Increasing Antimicrobial Resistance of Vibrio cholerae O1 Biotype El Tor Strains Isolated in a Tertiary-care Centre in India

The antimicrobial susceptibility patterns are on constant change with the recent emergence of multidrugresistant strains of most bacteria. Results of recent studies in India showed that most isolates of Vibrio cholerae O1 were resistant to the commonly-used antibiotics. The study was conducted to determine the antibiotic susceptibility patterns of V. cholerae O1 isolated during 2008-2010 at the hospital of the Jawaharlal Nehru Institute of Post Graduate Medical Education and Research, Puducherry, India. In total, 154 strains of V. cholerae O1 from 2,658 stool specimens were reported during January 2008\u2013December 2010\u201434 in 2008, 2 in 2009, and 118 in 2010. The isolates of V. cholerae O1 were subjected to antimicrobial susceptibility testing using the Kirby-Bauer method. The antibiotic disks tested were tetracycline (30 \u3bcg), furazolidone (100 \u3bcg), ampicillin (10 \u3bcg), ceftriaxone (30 \u3bcg), and ciprofloxacin (5 \u3bcg). Escherichia coli ATCC 25922 was used as the control organism. The minimum inhibitory concentrations (MICs) of ceftriaxone, ciprofloxacin, and tetracycline were determined using the agar dilution method for all the strains. The E-test method was used for the strains which had either intermediate resistance or were resistant to the antibiotics by the agar dilution method. The results of the agar dilution corroborated the results of the E-test. The MIC of ceftriaxone in 151 strains was <2 \u3bcg/mL while it was 16 \u3bcg/mL in three strains; the latter three strains were resistant to ceftriaxone by the disc-diffusion test. The MIC of ciprofloxacin in 150 strains was <0.5 \u3bcg/mL while the MIC of tetracycline was <1 \u3bcg/mL. In the remaining four strains, the MIC of tetracycline was >32 \u3bcg/mL, and the MIC of ciprofloxacin was >8 \u3bcg/mL. These four strains were resistant to both tetracycline and ciprofloxacin by the disc-diffusion test and were exclusive of the three ceftriaxone-resistant strains. The majority of the isolates were obtained from children aged 0-5 year(s)\u201470.3% (83 of 118) and 41.2% (14 of 34) were reported in 2010 and 2008 respectively. Since treating severe cases of cholera with antibiotics is important, the continuing spread of resistance in V. cholerae to the most important agents of therapy is a matter of concern. Also, chemoprophylaxis with antimicrobial agents is likely to become even more difficult

Clinical and Microbiological Profiles of Shigellosis in Children

Shigellosis presents with varied clinical features are dictated by the species involved, virulence factors of the strain, and the host immune status. We studied the species, virulence genes, and antibiotic susceptibility pattern of the Shigella strains isolated from 33 children aged less than 12 years, with clinical features of shigellosis. Identification and antibiotic sensitivity of Shigella species were done using disc diffusion and E-test. Multiplex PCR was done for the detection of virulence genes (ipaH, ial, set1A, set1B, sen, and stx) and ESBL genes. Parents of the children were interviewed using structured questionnaire to assess the severity of the disease; 26 (79%) of the isolates were Shigella flexneri . Ciprofloxacin and ceftriaxone resistance was seen in 23 (69%) and 3 (9%) Shigella isolates respectively. Two ceftriaxone-resistant strains were found to harbour blaCTX gene and the third blaTEM gene. Virulence gene ipaH was detected in 100% of strains while ial, sen, set1A, and set1B were detected in 85%, 61%, 48%, and 48% respectively

Albiglutide and cardiovascular outcomes in patients with type 2 diabetes and cardiovascular disease (Harmony Outcomes): a double-blind, randomised placebo-controlled trial

