Impact of GSTM1, GSTT1 and GSTP1 gene polymorphism and risk of ARV-associated hepatotoxicity in HIV-infected individuals and its modulation.

Glutathione S-transferase (GST) family is involved in a two-stage detoxification process of a wide range of environmental toxins, carcinogen and antiretroviral (ARV) therapy (ART) drugs. The aim of this study is to describe the impact of genetic polymorphisms of GSTM1, GSTT1 and GSTP1-313A/G in the risk of ARV-associated hepatotoxicity in HIV-infected individuals and its modulation in hepatotoxic patients. We enrolled a total of 34 patients with hepatotoxicity, 131 HIV-infected individuals without hepatotoxicity under non-nucleoside reverse transcriptase inhibitor containing ART and 153 unrelated healthy individuals. With a case-control design, polymorphisms of GSTM1, GSTT1 and GSTP1-313A/G gene were genotyped by PCR and restriction enzyme-length polymorphism. Genotypes of GSTT1 null were significantly higher in HIV-infected individuals as compared with healthy controls (P=0.01, odds ratio (OR)=1.54). HIV-infected individuals with GSTM1-null genotype showed higher risk (P=0.09, OR=1.37) for hepatotoxicity, but risk was not significant. On evaluating gene-gene interaction models, GSTM1 null and GSTT1 null showed significant association with the risk of hepatotoxicity in HIV-infected individuals (P=0.004, OR=2.67) owing to synergistic effect of these genes. Individuals with GSTT1-null and GSTM1-null genotypes showed higher risk of hepatotoxicity with advanced stage of (CD4<200) of HIV infection (P=0.18, OR=1.39; P=0.63, OR=1.13). In case-only analysis, GSTT1-null genotype among alcohol users showed elevated risk of hepatotoxicity in HIV-infected individuals (P=0.12, OR=1.36, 95% confidence interval (CI): 0.94-1.97) as compared with GSTT1 genotypes. The carriers GSTM1-null+GSTT1-null genotype among nevirapine user showed prominent risk of hepatotoxicity in HIV-infected individuals (P=0.12, OR=4.21, 95% CI: 0.60-29.54). Hence, we can conclude that GSTT1-null and GSTM1-null genotypes alone and in combination may predict the acquisition of hepatotoxicity.

A tumor deconstruction platform identifies definitive end points in the evaluation of drug responses.

Tumor heterogeneity and the presence of drug-sensitive and refractory populations within the same tumor are almost never assessed in the drug discovery pipeline. Such incomplete assessment of drugs arising from spatial and temporal tumor cell heterogeneity reflects on their failure in the clinic and considerable wasted costs in the drug discovery pipeline. Here we report the derivation of a flow cytometry-based tumor deconstruction platform for resolution of at least 18 discrete tumor cell fractions. This is achieved through concurrent identification, quantification and analysis of components of cancer stem cell hierarchies, genetically instable clones and differentially cycling populations within a tumor. We also demonstrate such resolution of the tumor cytotype to be a potential value addition in drug screening through definitive cell target identification. Additionally, this real-time definition of intra-tumor heterogeneity provides a convenient, incisive and analytical tool for predicting drug efficacies through profiling perturbations within discrete tumor cell subsets in response to different drugs and candidates. Consequently, possible applications in informed therapeutic monitoring and drug repositioning in personalized cancer therapy would complement rational design of new candidates besides achieving a re-evaluation of existing drugs to derive non-obvious combinations that hold better chances of achieving remission.

Occurrence of carcinoma prostate in the Pune Metropolitan Region: A 5-year study from 2007 to 2011.

BACKGROUND: A retrospective study was carried out to know only the occurrence of carcinoma prostate (CAP) in the Pune Metropolitan Region (PMR) over a period of five years (January 01, 2007 to December 31, 2011). All the histopathological (HPE) reports of all prostate specimens were collected from 23 medical colleges, private institutions and stand-.alone HPE laboratories in PMR. MATERIALS AND METHODS: Four types of prostate specimens were examined - endoscopic resection, open prostatectomy, transrectal needle biopsy of prostate and prostate from the cystoprostatectomy specimen (surgery carried out for primary carcinoma bladder). Specimens of radical prostatectomy were excluded as the biopsy was carried out earlier. RESULTS: A total of 5006 reports of the prostate specimens were examined out of which 779 showed the presence of CAP. Analysis of annual occurrence of CAP revealed that there was no significant variation in the CAP cases. Thus giving an average of CAP cases of 155.8 per year in PMR. Population data of the PMR was obtained from the official Government of India Census department for the year 2011. Total population of PMR in 2011 was 5,049,968 out of which the male population was 2,659,484. Thus the occurrence of CAP in the PMR works out to 5.86/100,000 male population. Results were compared with the published reports of CAP by the Indian Council of Medical Research (ICMR) of 7.2/100,000 and Globocon of 4.2/100,000 males. CONCLUSIONS: Occurrence of CAP in PMR is low at 5.86/100,000 male population is comparable with published figures of ICMR & Globocon 2012.

