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Immune imprinting is a phenomenon in which prior antigenic experiences influence responses to subsequent infection or vaccination 1,2 . The effects of immune imprinting on serum antibody responses after boosting with variant-matched SARS-CoV-2 vaccines remain uncertain. Here we characterized the serum antibody responses after mRNA vaccine boosting of mice and human clinical trial participants. In mice, a single dose of a preclinical version of mRNA-1273 vaccine encoding Wuhan-1 spike protein minimally imprinted serum responses elicited by Omicron boosters, enabling generation of type-specific antibodies. However, imprinting was observed in mice receiving an Omicron booster after two priming doses of mRNA-1273, an effect that was mitigated by a second booster dose of Omicron vaccine. In both SARS-CoV-2-infected and uninfected humans who received two Omicron-matched boosters after two or more doses of the prototype mRNA-1273 vaccine, spike-binding and neutralizing serum antibodies cross-reacted with Omicron variants as well as more distantly related sarbecoviruses. Because serum neutralizing responses against Omicron strains and other sarbecoviruses were abrogated after pre-clearing with Wuhan-1 spike protein, antibodies induced by XBB.1.5 boosting in humans focus on conserved epitopes targeted by the antecedent mRNA-1273 primary series. Thus, the antibody response to Omicron-based boosters in humans is imprinted by immunizations with historical mRNA-1273 vaccines, but this outcome may be beneficial as it drives expansion of cross-neutralizing antibodies that inhibit infection of emerging SARS-CoV-2 variants and distantly related sarbecoviruses., (© 2024. The Author(s), under exclusive licence to Springer Nature Limited.)

Long non-coding RNAs (lncRNAs) are significant contributors in maintaining genomic integrity through epigenetic regulation. LncRNAs can interact with chromatin-modifying complexes in both cis and trans pathways, drawing them to specific genomic loci and influencing gene expression via DNA methylation, histone modifications, and chromatin remodeling. They can also operate as building blocks to assemble different chromatin-modifying components, facilitating their interactions and gene regulatory functions. Deregulation of these molecules has been associated with various human diseases, including cancer, cardiovascular disease, and neurological disorders. Thus, lncRNAs are implicated as potential diagnostic indicators and therapeutic targets. This review discusses the current understanding of how lncRNAs mediate epigenetic control, genomic integrity, and their putative functions in disease pathogenesis., Competing Interests: The author declares there are no competing interests.

Few seroprevalence studies have been conducted on coronavirus disease (COVID-19) in Nepal. Here, we aimed to estimate seroprevalence and assess risk factors for infection in the general population of Nepal by conducting two rounds of sampling. The first round was in October 2020, at the peak of the first generalized wave of COVID-19, and the second round in July-August 2021, following the peak of the wave caused by the delta variant of SARS-CoV-2. We used cross-sectional probability-to-size (PPS)-based multistage cluster sampling to estimate the seroprevalence in the general population of Nepal at the national and provincial levels. We tested for anti-SARS-CoV-2 total antibody using the WANTAI SARS-CoV-2 Ab ELISA kit. In Round 1, the overall national seroprevalence was 14.4%, with provincial estimates ranging from 5.3% in Sudurpaschim to 27.3% in Madhesh Province. In Round 2, the estimated national seroprevalence was 70.7%, with the highest in the Madhesh Province (84.8%) and the lowest in the Gandaki Province (62.9%). Seroprevalence was comparable between males and females (Round 1, 15.8% vs. 12.2% and Round 2, 72.3% vs. 68.7%). The seroprevalence in the ecozones-Terai, hills, and mountains-was 76.3%, 65.3%, and 60.5% in Round 2 and 17.7%, 11.7%, and 4.6% in Round 1, respectively. In Nepal, COVID-19 vaccination was introduced in January 2021. At the peak of the first generalized wave of COVID-19, most of the population of Nepal remained unexposed to SARS-CoV-2. Towards the end of the second generalized wave in April 2021, two thirds of the population was exposed., (© 2023 The Authors. Influenza and Other Respiratory Viruses published by John Wiley & Sons Ltd.)

Mpox virus (MPXV) caused a global outbreak in 2022. Although smallpox vaccines were rapidly deployed to curb spread and disease among those at highest risk, breakthrough disease was noted after complete immunization. Given the threat of additional zoonotic events and the virus's evolving ability to drive human-to-human transmission, there is an urgent need for an MPXV-specific vaccine that confers protection against evolving MPXV strains and related orthopoxviruses. Here, we demonstrate that an mRNA-lipid nanoparticle vaccine encoding a set of four highly conserved MPXV surface proteins involved in virus attachment, entry, and transmission can induce MPXV-specific immunity and heterologous protection against a lethal vaccinia virus (VACV) challenge. Compared with modified vaccinia virus Ankara (MVA), which forms the basis for the current MPXV vaccine, immunization with an mRNA-based MPXV vaccine generated superior neutralizing activity against MPXV and VACV and more efficiently inhibited spread between cells. We also observed greater Fc effector T H 1-biased humoral immunity to the four MPXV antigens encoded by the vaccine, as well as to the four VACV homologs. Single MPXV antigen-encoding mRNA vaccines provided partial protection against VACV challenge, whereas multivalent vaccines combining mRNAs encoding two, three, or four MPXV antigens protected against disease-related weight loss and death equal or superior to MVA vaccination. These data demonstrate that an mRNA-based MPXV vaccine confers robust protection against VACV.

The spotted fever group (SFG) of Rickettsia are zoonotic disease-causing pathogens, commonly transmitted by hard ticks to a wide range of hosts, including humans. Rickettsia conorii is the common SFG recognised in India, whereas most of the infections due to other group species go undifferentiated at the species level. Hence, this study was conducted to screen host-seeking ticks in the Western Ghats region, India, for the DNA of SFG Rickettsia. The ticks were collected from Kerala, Goa, and Maharashtra states of India during a survey conducted between November 2017 and January 2018. In total, 288 tick pools were screened for Rickettsia spp. DNA using pan-Rickettsia real-time PCR, and conventional PCR targeting the gltA, OmpA and 17-kDa protein-coding genes. Nucleotide sequences were subjected to phylogenetic analysis using the NCBI BLAST tool to identify submitted sequences with higher homology. Neighbour-joining trees were constructed using the reference sequences of the GenBank database. Overall, Rickettsia spp. DNA was detected in 27.2% (62/228 pools) of host-seeking ticks across the Western Ghats region, with an estimated minimum infection rate of 0.057. Upon phylogenetic analysis, it was identified that the detected sequences were highly similar (> 99% sequence homology) to R. africae, Candidatus R. laoensis and an un-categorised Rickettsia species, and they were widely carried by Haemaphysalis ticks. The current study is the first report of R. africae and Candidatus R. laoensis in ticks in India. Although the pathogenicity of these species is not well documented, they may pose a potential threat to both animal and the human population in this geographical region., (© 2023. The Author(s).)

Metal-organic frameworks (MOFs) with their tunable topology, functionality and coordination environment have been considered as potential materials for various applications including electrocatalysis. Herein, we have synthesised a water coordinated nickel based 2D metal-organic framework (Ni-MOF) and a coordination complex (Ni-C) and investigated their electrocatalytic OER activity. The Ni-MOF showed a 2D sheet structure with one water coordination whereas a four water molecule coordinated charged complex was formed in the Ni-C. Thermogravimetric analysis (TGA) confirmed their water coordination and good thermal stability. Interestingly, electrocatalytic OER studies showed strongly enhanced activity for the Ni-MOF and that it required a low overpotential (194 mV) to produce a geometric current density of 10 mA cm -2 . The Ni-C required 225 mV to produce 10 mA cm -2 . The post-catalytic analysis suggested that the Ni-MOF and Ni-C are converted to nickel hydroxides/oxyhydroxides during electrocatalysis and acted as the catalytic centre. The low Tafel slope and charge transfer resistance further supported the higher activity of Ni-MOF based nickel hydroxides/oxyhydroxides. Chronoamperometric studies revealed the excellent stability of the Ni-MOF based catalyst over 72 h. The present study revealed the potential of developing highly active electrocatalysts based on Ni-MOFs by optimizing the topology and coordination environment.

