Multiple locus sequence typing of Salmonella Typhi, isolated in north India - a preliminary study.

BACKGROUND & OBJECTIVES: In India enteric fever is a major public health problem and Salmonella Typhi is the most common aetiologic agent. Any control strategy for such infections depends to a large extent on the understanding of the disease and relatedness of strains across the world. Multi locus sequence typing (MLST) is one such method of genotyping of bacteria based upon housekeeping genes of known function and chromosome position. MLST data of pathogens are important to determine the molecular evolution by a stable and reproducible method. This study was undertaken to determine the sequence types of representatives S. Typhi isolates obtained from enteric fever patients in a tertiary care centre in north India, over a period of 20 years (1990-2010). METHODS: A total of 30 representative isolates of S. Typhi identified by biochemical and serological tests were subjected to multi locus sequence typing (MLST). Seven housekeeping genes of known function and chromosome position were used for the typing by MLST. Sequencing was carried out by using an automated DNA sequencer and results were analyzed to generate phylogenetic tree. RESULTS: MLST pattern grouped S. Typhi into two sequence types- ST1 and ST2. ST1 was predominantly present followed by ST2. INTERPRETATION & CONCLUSIONS: By MLST the presence of both sequence types, ST1 and ST2, was found in S. Typhi isolates in our region. Predominately ST1 was present followed by ST2. These preliminary results corroborate the global distribution of both sequence types of S. Typhi and also emphasize for the continuous screening of S. Typhi.

Short-term exposure to air pollution and emergency room visits for acute respiratory symptoms among adults.

OBJECTIVE: To examine the short-term effect of ambient air pollution on daily acute respiratory emergency room visits among adults.METHODS: A time-series study (June 2017-February 2019) was carried out among adults (≥18 years) visiting a multi-specialty hospital in Delhi. We evaluated the association between the daily levels of particulate matter (PM) <2.5 µm in diameter (PM2.5) and PM <10 µm in diameter (PM10), ozone (O3), nitrogen dioxide (NO2), carbon monoxide (CO) and sulphur dioxide and daily count of emergency room (ER) visits for acute respiratory symptoms. Generalised additive model (GAM) was used with the Poisson link function to analyse the associations for 0-1 to 0-7 lag days.RESULTS: A total of 69,400 ER visits were recorded, of which 2,669 were by adults due to acute respiratory symptoms. At 0-7 lag days, an increment of 1 standard deviation in NO2 and PM2.5 concentration was associated with a percentage increase in acute respiratory ER visits of respectively 53.0% (95% CI 30.84-78.97) and 19.5% (95% CI 4.53-36.65). During 0-7 lag days, a positive trend was observed at higher concentrations of CO (>1.86-3.28 mg/m³), while a negative significant association was observed at low concentrations of CO (<1.171 mg/m³).CONCLUSION: Short-term exposure to ambient NO2 and PM2.5 was associated with acute respiratory emergency visits of adults at lag 0-7 days.

Antibodies that cross-neutralize the tier-2 pseudoviruses are produced in antiretroviral-naïve HIV-1-infected children from northern India.

In this cross-sectional study, we evaluated the efficiency of the plasma of 38 antiretroviral-naïve HIV-1-infected children from northern India against a standard panel of pseudoviruses (3 clade C and 3 clade B) by TZM-bl assay. Neutralization potential was observed to a variable extent, with a potency ranging up to reciprocal ID(50) titers of 1967. Cross-neutralization was observed in 28.9 % (11/38) of the children. There was a significant positive correlation between viremia and neutralization efficiency against two of the viruses studied (Du172 r = 0.49; p = 0.007 and RHPA r = 0.47; p = 0.01), suggesting that persistent antigenic stimulation is necessary for the generation of broadly neutralizing antibody responses in these children. Further mapping of the epitope specificities of the neutralization determinants in the polyclonal plasma would provide important information for immunogen design.

Scrub typhus in patients with acute febrile illness: a 5-year study from India.

