Reference Ranges of Different Lymphocyte Subsets in Indian Children: A Multi-Centric Study.

OBJECTIVE: To determine the reference ranges of various lymphocyte subsets in healthy Indian children. DESIGN: Descriptive cross-sectional study. SETTING: Four centers in India representing four geographical regions. PARTICIPANTS: 1104 children from neonatal age to 18 years of age. MEASUREMENT: One time measurement of absolute count and percentages of different lymphocyte subsets i.e. T lymphocytes (CD3+T, CD4+T, CD8+T cells), B lymphocytes (CD19+B cells) and Natural Killer lymphocytes (CD15/16+NK cells) in whole blood using multicolor flow cytometry. RESULTS: The absolute cell counts of various lymphocytes were found to increase from newborn to 10 months of age, followed by gradual decline until 18 years; however, the proportion of immune cells remained largely similar. Gender did not have a significant impact on the reference ranges, whereas counts were found to vary as per the geographical locations. CONCLUSIONS: These reference ranges will be useful to monitor and predict the immune status in pediatric population. The variation in region wise ranges could be confirmed by testing more number of samples in the specific age groups.

Evaluating the efficacy of a multistrain probiotic supplementation for prevention of neonatal sepsis in 0-2-month-old low birth weight infants in India-the "ProSPoNS" Study protocol for a phase III, multicentric, randomized, double-blind, placebo-controlled trial.

BACKGROUND: Progress has been made in the reduction of under-five mortality in India; however, neonatal mortality is reducing at a slower rate. Efforts are required to bring down neonatal mortality in order to attain the Sustainable Development Goal-3. Prevention of sepsis among the high-risk, vulnerable low birth weight neonates by a newer intervention with probiotic supplementation is promising. METHODS: A phase III, multicenter, randomized, double-blind, placebo-controlled study is being conducted at six sites in India. A total of 6144 healthy low birth weight (LBW) infants fulfilling the eligibility criteria would be enrolled within the first week of life, after obtaining written informed consent from the parents of the infant. Randomization in 1:1 ratio, stratified by site, sex, and birth weight, would be done through an interactive web response system (IWRS) using a standard web browser and email service. Vivomixx®, a probiotic containing a mix of 8 strains of bacteria, in a suspension form standardized to deliver 10 billion CFU/ml, or an organoleptically similar placebo would be fed to enrolled infants in a 1-ml/day dose for 30 days. The follow-up of enrolled infants for 60 days would take place as per a pre-specified schedule for recording morbidities and outcome assessments at the six participating sites. Screening for morbidities would be conducted by trained field workers in the community, and sick infants would be referred to designated clinics/hospitals. A physician would examine the referred infants presenting with complaints and clinical signs, and blood samples would be collected from sick infants for diagnosis of neonatal sepsis by performing sepsis screen and blood culture. Appropriate treatment would be provided as per hospital protocol. The study would be implemented as per the MRC guideline for the management of Global Health Trials in accordance with ICH-GCP and Indian Regulatory guidelines. A contract research organization would be engaged for comprehensive monitoring and quality assurance. The final analysis would be conducted in a blinded manner as per the statistical analysis plan (SAP) to estimate the primary outcomes of sepsis, possible serious bacterial infection (PSBI), and secondary outcomes. The codes will be broken after DMC permission. The protocol has been reviewed by the Research Ethics Committee of the Liverpool School of Tropical Medicine (REC-LSTM), from Research Ethics Committees of the six subject recruitment participating sites. DISCUSSION: This adequately powered and well-designed trial would conclusively answer the question whether probiotics can prevent neonatal sepsis in the high-risk group of low birth weight infants as indicated by a pilot study in 1340 LBW infants, evidence from systematic reviews of hospital-based studies, and a primary study on healthy newborns in Orissa. Results of the study would be generalizable to India and other low-middle-income countries. TRIAL REGISTRATION: Clinical Trial Registry of India (CTRI) CTRI/2019/05/019197 . Registered on 16 May 2019.

Role of probiotics VSL#3 in prevention of suspected sepsis in low birthweight infants in India: a randomised controlled trial.