The use of metal-organic compounds as electrocatalysts for water splitting reactions has gained increased attention; however, a fundamental understanding of the structural requirement for effective catalytic activity is still limited. Herein, we synthesized water-coordinated mono and bimetallic copper complexes (CuPz-H 2 O·H 2 O, CuPz-H 2 O, CuBipy-H 2 O·H 2 O, and CuMorph-H 2 O) with varied intermetallic spacing (pyrazine/4,4'-bipyridine) and explored the structure-dependent oxygen evolution reaction (OER) activity in alkaline medium. Single crystal structural studies revealed water-coordinated monometallic complexes (CuMorph-H 2 O) and bimetallic complexes (CuPz-H 2 O·H 2 O, CuPz-H 2 O, CuBipy-H 2 O·H 2 O). Further, CuPz-H 2 O·H 2 O and CuBipy-H 2 O·H 2 O contained lattice water along with coordinated water. Interestingly, the bimetallic copper complex with lattice water and shorter interspacing between the metal centres (CuPz-H 2 O·H 2 O) showed strong OER activity and required an overpotential of 228 mV to produce a benchmark current density of 10 mA cm -2 . Bimetallic copper complex (CuPz-H 2 O) without lattice water but the same intermetallic spacing and bimetallic complex with increased interspacing but with lattice water (CuBipy-H 2 O·H 2 O) exhibited relatively lower OER activity. CuPz-H 2 O and CuBipy-H 2 O·H 2 O required an overpotential of 236 and 256 mA cm -2 , respectively. Monometallic CuMorph-H 2 O showed the lowest OER activity (overpotential 271 mV) compared to bimetallic complexes. The low Tafel slope and charge transfer resistance of CuPz-H 2 O·H 2 O facilitated faster charge transfer kinetics at the electrode surface and supported the enhanced OER activity. The chronoamperometric studies indicated good stability of the catalyst. Overall, the present structure-electrocatalytic activity studies of copper complexes might provide structural insight for designing new efficient electrocatalysts based on metal coordination compounds., Competing Interests: There are no conflicts to declare., (This journal is © The Royal Society of Chemistry.)

Objective: To report genotype data of the patients with Wilson disease (WD) hailing from across several parts of India to add to the available spectrum of causative variants in ATP7B gene (ATPase copper transporting beta polypeptide gene) and associated phenotypes in the Indian population., Methods: The entire ATP7B gene was sequenced in 58 patients with WD and additional testing was also done by MLPA to look for intragenic deletions duplications and exome sequencing to rule out genetic variations with similar phenotypic overlap., Results: Of all patients, 37 patients had a total of 33 distinct pathogenic variations, including 29 in the exonic regions and 4 at intronic splice sites. Of the variations identified, six were novel. The underlying genomic variations could be identified in nearly two-thirds of the patients by sequencing the entire gene., Conclusions: This study reports the genotype-phenotype data to add to the available spectrum of causative variants in ATP7B gene. The inability to detect a pathogenic variation in some patients and the existence of phenotypic variations in individuals with the same variation suggest that additional factors or genes may play a role in causation of the disease. Further, a marked genetic heterogeneity was found in the study patients, indicating ethnic diversity of the Indian population., (© 2022. The Author(s), under exclusive licence to Dr. K C Chaudhuri Foundation.)

Hand, foot, and mouth disease (HFMD) is a common childhood infection caused by human enteroviruses and is clinically characterised by fever with vesicular rash on the hands, feet, and mouth. While enterovirus A71 (EV-A71) and coxsackievirus A16 (CVA16) were the major etiological agents of HFMD in India earlier, the data on recently circulating enteroviruses associated with HFMD are sparse. Here, we describe the molecular epidemiology of enteroviruses associated with HFMD in South India from 2015 to 2017. We used archived enterovirus real-time reverse transcription (RT) PCR-positive vesicle swab and/or throat swab specimens from clinically suspected HFMD cases collected from four secondary-care hospitals in South India between July 2015 and December 2017. PCR amplification and sequencing were done based on the 5'VP1, 3'VP1, VP2, or 5´NCR regions to identify enterovirus types. Genetic diversity among enteroviruses was inferred by phylogenetic analysis. Of the 107 enterovirus RNA real-time RT-PCR-positive HFMD cases, 69 (64%) were typed as CVA6, 16 (15%) were CVA16, and one (1%) was CVA10, whereas in 21 (20%) cases, the virus was not typeable by any of the methods used in the study. The majority of HFMD cases (89, 83%) were in children less than five years old, while 11 (10.3%) were in adults. 5'VP1 yielded the maximum number of enteroviruses genotyped, and phylogenetic analysis showed that the CVA6 strains belonged to subclade D3, while the subclades of CVA16 and CVA10 were B1c and D, respectively. The predominant etiological agent of HFMD in South India during 2015-2017 was CVA6, followed by CVA16 and CVA10., (© 2022. The Author(s), under exclusive licence to Springer-Verlag GmbH Austria, part of Springer Nature.)

Lung cancer is a potentially lethal illness. Cancer detection continues to be a challenge for medical professionals. The true cause of cancer and its complete treatment have still not been discovered. Cancer that is caught early enough can be treated. Image processing methods such as noise reduction, feature extraction, identification of damaged regions, and maybe a comparison with data on the medical history of lung cancer are used to locate portions of the lung that have been impacted by cancer. This research shows an accurate classification and prediction of lung cancer using technology that is enabled by machine learning and image processing. To begin, photos need to be gathered. In the experimental investigation, 83 CT scans from 70 distinct patients were utilized as the dataset. The geometric mean filter is used during picture preprocessing. As a consequence, image quality is enhanced. The K -means technique is then used to segment the images. The part of the image may be found using this segmentation. Then, classification methods using machine learning are used. For the classification, ANN, KNN, and RF are some of the machine learning techniques that were used. It is found that the ANN model is producing more accurate results for predicting lung cancer., Competing Interests: The authors declare that they have no conflict of interest., (Copyright © 2022 Sharmila Nageswaran et al.)

Background: Kyasanur Forest disease (KFD) was first reported in 1957 and became an emerging tick-borne viral disease of public health importance in India. However, very little is known about the host-virus interaction and pathogenesis of KFD in humans. This study described the presence, duration, and kinetics of KFDV RNA in body fluids in infected human cases., Methodology: We enrolled 76 laboratory-confirmed KFD individuals and followed them up in the study. We obtained serial samples of blood, throat swabs in viral transport medium (VTM), urine, stool, and semen during the acute and convalescent phase of KFD illness. In addition, specimens were inactivated, and nucleic acid was extracted and tested for KFDV real-time reverse transcriptase -PCR. Clinical data was also obtained from the subjects., Result: The study provides evidence of KFD virus RNA in different biological body fluids of humans. The percentage positivity of KFDV RNA in blood was 100% during the first four days of illness. PCR became negative in most cases by 7-8 days; a subset of cases (14%) had prolonged viremia for up to 15 days post-onset of illness. Relatively low copies of KFDV RNA were also detected in throat swabs and urine in the first week of illness. In addition, we detected KFDV RNA in stool samples of cases of those who had diarrhea at an early stage of infection., Conclusion: The study provides evidence of KFDV RNA in different biological body fluids, which will help understand the pathogenesis, transmission pattern and develop diagnostic algorithms of KFDV in humans. In Kyasanur Forest disease infection, the blood has more RNA copies/ml than other body fluids, and viremia may last up to two weeks post-infection., Competing Interests: Declaration of Competing Interest Both the authors report no conflicts of interest. Both the authors have submitted the ICMJE Form for Disclosure of Potential Conflicts of Interest. Conflicts that the editors consider relevant to the manuscript's content have been disclosed., (Copyright © 2022 The British Infection Association. Published by Elsevier Ltd. All rights reserved.)

