Prevalence of high-risk human papilloma virus types and cervical smear abnormalities in female sex workers in Chandigarh, India.

Purpose: Cervical cancer is the most common cancer among women in developing nations. Nearly 90% of the cases have been linked to the presence of high-risk human papillomavirus (hrHPV) types 16 and 18. The risk of cervical cancer may be high in female sex workers (FSWs) due to multiple sexual partners. This study aimed to determine the prevalence of cytological abnormalities and hrHPV types 16 and 18 in FSWs in Chandigarh, North India using the liquid-based cytology (LBC) approach. Materials and Methods: The cervical brush samples were collected from 120 FSW and 98 age-matched healthy controls (HCs). These were subjected to pap smear using conventional method, LBC and the detection of hrHPV types 16 and 18 was carried out using polymerase chain reaction. Results: The LBC samples showed better cytological details and also reduced the number of unsatisfactory smears from 11% in Pap to 1.5% in the LBC. A significantly higher number of inflammatory smears were reported in FSWs (51.7% vs. 34.7%, P = 0.01). The hrHPV types 16/18 were detected in 33/120 (27.5%) FSW versus 23/98 (23.5%) HCs. The risk of acquiring hrHPV was higher in FSWs, who had age at first sex ≤25 years, higher income and the habit of smoking. Conclusion: The high prevalence of hrHPV among FSWs and HCs suggests the need for the implementation of effective National Screening Programme for early detection of hrHPV types to decrease the burden of cervical cancer, especially in high-risk population.

Evaluation of Pathogenic Potentialities and Transmissibility of Aerobic and Anaerobic Bacteria in Sexually Transmitted Diseases.

Aims: Sexually transmitted diseases (STDs) caused by various aerobic and anaerobic bacteria have been reported from many developed and developing countries of the world. However, there is limited data available on the association of these pathogens with STDs on the Indian sub-continent. Therefore, the aim of this is to the presence of anaerobic and aerobic bacteria in sexually transmitted infections. Study Design: Patients attending the Department of Dermatology, Venereology & Leprology, Calcutta Medical College & Hospital, suspected to be suffering from STDs, were thoroughly examined and those having typical lesions of chancroid were excluded Original Research Article British Journal of Medicine & Medical Research, 4(27): 4533-4541, 2014 4534 from further work. The prevalence of different aerobic and anaerobic bacteria was determined from among the remaining patients after ruling out cases of chancres. Place and Duration of Study: This work was carried out in the Department of Oral Medicine, R. Ahmed Dental College; Department of Dermatalogy, Venereology & Leprology, Calcutta Medical College and Hospital; Division of Microbiology, Department of Pharmaceutical Technology, Jadavpur University and Department of Microbiology, Herbicure Healthcare Bio-Herbal Research Foundation, Kolkata, for a period of seven months from April 1 to October 31, 2012. Methodology: At least two smears were prepared from the infected ulcers of each of the patients who were not diagnosed as of chancroid. One smear was for dark ground microscopy and the other for Gram’s staining. Confirmation of chancre was by serological testing, while aerobes/anaerobes were identified following standard procedures. Results: The organisms isolated were Staphylococcus aureus, Shigella flexneri, Shigella sonnei, Gardenerella vaginalis, Actinomyces spp, Veillonella purvula, Peptococcus heliotrinreducens, Peptostreptococcus magnus and Peptostreptococcus hydrogenalis. These were subjected to tests for their antibiotic sensitivity pattern which was followed by successful specific therapy. Conclusion: Various Gram positive and Gram negative aerobes and anaerobes were found to be associated with STDs and these were transmissible among homosexual and heterosexual partners.

Evaluating the role of low-speed centrifugation towards transfecting human peripheral blood mononuclear cell culture.

The conventional method of transfection of suspension cells by chemical has proven to be very difficult. We present a new transfection protocol, wherein, low‑speed centrifugation of cell culture plates immediately after adding the lipid: DNA complex significantly enhances the transfection efficiency. Peripheral blood mononuclear cells (PBMCs) were transfected with BLOCK‑iT™ Fluorescent Oligo (scrambled siRNA) and lipofectamine complex using conventional and low‑speed centrifugation modified transfection protocols. The efficiency of transfection was determined using flowcytometer and cell viability was checked using MTT assay. Incorporation of low‑speed centrifugation significantly enhances the transfection efficiency of BLOCK‑iT™ in the suspension culture of PBMCs as compared to conventional transfection method (99.8% vs 28.3%; P < 0.0001), even at a low concentration of 40 picomoles without affecting the cell viability. Centrifugation enhanced transfection (CET) technique is simple, time‑saving and novel application without compromising the cell viability in the context of recently popular RNA interference in suspension cultures of PBMCs. This undemanding modification might be applicable to a wide variety of cell lines and solve crucial problem of researchers working with RNA interference in suspension cultures.

Evaluation of antigenicity and cell mediated immunity of Hepatitis E virus patients: Using non radioactive MTT assay.

Hepatitis E virus (HEV) is an important cause of hepatitis in developing nations. Disease spans from asymptomatic infection to acute viral hepatitis (AVH) and acute liver failure (ALF). Cell-mediated immunity (CMI) is less studied. Studies document CMI in HEV patients using [ 3 H]-thymidine incorporation (radioactive in nature). The aim of this study was to evaluate the antigenicity of recombinant HEV ORF 2 peptide (452-617 a.a) (pORF2) by non-radioactive MTT assay and detecting the proliferation indices of primary PBMC culture. A total of 27 laboratory confirmed HEV patients (16 AVH and 11 ALF) and 20 apparently healthy individuals (HC) were included. PBMCs were isolated, plated and stimulated with pORF2. After an incubation of 4 days, cells were looked for blastogenic transformation and subjected to MTT assay. PI of AVH, ALF and healthy controls were found to be 3.249 ± 0.219, 1.748 ± 0.076 and 0.226 ± 0.017, respectively. PI of AVH Vs HC, ALF Vs HC and AVH Vs ALF were found to be significantly higher ( P < 0.0001). This study demonstrates MTT to be an adaptable technique to evaluate CMI in HEV patients. Recombinant pORF2 was found to be antigenic in nature and PBMCs from AVH patients were immunologically more reactive than ALF patients.

