Detection of recombinant dengue virus 2 NS1 protein in Aedes aegypti mosquitoes using commercial Dengue NS1 ELISA kit

Dengue, a vector-borne disease remains as one of the most serious public health problems globally. Incidence of this disease is on an increasing trend and currently over a billion people in tropical and subtropical regions are at risk. In the absence of an operational vaccine, prevention of dengue virus (DENV) is primarily focused upon controlling mosquito vectors. Mosquito vector surveillance programmes require simple and rapid tools to detect mosquitoes infected with DENV. Here, we tested the commercially available DENV Detect™ NS1 ELISA kit (InBios International, Inc.) for detection of recombinant DENV-NS1 protein in Aedes mosquito samples. The kit was evaluated to find out the minimum detection limit of recombinant DENV-2 NS1 protein following the manufacturer’s instructions. Initially, the NS1 protein detection threshold of the kit was determined and later the assay was standardized for detection of NS1 protein in Aedes aegypti mosquito pools containing 5, 10 and 25 mosquitoes. The ELISA kit displayed high sensitivity towards detection of recombinant dengue virus-2 NS1 protein in mosquito pools (up to 25 mosquitoes per pool) at 25 pico gram concentration. Since the commercial NS1 ELISA is highly sensitive and follows a very simple procedure, it could be employed for DENV surveillance in Aedes aegypti mosquitoes, after carrying out laboratory and field bioassays with DENV infected specimens.

Single use versus reuse of endoscopy biopsy forceps: A survey of patient preference.

Background. Although there are no confirmatory data on this, we suspect that most endoscopy centres in India reuse single-use (‘disposable’) endoscopic biopsy forceps due to the cost of these forceps and the perceived low risk of infection transmission on reuse. Low-cost single-use biopsy forceps are now available in India, bringing into question the justification for such a practice. We aimed to determine the type of forceps (single-use or reused) patients would prefer during endoscopy for themselves, whether this is dependent on cost, and what cost would be acceptable to them. Methods. Among patients (conveniently selected from indoor or outdoor) reporting for endoscopy at the division of gastroenterology at a private tertiary-level hospital, we distributed an information sheet about the survey 30–45 minutes before the procedure. After they completed reading the sheet, an endoscopy nurse and/or doctor explained the study. The patient then completed a questionnaire of multiple choices with tick boxes. Results. Of 151 patients approached, 4 declined to participate. Of 147 patients surveyed (age range 16–83 years; 82 men), 127 (86.4%) preferred single-use forceps, 16 (10.9%) preferred reused forceps, and 4 (2.7%) could not decide and left the decision to the physician. When informed that single-use forceps may be available for about `1000 (approximately US$ 15), 131 patients (89.1%) preferred these forceps, 11 (7.4%) preferred reused forceps, and 5 (3.4%) could not decide. Forty-four patients (33.1%) stated that an acceptable cost for a forceps for them would be `500 (approximately US$ 8), for 65 patients (48.9%) patients it was `1000, and for 24 (18.1%) it was `1500. Conclusion. About 90% of patients in this survey preferred single-use forceps; a cost of `1000 for single-use forceps was acceptable to over two-thirds of them. Natl Med J India 2016;29:205–6

Video Review : Publication at any cost.

The Centre for Biomedical Ethics and Culture, Sindh University of Urology and Transplantation, Karachi, Pakistan, periodically uploads on its website videos that depict ethically challenging situations in a doctor’s life. Their latest upload is one titled “Publish or Perish”. The title is a catchphrase for practising doctors and needs no explanation. The story is about two busy young doctors who fell behind a colleague in the promotion queue simply because he had the advantage of some publications.

Pulmonary manifestations in inflammatory bowel disease: a prospective study.