Aging is an inexorable event occurring universally for all organisms characterized by the progressive loss of cell function. However, less is known about the key events occurring inside the nucleus in the process of aging. The advent of chromosome capture techniques and extensive modern sequencing technologies have illuminated a rather dynamic structure of chromatin inside the nucleus. As cells advance along their life cycle, chromatin condensation states alter which leads to a different epigenetic landscape, correlated with modified gene expression. The exact factors mediating these changes in the chromatin structure and function remain elusive in the context of aging cells. The accumulation of DNA damage, reactive oxygen species and loss of genomic integrity as cells cease to divide can contribute to a tumor stimulating environment. In this review, we focus on genomic and epigenomic changes occurring in an aged cell which can contribute to age-related tumor formation., Competing Interests: The authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest., (Copyright © 2022 Sikder, Arunkumar, Melters and Dalal.)

Background: This study was carried out to understand the circulating genotypes of Hepatitis A virus (HAV) in South West, East and North East India during the period 2017-2018 as a part of acute febrile illness surveillance at the Manipal Institute of Virology., Methods: Archived serum samples of 48 Hepatitis A confirmed cases were subjected to RNA extraction using QIAamp® Viral RNA Mini Kit (QIAGEN, Germany). The samples with molecular confirmation for HAV by reverse transcriptase real-Time PCR (Real Star® HAV RT-PCR Kit 2.0, Altona Diagnostics, GmbH, Hamburg, Germany) were further subjected to nested conventional PCR targeting the 5' UTR region. The purified PCR products were sequenced using Big Dye Terminator Kit (Applied Biosystems, USA), in a 3500 XL genetic analyzer (Applied Biosystems, USA). The edited sequences by means of MEGA X (MEGA version 10.1) were compared with reference sequences in the NCBI nucleotide database., Results: From states of Assam, Goa, Gujarat, Karnataka, Kerala, Maharashtra, Odisha, Tamil Nadu and Tripura, 139 Hepatitis A and 33 Hepatitis E cases were reported during the study period. The median age of the acute Hepatitis A cases was 19 years (IQR 12.8-24) and most of the affected individuals were students between 10 and 19 years (52.5%). In the present study, 14 samples from Assam, Goa, Gujarat, Karnataka, Odisha, Kerala, Maharashtra and Tamil Nadu were genotyped as genotype IIIA by nested conventional polymerase chain reaction., Conclusion: The circulating HAV genotype in South West, North East and East India between 2017 and 2018 was IIIA., Competing Interests: The authors have none to declare., (© 2022 Director General, Armed Forces Medical Services. Published by Elsevier, a division of RELX India Pvt. Ltd.)

Chromosome instability is a critical event in cancer progression. Histone H3 variant CENP-A plays a fundamental role in defining centromere identity, structure, and function but is innately overexpressed in several types of solid cancers. In the cancer background, excess CENP-A is deposited ectopically on chromosome arms, including 8q24/ cMYC locus, by invading transcription-coupled H3.3 chaperone pathways. Up-regulation of lncRNAs in many cancers correlates with poor prognosis and recurrence in patients. We report that transcription of 8q24-derived oncogenic lncRNAs plays an unanticipated role in altering the 8q24 chromatin landscape by H3.3 chaperone-mediated deposition of CENP-A-associated complexes. Furthermore, a transgene cassette carrying specific 8q24-derived lncRNA integrated into a naïve chromosome locus recruits CENP-A to the new location in a cis-acting manner. These data provide a plausible mechanistic link between locus-specific oncogenic lncRNAs, aberrant local chromatin structure, and the generation of new epigenetic memory at a fragile site in human cancer cells.

Water pollution by organic dyes poses great challenge to the environment and living organism. Hence effective removal of organic dyes by cost effective methods have received significant attention in recent years. Herein, we report the complete removal of organic dyes (rhodamine B), methylene blue) and eosin yellow) from water via effective adsorption by MoO 3 catalyst. Hydrothermally synthesised MoO 2 (1) and amorphous MoS x (2) using ammonium molybdate without and with thiourea exhibited low dye adsorption. In contrast, crystalline micro/nanoplates of MoO 3 (3 and 4) obtained from calcination of 1 and 2 showed highly enhanced dye adsorption. Particularly 4 showed higher dye adsorption compared to 3. UV-Visible absorption studies confirmed complete removal of organic dyes upon stirring with MoO 3 catalyst. Dye removal studies further revealed that cationic dyes are adsorbed faster than anionic dye that could be attributed to the surface charge of MoO 3 . Interestingly, the adsorbed dyes were not released from MoO 3 for more than 50 days. The exhausted MoO 3 catalyst can be recovered by annealing at 400 °C. MoO 3 catalyst has also been used as packing materials in dropper column and demonstrated effective removal of dyes by passing through dyes separately as well as mixture., (Copyright © 2021 Elsevier Ltd. All rights reserved.)

Purpose: Pediatric acute lymphoblastic leukemia (pALL) patients have better overall survival and methotrexate (MTX) is an effective drug used in their treatment. However, the treatment-related adverse effects (TRAEs) have a bigger impact on the therapy. In this study, we have evaluated the association of polymorphisms in genes encoding proteins engaged in MTX metabolism, and the cytogenetic aberrations with TRAEs., Methods: A total of 115 patients between the age of 1 and 18 years (average: 6.6) under maintenance therapy were selected for the study. SLC19A1 (c.80G > A), MTHFR (c.677C > T; c.1298A > C), and TYMS (c.*450&#95;*455del) genotypes were determined using PCR techniques and Sanger sequencing. Cytogenetic and SNP findings were analyzed for any association with the reported toxicities using odds ratio, chi-square test, multifactor dimensionality reduction (MDR) analysis for synergistic effect and, multinomial logistic regression analysis for the likelihood of adverse events., Results: Among the evaluated genetic variations, SLC19A1 (c.80G > A) was significantly associated with TRAEs (OR = 5.71, p = 0.002). Multinomial logistic regression analysis (chi-sq = 16.64, p < 0.001) and MDR analysis (chi-sq = 10.51 p < 0.001) confirmed the finding. On the other hand, no significant association was observed between adverse events and any specific cytogenetic aberration., Conclusion: SLC19A1 facilitates the import of cyclic dinucleotides and reduced folates, evaluating genotypes in this gene can help in better management of patients on methotrexate treatment. Assessing a broader gene panel can help in finding more associated markers and delivering personalized medicine to the patients., (© 2022. The Author(s), under exclusive licence to Springer-Verlag GmbH Germany, part of Springer Nature.)

Objective: The term ''Human Papillomavirus'' or ''HPV'' has become synonymous with uterine cervical cancer leading to feminisation of all the preventive measures, especially immunisation. Taking into consideration the rising number of HPV associated cancers among men in many developed countries and the risk of transmission to women, male HPV infection is a serious concern. A systematic review and meta-analysis of literature was performed to determine the global prevalence of HPV among men with oropharyngeal and anogenital cancers., Methods: A systematic review and meta-analysis of literature was performed searching electronic databases for published articles in English between January 1984- April 2020 based on standard systematic review guidelines. The meta-analysis component was modified appropriately for the synthesis of prevalence study results. National Institutes of Health checklist for observational, cohort and cross-sectional studies was used to assess the quality of the studies selected after the abstract and content review. The meta-analysis was performed in STATA version 13.0 (College Station, Texas 77,845 USA) and the forest plots were constructed using metan package in STATA., Results: Through the electronic search of databases, 3486 original articles were screened for eligibility. Fifty-eight articles were systematically reviewed and 42 articles were qualified for meta-analysis including 4,250 men with oropharyngeal, penile and prostate cancers. The pooled prevalence of HPV DNA in oropharyngeal cancers was 45% (95%CI 24.0%-66.0%). Meanwhile the pooled prevalence rates of 48% (CI 40.0%- 57.0%) and 19% (CI 10.0%-29.0%) were observed in penile and prostate cancers respectively. Even though, articles regarding HPV prevalence in anal cancers were systematically reviewed, none of the studies were qualified for meta-analysis., Conclusion: Higher pooled prevalence of HPV DNA was observed among men with oropharyngeal and penile cancers. Multicentric molecular studies investigating the prevalence of HPV in prostate cancers have to be planned in future.