Background: Glucagon-like peptide 1 receptor agonists differ in chemical structure, duration of action, and in their effects on clinical outcomes. The cardiovascular effects of once-weekly albiglutide in type 2 diabetes are unknown. We aimed to determine the safety and efficacy of albiglutide in preventing cardiovascular death, myocardial infarction, or stroke. Methods: We did a double-blind, randomised, placebo-controlled trial in 610 sites across 28 countries. We randomly assigned patients aged 40 years and older with type 2 diabetes and cardiovascular disease (at a 1:1 ratio) to groups that either received a subcutaneous injection of albiglutide (30–50 mg, based on glycaemic response and tolerability) or of a matched volume of placebo once a week, in addition to their standard care. Investigators used an interactive voice or web response system to obtain treatment assignment, and patients and all study investigators were masked to their treatment allocation. We hypothesised that albiglutide would be non-inferior to placebo for the primary outcome of the first occurrence of cardiovascular death, myocardial infarction, or stroke, which was assessed in the intention-to-treat population. If non-inferiority was confirmed by an upper limit of the 95% CI for a hazard ratio of less than 1·30, closed testing for superiority was prespecified. This study is registered with ClinicalTrials.gov, number NCT02465515. Findings: Patients were screened between July 1, 2015, and Nov 24, 2016. 10 793 patients were screened and 9463 participants were enrolled and randomly assigned to groups: 4731 patients were assigned to receive albiglutide and 4732 patients to receive placebo. On Nov 8, 2017, it was determined that 611 primary endpoints and a median follow-up of at least 1·5 years had accrued, and participants returned for a final visit and discontinuation from study treatment; the last patient visit was on March 12, 2018. These 9463 patients, the intention-to-treat population, were evaluated for a median duration of 1·6 years and were assessed for the primary outcome. The primary composite outcome occurred in 338 (7%) of 4731 patients at an incidence rate of 4·6 events per 100 person-years in the albiglutide group and in 428 (9%) of 4732 patients at an incidence rate of 5·9 events per 100 person-years in the placebo group (hazard ratio 0·78, 95% CI 0·68–0·90), which indicated that albiglutide was superior to placebo (p<0·0001 for non-inferiority; p=0·0006 for superiority). The incidence of acute pancreatitis (ten patients in the albiglutide group and seven patients in the placebo group), pancreatic cancer (six patients in the albiglutide group and five patients in the placebo group), medullary thyroid carcinoma (zero patients in both groups), and other serious adverse events did not differ between the two groups. There were three (<1%) deaths in the placebo group that were assessed by investigators, who were masked to study drug assignment, to be treatment-related and two (<1%) deaths in the albiglutide group. Interpretation: In patients with type 2 diabetes and cardiovascular disease, albiglutide was superior to placebo with respect to major adverse cardiovascular events. Evidence-based glucagon-like peptide 1 receptor agonists should therefore be considered as part of a comprehensive strategy to reduce the risk of cardiovascular events in patients with type 2 diabetes. Funding: GlaxoSmithKline

ANTIDIABETIC ACTIVITY OF SENNA SURATTENSIS IN ALLOXAN-INDUCED DIABETIC RATS

 Objective: Senna surattensis is a shrub plant which has been known for its diverse biological and pharmacological properties. This study is aimed to evaluate the antidiabetic activity of ethanolic extracts of S. surattensis (EESS) leaves in alloxan-induced diabetic rats.Methods: Experimental diabetes was induced by injection of a single dose of alloxan (120 mg/kg, intraperitoneal). Adult male Wistar albino rats were divided into five groups; normal control, diabetic control, diabetic EESS (200 mg/kg body weight (bw), diabetic EESS (400 mg/kg bw), and diabetic glibenclamide (5 mg/kg bw). Extracts were treated concurrently for 21 days. Blood samples were collected and centrifuged for estimation of fasting blood glucose (FBG), bw, serum biomarkers, lipid profile, total protein, albumin, and glycosylated hemoglobin (HbA1C) contents.Results: The increase in FBG, bw, liver biomarkers serum glutamic oxaloacetic transaminase, serum glutamic pyruvic transaminase, alkaline phosphatase, free fatty acid, phospholipids (PL), triglycerides, total cholesterol, low-density lipoprotein cholesterol, high-density lipoprotein cholesterol, total protein, albumin, and HbA1C content were recorded in diabetic control rats. Daily oral administration of EESS treatment significantly (p<0.01) reverted the levels of serum biomarkers and enzymes activities to near normal values. A similar reduction was produced in FBG after 21 days of extract administration which compared significantly (p<0.01) with the control group and glibenclamide treated groups.Conclusion: The results suggest that EESS has anti-diabetic activity in diabetic rats, thereby justifying its traditional claim and augmenting it into the present system of medicine

