base fractures is reported here along with a brief review of literature. A 35 years male was referred from a peripheral institute following head trauma with endotracheal tube and nasogastric tube in situ. A CT scan of the brain showed multiple skull base fractures and a high parietal extradural hematoma. It also revealed that the nasogastric tube had inadvertently found its way into the brain through the lamina cribrosa of the ethmoid bone. The tube was removed under aseptic conditions in the operation theatre but the patient expired on day 2 of admission due to the head injuries sustained.

Inverse Marcus Gunn phenomenon.

Inverse Marcus-Gunn phenomenon is very rare. It is usually acquired. We report a young male patient presenting with congenital ptosis and inverse Marcus-Gunn phenomenon.

Turritella attenuata (Kasinathan): as biological indicator of marine pollution--a trace metal analytical study.

A study to monitor marine pollution with reference to trace elements (Fe, Zn, Mn and Cu) on T. attenuata, commonly called as screw shell over a period of one year on the whole body and various organs, viz. digestive diverticula, foot, mantle and ovary was conducted from the sandy beach of Porto Novo Coast (Lat 11 degrees 29' N Long: 79 degrees 46' E) of Peninsular India using Atomic Absorption Spectrophotometer (AAS). Higher concentration of all the four trace metals analysed were recorded in the digestive diverticula, whereas lower concentration of zinc and manganese were recorded in the ovary during the monsoon period. The higher level of trace metal concentration in the monsoon period may be due to the presence of these pollutants in large amounts in water. The accumulation of selected trace metals varies in different seasons according to the extent of pollution load in the marine environment.