Isozymic variations in specific and nonspecific esterase and its thermostability in silkworm, Bombyx mori L.

Esterase isozymic variations were documented in the haemolymph of developed multivoltine and bivoltine silkworm breeds during unfavorable seed crop seasons of May – September using α- and β- napthylacetate separately to identify specific and nonspecific esterase having thermotolerant potentiality. Variations existed in the isozyme pattern with three bands (Est-2, 3 and 4) in pure Nistari race and other developed multivoltine and bivoltine breeds. Est-2 and Est-3 were non-specific esterases as they were observed when both α- and β-napthylacetate was used as substrates separately. Est- 4 band was observed only with α-napthylacetate as substrate and was therefore confirmed to be specific α-esterase band in the haemolymph of silkworm, Bombyx mori L. Zymograms showed that the non-specific esterase band (Est-3) with Rf of 0.43 and specific α-esterase band (Est-4) with Rf of 0.32 predominately withstood a temperature of 70 ± 2oC for a duration of 10 min and were confirmed as thermostable esterases in haemolymph of silkworm, Bombyx mori L. This also categorized the presence of thermostable esterases in developed multivoltine and bivoltine breeds of silkworm, even though the qualitative activity was more in the former than the latter. The qualitative presence of thermostable esterases and their activity could be adopted as an indicative biochemical marker in relation to thermotolerance in silkworm.

Prolonged jaundice attributed to super infection of hepatitis E virus in a case of resolving leptospirosis.

India is endemic for both Leptospira and hepatitis E virus (HEV). The clinical presentations of these diseases have overlapping features. We report a case of superinfection of HEV in a patient with resolving leptospirosis with underlying Hodgkin lymphoma. The diagnosis of HEV in our case was established by HEV-RNA PCR as our patient was immunosuppressed. The present study highlights the need for molecular diagnosis in the case of HEV infection with strong clinical suspicion and negative serological results.

Mycetoma caused by a new red grain mycetoma agent in two members of a family.

An 18-year-old woman from rural West Bengal was affected with mycetoma involving her neck, back, and chest. After an interval of eight years, her younger brother developed mycetoma on his left arm. No history of trauma or immune deficiency was present in either case. By microscopic examination of sinus-discharged materials from both the cases, identical rusty red, hard grains were demonstrated. Soluble red pigment-producing colonies grew in Sabouraud dextrose-agar medium. Isolates were positive for casein hydrolysis and negative for hydrolysis test of xanthine, hypoxanthine, tyrosine, and nitrate reduction. Thus it differed from the only known red grain mycetoma agent, Actinomadura pelletieri and was provisionally identified as Actinomadura vinacea. Familial affection in mycetoma, that too caused by a new agent, is reported here for its uniqueness.

Three years of cadaveric renal transplantation.

AIM: To study patient and graft outcome and donor characteristics in cadaveric renal transplantation. MATERIAL AND METHODS: Fifty nine patients (35 males and 24 females) who underwent cadaveric renal transplantation between Oct'95 and Nov'98 were analysed. The recovery of renal function was correlated with the type and duration of dialysis. The number of rejections, donor characteristics and patient and graft survival was studied. RESULTS: Fifty nine patients were followed for a mean period of 14.58 months (SD 10.73), their age ranged from 17 to 64 years. Ten patients had primary graft failure, out of which four died and six are on maintenance dialysis. Seven died of other causes. Twelve patients had been on CAPD and 47 were on haemodialysis. Recovery of renal function was earlier in the haemodialysis group (mean 19.29 days SD 16.97) than in the CAPD group (31.6 days SD 32.3 days). But the difference was not statistically significant (p=0.36). The age of the donors ranged from 2 to 72 years (mean 42.8, SD 17.3). Seventeen died of CVA, 16 died of head injury, and one died of sedative overdose. Dual kidney transplant was done in nine patients of whom five have adequate graft function. CONCLUSION: Follow up for a mean period of 14.58 months shows patient survival of 81.3% and graft survival 72.8% with no significant difference in recovery of renal function between CAPD and HD groups. Poor results are due to HLA mismatch leading to rejection. The majority of the deaths were related to septicaemia.

Computer simulation of the kinetics of the sodium pump.

A simple kinetic model for the membrane-bound enzyme, 'Na+, K(+)-ATPase', is presented. An equation for the efflux of Na+ and influx of K+ is derived. This also explains multiple ligand binding of homospecies of these cations and competition between heterospecies of these cations. The equation is computed by a small BASIC program and its validity is sought by comparing the results with a few experimental data.

Survival of orally administered isolated intestinal Lactobacillus acidophilus in different parts of gastrointestinal tract of mice.

A strain of Lactobacillus acidophilus (Strain HF) was isolated from human faeces. A chloramphenicol resistant strain (HFCm) and a strain (HFCmSm chloramphenicol and streptomycin were developed from the isolated strain (HF). All the ) restant to both Kyd Street, Calcutta 700 016 three strains showed similar in vitro susceptibility against host defence factors like gastric acid, bile salts and volatile as well as non-volatile fatty acids. In vivo tests were done by feeding these strains to mice. When the resistant strains were orally administered along with the antibiotic(s) they were stable up to 72 h.