BACKGROUND: Scrub typhus was once thought to be a disease of rural origin and was confined to specific pockets in South Asia. Early diagnosis and treatment is extremely important as it is associated with high mortality if left untreated. AIM: To delineate the clinical and molecular epidemiology of scrub typhus in patients presenting with acute febrile illness from various parts of India. METHODS: During the study period of 5 years (October 2013 to October 2018), a total of 1742 patients with acute febrile illness <15 days were enrolled after taking informed consent. Patients were diagnosed using IgM Enzyme-linked immunosorbent assay (ELISA) based on the pre-determined region specific cut offs. Patients with positive IgM ELISA were also subjected to IgM Immunofluorescence assay and nested polymerase chain reaction (PCR) assay. The demographic and relevant clinical details of the patients were documented and analyzed. RESULTS: A total of 210 (12.1%) patients were diagnosed with scrub typhus. Of these, nested PCR was positive in only 85 patients. Sequencing and phylogenetic analysis showed that the predominant circulating genotypes were Gilliam and Karp. On multivariate analysis, acute respiratory distress syndrome, myocarditis, encephalitis/encephalopathy, jaundice and splenomegaly were significantly more common in those patients who were diagnosed with scrub typhus. A total of 14 patients diagnosed with scrub typhus succumbed to the illness. CONCLUSION: Patients with fever, headache, pulmonary manifestations, CNS manifestations, myocarditis, transaminitis or thrombocytopenia presenting in the monsoon and post-monsoon season should be evaluated for scrub typhus irrespective of the geographical location in India.

Point-of-care urine lipoarabinomannan antigen detection for diagnosis of tuberculosis in children.

Diagnosis of tuberculosis (TB) in children remains challenging due to the paucibacillary nature of the disease. Detection of TB using urine lipoarabinomannan (LAM) antigen was evaluated in children with presumed TB. Children with presumed intrathoracic tuberculosis (ITTB) and lymph node TB (LNTB) were enrolled. Expectorated or induced sputum or gastric aspirates from ITTB patients and fine-needle cytological aspirates from LNTB patients were subjected to Ziehl-Neelsen staining, MGIT™960™ culture and Xpert® MTB/RIF testing. Urine samples were tested to detect LAM, and the sensitivity and specificity calculated. Of 280 children with presumed ITTB and 101 with presumed LNTB, respectively 71 (25.3%) and 25 (24.7%) were categorised as 'confirmed TB', 70 (25%) and 33 (32.7%) as 'unconfirmed TB', and 139 (49.6%) and 43 (42.5%) as 'unlikely TB'. Respectively 8 (2.8%) children with ITTB and 3 (2.9%) with LNTB were positive on smear, 56 (20.0%) and 23 (22.7%) on Xpert, and 50 (17.8%) and 9 (8.9%) on culture. LAM assay sensitivity was 73.2% in confirmed ITTB cases, and 76% in confirmed LNTB cases; LAM assay specificity in children with ITTB and those with LNTB initiated on anti-tuberculosis treatment was respectively 92% and 93%. Detection of TB using the LAM assay was significantly better than detection using Xpert (P < 0.05 vs. P < 0.002). Urinary LAM testing showed high specificity and sensitivity, was detected in more cases initiated on treatment than reference tests, and improved disease detection by 38.5% in ITTB patients and by 41.6% in LNTB patients. .

Emerging multidrug resistance isolates of hospital-acquired bacterial meningitis in a tertiary care centre in North India.