EV-D68 is an emerging enterovirus infection associated with severe acute respiratory illness (SARI), acute flaccid myelitis (AFM) and acute flaccid paralysis (AFP). While EV-D68 outbreaks and sporadic cases are reported globally, a single case has been reported from India. The present study aims to investigate the molecular epidemiology and clinical characteristics of EV-D68-associated SARI cases from South India. We screened influenza-negative archived throat swab specimens from Influenza-Like Illness (ILI) and SARI cases (n=959; 2016 to 2018 period) for enteroviruses by pan-enterovirus real-time RT-PCR. Thirteen samples positive for enteroviruses were typed by PCR and sequencing based on VPI, VP2 and/or 5'NCR regions. One EV-D68 RNA sample was subjected to next-generation sequencing for whole genome characterisation. Among 13 enterovirus cases, four were ECHO-11, three EV-D68, two CV-A16 and one each EV-71, CV-B1, CV-B2 and CV-A9. All three cases of EV-D68 infection were reported in children below 2 years of age from Kerala state of South India during June and July 2017. The patients developed pneumonia without any neurological complications. Sequencing based on VPI and 5'NCR regions showed that EV-D68 strains belong to the novel subclade B3. The EV-D68 complete genome identified with two unique amino acid substitutions in VP1 (T-246-I) and 3D (K-344-R) regions. This study reiterates the EV-D68 novel subclade B3 circulation in India and indicates the urgent need for structured EV-D68 surveillance in the country to describe the epidemiology.

The pathogenesis of dengue virus infection is attributed to complex interplay between virus, host genes and host immune response. Host factors such as antibody-dependent enhancement (ADE), memory cross-reactive T cells, anti-DENV NS1 antibodies, autoimmunity as well as genetic factors are major determinants of disease susceptibility. NS1 protein and anti-DENV NS1 antibodies were believed to be responsible for pathogenesis of severe dengue. The cytokine response of cross-reactive CD4+ T cells might be altered by the sequential infection with different DENV serotypes, leading to further elevation of pro-inflammatory cytokines contributing a detrimental immune response. Fcγ receptor-mediated antibody-dependent enhancement (ADE) results in release of cytokines from immune cells leading to vascular endothelial cell dysfunction and increased vascular permeability. Genomic variation of dengue virus and subgenomic flavivirus RNA (sfRNA) suppressing host immune response are viral determinants of disease severity. Dengue infection can lead to the generation of autoantibodies against DENV NS1antigen, DENV prM, and E proteins, which can cross-react with several self-antigens such as plasminogen, integrin, and platelet cells. Apart from viral factors, several host genetic factors and gene polymorphisms also have a role to play in pathogenesis of DENV infection. This review article highlights the various factors responsible for the pathogenesis of dengue and also highlights the recent advances in the field related to biomarkers which can be used in future for predicting severe disease outcome.

Objective: To study the clinical characteristics and factors associated with mortality among children with Shigella encephalopathy., Methods: The data collection was done prospectively from January, 2018 to May, 2019 with retrospective data from June, 2016 to December, 2017. The study cohort consisted of 58 children <12 years of age with Shigella encephalopathy admitted to the pediatric intensive care unit. Shigella encephalopathy was confirmed if culture or real time polymerase chain reaction (PCR) of a stool sample or rectal swab was positive, with temporal association of diarrhea with seizures, altered sensorium or both. Association of mortality with risk factors was tested using chi square test, and the strength of association was estimated in terms of relative risk (RR) and 95% CI., Results: Seizures and altered sensorium were the predominant neurological symptoms. Shock occurred in 32 (55%) children, while blood in stools was a feature in only 6 (10%) children. S. sonnei was the commonest species identified on stool culture (19;33%). On univariate analysis, prolonged seizures, shock, prolonged altered sensorium, multi-organ dysfunction, lymphocytopenia at admission and need for mechanical ventilation were significantly associated with mortality. On multivariate regression, delayed presentation (presentation to the hospital 48 hours after the onset of symptoms) and prolonged altered sensorium (>12 hours) were found to be independently associated with mortality., Conclusion: Recognition of factors associated with mortality in Shigella encephalopathy may assist in better monitoring of sicker children and improved outcomes.

Background: The majority of dog-mediated human rabies as well as rabies-related human deaths are reported from low-income countries of Asia and Africa where access to appropriate postexposure prophylaxis is limited or nonexistent. At present, India is second in position after China in terms of having the highest number of mobile phone users surpassing the United States., Objective: In this context, we decided to develop a user-friendly, technically less demanding, mobile App for health-care professionals, which is accessible even without Internet facility., Methodology: The current study was conducted in four phases, namely assemblage of informational contents on rabies, development of the software, assessment of the reliability of the questionnaire tool and evaluation of the mobile App. The evaluation of App was conducted among physicians and nursing staffs in a tertiary care referral hospital., Results: The information content was prepared referring national and international guidelines. The App was designed with Hypertext Markup Language 5 for presentation on the World Wide Web and was coined the name of "RabiApp." This is a hybrid App of the native App and web App, allowing the information to be stored in the local server. The mobile App was assessed using a validated and reliable questionnaire after confirming the internal consistency by means of Cronbach's alpha. The overall Cronbach's alpha for the main scale was 0.788, which was a respectable score., Conclusion: The developed App is a user-friendly, easily accessible platform, which can help health-care professionals in making decisions regarding rabies wound management, treatment, and prophylaxis., Competing Interests: There are no conflicts of interest., (Copyright: © 2020 Indian Journal of Community Medicine.)

Kyasanur Forest disease (KFD) is a tick-borne, acute, febrile viral illness endemic in southern India. No major studies have been done to understand the adaptive immune response during KFDV infection in humans. In this study, KFDV-positive patients were prospectively enrolled, and repeated peripheral blood collections were performed. Clinical and virologic characterization of these samples is reported along with phenotypic analysis of cellular immunity and quantitation of humoral immunity. We noted robust T and B cell responses, particularly of CD8 T cells, during KFDV infection in most of the patients. Virus clearance from the blood coincided with peak CD8 T cell activation and the appearance of KFDV-specific IgG. Increased frequency of plasmablasts and very few activated B cells were observed in the acute phase of KFD infection. Notably, only humoral immunity and activated B cell frequency in the acute phase correlated with prior KFDV vaccination, and only with 2 or more doses. This novel work has implications in KFD vaccine research as well as in understanding the pathogenesis.

Human Endogenous Retrovirus W Envelope (HERV-W ENV) mRNA or protein can be found in peripheral blood mononuclear cells (PBMCs) and exocrine pancreas of patients with type 1 diabetes (T1D). Further, previous observations have shown an association between enteroviral infection and development of T1D; specifically, coxsackievirus-B (CV-B) has been detected in the blood and pancreas of patients with T1D. Notably, viruses can activate HERV-W expression. Hence, we evaluated the effect of CV-B4 infection on HERV-W ENV mRNA expression. Primary human pancreatic ductal cells were obtained from five brain-dead donors. In the pancreatic cells of three donors, the HERV-W ENV mRNA level measured using RT-qPCR was upregulated upon CV-B4 infection. The HERV-W ENV protein was detected in the infected cells using the immunoblot assay. In human PBMCs inoculated with CV-B4 or when CV-B4 was incubated with an enhancing serum, the HERV-W ENV mRNA level was higher than the background RNA level. In monocyte-derived macrophages obtained from 5 of 13 donors, the HERV-W ENV mRNA level was higher in cultures inoculated with CV-B4 than in the control. Therefore, CV-B4 can upregulate or induce the transcription of a certain HERV-W ENV copy (or copies) in primary cell cultures, such as monocytes, macrophages, and pancreatic cells.