Removal of migrated metallic prostatic stent by holmium laser.

A 90-year-old male with prostatic hyperplasia with a history of ischemic heart disease and right-sided hemiplegia had undergone a Urolume stent placement because of acute urinary retention 9 months earliar. The stent had migrated into the bladder causing dysuria and a poor stream of urine. We fragmented the prostatic stent by Holmium (HO: YAG) laser followed by a laser prostatectomy. After the procedure, the patient voided satisfactorily.

Is stenting required before retrograde intrarenal surgery with access sheath.

BACKGROUND: Flexible ureterorenoscopies continue to assume an increasing role in the armamentarium of the endourologist. In many centers around the world, prior stenting is carried out before retrograde intrarenal surgery (RIRS) to passively dilate the ureter, which facilitates passage of a flexible ureteroscope with or without an access sheath. In our series, the first stage of passive dilatation with prior stenting was totally avoided without compromising the success of the procedure. MATERIALS AND METHODS: From January 2004 to December 2007, 54 patients with 55 renal units underwent RIRS. The patients were between 28 and 65 years old. All patients had renal stones ranging in size from 8 mm to 22 mm. The mean serum creatinine level was 1.1 mg%. The lower ureter was dilated under 'C - arm' fluoroscopy guidance up to 14 FR. An access sheath of 10/12 Fr was passed over the working guide wire. RIRS (7.5/9.3 Fr) was introduced into the access sheath. The stones were fragmented using a holmium laser. The mean operating time was 85 mins (45-130 mins). RESULTS: In 52 out of 55 renal units (94.5%), a flexible ureteroscope could be passed successfully into the kidney through an access sheath. In 3 of the cases (5.4%), the lower ureter could not be dilated. In these patients, the procedure was staged after passing a 6/26 JJ stent. An X-ray KUB was done at the 3-month follow-up visit. A total of 50 renal units (94.3%) were stone free at the 3-month follow-up visit. CONCLUSION: In a majority of the cases, RIRS could be accomplished successfully during the first sitting. Single stage RIRS did not alter the subsequent stone clearance or increase the incidence of morbidity or complications.

Evolution of cancer stem cells.

Cancer as a disease driven by cancer stem cells is a concept that has emerged over the last few years. However, several issues relating to this phenomenon as yet remain unaddressed. A fundamental question is one relating to the identification of events leading to transformation of a normal tissue stem cell to a cancer stem cell. Complete knowledge of this evolutionary process may be crucial for the development of novel effective therapies that influence patient prognosis. The scope of this review is to discuss reports that have begun to elucidate stem cell transformation either as an isolated event or as a progression as an attempt towards understanding some of the critical events involved in the process.

Use of continuous positive airway pressure during spinal anaesthesia in a patient with severe chronic obstructive pulmonary disease.

We report the case of a patient with severe chronic obstructive pulmonary disease who underwent local resection of a carcinoma of the rectum under spinal anaesthesia. Although the patient was keen to avoid general anaesthesia and to have the operation under a spinal anaesthetic, pre-operative assessment showed that he could not lie flat. As the surgical procedure required the patient to be in the lithotomy position, ideally with a head-down tilt, it was hoped that continuous positive airway pressure with a facemask during spinal anaesthesia might help him to tolerate the position comfortably. Continuous positive airway pressure at 7.5 cmH(2)O was successfully used to facilitate breathing during surgery under spinal anaesthesia. A combination of regional anaesthesia and continuous positive airway pressure via a facemask is easy to use and may be a useful option in the management of these challenging patients.

Nuclear-mitochondrial genomic profiling reveals a pattern of evolution in epithelial ovarian tumor stem cells.

Analyses of genome orthologs in cancer on the background of tumor heterogeneity, coupled with the recent identification that the tumor propagating capacity resides within a very small fraction of cells (the tumor stem cells-TSCs), has not been achieved. Here, we describe a strategy to explore genetic drift in the mitochondrial genome accompanying varying stem cell dynamics in epithelial ovarian cancer. A major and novel outcome is the identification of a specific mutant mitochondrial DNA profile associated with the TSC lineage that is drastically different from the germ line profile. This profile, however, is often camouflaged in the primary tumor, and sometimes may not be detected even after metastases, questioning the validity of whole tumor profiling towards determining individual prognosis. Continuing mutagenesis in subsets with a mutant mitochondrial genome could result in transformation through a cooperative effect with nuclear genes - a representative example in our study is a tumor suppressor gene viz. cAMP responsive element binding binding protein. This specific profile could be a critical predisposing step undertaken by a normal stem cell to overcome a tightly regulated mutation rate and DNA repair in its evolution towards tumorigenesis. Our findings suggest that varying stem cell dynamics and mutagenesis define TSC progression that may clinically translate into increasing tumor aggression with serious implications for prognosis.