Purpose. Acute bacterial meningitis continues to be a potentially life threatening condition. Hospital-acquired meningitis is rapidly increasing and adding an immense burden to the health system due to the emergence of multidrug resistance isolates. The purpose of this study is to find the antibiotic susceptibility pattern of the bacteria detected from hospital- and community-acquired meningitis.Methodology. A total of 400 Cerebrospinal fluid (CSF) samples from the suspected meningitis cases were collected and processed for cell count, biochemical examination, Gram staining, latex agglutination and culture. Bacteria grown on blood, chocolate and Mac-conkey agar were identified by matrix-assisted laser desorption/ionization-time of flight. Antibiotic susceptibility tests were performed as per Clinical and Laboratory Standard Institute guidelines.Results. Of the isolates, most prevalent Gram negative organisms in hospital-acquired bacterial meningitis were Escherichia coli 13 (27.08 %), Acinetobacter baumannii 12 (25 %), Klebsiella pneumoniae 5 (10.42 %), Pseudomonas aeruginosa 4 (8.33 %) and Gram positive organisms were Staphylococcus aureus 4 (8.33 %), Enterococcus faecium 3 (6.25 %) and CONS 2 (4.16 %). Streptococcus pneumoniae 3 (6.25 %) was the predominant organism in community-acquired bacterial meningitis. All the Gram negative isolates were multidrug resistance. Only colistin and imipenem were effective antibiotics against them. Likewise Gram positive organisms were susceptible to most of the antibiotics tested. However, E. faecium was only susceptible to Vanco+Teicoplanin.Conclusion. In hospital-acquired bacterial meningitis, multidrug resistance Gram negative bacteria are a huge challenge for the treatment of patients. Hence, antimicrobial stewardship should be followed to counteract with the emerging multidrug resistance isolates.

Genetic variations in olfactory receptor gene OR2AG2 in a large multigenerational family with asthma.

It is estimated from twin studies that heritable factors account for at-least half of asthma-risk, of which genetic variants identified through population studies explain only a small fraction. Multi-generation large families with high asthma prevalence can serve as a model to identify highly penetrant genetic variants in closely related individuals that are missed by population studies. To achieve this, a four-generation Indian family with asthma was identified and recruited for examination and genetic testing. Twenty subjects representing all generations were selected for whole genome genotyping, of which eight were subjected to exome sequencing. Non-synonymous and deleterious variants, segregating with the affected individuals, were identified by exome sequencing. A prioritized deleterious missense common variant in the olfactory receptor gene OR2AG2 that segregated with a risk haplotype in asthma, was validated in an asthma cohort of different ethnicity. Phenotypic tests were conducted to verify expected deficits in terms of reduced ability to sense odors. Pathway-level relevance to asthma biology was tested in model systems and unrelated human lung samples. Our study suggests that OR2AG2 and other olfactory receptors may contribute to asthma pathophysiology. Genetic studies on large families of interest can lead to efficient discovery.

Salmeterol vs. formoterol: a comparison of rapid bronchodilator effect in a randomized controlled trial.

We conducted this double blind randomized controlled trial to compare the rapid bronchodilator effect of salmeterol and formoterol in 60 children with stable asthma. Participants were randomized to receive either salmeterol (50 microg) (n=31) or formoterol (24 microg) (n=29) by metered dose inhaler and spacer. Spirometry was performed at baseline, at 30 minutes, and at 60 minutes. Bronchodilatation was assessed by changes in FEV(1) at 30 and 60 minutes. Baseline parameters were comparable in the two groups. There was no significant difference in the FEV(1) at 30 and 60 minutes between two groups. We conclude that salmeterol and formoterol both cause bronchodilator response at end of 60 minutes and are not different with regards to their rapid bronchodilator response.

Cystic fibrosis in India.

Cystic fibrosis (CF) was considered to be non-existent in Indian subcontinent. Reports in last one decade have suggested that cystic fibrosis occurs in India but its precise magnitude is not known. Studies on migrant Indian population in United States and United Kingdom estimate frequency of CF as 1:10,000 to 1:40,000. The clinical features are similar to that reported in Caucasian population. CF in Indian children is usually diagnosed late and in advanced stage. Children are more malnourished and may have clinically evident deficiency of fat soluble vitamins. The frequency of clubbing, colonization with Pseudomonas, and laboratory evidence of pseudo-Bartter syndrome is relatively more at the time of diagnosis. Diagnostic facilities in form of sweat chloride estimation and genetic studies are not available readily. Mutation profile is different. The frequency of common mutation F508del in Indian children is between 19% and 34%. Other mutations are heterogeneous. Management of CF in India is difficult due to less number of trained manpower, limited availability, and high cost of pharmacologic agents. The determinants of early death include: severe malnutrition and colonization with Pseudomonas at the time of diagnosis, more than four episodes of lower respiratory infection per year and age of onset of symptoms before 2 months of age. To conclude, CF does occur in India; however, precise magnitude of problem is not known. There is need to create awareness amongst pediatricians, developing diagnostic facilities, and management protocols based on locally available resources.