Human influenza A and B viruses are highly contagious and cause similar illnesses and seasonal epidemics. Currently available antiviral drugs have limited efficacy in humans with compromised immune systems; therefore, alternative strategies for protection are needed. Here, we investigated whether monoclonal antibodies (MAbs) targeting hemagglutinin (HA) and/or neuraminidase (NA) proteins would protect immunosuppressed mice from severe infections with influenza B virus. Pharmacologically immunosuppressed BALB/c mice were inoculated with B/Brisbane/60/2008 (BR/08) influenza virus and were treated with a single dose of 1, 5, or 25 mg/kg of body weight per day of either an anti-HA MAb (1D2) or an anti-NA MAb (1F2) starting at 24 hours postinoculation (hpi). Monotherapy with 1D2 or 1F2 MAbs provided dose-dependent protection of mice, with decreased BR/08 virus replication and spread in the mouse lungs, compared with those of controls. Combination treatment with 1D2 and 1F2 provided greater protection than did monotherapy, even when started at 48 hpi. Virus spread was also efficiently restrained within the lungs, being limited to 6%, 10%, and 10% of that seen in active infection when treatment was initiated at 24, 48, and 72 hpi, respectively. In most cases, the expression of cytokines and chemokines was altered according to when treatment was initiated. Higher expression of proinflammatory IP-10 and MCP-1 in combination-treatment groups, but not in monotherapy groups, to some extent, promoted better control of virus spread within the lungs. This study demonstrates the potential value of MAb immunotherapy in treating influenza in immunocompromised hosts who are at increased risk of severe disease., (Copyright © 2020 American Society for Microbiology.)

In most species, the centromere is comprised of repetitive DNA sequences, which rapidly evolve. Paradoxically, centromeres fulfill an essential function during mitosis, as they are the chromosomal sites wherein, through the kinetochore, the mitotic spindles bind. It is now generally accepted that centromeres are transcribed, and that such transcription is associated with a broad range of functions. More than a decade of work on this topic has shown that centromeric transcripts are found across the eukaryotic tree and associate with heterochromatin formation, chromatin structure, kinetochore structure, centromeric protein loading, and inner centromere signaling. In this review, we discuss the conservation of small and long non-coding centromeric RNAs, their associations with various centromeric functions, and their potential roles in disease.

Epithelial to mesenchymal transition (EMT) is a complex plastic and reversible cellular process that has critical roles in diverse physiological and pathological phenomena. EMT is involved in embryonic development, organogenesis and tissue repair, as well as in fibrosis, cancer metastasis and drug resistance. In recent years, the ability to edit the genome using the clustered regularly interspaced palindromic repeats (CRISPR) and associated protein (Cas) system has greatly contributed to identify or validate critical genes in pathway signaling. This review delineates the complex EMT networks and discusses recent studies that have used CRISPR/Cas technology to further advance our understanding of the EMT process.

Background: An outbreak of Nipah virus (NiV) disease occurred in the Kozhikode district of Kerala State in India in May 2018. Several cases were treated at the emergency medicine department (ED) of the Government Medical College, Kozhikode (GMCK). The clinical manifestations and outcome of these cases are described., Methods: The study included 12 cases treated in the ED of GMCK. Detailed clinical examination, laboratory investigations, and molecular testing for etiological diagnosis were performed., Results: The median age of the patients was 30 years and the male to female ratio was 1.4:1.0. All the cases except the index case contracted the infection from hospitals. The median incubation period was 10 days, and the case fatality ratio was 83.3%. Ten (83.3%) patients had encephalitis and 9 out of 11 patients whose chest X-rays were obtained had bilateral infiltrates. Three patients had bradycardia and intractable hypotension requiring inotropes. Encephalitis, acute respiratory distress syndrome, and myocarditis were the clinical prototypes, but there were large overlaps between these. Ribavirin therapy was given to a subset of the patients. Although there was a 20% reduction in NiV encephalitis cases treated with the drug, the difference was not statistically significant. The outbreak ended soon after the introduction of total isolation of patients and barrier nursing., Conclusion: The outbreak of NiV disease in Kozhikode in May 2018 presented as encephalitis, acute respiratory distress and myocarditis or combinations of these. The CFR was high. Ribavirin therapy was tried but no evidence for its benefit could be obtained., (© The Author(s) 2019. Published by Oxford University Press for the Infectious Diseases Society of America. All rights reserved. For permissions, e-mail: journals.permissions@oup.com.)

Influenza viruses are known to cause acute respiratory illness, sometimes leading to high mortality rates. Though there are approved influenza antivirals available, their efficacy has reduced over time, due to the drug resistance crisis. There is a perpetual need for newer and better drugs. Drug screening based on the interaction dynamics with different viral target proteins has been a preferred approach in the antiviral drug discovery process. In this study, the FDA approved drug database was virtually screened with the help of Schrödinger software, to select small molecules exhibiting best interactions with the influenza A virus endonuclease protein. A detailed cytotoxicity profiling was carried out for the two selected compounds, cefepime and dolutegravir, followed by in vitro anti-influenza screening using plaque reduction assay. Cefepime showed no cytotoxicity up to 200 μM, while dolutegravir was non-toxic up to 100 μM in Madin-Darby canine kidney cells. The compounds did not show any reduction in viral plaque numbers indicating no anti-influenza activity. An inefficiency in the translation of the molecular interactions into antiviral activity does not necessarily mean that the molecules were inactive. Nevertheless, testing the molecules for endonuclease inhibition per se can be considered a worthwhile approach., (© FEMS 2020.)

A single nucleotide polymorphism (SNP) rs2853669 (A>G) in the telomerase reverse transcriptase (TERT) promoter has recently been reported in chr5:1,295,349 T>C (T349C), and was shown to be associated with increased cancer risk and poor survival in a specific population. However, at present, the role of this particular SNP with TERT promoter driver mutations and its genetic association with human papilloma virus (HPV) in patients with cervical cancer has not been determined. In the present study, the genetic association of the functional SNP rs2853669 in the presence/absence of TERT promoter hotspot mutations and HPV in patients with cervical cancer of South Indian origin was evaluated. To understand and compare the frequency of the variant allele and its risk association in different cancer types of various populations, the SNP was genotyped in 257 cervical cancer samples and 295 controls, and its associations with TERT promoter hotspot mutations and HPV were analyzed. Furthermore, an extensive search of previously published articles in PubMed, Embase and Web of Science was conducted; a meta-analysis was carried out to elucidate the association of the SNP with different cancer types in global populations. The SNP analysis showed significantly high frequency (41%) of homozygous variant allele rs2853669 (GG) in patients with cervical cancer compared with control samples [Recessive allele model odds ratio (OR)=1.71; 95% CI=1.20-2.43; P=0.003]. No significant interaction was observed between the TERT SNP rs2853669 and HPV status as well as other hotspot TERT promoter (C228T and C250T) mutations determined in our previous study. In addition, the overall meta-analysis revealed a significant association of the SNP rs2853669 with other cancer types in different ethnic populations (OR=1.09; 95% CI=1.03-1.16; P=0.004). The present results suggested that the TERT SNP rs2853669 could play an important role in the risk of cervical cancer in a South Indian population., (Copyright © 2019, Spandidos Publications.)

Kyasanur Forest disease virus (KFDV) and Alkhurma hemorrhagic fever virus (AHFV) are enveloped, positive-stranded RNA viruses of clinical importance with complex enzootic life cycles involving hematophagous ticks which feed on small and large mammals. Humans and monkeys are dead-end hosts for these viruses. Recent trends in epidemiological data suggest both virus incidences are steadily increasing and their geographical distribution expanding out of previously known circulation regions. For the detection and discrimination of these two closely related flaviviruses, we have developed quantitative real-time RT-PCR assays with 100 % sensitivity and 100 % specificity for KFDV, 100 % sensitivity and 99.4 % specificity for AHFV as determined using 550 clinical samples collected between 2015-2018 from Western Ghats region of India. This rapid and sensitive assay will enable researchers to accurately diagnose the presence of the virus during viremia in human and animal blood samples, and also from tick specimens. Incorporation of these new tests into a routine diagnosis will help in the diagnosis of KFDV as well as AHFV in the endemic areas and also would provide an early warning of the spread of this virus to newer regions with similar epidemiology., (Copyright © 2020 Elsevier GmbH. All rights reserved.)