Background Although pulmonary abnormalities have been recognized in patients with inflammatory bowel diseases (IBD), their prevalence and clinical significance are not known. Aim To study the prevalence and clinical significance of pulmonary abnormalities in patients with IBD. Methods Ninety-five non-consecutive patients with IBD (12 Crohn’s disease, 83 ulcerative colitis; mean age 41.9 [SD 13] years; 47 women) were prospectively studied from January 2007 to March 2010. Pulmonary function tests (PFT) and high-resolution CT (HRCT) chest were performed in them. PFT were compared to those in 270 healthy (control) subjects matched for age, sex and smoking status. Results Twenty-seven (28.5%) patients and 11 (4%) control subjects had abnormal PFT (p<0.0001). Small airway obstruction was seen in 18 patients, restrictive defect in six and mixed defect in three. Twenty-one (22%) patients had abnormal HRCT findings – bronchiectasis and nodules (nine patients each, including one with nodules who later developed active tuberculosis after infliximab therapy), parenchymal bands (8), mediastinal lymphadenopathy (five, including two with tuberculosis on histology and culture), emphysema (5), brochiolitis (2), pleural effusion or thickening (2), pericardial effusion (2), patchy consolidation (1), ground-glass opacities (1) and lung metastasis (1). Three patients had symptoms (one asthma, two cough). Conclusion PFT and HRCT chest showed abnormality in about one-quarter of patients with IBD. A majority of patients with these abnormalities were asymptomatic.

Epidemiology and symptom profile of gastroesophageal reflux in the Indian population: Report of the Indian Society of Gastroenterology Task Force.

Background Gastroesophageal reflux disease (GERD) and its complications are thought to be infrequent in India; there are no data from India on the prevalence of and risk factors for GERD. The Indian Society of Gastroenterology formed a task force aiming to study: (a) the frequency and profile of GERD in India, (b) factors including diet associated with GERD. Methods In this prospective, multi-center (12 centers) study, data were obtained using a questionnaire from 3224 subjects regarding the frequency, severity and duration of heartburn, regurgitation and other symptoms of GERD. Data were also obtained regarding their dietary habits, addictions, and lifestyle, and whether any of these were related or had been altered because of symptoms. Data were analyzed using univariate and multivariate methods. Results Two hundred and forty-five (7.6%) of 3224 subjects had heartburn and/or regurgitation at least once a week. On univariate analysis, older age (OR 1.012; 95% CI 1.003–1.021), consumption of non-vegetarian and fried foods, aerated drinks, tea/coffee were associated with GERD. Frequency of smoking was similar among subjects with or without GERD. Body mass index (BMI) was similar in subjects with and without GERD. On multivariate analysis, consumption of non-vegetarian food was independently associated with GERD symptoms. Overlap with symptoms of irritable bowel syndrome was not uncommon; 21% reported difficulty in passage of stool and 9% had mucus in stools. About 25% of patients had consulted a doctor previously for their gastrointestinal symptoms. Conclusion 7.6% of Indian subjects have significant GERD symptoms. Consumption of non-vegetarian foods was an independent predictor of GERD. BMI was comparable among subjects with or without GERD.

Prevalence and clinical course of Clostridium difficile infection in a tertiary-care hospital: a retrospective analysis.

The spectrum of Clostridium difficile-associated diarrhea (CDAD) is changing. Apart from antibiotic use, other risk factors such as use of proton pump inhibitors (PPI) and immunosuppressive agents, intensive care unit (ICU) stay and inflammatory bowel disease are being recognized. We retrospectively analyzed data on patients whose stool samples were tested for C. difficile toxin (CDT) by enzyme linked immunosorbent assay between June 2006 and May 2008. Demographic and clinical data, and risk factors (antibiotic use, underlying malignancy, chemotherapy, use of PPI, ICU stay) were noted. The details of treatment for CDAD, response, complication and follow up were recorded. Patients whose stool samples were CDT-positive were grouped as study subjects and those with negative stool samples were included in the control group. Of the 99 patients (mean age 46.7 years; 58 men) whose stool samples were tested during this period, 17 (17%) were positive for CDT. As compared with control subjects (n=82), study subjects were more likely to have fever, prolonged ICU stay, underlying malignancy, and exposure to immunosuppressive and chemotherapeutic agents. On multivariate analysis, exposure to immunosuppressive agents was the only risk factor associated with CDAD. Fifteen patients were treated with metronidazole and two with vancomycin. Two patients did not respond to metronidazole but responded to vancomycin. No patient developed any complication. The prevalence of C. difficile toxin in diarrheal stools sent for C. difficile toxin testing was 17%. Exposure to immunosuppressive agents was a risk factor for the infection. Metronidazole was effective in a majority of patients.