Increasing Antimicrobial Resistance of Vibrio cholerae O1 Biotype El Tor Strains Isolated in a Tertiary-care Centre in India

The antimicrobial susceptibility patterns are on constant change with the recent emergence of multidrugresistant strains of most bacteria. Results of recent studies in India showed that most isolates of Vibrio cholerae O1 were resistant to the commonly-used antibiotics. The study was conducted to determine the antibiotic susceptibility patterns of V. cholerae O1 isolated during 2008-2010 at the hospital of the Jawaharlal Nehru Institute of Post Graduate Medical Education and Research, Puducherry, India. In total, 154 strains of V. cholerae O1 from 2,658 stool specimens were reported during January 2008–December 2010—34 in 2008, 2 in 2009, and 118 in 2010. The isolates of V. cholerae O1 were subjected to antimicrobial susceptibility testing using the Kirby-Bauer method. The antibiotic disks tested were tetracycline (30 μg), furazolidone (100 μg), ampicillin (10 μg), ceftriaxone (30 μg), and ciprofloxacin (5 μg). Escherichia coli ATCC 25922 was used as the control organism. The minimum inhibitory concentrations (MICs) of ceftriaxone, ciprofloxacin, and tetracycline were determined using the agar dilution method for all the strains. The E-test method was used for the strains which had either intermediate resistance or were resistant to the antibiotics by the agar dilution method. The results of the agar dilution corroborated the results of the E-test. The MIC of ceftriaxone in 151 strains was <2 μg/mL while it was 16 μg/mL in three strains; the latter three strains were resistant to ceftriaxone by the disc-diffusion test. The MIC of ciprofloxacin in 150 strains was <0.5 μg/mL while the MIC of tetracycline was <1 μg/mL. In the remaining four strains, the MIC of tetracycline was >32 μg/mL, and the MIC of ciprofloxacin was >8 μg/mL. These four strains were resistant to both tetracycline and ciprofloxacin by the disc-diffusion test and were exclusive of the three ceftriaxone-resistant strains. The majority of the isolates were obtained from children aged 0-5 year(s)—70.3% (83 of 118) and 41.2% (14 of 34) were reported in 2010 and 2008 respectively. Since treating severe cases of cholera with antibiotics is important, the continuing spread of resistance in V. cholerae to the most important agents of therapy is a matter of concern. Also, chemoprophylaxis with antimicrobial agents is likely to become even more difficult

Comparison of nested-multiplex, Taqman & SYBR Green real-time PCR in diagnosis of amoebic liver abscess in a tertiary health care institute in India

Background & objectives: Amoebiasis is a common parasitic infection caused by Entamoeba histolytica and amoebic liver abscess (ALA) is the most common extraintestinal manifestation of amoebiasis. The aim of this study was to standardise real-time PCR assays (Taqman and SYBR Green) to detect E. histolytica from liver abscess pus and stool samples and compare its results with nested-multiplex PCR. Methods: Liver abscess pus specimens were subjected to DNA extraction. The extracted DNA samples were subjected to amplification by nested-multiplex PCR, Taqman (18S rRNA) and SYBR Green real-time PCR (16S-like rRNA assays to detect E. histolytica/E. dispar/E. moshkovskii). The amplification products were further confirmed by DNA sequence analysis. Receiver operator characteristic (ROC) curve analysis was done for nested-multiplex and SYBR Green real-time PCR and the area under the curve was calculated for evaluating the accuracy of the tests to dignose ALA. Results: In all, 17, 19 and 25 liver abscess samples were positive for E. histolytica by nested-multiplex PCR, SYBR Green and Taqman real-time PCR assays, respectively. Significant differences in detection of E. histolytica were noted in the real-time PCR assays evaluated ( P<0.0001). The nested-multiplex PCR, SYBR Green real-time PCR and Taqman real-time PCR evaluated showed a positivity rate of 34, 38 and 50 per cent, respectively. Based on ROC curve analysis (considering Taqman real-time PCR as the gold standard), it was observed that SYBR Green real-time PCR was better than conventional nested-multiplex PCR for the diagnosis of ALA. Interpretation & conclusions: Taqman real-time PCR targeting the 18S rRNA had the highest positivity rate evaluated in this study. Both nested multiplex and SYBR Green real-time PCR assays utilized were evaluated to give accurate results. Real-time PCR assays can be used as the gold standard in rapid and reliable diagnosis, and appropriate management of amoebiasis, replacing the conventional molecular methods