Snail and Slug are major determinants of ovarian cancer invasiveness at the transcription level.

OBJECTIVES: The transcriptional factors Snail and Slug have been reported to be important in cell migration during development and also during tumor metastasis. Their expression and role in ovarian cancer, hitherto unexplored, was examined to understand the molecular events in ovarian cancer metastases since the latter is responsible for the high degree of mortality associated with the disease. METHODS: Ectopic expression of mSnail and mSlug in the epithelial ovarian cancer cell line SKOV3 was carried out and stable clones were selected. These were used to examine specific repression of the adherens, tight and desmosomal junction components by the two transcription factors. Furthermore, functional implications with respect to enhanced migration of cells, tumorigenecity and metastasis were also studied. RESULTS: The ectopic expression of Snail or Slug resulted in epithelial-mesenchymal transition (EMT), enhanced motility, invasiveness and tumorigenecity in the cell line SKOV3. In addressing the mechanism by which Snail and Slug lead to loss of intercellular adhesion, specific repression of adherens junction components (E-cadherin and betacatenin), tight junction components (Occludin and ZO-1) and desmosomal junction components (Dsg2) were observed. Snail suppresses expression of adherens and tight junction components, while Slug suppresses expression of all the three junction components; concertedly, bringing down the intercellular adhesion between cells. Further activation of these transcriptional factors in hypoxic conditions revealed a rapid upregulation of Slug expression as an immediate reaction that probably triggers off a signaling cascade leading to Snail expression. CONCLUSIONS: These results indicate distinct roles of the transcriptional factors Snail and Slug during ovarian cancer metastasis and cell survival through mediation of EMT.

A study of sensitivity and resistance of pathogenic micro organisms causing UTI in Kathmandu valley.

UNLABELLED: A retrospective study was conducted in five hospitals to observe the prevalence of organisms causing UTI and their sensitivity to antibiotics. METHODOLOGY: Altogether, data from five hundred samples of urine from five hospitals in Kathmandu was collected for this study from January 2005 to April 2005. RESULT: A total of 244 samples were found to be positive. Altogether six types of organisms were isolated as the causative factors. E. coli (49%), S. aureus, (coagulase positive) (23%), Proteus species (3.6%), Klebsiella (9.71%), Pseudomonas (0.8%) and Citrobacter (2.8%). Analysis of the samples showed that UTI was more common in females of younger age group as compared to males. The common age group for females was 21-30 years, whereas that for males was 31-40 years in all the hospitals except in hospital A, where the maximum number of females was from 31-40 years and males were between 71-80 years. The most common organism to cause UTI was found to be E. coli (49%), followed by S. aureus (23%) and Klebsiella (9.71%). All the organisms causing UTI were sensitive to nitrofurantoin and amoxycillin and ciprofloxacin was found to be least effective. Similarly, in three hospitals, B (88.2%), D (64.7%) and E (65.3%), amoxycillin was found to be most effective, amikacin and gentamycin (92.5%) was most effective in hospital C, and nitrofurantoin in hospital A (78%). The second commonest organism, i.e., S. aureus (23%) was most sensitive to cephalosporin (88.8%) of second generation, followed by nitrofurantoin (77.7%), amikacin (80.6%) and norfloxacin (65.5%). The third common organism, Klebsiella (9.71%) was most sensitive to norfloxacin (75%) and nitrofurantoin (75%). Lastly, Pseudomonas was resistant to all the antibiotics in hospital A, D and E, nil in hospital B and sensitive to amikacin (100%) in hospital C.

Medical education - changing perspectives.

Various aspects of medical education have been reviewed with special reference to medical institutions in Nepal. The newer trends in teaching methodology like audiovisual and computer aided methods are being followed in most of the institutions of Nepal. Similarly, attempts are being made to implement integrated teaching which, though not perfect, differs from institution to institution. The attempts seem to be more or less satisfactory with awareness amongst most of the teachers and planners. However, the methods of assessment of the students at every level, call for changes and improvement, in the light of modern trends. Similarly, the process of selection in medical colleges needs change. The replies to the questionnaire given to the students of two batches were very interesting and it is worthwhile to undertake such studies in other institutions as well. Valid suggestions opined by them should be implemented.

Stem cell pharmacogenomics.