A review on peripheral blood CD4+ T lymphocyte counts in healthy adult Indians.

The CD4+ T lymphocytes are the crucial cells in the cascade of events in forming immune response to the foreign antigen and hence monitoring the CD4+ T cell counts to understand the extent of immune deficiency is a common practice. CD4+ T cells are also the primary target cells for human immunodeficiency virus (HIV). Hence CD4+ T lymphocyte count is the most important marker of immune dysfunction in HIV disease progression. The estimation of CD4+ T cell counts is used to decide the initiation of anti retroviral therapy (ART), to monitor the efficacy of ART and to start treatment for opportunistic infections (OIs). To develop the threshold levels of CD4+ T cell counts, data from western countries are being used in India. The CD4+ T cell counts are known to be influenced by race and environmental factors. Hence it is important to establish the reference ranges for the CD4+ T cell counts in the target population to understand the immune dysfunction. The information on the lower limits of the CD4+ T cells count is necessary to decide the initiation and monitoring of ART. The published data on the CD4+ T cells count in healthy Indian adult population have been reviewed, analyzed and discussed in this review article. The requirement of establishment of reference ranges in Indian population is discussed.

Comparison of Mycobacterium tuberculosis culture using liquid culture medium and Lowenstein Jensen medium in abdominal tuberculosis.

Background Traditionally, the Lowenstein Jensen (LJ) medium has been used for culturing Mycobacterium tuberculosis. In abdominal tuberculosis (TB), the reported yield from tissue culture is between 20% and 60%. Liquid cultures are reported to give a higher yield but there is little data available in abdominal TB. Aim To compare the yield of TB culture with BACTEC 460TB liquid medium and LJ medium for patients with suspected abdominal TB and determine cost effectiveness. Methods This prospective study was done in consecutive cases with clinical, radiological, endoscopic/surgical, and histological suspicion of abdominal TB. Tissue biopsies obtained at colonoscopy or surgery were processed and plated on LJ medium as well as the BACTEC 460TB system. NAP (ρ-nitro-α-acetylamino-β-hydroxy-propiophenone) differentiation was carried out to determine species. The cost of each method and cost per yield were calculated. Results Of the 29 cases, 22 cases (76%) were positive on BACTEC 460TB culture while 14 (48%) were positive on LJ medium giving a 64% increment in yield. However, the culture of one patient grew on LJ medium, where the BACTEC 460TB was negative. The additional cost of BACTEC 460TB is Rs. 460 and LJ is Rs. 40. Conclusions Samples from patients with abdominal TB should be processed on both liquid and LJ medium. For high yield, the use of a liquid culture medium system is essential.

Self-reported treatment adherence in inflammatory bowel disease in Indian patients.

Introduction Non-adherence to medical therapy is emerging as an important determinant of relapse in patients with inflammatory bowel disease (IBD). Aim To find the prevalence of and reasons for nonadherence to medical therapy in Indian patients with IBD and its correlation with disease outcome. Methods In this cross-sectional study, we checked for adherence to treatment in 127 patients with IBD (117 ulcerative colitis and 10 Crohn’s disease) using a questionnaire that inquired into frequency of missed doses, causes for missed doses, and its relation to relapse of disease. Results Of the 127 patients (mean age 42.8 years; 68 women), 103 (81%) were non-adherent to treatment, defined as taking 80% or less of the dose advised. The reasons for non-adherence (not mutually exclusive) were: forgetfulness– 98 patients (77%), felt better–18 (14.2 %), high frequency of doses–13 (10.1%), no effect of medications–10 (7.87%), non-availability of medications–3 (2.3%). Non-adherent patients were three times more likely to develop a relapse as compared to those with adherence (OR 3.389, 95% CI 1.29–8.88, p=0.012). Conclusions Over 80% of patients with IBD in this survey were non-adherent to medical treatment; forgetfulness was mentioned as the most common cause. Non-adherent patients were more likely to relapse. Patients need to be educated regarding the need for adherence to treatment in IBD.