Comparison of nested-multiplex, Taqman & SYBR Green real-time PCR in diagnosis of amoebic liver abscess in a tertiary health care institute in India

Background & objectives: Amoebiasis is a common parasitic infection caused by Entamoeba histolytica and amoebic liver abscess (ALA) is the most common extraintestinal manifestation of amoebiasis. The aim of this study was to standardise real-time PCR assays (Taqman and SYBR Green) to detect E. histolytica from liver abscess pus and stool samples and compare its results with nested-multiplex PCR. Methods: Liver abscess pus specimens were subjected to DNA extraction. The extracted DNA samples were subjected to amplification by nested-multiplex PCR, Taqman (18S rRNA) and SYBR Green real-time PCR (16S-like rRNA assays to detect E. histolytica/E. dispar/E. moshkovskii). The amplification products were further confirmed by DNA sequence analysis. Receiver operator characteristic (ROC) curve analysis was done for nested-multiplex and SYBR Green real-time PCR and the area under the curve was calculated for evaluating the accuracy of the tests to dignose ALA. Results: In all, 17, 19 and 25 liver abscess samples were positive for E. histolytica by nested-multiplex PCR, SYBR Green and Taqman real-time PCR assays, respectively. Significant differences in detection of E. histolytica were noted in the real-time PCR assays evaluated ( P<0.0001). The nested-multiplex PCR, SYBR Green real-time PCR and Taqman real-time PCR evaluated showed a positivity rate of 34, 38 and 50 per cent, respectively. Based on ROC curve analysis (considering Taqman real-time PCR as the gold standard), it was observed that SYBR Green real-time PCR was better than conventional nested-multiplex PCR for the diagnosis of ALA. Interpretation & conclusions: Taqman real-time PCR targeting the 18S rRNA had the highest positivity rate evaluated in this study. Both nested multiplex and SYBR Green real-time PCR assays utilized were evaluated to give accurate results. Real-time PCR assays can be used as the gold standard in rapid and reliable diagnosis, and appropriate management of amoebiasis, replacing the conventional molecular methods

Detailed Pharmacognostical Standardization Studies on <em>Calotrophis Procera</em> (Aiton) Dryand Fruit

Calotropis procera (Aiton) Dryand (Asclepiadaceae) is very well-known latex wild plant, it is covered in tropical Asia and Africa, traditionally as well as medicinally the plant has been used for insecticidal, antimalarial, antiviral, antimicrobial, analgesic, antifertility, antitumor, antihyperglycemic, hepatoprotective, anti-inflammatory, antidiarrhoeal, anticonvulsant, oestrogenic, antidiabetic and anthelmintic activity. For this reason, the main theme of research work was carried out on detailed pharmacognistical studies for quality control and standardization and this study provides the basis for the herbal remedy and provide physiological information about plant species. This research is helpful to identify and estimate the purity of the drug and can also be used to screen for adulteration and gives a drug quality. This comprehensive research work is for identification, collection, authentication and it is easy to identify the adulterants. Detailed macroscopical, T.S, L.S, Powder microscopy, Physicochemical parameters, Extractive values, Fluorescence, Preliminary Phytochemical studies were performed as per the standard. It is a very important research work and the author had has been established the evaluation standards for a medicinal plant. This evaluation is for the authentication of a fruit plant of Calotropis procera and the fruit physiology is useful for both qualitative and quantitative estimation of the medicinal herbal drugs