Therapeutic stem cell applications represent a newly evolving approach for the treatment of several genetic and degenerative diseases. The advent of pharmacogenomics too, holds promise for an individualized, optimal treatment regime for a large variety of medical conditions. A combination of the benefits of these two technologies creates a new niche in therapeutic medicine research viz. that of stem cell pharmacogenomics (SCP). The development of this approach requires the application of existing technologies in genomics, proteomics and bioinformatics to resolve the various issues involved in advancing the therapeutic applications of stem cell medicine. In this brief overview of the subject, we attempt to provide fresh insights into the exclusive niche of stem cell pharmacogenomics and discuss some of the priority issues that need to be targeted, based on the existing principles of pharmacogenomics, stem cell characteristics and transplantation medicine. Advances in these areas are imperative in realizing the dream of stem cell therapies contributing towards the improvisation of the quality of human life.

Percutaneous nephrolithotomy for complex pediatric renal calculus disease.

BACKGROUND AND PURPOSE: Pediatric renal calculus disease has been a management dilemma in view of the concern about the effects of the various treatment modalities on the growing kidney, the significant recurrence rate, and the long-term outcome. We report our experience with percutaneous nephrolithotomy (PCNL) monotherapy in staghorn or complex pediatric renal calculi. PATIENTS AND METHODS: We retrospectively analyzed the case records of 116 patients younger than 15 years who underwent PCNL. The stones included 56 complex calculi. We defined complex calculi as either staghorn (complete or partial) or those with a large bulk and involving more than one calix, the upper ureter, or both. RESULTS: Complete clearance was achieved in 50 patients (89.8%). Of these, 22 (39%) required a single tract, while 34 (61%) required multiple tracts. With subsequent SWL, the clearance rate increased to 96%. The average hemoglobin drop was 1.9 g/dL. Assessing the factors affecting the hemoglobin drop, the number of tracts and the size of tracts were found to be significant (P<0.01). The average change in the serum creatinine concentration between the preoperative and postoperative measurements was +0.03 mg/dL and was not different in patients with a single tract and those with multiple tracts (+0.02 and +0.04 mg/dL, respectively; P=NS). Intravenous urography done in 36 renal units postoperatively revealed good function in all. A DMSA renal scan in six children showed no scar. CONCLUSIONS: Monotherapy with PCNL is safe and effective in the management of staghorn and complex renal calculi in single hospital stay. Ultrasound-guided peripheral caliceal puncture and limiting the tract dilatation to 22F are important factors in reducing the blood loss. Multiple tracts increase the hemoglobin drop but are not associated with an increased risk of complications (bleeding, postoperative infection, and prolonged urinary leak). Also, there is no deterioration in renal function after either single- or multiple-tract PCNL.

Peroxynitrite activates mitogen-activated protein kinase (MAPK) via a MEK-independent pathway: a role for protein kinase C.

In this study we show that phosphorylation of extracellular signal-regulated kinase (ERK1/2; also known as p44/42MAPK) following peroxynitrite (ONOO(-)) exposure occurs via a MAPK kinase (MEK)-independent but PKC-dependent pathway in rat-1 fibroblasts. ONOO(-)-mediated ERK1/2 phosphorylation was not blocked by MEK inhibitors PD98059 and U0126. Furthermore, no increase in MEK phosphorylation was detected upon ONOO(-) treatment. Staurosporine was used to investigate whether protein kinase C (PKC) is involved. This was confirmed by down-regulation of PKC by phorbol-12,13-dibutyrate, which resulted in significant reduction of ERK1/2 phosphorylation by ONOO(-), implying that activation of ERK by ONOO(-) depends on activation of PKC. Indeed, PKCalpha and epsilon were activated upon ONOO(-) exposure. When cells were treated with ONOO(-) in a calcium-free buffer, no activation of PKCalpha was detected. Concomitantly, a reduction of ERK1/2 phosphorylation was observed suggesting that calcium was required for translocation of PKCalpha and ERK phosphorylation by ONOO(-). Indeed, ONOO(-) exposure resulted in increased cytosolic calcium, which depended on the presence of extracellular calcium. Finally, data using Gö6976, an inhibitor of calcium-dependent PKC activation, implied that ONOO(-)-mediated ERK1/2 phosphorylation depends on activation of a calcium-dependent PKC.

Neonatal myiasis.

This case involves an orphan female neonate-abandoned in a dustbin in Poona, India-who was infected by the larval forms of the blowfly. The blowfly causing this infestation belonged to the family Calliphoridae and genus Calliphora. The fly of this genus is of importance in Indian veterinary science and is found abundantly around decaying matter in Poona. The larvae occurring in carrion, flesh, etc, usually infest open wounds of animals and rarely infest humans.