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BACKGROUND: Cross-species horizontal gene transfer (HGT) involves the transfer of genetic material between different species of organisms. In recent years, mounting evidence has emerged that cross-species HGT does take place and may play a role in the development and progression of diseases. METHODS: Transcriptomic data obtained from patients with gallbladder cancer (GBC) was assessed for the differential expression of antisense RNAs (asRNAs). The Basic Local Alignment Search Tool (BLAST) was used for cross-species analysis with viral, bacterial, fungal, and ancient human genomes to elucidate the evolutionary cross species origins of these differential asRNAs. Functional enrichment analysis and text mining were conducted and a network of asRNAs targeting mRNAs was constructed to understand the function of differential asRNAs better. RESULTS: A total of 17 differentially expressed antisense RNAs (asRNAs) were identified in gallbladder cancer tissue compared to that of normal gallbladder. BLAST analysis of 15 of these asRNAs (AFAP1-AS1, HMGA2-AS1, MNX1-AS1, SLC2A1-AS1, BBOX1-AS1, ELFN1-AS1, TRPM2-AS, DNAH17-AS1, DCST1-AS1, VPS9D1-AS1, MIR1-1HG-AS1, HAND2-AS1, PGM5P4-AS1, PGM5P3-AS1, and MAGI2-AS) showed varying degree of similarities with bacterial and viral genomes, except for UNC5B-AS1 and SOX21-AS1, which were conserved during evolution. Two of these 15 asRNAs, (VPS9D1-AS1 and SLC2A1-AS1) exhibited a high degree of similarity with viral genomes (Chikungunya virus, Human immunodeficiency virus 1, Stealth virus 1, and Zika virus) and bacterial genomes including (Staphylococcus sp., Bradyrhizobium sp., Pasteurella multocida sp., and, Klebsiella pneumoniae sp.), indicating potential HGT during evolution. CONCLUSION: The results provide novel evidence supporting the hypothesis that differentially expressed asRNAs in GBC exhibit varying sequence similarity with bacterial, viral, and ancient human genomes, indicating a potential shared evolutionary origin. These non-coding genes are enriched with methylation and were found to be associated with cancer-related pathways, including the P53 and PI3K-AKT signaling pathways, suggesting their possible involvement in tumor development.

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Background: The cancer registry provides reliable data from the population. In this article, we provide cancer burden and its patterns from the Varanasi district. Methods: The method adopted by the Varanasi cancer registry is community interaction along with regular visits to more than 60 sources to collect data on cancer patients. The cancer registry was established by the Tata Memorial Centre, Mumbai, in 2017 covering 4 million population (57% rural and 43% urban population). Results: The registry has recorded 1,907 incidence cases (1,058 male and 849 female). The age-adjusted incidence rate per 100,000 population in male and female of Varanasi district is 59.2 and 52.1, respectively. One in 15 male and one in 17 female are at risk of developing the disease. Mouth and tongue cancers are the predominant cancers in male, whereas breast, cervix uteri, and gallbladder are the leading cancer sites among the female. In female, cervix uteri cancer is significantly higher (double) in rural areas when compared with urban areas (rate ratio [RR] 0.5, 95% confidence interval [CI; 0.36, 0.72]), whereas in male, mouth cancer is higher in urban areas when compared with rural areas (RR 1.4, 95% CI [1.11, 1.72]). More than 50% of cancer cases in male are due to tobacco consumption. There may be underreporting of the cases. Conclusion: The results of the registry warrant policies and activities related to early detection services for the mouth, cervix uteri, and breast cancers. The Varanasi cancer registry is the foundation for cancer control and will play an important role in the evaluation of the interventions.

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Marjolin's ulcer (MU) with rapid progression and multiple distant metastases is rare. We report a case of an MU of the thigh developing in a postburn scar, which after successful R0 resection developed multiple bony and visceral metastasis to femur, vertebra, skull, lung, and liver within 3 months of excision. We highlight the "hurricane-like" recurrence in an MU, which is unusual for the metachronous disease. We stress that MUs, unlike conventional wisdom, need a thorough metastatic workup at presentation, and prognostication of a possible rapidly progressive course after surgery which may lead to grave prognosis and mortality.