Objective: The causal association of human papillomavirus (HPV) in uterine cervical cancer was well established and this oncogenic virus was reported to be a biomarker for overall recurrence and central pelvic recurrence. The objective of the present systematic review and meta-analysis was to assess the role of HPV DNA testing in early detection of recurrence among cervical cancer survivors after radiotherapy., Methods: We performed a systematic review and meta-analysis by means of searching electronic databases for published articles between January 1984 and June 2018, on the basis of standard systematic review guidelines prescribed by major agencies namely Cochrane Collaboration (https://www.cochrane.org) and Campbell Collaboration (https://www.campbellcollaboration.org). The meta-analysis component was further modified appropriately for the synthesis of sensitivity and specificity results., Results: A total of 1,055 cervical cancer cases who had received pelvic radiation with or without chemotherapy from ten cohort studies were evaluated. The overall pooled sensitivity and specificity of HPV DNA testing was 0.84 (95% confidence interval [CI]= 0.66-0.94) and 0.35 (95% CI=0.20-0.54) respectively. The positive likelihood ratio was 1.3 (95% CI=1.0-1.7) and the negative likelihood ratio was 0.45 (95% CI=0.18-1.10) with an estimated diagnostic odds ratio of 3 (95% CI=1-9)., Conclusion: The screening for HPV DNA testing during follow-up facilitates early detection of recurrence after radiotherapy., Competing Interests: No potential conflict of interest relevant to this article was reported., (Copyright © 2020. Asian Society of Gynecologic Oncology, Korean Society of Gynecologic Oncology.)

Chikungunya fever is a viral disease transmitted to humans by the bite of infected mosquitoes. The disease is characterized by fever, headache, rash, severe joint, and muscle pain. To evaluate the disease burden in the population and the effectiveness of public health measures, periodic seroprevalence surveys are essential. Chikungunya outbreaks were reported from many Asian countries since 2005, after more than three decades of disappearance. The study aimed to estimate the seroprevalence of the chikungunya virus in southern parts of Karnataka state, through demonstrating chikungunya virus-specific neutralizing antibodies. A cross-sectional study was carried out using 509 archived blood samples from a hospital-based acute febrile illness surveillance project, representative of the period between June 2014 and 2018. The study reported a 3.7% seroprevalence of chikungunya virus-neutralizing antibodies in Thirthahalli and Hosanagara taluks of South Karnataka. The low prevalence of chikungunya-neutralizing antibodies indicates that a major population is unexposed and prone to future outbreaks., (© 2019 Wiley Periodicals, Inc.)

Women with persistent human papillomavirus (HPV) infections have a high risk of developing cervical cancer (CaCx). HPV-16 alone accounts for more than 60% of CaCx worldwide. Most of the HPV infections are transient and only a subset of women develop persistent HPV-16 infection. Many studies have shown associations of different human leukocyte antigen (HLA) alleles with HPV-mediated CaCx, but there are only a few studies globally that relate to persistent HPV-16 infection. Furthermore, such studies from India are sparse. Hence, we investigated the association of HLA-A, B, DRB, and DQB alleles with persistent HPV-16 infection and HPV-16-positive CaCx in south India (Tamil Nadu). HPV-16 persistent infection was observed in 7% of normal women. A total of 50 women with HPV-16-positive CaCx, 21 women with HPV-16 persistent infection, and 74 HPV-16-negative normal women were recruited for this study. Low-resolution typing of HLA-A, B, DRB, and DQB alleles was performed. HLA-B*44 and DRB1*07 showed a significant association with persistent HPV-16 infection (odds ratio, p -value = 26.3, 0.03 and 4.7, 0.01, respectively). HLA-B*27 and DRB1*12 were significantly associated with both HPV-16 + CaCx and persistent HPV-16 infection (23.8, 0.03; 52.9, 0.01; 9.8, 0.0009; and 13.8, 0.009; respectively). HLA-B*15 showed a negative association with HPV-16-positive CaCx (0.1, 0.01), whereas DRB1*04 exhibited protection to both HPV-16-positive CaCx and persistent HPV-16 infection (0.3, 0.0001 and 0.1, 0.0002, respectively). Thus, we show HLA allelic association with HPV-16 infection in Tamil Nadu. Larger studies on high-resolution HLA typing coupled with HPV-16 genome diversity will offer further insights into host/pathogen genome coevolution.

Background: The Brokpas are an isolated tribal population of the Dah-Hanu villages of the Leh district of India. They speak Dardic, a sub-branch of the Indo-European language family, and are putatively identified as "pure Aryan," a hegemonic impression perpetuated by foreign tourism. Aim: To determine if the above is true by looking for an appreciable frequency of NRY-HG-R1a1(M17) signatures which are common to Indo-European language speakers of mainland India and elsewhere. Subjects and methods: We studied 75 random Brokpa males from the Dah-Hanu region, on the northern bank of the Indus river. Results: Interestingly, the Brokpa males possessed a high proportion of NRY-HG-L1a2(M357) (62.7%) that are found sporadically in India and her neighbourhood. A global analysis of this clade (present study, 214 of 3327 men from 63 populations; from the literature 56 of 873) suggested that they originated from southern India. Conclusion: The Y chromosomal studies suggest the Brokpa to be pre-Vedic settlers of the Himalayas, 9000 ybp, with an isolated evolution. The mtDNA profile shows a predominance of mtDNA HG A4 that must have arrived from outside the Indian subcontinent.

Background: Nipah virus (NiV) is 1 of 10 potential causes of imminent public health emergencies of international concern. We investigated the NiV outbreak that occurred in May 2018 in Kerala, India. Here we describe the longitudinal characteristics of cell-mediated and humoral immune responses to NiV infection during the acute and convalescent phases in 2 human survivors., Methods: Serial blood samples were obtained from the only 2 survivors of the NiV outbreak in Kerala. We used flow cytometry to determine the absolute T-lymphocyte and B-lymphocyte counts and the phenotypes of both T and B cells. We also detected and quantitated the humoral immune response to NiV by virus-specific immunoglobulin M (IgM) and immunoglobulin G (IgG) enzyme-linked immunosorbent assay., Results: Absolute numbers of T lymphocytes remained within normal limits throughout the period of illness studied in both survivors. However, a marked elevation of activated CD8 T cells was observed in both cases. More than 30% of total CD8 T cells expressed Ki67, indicating active proliferation. Proliferating (Ki-67+) CD8 T cells expressed high levels of granzyme B and PD-1, consistent with the profile of acute effector cells. Total B-lymphocyte, activated B-cell, and plasmablast counts were also elevated in NiV survivors. These individuals developed detectable NiV-specific IgM and IgG antibodies within a week of disease onset. Clearance of NiV RNA from blood preceded the appearance of virus-specific IgG and coincided with the peak of activated CD8 T cells., Conclusions: We describe for the first time longitudinal kinetic data on the activation status of human B- and T-cell populations during acute NiV infection. While marked CD8 T-cell activation was observed with effector characteristics, activated CD4 T cells were less prominent., (© The Author(s) 2019. Published by Oxford University Press for the Infectious Diseases Society of America. All rights reserved. For permissions, e-mail: journals.permissions@oup.com.)

There are inconsistent reports regarding the role of HPV in the origin and progression of oral squamous cell carcinoma (OSCC). The observed heterogeneity was mainly attributed to the social and cultural habits of the enrolled cases, discrepancies in the nature of samples procured and varying sensitivity of the assays employed for detection of HPV. The objective of this study was to assess the prevalence of HPV in OSCC in South West India. This was a cross sectional study conducted over a period extending from October 2015 to June 2017. This study involved Department of ENT-Head and Neck Surgery and Department of Virology, Manipal Academy of Higher Education. Fifty histologically confirmed oral squamous cell carcinoma patients undergoing wide local excision of the tumour were enrolled for the study. Intraoperatively 4-5 mm of tissue samples were transported in sterile normal saline at 4-80 °C. The primary screening of tissue samples was performed by nested PCR using PGMY09/11 consensus primers and GP5+/6+ consensus primers and TaqMan based real time multiplex PCR for HPV-16, HPV-18, HPV-31 and HPV-45. All samples tested negative for HPV DNA by conventional nested PCR and TaqMan based real-time Multiplex PCR ruling out four common HPV subtypes such as HPV-16, 18, 31, 45. We did not find presence of HPV DNA in the tissues of patients with OSCC from southwest India. However, studies with more geographic representation from other parts of India are required before generalising our findings., (© Association of Otolaryngologists of India 2018.)

