Rotavirus is the most common cause of acute gastroenteritis in infants and children worldwide. The functional correlation of B-and T-cells to long-lasting immunity against rotavirus infection in the literature is limited. In this work, a series of computational immuno-informatics approaches were applied and identified 28 linear B-cells, 26 conformational B-cell, 44 T C cell and 40 T H cell binding epitopes for structural and non-structural proteins of rotavirus. Further selection of putative B and T cell epitopes in the multi-epitope vaccine construct was carried out based on immunogenicity, conservancy, allergenicity and the helical content of predicted epitopes. An in-silico vaccine constructs was developed using an N-terminal adjuvant (RGD motif) followed by T C and T H cell epitopes and B-cell epitope with an appropriate linker. Multi-threading models of multi-epitope vaccine construct with Band T-cell epitopes were generated and molecular dynamics simulation was performed to determine the stability of designed vaccine. Codon optimized multi-epitope vaccine antigens was expressed and affinity purified using the E. coli expression system. Further the T cell epitope presentation assay using the recombinant multi-epitope constructs and the T cell epitope predicted and identified in this study have not been investigated. Multiepitope vaccine construct encompassing predicted B-and T-cell epitopes may help to generate long-term immune responses against rotavirus. The computational findings reported in this study may provide information in developing epitope-based vaccine and diagnostic assay for rotavirus-led diarrhea in children's.

Background: Approximately 85% of all term and most preterms develop clinical jaundice. Hyperbilirubinrmia is defined as a total bilirubin (TB) >95th percentile on the hour-specific Bhutani normogram. Pathological jaundice implies the onset of jaundice before 24 hours of age, rate of rise in TSB of >0.2 mg/dl/hour and jaundice persisting after 14 days in term and 21 days in late-preterms. The aim of this study was to determine the incidence, progression and the predictors of pathological jaundice among the late preterm infants admitted in Paediatrics ward of a tertiary care centre, Imphal.Methods: A hospital based prospective cohort study was carried out in paediatrics department, RIMS during a period of 2 years (September 2019 to August 2022) with approval from research ethics board. Sample size was 100 based on consecutive sampling.Results: Pathological jaundice developed among 65.0% (95% CI: 54.7-74.1%) neonates. The median duration of onset was 47.0 hours. The mean bilirubin at the time of diagnosis of jaundice was 15.5 mg/dl. Three neonates underwent exchange transfusion. Of these 3, one had pre-exchange bilirubin encephalopathy. Male gender, breast feeding, sepsis, infants blood group (B +ve), jaundice among the siblings and birth trauma (birth asphyxia/cephalhematoma) were significantly associated with the development of neonatal hyperbilirubinemia.Conclusions: Jaundice is condition that is often present and constitutes one of the major risks for neurodevelopmental issues in later life and the risk is further compounded by prematurity. Hence further studies with a larger sample size on a multicentric level could add robustness to our study thereby helping in better understanding and management of the condition.

Introduction:Incidence of Rh negative pregnancy in western countries is 15%, but in India it varies from 3% to 5.7%. It is a high risk pregnancy, as it may cause antigen-antibody reaction and haemolysis. However, it can be prevented by adequate measures. Aims and objectives: To determine feto-outcome in Rh negative pregnant women. Materials and methods: Across sectional study was carried out in the department of Obstetrics & Gynaecology and Paediatrics, Regional Institute of Medical Sciences, Imphal, Manipur for the duration of 2 years. The total of 38 Rh negative pregnant women were included in the study after applying inclusion and exclusion criteria. Results: In the present study among 38 patients, 31.6% were delivered through Elective LSCS, 26.31% were vaginally delivered and 21.1% were Emergency LSCS. Among 38 patients, 5.3% were preterm birth, 92.1% were term birth and 2.6% were post term birth. In this study, 7.9% of patients in the study group were indirect coomb' test (ICT) positive, 92% were ICT negative. Out of 38 patients, 20 were booked who received antenatal anti-D prophylaxis in contrast to 18 unbooked patients. 35 out of 38 patients received post-natal prophylaxis with 300 mcg anti-D. Perinatal outcome in 38 patients was 73.7% were healthy, NICU admission were 15.8%, intrauterine death (IUD) were 5.3% and 2.6% were neonatal death. The conventional treatment measures are appropriate hydration, phototherapy and exchange transfusion. Conclusions: Rh haemolytic disease of the new-born forms common and preventable cause of maternal and perinatal morbidity. Anti-D IgG administration and proper fetal and neonatal monitoring when required decreases the burden of disease