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Arginase expression has been recently shown to increase in numerous disease states like neurodegeneration, inflammation, and malignancies. Although it has been found to be functionally important in various disease pathologies, little is known about its role in wound healing. Here, we look at the expression of arginase and its isoforms in chronic non-healing wounds and also study the expression of nitric oxide synthase (NOS) and oxidative stress enzymes in them. Wound tissues and blood samples were collected at the time of index presentation and follow-up from 61 chronic non-healing wound cases. The expression patterns of arginase isoenzymes, NOS, superoxide dismutases (SOD), lactic acid dehydrogenase (LDH), and catalase were examined by using enzyme-linked immunosorbent assay, immunohistochemistry, and western blot analysis at the transcript and protein level. We reported a significant decrease of serum arginase levels in chronic nonhealing wounds in the progress of wound healing. Interestingly, tissue arginase levels were found to be increased with improved wound condition at follow-up. Tissue NOS, LDH, and catalase activity were also found to be increased with the progress of healing, whereas SOD levels were downregulated. Our findings reported increased expression at the transcript level of arginase-I and arginase-II in chronic non-healing wounds for the first time. In conclusion, we observed decreased serum arginase levels in completely healed patients as compared to non-healed cases. Our study findings support the hypothesis that inhibition of the activity of arginase delays wound healing. Arginase and iNOS may also find their place in the future as possible biomarkers for wound healing.

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Fistula-in-ano though not a life-threatening condition, yet its symptoms often significantly impact patients' social, intimate, and work lives. There is an established role of bacterial microflora in acute infections. However, we proposed that biofilm-forming organisms might be present in the microflora of anal fistula of prolonged duration. This aspect has rarely been studied earlier. Therefore, the study describes the microbiology of anal fistula and the biofilm-forming capacity of the isolated organisms. A total of 30 patients were included in the study as per the criteria. Anal fistula tissue sample, tissue fluid, and blood samples were collected from each individual. The collected specimens were detected for the presence of aerobic and anaerobic microflora through standard microbiological method and polymerase chain reaction. Furthermore, the role of biofilm formation by microtitre plate assay and serum matrix metalloproteinases-9 was also studied. The result showed significant predominance of gut-derived microflora with high-to-moderate biofilm-producing ability in anal fistulas of prolonged duration. The study emphasises the presence of biofilm-forming bacteria in chronic, non-healing fistula.

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Nonhealing ulcers are a great challenge to surgeons as they may occasionally culminate in amputation of the affected part. Mostly nonhealing of wounds results due to infection by antibiotic-resistant bacteria and subsequent biofilm formation. However, customized bacteriophage therapy may take care of both of the above-mentioned hurdles. A total of 48 study subjects of age group 12 to 70 years, having minimum one eligible full-thickness wound and failed to heal in 6-week duration with conventional therapy, were included in this exploratory prospective study. Patients with systemic diseases, that is, burn, malignancy, dermatological disorders, and ulcers with leprosy or tuberculosis, were excluded. However, subjects having diabetes and hypertension were included in the study. The customized monophage for single bacterial infection and cocktail of phages specific to 2 or more infecting bacteria were applied on an alternate day over the wound surface. A total of 5 to 7 applications were made till the wound became free of infecting bacteria. The study period extended from August 2018 to May 2019. The study subjects were followed for 3 months since the start of therapy. A cure rate of 81.2% could be obtained, of which 90.5% (19/21) patients were nondiabetic and 74.1% (20/27) diabetic. The wounds infected with Klebsiella pneumoniae were observed with relatively delayed healing. Post phage therapy, the mean hemoglobin level and percentage of lymphocytes increased significantly. The customized local phage therapy is very promising in nonhealing ulcers.