Purpose: Hepatitis E virus (HEV) is an emerging pathogen causing acute viral hepatitis worldwide. Clinical manifestations often occur in young adults with an increased mortality rate among pregnant women. HEV genotypes 1 and 4 are mainly reported among humans and swines, respectively., Aims: The aim was to study the currently circulating genotypes of HEV in India., Materials and Methods: A retrospective cross-sectional study was carried out at Manipal Institute of Virology to know the circulating genotypes of hepatitis E, spanning over 5 years from August 2014 to September 2018. The serum samples screened serologically positive and confirmed positive for active infection by real-time reverse transcriptase-polymerase chain reaction (PCR) (Real Star ® HEV RT-PCR Kit 2.0, Altona Diagnostics, GmbH, Hamburg, Germany) were further subjected to nested conventional PCR targeting the RdRp gene of non-structural ORF1 region. The purified PCR product was sequenced in BigDye Terminator v3.1 Cycle Sequencing Kit (Life Technologies, Thermo Fisher Scientific, USA). The chromatograms obtained by sequencing were analysed using Sequencher 5.4.6, and HEV FASTA sequences were compared with reference sequences for HEV in GenBank Nucleotide Blast., Results: During the study period, there were 317 cases of laboratory-confirmed cases of acute viral hepatitis comprising 202, 70, 43 and 2 cases of hepatitis A, E, B and C, respectively. Serum samples of 70 acute hepatitis cases were positive for anti-hepatitis E IgM. According to the clinical case classification, there were 66 cases of acute viral hepatitis and four cases of fulminant hepatic liver failure. The mean age of the patients was 30.3 years (standard deviation = 12.5). The samples from various parts of India were genotyped as 1a., Conclusion: The HEV genotypes 1a was observed to be the currently circulating strain in the regions studied., Competing Interests: None

Seasonal influenza virus infections cause significant morbidity and mortality every year. Annual influenza virus vaccines are effective but only when well matched with circulating strains. Therefore, there is an urgent need for better vaccines that induce broad protection against drifted seasonal and emerging pandemic influenza viruses. One approach to design such vaccines is based on targeting conserved regions of the influenza virus hemagglutinin. Sequential vaccination with chimeric hemagglutinin constructs can refocus antibody responses towards the conserved immunosubdominant stalk domain of the hemagglutinin, rather than the variable immunodominant head. A complementary approach for a universal influenza A virus vaccine is to induce T-cell responses to conserved internal influenza virus antigens. For this purpose, replication deficient recombinant viral vectors based on Chimpanzee Adenovirus Oxford 1 and Modified Vaccinia Ankara virus are used to express the viral nucleoprotein and the matrix protein 1. In this study, we combined these two strategies and evaluated the efficacy of viral vectors expressing both chimeric hemagglutinin and nucleoprotein plus matrix protein 1 in a mouse model against challenge with group 2 influenza viruses including H3N2, H7N9 and H10N8. We found that vectored vaccines expressing both sets of antigens provided enhanced protection against H3N2 virus challenge when compared to vaccination with viral vectors expressing only one set of antigens. Vaccine induced antibody responses against divergent group 2 hemagglutinins, nucleoprotein and matrix protein 1 as well as robust T-cell responses to the nucleoprotein and matrix protein 1 were detected. Of note, it was observed that while antibodies to the H3 stalk were already boosted to high levels after two vaccinations with chimeric hemagglutinins (cHAs), three exposures were required to induce strong reactivity across subtypes. Overall, these results show that a combinations of different universal influenza virus vaccine strategies can induce broad antibody and T-cell responses and can provide increased protection against influenza., (Copyright © 2019 The Authors. Published by Elsevier Ltd.. All rights reserved.)

Chikungunya is a mosquito-borne viral illness associated with chronic arthritic symptoms that persist for months. The IgM antibody appears within a week post any infection and declines at 2-3 months. The present study was aimed to demonstrate the presence of specific IgM antibody among chikungunya confirmed cases. Blood samples were collected from chikungunya PCR positive patients at the time of diagnosis, at 1-week, 1, 8, 10 and 12 months post infection. All acute and follow-up serum samples were evaluated for chikungunya virus-specific IgM antibodies using ELISA technique. Our findings indicate the persistence of anti-chikungunya IgM up to 10-months post-infection in a majority of chikungunya virus infected persons. Interpretation of results should be carefully done as only IgM ELISA is used to diagnose acute infection, especially post chikungunya outbreak. The presence of IgM antibody does not rule out the absence of any other diagnosis due to its persistence. Thus, we hypothesize that real-time PCR is more reliable for the detection of acute chikungunya cases in endemic areas while IgM detection may be useful in identifying exposure to this disease., Competing Interests: Conflict of interestThe authors declare that they have no conflict of interest., (© Indian Virological Society 2019.)

Proper vascular function is important for well-being of mother and growing fetus. VEGF TOTAL , and VEGF 165b levels and its vascular endothelial complications in gestational diabetes mellitus (GDM) together with the association of inflammation and advanced glycation end products (AGEs) are less studied. VEGF 165b /VEGF TOTAL (VEGF RATIO) in GDM pregnant women was investigated in this study. Plasma VEGF TOTAL was lower in GDM (17.68 ± 1.30 pg/mL) compared to non-GDM (25.69 ± 1.40 pg/mL). VEGF 165b , ICAM-1, and AGEs were higher in GDM (9.9 ± 1.4 pg/mL, 201.04 ± 7.85 µg/mL, and 10.40 ± 0.98 µg/mL, respectively) and lower in non-GDM (6.47 ± 0.70 pg/mL, 174.1 ± 7.11 µg/mL, and 4.71 ± 0.39 µg/mL, respectively). Compared to non GDM (0.25 ± 0.02), VEGF RATIO was higher in GDM (0.45 ± 0.04) and correlated with -ICAM-1 ( r = 0.375, p < .001) and AGEs ( r = 0.199, p < .05). Tertile stratification of VEGF RATIO implied that frequency of GDM increases with increasing tertiles of VEGF RATIO ( p for trend <.001). Association of VEGF RATIO with GDM was significant even after adjusting for AGEs (OR = 1.279, CI = 1.118-1.462, p < .0010) but it lost its significance when adjusted for ICAM-1 (OR = 1.006, CI = 0.995-1.017, p = .308). VEGF RATIO plays an important role in GDM in association with vascular inflammation.

Objective: To design and implement a locally relevant competency- based MPH programme., Methods: The demand for trained public health professionals in South Asia is enormous and growing, which created a unique opportunity for a Fogarty International Center-funded University of Alabama at Birmingham-South Asia [Aga Khan University, Pakistan; Manipal Academy of Higher Education, India; and University of Kelaniya, Sri Lanka] international research training in environmental and occupational health (ITREOH) programme. In 2009, a Master of Public Health (MPH) degree programme was designed using a combination of competencies developed by the Association of School of Public Health, the World Health Organization and the Centers for Disease Control and Prevention., Results: A competency- based curriculum was developed with two specialty tracks in applied epidemiology and environmental and occupational health, emphasising applied practice and research., Conclusions: This is the most comprehensive skill-based MPH programme in the region, which positions each institution as a regional leader in public health training. The success of the programme has been amply demonstrated by placements of graduated MPH students in leadership roles in public, private and academic sectors within their countries.

Seasonal influenza viruses cause significant morbidity and mortality in the global population every year. Although seasonal vaccination limits disease, mismatches between the circulating strain and the vaccine strain can severely impair vaccine effectiveness. Because of this, there is an urgent need for a universal vaccine that induces broad protection against drifted seasonal and emerging pandemic influenza viruses. Targeting the conserved stalk region of the influenza virus hemagglutinin (HA), the major glycoprotein on the surface of the virus, results in the production of broadly protective antibody responses. Furthermore, replication deficient viral vectors based on Chimpanzee Adenovirus Oxford 1 (ChAdOx1) and modified vaccinia Ankara (MVA) virus expressing the influenza virus internal antigens, the nucleoprotein (NP) and matrix 1 (M1) protein, can induce strong heterosubtypic influenza virus-specific T cell responses in vaccinated individuals. Here, we combine these two platforms to evaluate the efficacy of a viral vectored vaccination regimen in protecting ferrets from H3N2 influenza virus infection. We observed that viral vectored vaccines expressing both stalk-targeting, chimeric HA constructs, and the NP+M1 fusion protein, in a prime-boost regimen resulted in the production of antibodies toward group 2 HAs, the HA stalk, NP and M1, as well as in induction of influenza virus-specific-IFNγ responses. The immune response induced by this vaccination regime ultimately reduced viral titers in the respiratory tract of influenza virus infected ferrets. Overall, these results improve our understanding of vaccination platforms capable of harnessing both cellular and humoral immunity with the goal of developing a universal influenza virus vaccine.

