A novel reassortant avian influenza H4N6 virus isolated from an environmental sample during a surveillance in Maharashtra, India.

Background & objectives: Low pathogenic avian influenza (LPAI) viruses cause mild clinical illness in domestic birds. Migratory birds are a known reservoir for all subtypes of avian influenza (AI) viruses. The objective of the study was to characterize AI H4N6 virus isolated from an environmental sample during surveillance in Maharashtra, India. Methods: AI surveillance in wild migratory birds was conducted during the winter migratory bird season (2016-2017) in Pune, India. AI H4N6 virus was isolated from the faecal droppings of a wild migratory waterbird. Virological and molecular characterization of the isolated virus was carried out. Virus titration, haemagglutination inhibition assay, receptor specificity assay, intravenous pathogenicity index and neuraminidase inhibition assays were performed. Full genome sequencing, molecular and phylogenetic analyses were also conducted. Results: The virus was found to be of low pathogenicity, with avian type receptor specificity, and was susceptible to neuraminidase inhibitors. Phylogenetic and molecular analysis revealed that the present virus is a result of extensive reassortment with AI H8N4, H6N2, H4N3 and H3N6, predominantly as donor viruses among others. Interpretation & conclusions: This is the first report of the isolation and characterization of an LPAI H4N6 virus from an environmental sample from India. The present study showed that the H4N6 virus is a novel reassortant and divergent as compared with the reported H4N6 viruses from poultry in India, indicating independent introduction. This highlights the role of wild and migratory birds in the transmission of AI viruses and necessity of such studies at the human-animal interface.

Dosimetric comparison of O-ring versus conventional C-arm linear accelerator for volumetric modulated arc therapy (VMAT) of head and neck cancer and carcinoma cervix patients.

The objective of this study is dosimetric comparison between the O-ring Halcyon and C-arm Clinac iX linac for volumetric modulated arc therapy (VMAT) plans for head & neck (H&N) cancer and carcinoma cervix patients. Total 60 patients of H&N cancer and carcinoma cervix were enrolled prospectively from March 2021 to March 2023. VMAT plans with 6 MV photons for Halcyon and Clinac iX were generated and compared for each patient by dose volume histogram for planning target volume coverage and organ at risk (OAR) sparing. There were no differences in between both the linacs for PTV D2% and D98%, homogeneity index, conformity index, Dmax (maximum dose) and Dmean (mean dose) of OAR. Halcyon had significantly shorter treatment time compared to Clinac iX. Halcyon delivered higher integral dose and monitor units. O-ring Halcyon produces VMAT plans comparable to other C-arm linacs for H&N and carcinoma cervix patients.

Establishment of cell line from embryonic tissue of Pipistrellus ceylonicus bat species from India & its susceptibility to different viruses.

BACKGROUND & OBJECTIVES: Pipistrellus ceylonicus bat species is widely distributed in South Asia, with additional populations recorded in China and Southeast Asia. Bats are the natural reservoir hosts for a number of emerging zoonotic diseases. Attempts to isolate bat-borne viruses in various terrestrial mammalian cell lines have sometimes been unsuccessful. The bat cell lines are useful in isolation and propagation of many of the viruses harboured by bats. New stable bat cell lines are needed to help such investigations and to assist in the study of bat immunology and virus-host interactions. In this study we made an attempt to develop a cell line from P. ceylonicus bats. METHODS: An effort was made to establish cell line from embryo of P. ceylonicus species of bat after seeding to Dulbecco's modified eagle medium (DMEM) supplemented with 10 per cent foetal bovine serum; a primary cell line was established and designated as NIV-BtEPC. Mitochondrial DNA profile analysis was done using cyt-b and ND-1 gene sequences from the cell line. Phylogenetic tree was constructed using neighbour-joining algorithm for cyt-b and ND-1 genes with 1000-bootstrap replicates. RESULTS: NIV-BtEPC cell line was susceptible to Chandipura (CHPV) and novel adenovirus (BtAdv-RLM) isolated from Rousettus leschenaulti from India but did not support multiplication of a number of Bunyaviruses, Alphaviruses and Flavivirus. This might be useful for isolation of a range of viruses and investigation of unknown aetiological agents. INTERPRETATION & CONCLUSIONS: In this study, a new bat cell line was developed from P. ceylonicus. This cell line was successfully tested for the susceptibility to Chandipura and BtAdv-RLM virus isolated from bats. The approach developed and optimised in this study may be applicable to the other species of bats and this established cell line can be used to facilitate virus isolation and basic research into virus-host interaction.