Hypercoagulable state in idiopathic ulcerative colitis: role of hyperhomocysteinemia and hyperfibrinogenemia.

BACKGROUND: Previous reports on hypercoagulable factors in inflammatory bowel diseases involve heterogeneous populations and patients on various medications. AIMS: To determine the frequency of thrombotic complications in ulcerative colitis (UC); to evaluate for hyperhomocysteinemia and its relationship to vitamin B12 and folate levels and methylene tetrahydrofolate reductase (MTHFR) mutation; and to evaluate for hyperfibrinogenemia and factor V Leiden mutation. METHODS: Eighty-six adult patients with UC were seen during the study period; 28 of them underwent blood tests and constituted the study population. Patients who received medications that affect these factors were among the 58 excluded. Tests were obtained at baseline and after 2 months during remission. Patients received folic acid in addition to treatment for UC. RESULTS: Vascular thrombotic events were noted in 4 patients during follow up. Hyperhomocysteinemia was detected in 11 (39.3%) patients (controls 15/100, p=0.007). Heterozygous state for MTHFR C677T mutation was found in 5 (17.9%) patients (controls: 0.2% homozygous, 13.6% heterozygous, p>0.05). Plasma homocysteine did not correlate with extent, severity or duration of disease, or with MTHFR C677T heterozygous state, but correlated with serum folic acid level (p=0.003) and BMI (p=0.03). With folate supplementation, homocysteine decreased significantly in patients who had hyperhomocysteinemia at baseline. Hyperfibrinogenemia was detected in 3 patients (none in 100 controls). Plasma fibrinogen was not affected by duration, extent or severity of UC and did not decrease with remission of disease. Only one patient had heterozygous factor V Leiden mutation. CONCLUSION: Vascular thrombosis occurred in less than a fifth of the UC population studied. Hyperhomocysteinemia reversible by folate supplementation and hyperfibrinogenemia were observed, but their contribution and that of factor V Leiden mutation appear to be insignificant.

Epidemiological and clinical profile of irritable bowel syndrome in India: report of the Indian Society of Gastroenterology Task Force.

AIMS: To study the profile of irritable bowel syndrome (IBS), and the frequency of such symptoms among the general population, in India. METHODS: In this prospective, multi-center study, data were obtained from 2785 patients with chronic lower gastrointestinal symptoms (complainants) with no alarm feature and negative investigations for organic causes visiting physicians at 30 centers, and from 4500 community subjects (non-complainants), using separate questionnaires. RESULTS: Most complainants were middle-aged (mean age 39.4 years) and male (1891; 68%). The common symptoms were: abdominal pain or discomfort (1958; 70%), abdominal fullness (1951; 70%); subjective feeling of constipation (1404 of 2656; 53%), or diarrhea (1252 of 2656, 47%), incomplete evacuation (2134; 77%), mucus with stools (1506; 54%), straining at stools (1271; 46%), epigastric pain (1364; 49%) and milk intolerance (906; 32%). Median stool frequency was similar in patients who felt they had constipation or those who felt they had diarrhea. Information to subtype symptoms using standard criteria was available in 1301 patients; of these, 507 (39%) had constipation-predominant IBS ( 3 <or= stools/week), 50 (4%) had diarrhea-predominant IBS (>3 stools/day) and 744 (57%) had indeterminate symptoms. Among non-complainants, most subjects reported daily defecation frequency of one (2520 [56%]) or two (1535 [34%]). Among non-complainants, 567 (12.6%) reported abdominal pain, 503 (11%) irregular bowel, 1030 (23%) incomplete evacuation, 167 (4%) mucus and 846 (18%) straining at stools; a combination of abdominal pain or discomfort relieved by defecation, and incomplete evacuation was present in 189/4500 (4.2%) community subjects. CONCLUSIONS: Most patients with IBS in India are middle-aged men, and have a sense of incomplete evacuation and mucus with stools. Abdominal pain or discomfort is frequent but not universal. Importantly, stool frequency was similar irrespective of whether the patients felt having constipation or diarrhea. Most (90%) non-complainant subjects had 1 or 2 stools per day; symptoms complex suggestive of IBS was present in 4.2% of community subjects.