Antibiotics for community acquired pneumonia in children.

BACKGROUND: Pneumonia is the leading cause of mortality in children. In developing countries, pneumonia is usually caused by bacterial pathogens. The early administration of empirical antibiotics improves the patients' clinical outcomes. There are currently no systematic reviews of clinical trials on this subject. OBJECTIVES: To identify effective antibiotic drug therapy for community acquired pneumonia (CAP) in children by comparing various antibiotics. SEARCH STRATEGY: We searched the Cochrane Central Register of Controlled Trials (CENTRAL) (The Cochrane Library, Issue 4, 2005), MEDLINE (OVID) (1966 to January 2006) and EMBASE (WebSPIRS) (1990 to September 2005). There were no language restrictions. SELECTION CRITERIA: Randomized controlled trials (RCTs) in children of either sex, which compared at least two antibiotics for CAP in hospital or ambulatory settings. DATA COLLECTION AND ANALYSIS: Data from full articles of selected studies were independently extracted by two authors. MAIN RESULTS: The review of these studies suggests that for treatment of pneumonia, co-trimoxazole is inferior in efficacy to both amoxycillin (failure rates odds ratio (OR) 1.33; 95% CI 1.05 to 1.67) and procaine penicillin (cure rates OR 2.64; 95% CI 1.57 to 4.45). Penicillin in conjunction with gentamycin was better than chloramphenicol alone (re-hospitalization rates OR 1.61; 95% CI 1.02 to 2.55). Co-amoxyclavulanic acid was better than amoxycillin alone (cure rates OR 10.44; 95% CI 2.85 to 38.21). There was no differences between injectable penicillin and oral amoxycillin (failure rates OR 1.03; 95% CI 0.81 to 1.31); azithromycin and erythromycin (cure rates OR 1.17; 95% CI 0.70 to 1.95); cefpodoxime and amoxycillin (cure rates OR 0.69; 95% CI 0.18 to 2.60); or azithromycin and co-amoxyclavulanic acid (cure rates OR 1.02; 95% CI 0.54 to 1.95, failure rates OR 1.42; 95% CI 0.43 to 4.66). AUTHORS' CONCLUSIONS: There were many studies each investigating multiple antibiotics with different methodologies. For treatment of ambulatory patients with CAP, amoxycillin was better than co-trimoxazole; there was no difference between azithromycin and erythromycin, or between cefpodoxime and co-amoxyclavulanic acid. For hospitalized patients, procaine penicillin was better than co-trimoxazole; and the combination of penicillin and gentamycin was better than chloramphenicol alone. Injectable penicillin and oral amoxycillin had similar failure rates. For the rest of the antibiotics there were only single studies available. There is a need for more studies with large patient populations and similar methodologies in order to compare newer antibiotics.

Home visits by community health workers to improve identification of serious illness and care seeking in newborns and young infants from low- and middle-income countries.