Background: Diarrhoeal diseases are responsible for causing 3 million deaths worldwide every year especially among the children and also the commonest cause of morbidity and mortality in developing countries like India. Infective diarrhoea could be either bacterial, viral, parasitic or occasionally a combination of these.Methods: A cross-sectional study was carried out in children below 12 years with acute diarrhoea in theMicrobiology Department, RIMS, Imphal for a period of 2 years. Stool samples were subjected to routine microbiological examination, followed by culture and sensitivity. Data were collected in a predesigned data collection sheet.Results: Majority of the diarrhoeal cases were seen among the age group of 1-3 years (44.3%), predominantly among the male children (66.2%) and mostly in summer. Out of 210 culture positive stool samples, Escherichia coli(83.3%) was the predominant enteropathogen with followed by Shigella spp.(12.9%), Klebsiella spp. (2.9%) and Salmonella spp. (1%). Serotyping revealed thirty five enteropathogenic E. coli, eighteen Shigella flexneri, seven Shigellasonnei, two Shigella boydii and two Salmonella typhimurium. Majority of the isolates showed high resistance to amoxicillin, ampicillin, ciprofloxacin, ofloxacin and cotrimoxazole.Conclusions: Bacterial enteropathogens are an important cause of acute diarrhoea among children. Rehydration therapy remains the initial treatment. Though it is usually self-limiting, empirical and specific antimicrobial therapy can be considered in certain situations. Awareness of improving hygiene and infectious diseases may help reduce the burden of infection.

Background: Wheezing is common throughout infancy and childhood except in the neonatal period where it is relatively rare. By 10 years of age, about 19% of children experience wheezing with an average onset at 3 years of age. This study was aimed to identify the diverse factors associated with wheezing in children aged 2 months to 60 months and to study clinical profile along with short term outcome of the same.Methods: It was a hospital based cross-sectional study carried out in the Department of Paediatrics, Regional Institute of Medical Sciences Hospital (RIMS), Imphal, Manipur. The Study population consisted of randomly selected 131 children aged 2 months to 60 months who were admitted in Paediatrics ward with the symptom of wheezing.Results: Authors found that age below 12 months, male sex, low socioeconomic conditions and artificial breastfeeding practices were important risk factors for wheeze.Conclusions: Wheezing is accountable for a high demand of medical consultations and emergency care services with relatively high rates of hospitalization. Along with ARI, it plays an important role in infant mortality. In Manipur, it is being observed that increasing number of children with wheezing are attended by paediatricians in ED, OPD and ward, thereby proving an added burden to the younger age group. Therefore proper health education and counselling of parents, promotion of exclusive breast feeding and improvement of socioeconomic status can play a vital role in preventing occurrence of wheeze among the children.

BACKGROUND Perinatal asphyxia is an insult to the foetus or newborn due to lack of oxygen (hypoxia) and/or lack of perfusion (ischaemia) to various organs, of sufficient magnitude and duration to produce more than fleeting functional and/or biochemical changes. There are various studies available to indicate foetal asphyxia like intrapartum electronic foetal monitoring, foetal or umbilical cord pH measurement, Meconium-Stained Amniotic Fluid (MSAF), APGAR score, Hypoxic-Ischaemic Encephalopathy (HIE) and evidence of multiorgan dysfunction. But in a resource-limited institute a need for simpler and cheaper alternatives are sought. In this prospective analysis, we report our experience with a study on nucleated RBC (nRBC) count and its association with foetal asphyxia and its immediate neurological outcome in term newborns in RIMS Hospital, Imphal, Manipur. The objective is to study the association between the nucleated RBC count and foetal asphyxia and its immediate neurological outcome in term newborns. MATERIALS AND METHODS In this case-control study conducted over 2 years in a tertiary care referral hospital in Manipur, India, 50 asphyxiated (study group) and 50 non-asphyxiated (control group) neonatal cord blood samples were randomly selected and analysed. RESULTS nRBC counts were significantly higher in those with meconium-stained amniotic fluid (MSAF) (12.66 ± 6.17) than in normal amniotic fluid group (9.25 ± 5.88). nRBC/100 WBCs were also significantly elevated in asphyxiated newborns (14.10 ± 4.79) in comparison to non-asphyxiated group (5.44 ± 3.47). As the stage of Hypoxic-Ischaemic Encephalopathy (HIE) increased from grade I (9.4 ± 4.57) to grade III (21 ± 2.94), nRBC count also increased in frequency showing a direct proportion. CONCLUSION In limited health care settings, nRBC count can be an easy and economical alternative for detecting perinatal asphyxia and also the severity of HIE.