India's Multi-Sectoral Response to Oxygen Surge Demand during COVID-19 Pandemic: A Scoping Review.

Oxygen support became one of the rate-limiting steps for medical care during COVID-19 pandemic in India. The primary aim of this study was to appraise the manufacturing, supply, and distribution of medical oxygen during the pandemic. The secondary objectives were to highlight the coordination of various stakeholders to mitigate the oxygen surge and to present a critical analysis of India's response to the emergent situation. Using an analytic approach, we have delineated India's response to mitigate the medical oxygen surge during the distressing second peak between March-May 2021. In the pre-COVID-19 era, of the total 6900 MT of oxygen produced in India, only 1000 MT was available for medical usage, which was increased up to 19940 MT through the strengthening of in-house oxygen manufacturing, low-cost innovations, and enhanced storage facilities. High-burden states were identified, and transport was facilitated through departments of railways, defence and civil aviation. Real-time scrutiny of the oxygen supply was provided. Essential customs duties on importing oxygen and vital equipment were exempted, along with other swift decisions. National Oxygen Stewardship Program' was initiated to build the capacity of health care workers in oxygen therapy and rational use of surplus oxygen. The pandemic overwhelmed the health system. But a coordinated multi-stakeholder approach facilitated the fight against oxygen surge. However, a comprehensive pandemic response will need more than just oxygen. This resourceful utilization offers a silver lining and facilitates the improvement of health systems and health outcomes in the long term.

Ribosomal protein control of hematopoietic stem cell transformation through direct, non-canonical regulation of metabolism.

We report here that expression of the ribosomal protein, RPL22, is frequently reduced in human myelodysplastic syndrome (MDS) and acute myelogenous leukemia (AML); reduced RPL22 expression is associated with worse outcomes. Mice null for Rpl22 display characteristics of an MDS-like syndrome and develop leukemia at an accelerated rate. Rpl22-deficient mice also display enhanced hematopoietic stem cell (HSC) self-renewal and obstructed differentiation potential, which arises not from reduced protein synthesis but from increased expression of the Rpl22 target, ALOX12, an upstream regulator of fatty acid oxidation (FAO). The increased FAO mediated by Rpl22-deficiency also persists in leukemia cells and promotes their survival. Altogether, these findings reveal that Rpl22 insufficiency enhances the leukemia potential of HSC via non-canonical de-repression of its target, ALOX12, which enhances FAO, a process that may serve as a therapeutic vulnerability of Rpl22 low MDS and AML leukemia cells. Highlights: RPL22 insufficiency is observed in MDS/AML and is associated with reduced survivalRpl22-deficiency produces an MDS-like syndrome and facilitates leukemogenesisRpl22-deficiency does not impair global protein synthesis by HSCRpl22 controls leukemia cell survival by non-canonical regulation of lipid oxidation eTOC: Rpl22 controls the function and transformation potential of hematopoietic stem cells through effects on ALOX12 expression, a regulator of fatty acid oxidation.

Biodegradable nanoparticles are excellent vehicle for site directed in-vivo delivery of drugs and vaccines.

Biodegradable nanoparticles (NPs) are gaining increased attention for their ability to serve as a viable carrier for site specific delivery of vaccines, genes, drugs and other biomolecules in the body. They offer enhanced biocompatibility, superior drug/vaccine encapsulation, and convenient release profiles for a number of drugs, vaccines and biomolecules to be used in a variety of applications in the field of medicine. In this manuscript, the methods of preparation of biodegradable NPs, different factors affecting optimal drug encapsulation, factors affecting drug release rates, various surface modifications of nanoparticles to enhance in-vivo circulation, distribution and multimodal functionalities along with the specific applications such as tumor targeting, oral delivery, and delivery of these particles to the central nervous system have been reviewed.