Effect of liver disease on maternal and fetal outcome--a prospective study.

BACKGROUND: Liver disease in pregnancy can have serious consequences. Its prevalence, profile and effect on outcome of pregnancy have not been documented systematically in India. We prospectively determined the frequency, causes and outcome of liver disease in pregnant women. SETTING: Tertiary-care teaching hospital. METHODS: Consecutive pregnant women with liver disease attending the inpatient or outpatient services of the Departments of Gastro-enterology and Obstetrics and Gynecology between December 2002 and October 2004 were evaluated and followed up till 2 weeks postpartum or death. RESULTS: Liver disease was found in 107 (0.9%) of 12,061 pregnancies. Of these, fifty six (52.3%) had pregnancy-specific liver disorders (pregnancy-induced hypertension [PIH]-associated liver dysfunction 36--including HELLP syndrome 22 and pre-eclamptic liver dysfunction 14; intrahepatic cholestasis of pregnancy 10; hyperemesis gravidarum 7; acute fatty liver of pregnancy 3). Liver disorders not specific to pregnancy included hepatitis E (16), hepatitis B, non A-E hepatitis and chronic liver disease (5 each) and others (14); in 6 patients no cause could be found. Ninety-six patients completed follow up. Overall maternal and perinatal mortality rates were 19.7% and 35.4%, respectively. CONCLUSIONS: PIH-associated liver dysfunction was the most common cause of liver disease in pregnancy. This is associated with significant maternal and perinatal morbidity and mortality.

Celiac crisis with hypokalemic paralysis in a young lady.

Celiac crisis presents as severe acute diarrhea with life-threatening metabolic derangement in a patient with celiac disease. We report a 30-year-old lady who was admitted with one-month history of worsening small bowel-type diarrhea. She developed acute quadriparesis due to refractory hypokalemia. Celiac disease was diagnosed on the basis of positive serology and histological features. She improved with aggressive correction of hypokalemia and gluten-free diet. Celiac crisis is a rare presentation of this heterogeneous disease in adulthood.

Endoscopic biliary drainage in patients with amebic liver abscess and biliary communication.

BACKGROUND: Percutaneous drainage or surgery is required when amebic liver abscess (ALA) fails to respond to medical management. In some of these patients, non-response may be due to communication of ALA with the biliary tree. This report describes our experience with the use of endoscopic biliary draining in such patients. METHODS: Medical records of patients with ALA undergoing either needle aspiration or percutaneous pigtail drainage were retrieved; the indications for drainage were: abscess volume exceeding 250 mL, a thin rim of tissue (< 1 cm thick) around the abscess, systemic toxic features and failure to improve on medical treatment. Patients with abscess drain output >25 mL/day persisting for 2 weeks or presence of bile in the drain fluid underwent endoscopic biliary drainage. RESULTS: A total of 115 patients with ALA underwent percutaneous treatment. None of the 25 patients with needle aspiration needed any further treatment. Of the 90 who underwent catheter drainage, the catheter could be removed within one week in 77 patients; the remaining 13 patients (median age 42 years, range 24-65; all men) had an abscess-biliary communication. In them, the median catheter output was 88 mL/day (range 45-347) and 54 mL/day (28-177) at 2 days and 2 weeks after catheter placement. The drain fluid contained bile in all 13 patients and in addition contained pus in 10 patients. Eleven patients had a solitary abscess and two had multiple abscesses. Cholangiogram showed biliary communication in all 13 patients. All patients were treated with placement of 10F biliary endoprosthesis or 10F nasobiliary drain. Pigtail catheter was removed within 1 week in 11 of 13 patients. CONCLUSION: In patients with amebic liver abscess communicating with the biliary tree, biliary stenting may hasten clinical recovery and allow early removal of liver abscess catheter drain.

Primary malignant melanoma of right colon.

We report a 62-year-old lady who presented with abdominal lump and neurological deficit. Investigations showed ascending colon mass, subcutaneous nodule and right parietal lobe lesion. Histology and immunohistochemistry were suggestive of malignant melanoma. The search for primary site was negative.