OBJECTIVES: To assess the effect of the probiotic VSL#3 in prevention of neonatal sepsis in low birthweight (LBW) infants. DESIGN: Randomised, double-blind, placebo-controlled trial. SETTING: Community setting in rural India. PARTICIPANTS: LBW infants aged 3-7 days. INTERVENTIONS: Infants were randomised to receive probiotic (VSL#3, 10 billion colony-forming units (cfu)) or placebo for 30 days, and were followed up for 2 months. MAIN OUTCOME MEASURE: Possible serious bacterial infection (PSBI) as per the Integrated Management of Neonatal Childhood Illnesses algorithm, as diagnosed by fieldworkers/physicians. RESULTS: 668 infants were randomised to VSL#3 and 672 to placebo. By intention-to-treat analysis, the risk of PSBI among infants in the overall population of LBW infants was not statistically significant (RR 0.79 (95% CI 0.56 to 1.03)). Probiotics reduced median days of hospitalisation (6 days vs 3 days in probiotics) (p=0.018) but not the risk of hospitalisation (RR 0.66 (95% CI 0.42 to 1.04). The onset of PSBI in 10% of infants occurred on the 40th day in the probiotics arm versus the 25th day in the control arm (p=0.063). CONCLUSIONS: Daily supplementation of LBW infants with probiotics VSL#3 (10 billion cfu) for 30 days led to a non-significant 21% reduction in risk of neonatal sepsis. A larger study with sufficient power and a more specific primary end point is warranted to confirm the preventive effect of VSL#3 on neonatal sepsis in LBW infants. TRIAL REGISTRATION NUMBER: The study is registered at the Clinical Trial Registry of India (CTRI/2008/091/000049).

Gene-environment interaction in Alzheimer's disease.

OBJECTIVES: The aim was to examine the gene environment (GxE) interaction with reference to APO E genotypes, serum lipids and organochlorine pesticides (OCPs) as one of the factors in the etiology of Alzheimer's disease (AD). METHODS: A case control study was used to examine, APOE HhaI polymorphism by polymerase chain reaction (PCR)/PCRrestriction fragment length polymorphism method, serum lipids by autoanalyser and OCPs by gas chromatography (GC). RESULTS: APOE ∈4 allele frequency was significantly high (p=0.000, OR=5.73, CI=2.68-12.50) in AD as compared to controls. The serum cholesterol, ß- hexachlorocyclohexane and dieldrin are risk factors for AD independent of the APOE ∈4 risk allele, recording an odds ratio of 1.16, 11.38 and 10.45 respectively. CONCLUSION: GxE interactions exist with APOE ∈4 allele status that need to be considered for the study design and analysis of such data in future studies of AD.

Assessment of vaccine wastage during a pulse polio immunization programme in India.

A study to assess the wastage factor of oral polio vaccine (OPV) in the Pulse Polio Immunization (PPI) programme of the Government of India was undertaken by the Indian Council of Medical Research (ICMR) at approximately 31,000 immunization booths all over the country. The study was conducted through the network of 31 Human Reproduction Research Centres (HRRCs) and other ICMR institutes. Wastage at the point of administration of OPV was estimated to be 14.5% with a wastage factor of 1.17 which is well below the assumed wastage of 33% and the corresponding wastage factor of 1.5 in the PPI programme. The wastage and wastage factor as estimated in the present study were also less than the wastage of 25% and the wastage factor of 1.33 recommended by the World Health Organization. Minimum wastage (6.3%) at Kanchipuram and maximum wastage (22.1%) at Kanpur were observed. Further, the wastage of unopened vials and vials during use was also observed following colour changes on the vaccine vial monitor (VVM), indicating poor cold-chain maintenance at the immunization site. In total, 13 booths reported wastage of nine or more unopened vials, whereas 19 booths reported wastage of nine or more vials during use because of colour changes on VVM. Other reasons for wastage of vaccine were also observed from a sample of booths. The technology of introducing VVM on OPV vials for monitoring the cold-chain proved useful in situations in which mass vaccination programmes such as PPI are carried out.

An application of the technique of density estimation in geographical epidemiology of goitre in the Muzaffarpur district of Bihar state of India.

The Gangetic plains in the Sub-Himalayan belt of India are known endemic areas for goitre. The districts of Muzaffarpur and Sitamarhi in the eastern zone in the Bihar state of India showed a prevalence of over 30 per cent of goitre in a survey conducted by the Indian Council of Medical Research (ICMR) during 1983-1986. In the present paper, the technique of density estimation is employed to estimate the probability density functions of goitre-affected and normal (non-goitre) populations of the district of Muzaffarpur. The likelihood ratio is then plotted against distance of the villages from the Indo-Nepal border keeping their distances from the headquarters in the district of Sitamarhi at fixed level. Using the odds form of Bayes rule, the posterior odds of goitre are calculated and compared at two given points.