Molecular surveillance of influenza viruses is essential for early detection of novel variants. The aim of the present study was to analyze the hemagglutinin gene of influenza A(H1N1)pdm09 and A(H3N2) viruses circulating during the 2017 season. To investigate the genetic diversity of hemagglutinin gene of influenza A(H1N1)pdm09 and A(H3N2) viruses from 2017 season, ten samples from each subtype were sequenced and analyzed. The season was predominated by influenza A(H1N1)pdm09 viruses. Ten samples were sequenced from each subtype and all sequenced influenza A(H1N1)pdm09 and A(H3N2) viruses belonged to clades 6B.1 and 3C.2a, respectively. Sequence analysis of H1 gene in comparison to 2010-2016 vaccine strain showed mutations K166Q and S188T (K180Q and S202T here) that most likely resulted in antigenic drift and emergence of variant viruses. H3 gene substitutions N137K, N187K, I422V, and G500E that define clade 3C.2a1 were detected during analysis of sequences in comparison to 2017-2018 vaccine strain of northern hemisphere. These substitutions contributed to the change of WHO's recommendation of the 2018-2019 vaccine strain for northern hemisphere. The results of this study provide insights about the continuous genetic variability of the HA gene.

Background: WHO has recommended Visual Inspection with Acetic acid (VIA) or Human Papillomavirus (HPV) DNA testing if feasible, for cervical cancer screening in low income countries. However, the number of women undergoing screening is very low as a result of limited information, inadequate infrastructure and invasive nature of sampling. Methods: A cross sectional study was carried out comparing HPV DNA detection by Polymerase Chain Reaction (PCR) in paired cervical and urine samples procured from histologically confirmed cervical cancer cases. Results: Amongst the samples collected from 114 cervical cancer cases, HPV DNA was tested positive in cervical samples of 89 (78.1%) and urine samples of 55 (48.2%) patients. The agreement between the two sampling methods was 66.7% and the kappa value was 0.35 indicating a fair agreement. The sensitivity of HPV detection using urine samples was 59.6% (95% confidence interval 49.16%-69.15%) and the specificity was 92% (95% confidence interval 75.0%-97.8%). Conclusion: Even though not acceptable as an HPV DNA screening tool due to low sensitivity, the urine sampling method is inexpensive and more socially acceptable for large epidemiological surveys in developing countries to estimate the burden.

Background & Objectives: Dengue is a major public health problem in northeast India where the majority of the cases go unreported and undiagnosed. Even though all four dengue serotypes are reported, there is a dearth of information on genetic diversity. The present cross-sectional study was undertaken during 2016-17 to determine the genetic variance of dengue virus serotype 2 (DENV-2) based on the envelope (E) glycoprotein gene., Methods: The serum samples collected from the northeast parts of India, as a part of hospital-based acute febrile illness surveillance, were serotyped. Viral RNA was extracted from DENV-2 serum samples using QIAquick® RNA Extraction Kit. The E gene was amplified by conventional reverse-transcriptase polymerase chain reaction (RT-PCR) and the PCR products were sequenced., Results: The E glycoprotein gene of nine serum samples with high viral RNA concentration (Ct <25) was sequenced. The E gene sequences of eight DENV-2 strains from Assam and Meghalaya aligned with genotype IV (Cosmopolitan) and one strain from Tripura segregated with Asian-I genotype., Interpretation & Conclusion: Ongoing laboratory-based surveillance is mandatory to understand the transmission dynamics of dengue in endemic countries. This study concluded that in northeast India, presently two distinct genotypes of DENV-2, namely genotype IVb (Cosmopolitan) and Asian-I genotype are in circulation., Competing Interests: None

Background & Objectives: Influenza virological surveillance is an essential tool for the early detection of novel genetic variants of epidemiologic and clinical significance. This study was aimed to genetically characterize A(H1N1)pdm09 virus circulating in 2017 and to compare it with the global data., Methods: The regional/State Viral Research and Diagnostic Laboratories (VRDLs) provided influenza diagnosis for referred clinical samples and shared influenza A(H1N1)pdm09 positives with the Indian Council of Medical Research-National Institute of Virology (ICMR-NIV), Pune, India, for hemagglutinin (HA) gene phylogenetic analysis. Sites at Manipal, Jaipur and Dibrugarh performed the sequencing and shared the sequence data for analysis. The antiviral susceptibility of influenza viruses was assessed for known molecular marker H275Y at the ICMR-NIV, Pune., Results: All the eight VRDLs had well-established influenza diagnostic facilities and showed increased activity of influenza A(H1N1)pdm09 during 2017. Phylogenetic analysis showed that the viruses from the different regions of the country were similar to A/Michigan/45/2015 strain which was the 2017-2018 recommended vaccine strain and were clustered with the globally circulating clade 6B.1 with signature mutations S84N, S162N and I216T. The clade 6B.1 showed further subgrouping with additional mutations S74R, S164T and I295V; however, there was no significant association between the presence of these mutations and severity of disease due to influenza. All the study viruses were sensitive to oseltamivir., Interpretation & Conclusions: During the study period, all the study sites reported globally circulating A/Michigan/45/2015 vaccine strain of influenza A(H1N1)pdm09 viruses and remained sensitive to oseltamivir. Further genetic and antigenic characterization of influenza viruses is recommended to address public health concerns., Competing Interests: None

Influenza viruses are major respiratory pathogens that cause seasonal epidemics and occasional pandemics. Immune response to influenza viruses is majorly targeted against the hemagglutinin antigen. A laboratory-based retrospective cross-sectional study was conducted on 50 acute and 50 follow-up samples to assess the immune response to influenza A(H1N1)pdm09 virus after natural infection and detect the presence of pre-existing antibodies against influenza A(H3N2) and influenza B viruses. Two-fourfold rise in hemagglutination-inhibition (HAI) titer was observed in 100% of the follow-up samples for influenza A(H1N1)pdm09 virus. No change in HAI titers for influenza A(H3N2) and influenza B viruses was observed.

Background: Nipah Virus (NiV) is a highly fatal emerging zoonotic virus and a potential threat to global health security. Here we describe the characteristics of the NiV outbreak that occurred in Kerala, India, during May-June 2018., Methods: We used real-time reverse transcription polymerase chain reaction analysis of throat swab, blood, urine, and cerebrospinal fluid specimens to detect NiV. Further, the viral genome was sequenced and subjected to phylogenetic analysis. We conducted an epidemiologic investigation to describe the outbreak and elucidate the dynamics of NiV transmission., Results: During 2-29 May 2018, 23 cases were identified, including the index case; 18 were laboratory confirmed. The lineage of the NiV responsible for this outbreak was closer to the Bangladesh lineage. The median age of cases was 45 years; the sex of 15 (65%) was male. The median incubation period was 9.5 days (range, 6-14 days). Of the 23 cases, 20 (87%) had respiratory symptoms. The case-fatality rate was 91%; 2 cases survived. Risk factors for infection included close proximity (ie, touching, feeding, or nursing a NiV-infected person), enabling exposure to droplet infection. The public health response included isolation of cases, contact tracing, and enforcement of hospital infection control practices., Conclusion: This is the first recorded NiV outbreak in South India. Early laboratory confirmation and an immediate public health response contained the outbreak., (© The Author(s) 2018. Published by Oxford University Press for the Infectious Diseases Society of America. All rights reserved. For permissions, e-mail: journals.permissions@oup.com.)