The objectives of this review were to evaluate the effect of home visits by trained community health workers (CHWs) to successfully identify newborns and young infants (up to 59 days of age) with serious illness and improve care seeking from a health facility. The authors searched the Cochrane Central Register of Controlled Trials, MEDLINE and EMBASE. Abstracts of all articles were read by two authors independently and relevant articles selected. Data were extracted in a pretested questionnaire by two authors independently. Statistical analysis was performed using Review Manager software. A meta-analysis of included randomized controlled trials (RCTs) was carried out. Pooled estimates (risk ratios (RRs) with 95% confidence intervals (CIs)) of the evaluated outcome measures were calculated by the generic inverse variance method. Seven articles were identified for inclusion in the review. None of them compared the diagnosis of serious illness in young infants by health workers to a 'gold standard' diagnosis. Three studies were available for evaluating the ability of CHWs to identify seriously ill young infants/signs of serious illness. These studies suggest that sensitivity to identify serious illness ranged from 33.3 to 90.5% and specificity from 75.61 to 98.4%. For the outcome of improved care seeking from a health facility, after pooling the data from six RCTs with 4760 subjects in the intervention and 4398 subjects in the control arm, there was a significant improvement in care seeking in the home visit arm (RR=1.35; 95% CI=1.15 to 1.58). Moderate quality evidence indicated that home visits by trained CHWs were associated with improved care-seeking for sick young infants from health facilities by appropriate health care providers in resource-limited settings. However, there is a lack of data regarding successful identification of serious illness. Evidence from validation studies supports the implementation of home visits by trained CHWs for improving outcomes in sick newborns and young infants in resource-limited areas. Further well-designed studies evaluating the effect of home visits by CHWs on successful identification of seriously ill newborns and young infants should include verification by a 'gold standard'.

Pharmacokinetics of isoniazid, rifampicin, pyrazinamide and ethambutol in HIV-infected Indian children.

SETTING: Co-infection with the human immunodeficiency virus (HIV) may lead to inadequate plasma concentrations of anti-tuberculosis drugs in children with tuberculosis (TB). OBJECTIVE: To describe the influence of HIV infection on the pharmacokinetics of isoniazid, rifampicin, pyrazinamide and ethambutol in children. DESIGN: Prospective drug estimation study in two cohorts of children: HIV-infected (n = 24) and non-HIV-infected (n = 32) with TB. Dosages used were based on earlier World Health Organization recommendations. All four drugs were estimated simultaneously using liquid chromatography mass spectrometry. RESULTS: The HIV-TB co-infected children had a mean age of 105.9 months (standard deviation 43.1); there were 10 girls (41.7%). The maximum plasma concentration (Cmax), time taken to achieve Cmax, area under curve from 0-4 h and 2 h concentrations of isoniazid (INH), rifampicin (RMP) and pyrazinamide (PZA) were not affected by the HIV status of the children. Ethambutol (EMB) concentrations were lower in HIV-TB co-infected children. Inadequate 2 h concentrations of INH, RMP and EMB were found in the majority of the children in both groups. PZA concentrations were adequate in almost all children. Younger age and lower dose were associated with lower 2 h concentrations of INH and RMP. CONCLUSION: Inadequate concentrations of INH, RMP and EMB in both HIV-TB-infected and non-HIV-infected children provide support for the recently revised recommended doses of INH and RMP. EMB levels were lower in HIV-infected children; however, more studies are needed to validate this observation.

Mortality and Other Outcomes in Relation to First Hour Fluid Resuscitation Rate: A Systematic Review.

OBJECTIVE: To determine the effect of different regimen of first hour fluid administration rates on mortality and severe consequences of impaired circulation in 2 to 60 months old children with impaired circulation. DESIGN: Systematic review of randomized controlled trials. DATA SOURCES: Various databases including PubMed, Cochrane Library and EMBASE were searched. RESULTS: We found only two relevant trials; one was excluded as there was no comparator arm. Only one study (The FEAST Trial) compared boluses with maintenance fluid alone in children with severe febrile illness and one or more signs of impaired perfusion. The 48 hour mortality was more in the bolus group (RR 1.45, 95% CI 1.13,1.86). The quality of evidence is rated as moderate. For the children who met the WHO criteria for shock (severely impaired circulation) (n=65 children), those receiving boluses had higher mortality (RR 2.40, 95% CI 0.84, 6.88); the quality of evidence was rated as very low. CONCLUSION: A single large randomized controlled trial conducted in low-resource settings indicates that administration of fluid bolus is associated with higher mortality in comparison to the maintenance fluids alone in children with severe febrile illness and one or more signs of impaired perfusion. The findings are not generalizable to contexts with different severity of and different causes of shock and in centers with better facilities. There is urgent need for research in different settings to determine the optimal rate of fluid resuscitation in the first hour in children presenting with impaired circulation, particularly with severely impaired circulation.