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A cross-sectional study was conducted to assess the utility of online teaching for general surgery postgraduate residents at the end of 1 month of online teaching during the COVID-19 lockdown. A questionnaire related to different aspects of online teaching was developed on a 5-point Likert scale from "strongly agree" to "strongly disagree". Following a pilot study, the questionnaire was shared among surgery residents, and response collection was done. The residents who did not attend at least 90% of scheduled classes were excluded. A total of 55 resident doctors without prior experience of online teaching program were enrolled in the study. All the participants responded to all questions and submitted their responses within stipulated time. On the basis of validity testing, questions 1 and 2 were found invalid and hence deleted. The median value for the total cohort was 4 indicating that majority of residents gave affirmative response. On segregating the result with the year of residency program, similar results were found in the 1st year and 2nd year group; however, the 3rd year group showed a median of 3, thus eliciting that only 50% of 3rd year residents liked online teaching. From the present study, it can be concluded that the online teaching is a feasible alternative to the physical class among surgical postgraduates. The questionnaire utilized in the present study can be used for assessing similar online teaching protocols.

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CONTEXT: WHO in its development of the roadmap on access to medicines and vaccines 2019-2023 has emphasized that the greatest challenge in achieving Universal Health Coverage (UHC) stem from persistent barriers to accessing health services and to accessing affordable and quality assured health products. In this context, WHO introduced the concept of essential medicines in 1977, and since then, this concept, has been revised every 2 years and is accepted by many countries as guideline to develop National Drug Policy. The concept emphasizes that all aspects of drug management, including procurement, storage, distribution, and use, are easier if fewer essential medicines/items must be dealt with. Essential Medicines are those that satisfy the priority healthcare needs of the population. AIMS: During 2017-18, based on the principles of Essential Medicines Concept, we decided to develop and implement "Quality Procurement Management Policies'(QPMP) at Sir Sundar Lal Hospital, at Varanasi. The Pharmaco-econmics and Pharmacovigilance tools are also used to further maximize financial and therapeutic benefits. MATERIALS AND METHODS: This is evidence based, an observational, and retrospective study. Initially, the analysis of current data on sales of antibiotics during November 2017, December 2017, and January 2018, the procurement practices, selection of medicines, and pattern of use was studied and based on these findings reforms were designed and implemented through the Hospital Formulary Committee. RESULTS: This study revealed that- By the application of QPMP, both the Cost and Quality factors of Medication Management can be dealt with very effectively. And it is possible to make available Quality Assured Medicines at most economic prices, resulting in substantial savings. This study has further, shown that the confidence of physicians also improved as quality assured medicines are provided. CONCLUSION: The study at Sir Sundar Lal Hospital is thus worth emulating and shown that a doable roadmap with signposts at each and every turn can be designed and can be effectively implemented so that the goal of UHC can be achieved.

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BACKGROUND: A chronic wound usually results due to halt in the inflammatory phase of wound healing. Bacterial infections and biofilm formation are considered to be the basic cause of it. Chronic wounds significantly impair the quality of life. Antibiotics are now failing due to biofilm formation emergence of drug-resistant bacteria. OBJECTIVE: This study aims to see the effect of bacteriophage therapy in chronic nonhealing wound infected with the following bacteria: Escherichia coli, Staphylococcus aureus, and Pseudomonas aeruginosa. SUBJECT: Patients with chronic nonhealing wound not responding to conventional local debridement and antibiotic therapy were included in the study. The age of patients ranged between 12 and 60 years. METHOD: A total of 20 patients selected and tissue biopsies and wound swabs were taken for isolation of the bacteria. After confirmation of organism, a cocktail of customized bacteriophages was topically applied over the wound on alternate days till the wound surface became microbiologically sterile. Mean bacterial count and clinical assessment were done and compared at the time of presentation and after 3 and 5 doses of application. RESULTS: A significant improvement was observed in the wound healing, and there were no signs of infection clinically and microbiologically after 3 to 5 doses of topical bacteriophage therapy. Seven patients achieved complete healing on day21 during follow up while in others healthy margins and healthy granulation tissue were observed. CONCLUSION: Topical bacteriophage application may be quite effective therapy for the treatment of chronic nonhealing wounds.