Researchers around the world are developing more than 145 vaccines (DNA/mRNA/whole-virus/viral-vector/protein-based/repurposed vaccine) against the SARS-CoV-2 and 21 vaccines are in human trials. However, a limited information is available about which SARS-CoV-2 proteins are recognized by human B- and T-cell immune responses. Using a comprehensive computational prediction algorithm and stringent selection criteria, we have predicted and identified potent B- and T-cell epitopes in the structural proteins of SARS-CoV and SARS-CoV-2. The amino acid residues spanning the predicted linear B-cell epitope in the RBD of S protein (370-NSASFSTFKCYGVSPTKLNDLCFTNV-395) have recently been identified for interaction with the CR3022, a previously described neutralizing antibody known to neutralize SARS-CoV-2 through binding to the RBD of the S protein. Intriguingly, most of the amino acid residues spanning the predicted B-cell epitope (aa 331-NITNLCPFGEVFNATRFASVYAWNRK-356, 403-RGDEVRQIAPGQTGKIADYNYKLPD-427 and aa 437- NSNNLDSKVGGNYNYLYRLFRKSNL-461) of the S protein have been experimentally verified to interact with the cross-neutralizing mAbs (S309 and CB6) in an ACE2 receptor-S protein interaction independent-manner. In addition, we found that computationally predicted epitope of S protein (370-395) is likely to function as both linear B-cell and MHC class II epitope. Similarly, 403-27 and 437-461 peptides of S protein were predicted as linear B cell and MHC class I epitope while, 177-196 and 1253-1273 peptides of S protein were predicted as linear and conformational B cell epitope. We found MHC class I epitope 316-GMSRIGMEV-324 predicted as high affinity epitope (HLA-A*02:03, HLA-A*02:01, HLA-A*02:06) common to N protein of both SARS-CoV-2 and SARS-CoV (N317-325) was previously shown to induce interferon-gamma (IFN-γ) in PBMCs of SARS-recovered patients. Interestingly, two MHC class I epitopes, 1041-GVVFLHVTY-1049 (HLA-A*11:01, HLA-A*68:01, HLA-A*03:01) and 1202-FIAGLIAIV-1210 (HLA-A*02:06, HLA-A*68:02) derived from SARS-CoV S protein with epitope conservancy between 85 to 100% with S protein of SARS-CoV-2 was experimentally verified using PBMCs derived from SARS-CoV patients. We observed that HLA-A*02:01, HLA-A*02:03, HLA-A*02:06, HLA-A*11:01, HLA-A*30:01, HLA-A*68:01, HLA-A*68:02, HLA-B*15:01 and HLA-B*35:01 have been predicted to bind to the maximum number of MHC class I epitope (based on the criterion of allele predicted to bind more than 30 epitopes) of S protein of SARS-CoV-2. Similarly, we observed that HLA-A*02:06, HLA-A*30:01, HLA-A*30:02, HLA-A*31:01, HLA-A*32:01, HLA-A*68:01, HLA-A*68:02, HLA-B*15:01 and HLA-B*35:01 are predicted to bind to the maximum number of MHC class I epitope of N protein of SARS-CoV-2. We found that HLA-DRB1*04:01, HLA-DRB1*04:05, HLA-DRB1*13:02, HLA-DRB1*15:01, HLA-DRB3*01:01, HLA-DRB3*02:02, HLA-DRB4*01:01, HLA-DRB5*01:01, HLA-DQA1*04:01, DQB1*04:02, HLA-DPA1*02:01, DPB1*01:01, HLA-DPA1*01:03, DPB1*02:01, HLA-DPA1*01:03, DPB1*04:01, HLA-DPA1*03:01, DPB1*04:02, HLA-DPA1*02:01, DPB1*05:01, HLA-DPA1*02:01, and DPB1*14:01 are predicted to bind to the maximum number of MHC class II epitope of S protein of SARS-CoV-2. Alleles such as HLA-DRB1*04:01, HLA-DRB1*07:01, HLA-DRB1*08:02, HLA-DRB1*09:01, HLA-DRB1*11:01, HLA-DRB1*13:02, HLA-DRB3*02:02, HLA-DRB5*01:01, HLA-DQA1*01:02, DQB1*06:02, DPB1*05:01 and HLA-DPA1*02:01 are found to interact with the maximum number of MHC class II epitope of N protein of SARS-CoV-2. Using the IEDB tool we found the occurrence of HLA alleles with population coverage of around 99% throughout the world. The findings of computational predictions of mega-pool of B- and T-cell epitopes identified in the four main structural proteins of SARS-CoV-2 provides a platform for future experimental validations and the results of present works support the use of RBD or the full-length S and N proteins in an effort towards designing of recombinant protein-based vaccine and a serological diagnostic assay for SARS-CoV-2.

No abstract