Outcomes of Category II anti-tuberculosis treatment in Indian children.

SETTING: The emerging threat of multidrug-resistant tuberculosis (MDR-TB) has cast doubt on the efficacy of Category II anti-tuberculosis treatment in retreatment cases. Data on outcomes of treatment with the Category II regimen in children are scarce. OBJECTIVE: To study outcomes of Category II anti-tuberculosis treatment in Indian children. DESIGN: Charts belonging to patients registered between 2004 and 2012 at the Paediatric Tuberculosis Clinic, All India Institute of Medical Sciences, New Delhi, India, were reviewed, and children receiving Category II anti-tuberculosis treatment were included in the study. Outcomes were recorded as treatment success and poor outcome, which included treatment failure and default. RESULTS: A total of 125 children (mean age 101.6 months, standard deviation 42.9; girls 58 [46.4%]) were initiated on Category II anti-tuberculosis treatment, mainly due to worsening clinical conditions (36.8%) and relapse (36%). Treatment success, treatment failure and default were recorded in respectively 80 (64%), 20 (16%) and 25 (20%) children. Children who were non-adherent to previous treatment tended to default from Category II treatment as well (11.8% in previous non-defaulters vs. 37.5% in previous defaulters, P = 0.004). CONCLUSION: Category II anti-tuberculosis treatment was effective in approximately 60% of children who had failed or defaulted from the previous regimen. All efforts should be made to isolate Mycobacterium tuberculosis and perform drug susceptibility testing to identify MDR-TB in children.

Traditional Indian systems of medicine.

INTRODUCTION: A number of traditional systems of medicine exist in India of which Ayurveda is the most popular. Despite being in use for more than 3000 years, few properly designed trials have scientifically examined the clinical potential of Ayurvedic and other medications. METHODS: We reviewed the MEDLINE database to identify clinical trials conducted using traditional Indian medicines. Single case reports were excluded. RESULTS: Ayurvedic preparations have been successfully used for the treatment of bronchial asthma, ischaemic heart disease and hyperlipidaemia. Formulations containing curcumin were reported to reduce inflammation and disability in double-blind clinical trials on patients with rheumatoid arthritis. A number of products are reported to be useful in patients with acute viral hepatitis. A multicentric study by the Indian Council of Medical Research showed that a preparation from Pterocarpus marsupium was effective in reducing levels of blood glucose and glycosylated haemoglobin in patients with non-insulin-dependent diabetes mellitus. In another multicentric trial, patients with fistula-in-ano were randomised to surgery or application of medicated seton (Ksharsootra). Surgical treatment led to a faster cure but recurrence rates were lower with medicated seton. Administration of extract from Bacopa monnieri, to children with mental retardation, was reported to significantly improve short-term and long-term memory. CONCLUSIONS: Evidence-based studies on the efficacy and safety of traditional Indian medicines are limited. The essential ingredient in most formulations is not precisely defined. High quality studies are necessary to evaluate and compare the value of traditional Indian drugs to modern medicine.

Management of unresponsive asthma.

Difficult asthma is defined as asthma that is not controlled despite treatment with> 800 micro g budesonide or equivalent per day. Poor control is defined as the need for bronchodilators more than three times a week, school absence of more than five days a term, or one episode or more of wheezing each month. Common causes of poor response to treatment include; wrong diagnosis, inappropriate medications or improper inhalation technique, poor adherence to medications and co-morbidity. Steroid resistant asthma is uncommon and estimated to be 1 in 1000-10000 asthmatic patients. If there is no functional improvement to prednisolone 2 mg/kg/day for 2 weeks with adherence checked by measuring serum prednisolone and cortisol levels, a fibreoptic bronchoscopic examination with bronchoalveolar lavage and large airway biopsy should be considered. Eosinophilic inflammation identified on the biopsy in a child who is unresponsive to prednisolone may benefit from alternative anti-inflammatory treatments such as cyclosporin. Neutrophilic infiltration in biopsy may benefit with macrolide antibiotics, 5-lipogenase inhibitors or theophyllines.