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Salmonella enterica serovar Typhi faces several environmental stresses while going through the stomach (acidic pH) to the small intestine (basic pH) and intracellularly in macrophages (acidic pH) in humans. The acidic pH followed by alkaline pH in the small intestine might be responsible for expression of certain stress-induced genes, resulting in not only better survival but also induction of multiplication and invasion of the bacterium in the small intestine. Based on this hypothesis, we developed a process wherein we exposed the blood, urine, and stool specimens from 90 acute typhoid fever patients and 36 chronic typhoid carriers to acidic pH to see the effect on isolation rate of S. Typhi. About 5 g of freshly passed unpreserved stool, a centrifuged deposit of 15 ml of urine, and 5 ml of blood clot were subjected to 5 ml of Luria-Bertani (LB) broth (pH 3.5) for 20 min, followed by enrichment in bile broth-selenite F broth. When the combined isolation from all 3 specimens, i.e., blood, urine, and stool, after acid exposure was considered, a total of 77.7% of the acute typhoid patients were observed to be positive for the isolation of the S. Typhi serotype, compared to 8.8% by the conventional method. Similarly, 42% (15/36) of chronic carriers yielded positive for S. Typhi growth after acid exposure, compared to 5.5% (2/36) by the conventional method. It therefore can be concluded that acid shock triggers the multiplication of the bacteria, resulting in better isolation rates from blood clot, stool, and urine specimens.

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INTRODUCTION: Enteric fever is a systemic disease caused by Salmonella organism such as serotypes Typhi and ParaTyphi A, B, C. Salmonella ParaTyphi A contributes more than 50% of all the enteric fever cases and it has recently been projected as an emerging pathogen. MATERIALS AND METHODS: The present study was aimed to detect Salmonella Typhi and ParaTyphi A in urine, blood and stool specimens collected from cases of enteric fever (110), chronic typhoid carriers (46) and healthy controls (75) to explore the possibility of mixed infection by nested PCR. A new nested PCR primer was designed targeting putative fimbrial protein (stkG) gene which is one of the fimbrial gene families to Salmonella ParaTyphi A and for S. Typhi already reported primers targeting flagellin (fliC) gene. RESULTS: Large volume of urine specimens (15 ml) was found to be the best for detection of Salmonella serotypes. The urine sample was found to have mixed-infection by both the serotypes in 40.9% of the cases but lower in blood (27.3%) and stool (13.6%). CONCLUSION: The present study concludes that occurrence of mixed infection may be quite frequent in typhoid and chronic typhoid carriers' individuals, although the reported recent rise in ParaTyphi A incidence may not be real.

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BACKGROUND: Carcinoma of the gallbladder is the commonest malignancy of the biliary tract in northern India The etiologic relation of specific metals (heavy and trace) and their compounds to neoplasia has been a topic of investigation for some time but not adequately described for carcinoma of the gallbladder. The aim of the present study was to evaluate the relation of heavy and trace metals to this malignancy. METHODS: The levels of selenium, zinc, copper, manganese, cadmium, chromium, lead, and nickel were estimated in serum, bile, gallstones, and gallbladder tissue using atomic absorption spectrophotometry. The tests were carried out in 30 patients with gallbladder cancer and 30 sex-matched patients with cholelithiasis. RESULTS: Selenium and zinc levels were significantly reduced (p < 0.001) and copper concentration was found to be significantly higher (p < 0.001) in serum, bile, and gallbladder tissue from patients with carcinoma of the gallbladder. Lead, cadmium, chromium, and nickel levels were elevated in serum and bile in patients with carcinoma of the gallbladder. CONCLUSIONS: The present study demonstrated a significant decrease in serum, biliary, and tissue levels of selenium and zinc but increased copper, lead, cadmium, chromium, and nickel levels in patients with carcinoma of the gallbladder, indicating a strong relation between the concentrations of these metals and gallbladder cancer.

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INTRODUCTION: It is important to identify Salmonella Typhi infection quickly to treat acute fever patients and to prevent transmission by chronic typhoid carriers; therefore, a very specific and sensitive diagnostic technique is highly desirable, especially in endemic areas. The objective of this study was to develop a PCR protocol targeting the putative fimbrial staA gene of S. Typhi. This is a preferred target gene that is specifically amplified in the S. Typhi serotype compared to the commonly targeted fliC gene which may also be amplified from the non-typhoidal Salmonella Munchen serotype. METHODOLOGY: A new nested PCR primer methodology was designed to target the staA gene, which is a member of the fimbrial gene family specific to Salmonella Typhi only. RESULTS: The primers were found to be very specific as the desired amplicon (377 bp) could be generated exclusively from S. Typhi strains including the reference strain (MTCC 3216) and 78 clinical isolates . Restriction digestion with HinfI confirmed the identity of the amplified DNA fragment in clinical specimens of S. Typhi origin.  Furthermore, these primers were able to detect a minimum of three colony forming units per ml (1fg) in spiked blood samples. The detection sensitivity of the described primers is comparable to that of previously published primers targeting fliC gene sequences. CONCLUSIONS: This study indicates that the primers targeting the putative fimbrial staA gene are very specific to the Typhi serotype and may be a better alternative to fliC targeted amplification based diagnosis.

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OBJECTIVE: To assess the role of nicotine in gallbladder carcinoma and its association with the stage and degree of cancer differentiation. METHODS: Tissue samples from gallbladder were obtained from 20 patients with gallbladder cancer and 20 age- and gender-matched patients with cholelithiasis who served as the control group. Gallbladder tissue (2 g) was extracted and analyzed for nicotine content using capillary gas chromatography. Nitrogen was used as the carrier gas. Standard curves of nicotine in methanol were made by injecting the internal standards. RESULTS: A significantly higher tissue nicotine concentration was observed in the gallbladder carcinoma group than that in the control group (179.63 ng/mg vs 6.00 ng/mg, P < 0.001). The stage and degree of cancer differentiation did not seem to affect the nicotine levels. Gallbladder tissue contained a significantly higher nicotine concentration in smokers with cancer compared with those in the control group (1570.00 ng/mg vs 232.25 ng/mg, P < 0.001). Interestingly, non-smokers in cancer group also had a higher nicotine concentration than the control group (161.50 ng/mg vs 4.00 ng/mg, P = 0.002). CONCLUSION: Nicotine is selectively concentrated in malignant gallbladder tissue irrespective of smoking status, showing its strong association with gallbladder cancer.

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Direct evidence comes from research that directly compares the interventions in which we are interested when applied to the populations in which we are interested and measures outcomes important to patients. Evidence can be indirect in one of four ways. First, patients may differ from those of interest (the term applicability is often used for this form of indirectness). Secondly, the intervention tested may differ from the intervention of interest. Decisions regarding indirectness of patients and interventions depend on an understanding of whether biological or social factors are sufficiently different that one might expect substantial differences in the magnitude of effect. Thirdly, outcomes may differ from those of primary interest-for instance, surrogate outcomes that are not themselves important, but measured in the presumption that changes in the surrogate reflect changes in an outcome important to patients. A fourth type of indirectness, conceptually different from the first three, occurs when clinicians must choose between interventions that have not been tested in head-to-head comparisons. Making comparisons between treatments under these circumstances requires specific statistical methods and will be rated down in quality one or two levels depending on the extent of differences between the patient populations, co-interventions, measurements of the outcome, and the methods of the trials of the candidate interventions.

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This article deals with inconsistency of relative (rather than absolute) treatment effects in binary/dichotomous outcomes. A body of evidence is not rated up in quality if studies yield consistent results, but may be rated down in quality if inconsistent. Criteria for evaluating consistency include similarity of point estimates, extent of overlap of confidence intervals, and statistical criteria including tests of heterogeneity and I(2). To explore heterogeneity, systematic review authors should generate and test a small number of a priori hypotheses related to patients, interventions, outcomes, and methodology. When inconsistency is large and unexplained, rating down quality for inconsistency is appropriate, particularly if some studies suggest substantial benefit, and others no effect or harm (rather than only large vs. small effects). Apparent subgroup effects may be spurious. Credibility is increased if subgroup effects are based on a small number of a priori hypotheses with a specified direction; subgroup comparisons come from within rather than between studies; tests of interaction generate low P-values; and have a biological rationale.

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Splenic angiosarcoma is a rare neoplasm that often remains asymptomatic till the onset of massive intra abdominal bleeding that require emergency splenectomy. We report here a case of 60 year old male who presented with on and off fever and a splenomegaly was found on clinical examination suggesting a lymphoproliferative disorder. A contrast enhanced computerized tomography suggested splenic trauma with intracapsular bleed. A splenectomy was carried out that revealed splenic angiosarcoma. Splenomegaly and fever is a very rare presentation of angiosarcoma. Though rare it should be kept in mind when investigating pyrexia of unknown origin.

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AIM: Nowadays laparoscopic-assisted appendicectomy using the two-port technique is gaining popularity due to its certain benefits over the open version. General anesthesia with positive pressure ventilation is the preferred mode of anesthesia in this technique. We conducted a pilot study using the two-port technique in adult patients, with uncomplicated appendicitis under local anesthesia, to evaluate its feasibility. MATERIALS AND METHODS: In this prospective study 12 consecutive patients of ASA grade I and II, with a mean age of 22.5 years, suffering from acute appendicitis, were included. All the patients received ondansetron, diazepam, and pentazocine as premedication. Monitored anesthesia care was given. The site of trocar insertions were infiltrated with 1% lignocaine with adrenalin. The pneumoperitoneum was created using carbon-di-oxide. After exteriorization of the appendix using the trocar, appendicectomy was performed as in the open procedure. RESULTS: Eleven out of twelve patients were successfully operated using this method without converting it into an open method. Two cases required an extra port to free the appendix from the adhesion. There were no intra- or post-operative complications present. CONCLUSION: Two-port laparoscopic-assisted appendicectomy under local anesthesia is a safe and effective method for uncomplicated appendicitis in adults, and the procedure is suitable where limited set-up for anesthesia is present.

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Carcinoma of the gallbladder (CaGB) is the fifth commonest gastrointestinal tract cancer and is endemic in several countries. The interplay of genetic susceptibility, infections, and life style factors has been proposed to be responsible for carcinogenesis of gallbladder. Persistence of infection leading to chronic inflammation, and production of certain toxins and metabolites with carcinogenic potentials, by certain bacteria has been speculated to be involved in the transformation of the gallbladder epithelium. Therefore, any bacteria that have evolved to acquire both of the above carcinogenic mechanisms can cause cancer. Salmonella typhi has been found to be prominently associated with CaGB. Chronic typhoid carriage (persistence) and production of mediators of chronic inflammation and a genotoxic toxin (cytotoxic distending toxin, CdtB) are also known for this bacterium. Furthermore, the natural concentrating function of the gallbladder might amplify the carcinogenic effect of the mediators of carcinogenesis. In addition to S. typhi, certain species of Helicobacter (H. bilis and H. hepaticus) and Escherichia coli have also been implicated in carcinogenesis. As the isolation rate is very poor with the presently available culture techniques, the existence of bacteria in a viable but non-cultivable state is quite likely; therefore, sensitive and specific molecular techniques might reveal the etiological role of bacterial infection in gallbladder carcinogenesis. If bacteria are found to be causing cancers, then eradication of such infections might help in reducing the incidence of some cancers.

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