Use of silver nanoparticles increased inhibition of cell-associated HIV-1 infection by neutralizing antibodies developed against HIV-1 envelope proteins.

BACKGROUND: HIV/AIDS pandemic is a worldwide public health issue. There is a need for new approaches to develop new antiviral compounds or other therapeutic strategies to limit viral transmission. The envelope glycoproteins gp120 and gp41 of HIV are the main targets for both silver nanoparticles (AgNPs) and neutralizing antibodies. There is an urgency to optimize the efficiency of the neutralizing antibodies (NABs). In this study, we demonstrated that there is an additive effect between the four NABs and AgNPs when combined against cell-associated HIV-1 infection in vitro RESULTS: Four NABs (Monoclonal antibody to HIV-1 gp41 126-7, HIV-1 gp120 Antiserum PB1 Sub 2, HIV-1 gp120 Antiserum PB1, HIV-1 gp120 Monoclonal Antibody F425 B4e8) with or without AgNPs of 30-50 nm in size were tested against cell free and cell-associated HIVIIIB virus. All NABs inhibited HIV-1 cell free infection at a dose response manner, but with AgNPs an antiviral additive effect was not achieved Although there was no inhibition of infection with cell-associated virus by the NABs itself, AgNPs alone were able to inhibit cell associated virus infection and more importantly, when mixed together with NABs they inhibited the HIV-1 cell associated infection in an additive manner. DISCUSSION: The most attractive strategies to deal with the HIV problem are the development of a prophylactic vaccine and the development of effective topical vaginal microbicide. For two decades a potent vaccine that inhibits transmission of infection of HIV has been searched. There are vaccines that elicit NABs but none of them has the efficacy to stop transmission of HIV-1 infection. We propose that with the addition of AgNPs, NABs will have an additive effect and become more potent to inhibit cell-associated HIV-1 transmission/infection. CONCLUSIONS: The addition of AgNPs to NABs has significantly increased the neutralizing potency of NABs in prevention of cell-associated HIV-1 transmission/infection. Further exploration is required to standardize potentiation of NABs by AgNPs. It is also required to evaluate in vivo toxicity of AgNPs before AgNPs could be incorporated in any antiviral vaginal creams.

Silver nanoparticles are broad-spectrum bactericidal and virucidal compounds.

The advance in nanotechnology has enabled us to utilize particles in the size of the nanoscale. This has created new therapeutic horizons, and in the case of silver, the currently available data only reveals the surface of the potential benefits and the wide range of applications. Interactions between viral biomolecules and silver nanoparticles suggest that the use of nanosystems may contribute importantly for the enhancement of current prevention of infection and antiviral therapies. Recently, it has been suggested that silver nanoparticles (AgNPs) bind with external membrane of lipid enveloped virus to prevent the infection. Nevertheless, the interaction of AgNPs with viruses is a largely unexplored field. AgNPs has been studied particularly on HIV where it was demonstrated the mechanism of antiviral action of the nanoparticles as well as the inhibition the transmission of HIV-1 infection in human cervix organ culture. This review discusses recent advances in the understanding of the biocidal mechanisms of action of silver Nanoparticles.

Vitamin D levels in paediatric intensive care unit patients and its relation to severity of illness: An Indian experience.

Vitamin D deficiency is a common disorder that is associated with morbidity and mortality in the general population. We conducted a cross-sectional study of 384 children admitted to paediatric intensive care to determine its prevalence and association with severity of illness and outcome in critically ill children. The severity of illness was evaluated using the paediatric risk of mortality score (PRISM III), on admission, at 24 and 48 h. Vitamin D deficiency was observed in 175 children (45.6%) and was associated with higher severity of illness, need for mechanical ventilation and increased mortality.

Selection and application of biological safety cabinets in diagnostic and research laboratories with special emphasis on COVID-19.

The ongoing coronavirus disease (COVID-19) pandemic is a global public health emergency. Adherence to biosafety practices is mandatory to protect the user as well as the environment, while handling infectious agents. A biological safety cabinet (BSC) is the most important equipment used in diagnostic and research laboratories in order to safeguard the product, the person, and the environment. The World Health Organization has emphasized the use of validated BSCs in order to ensure quality of the results. There are different classes of BSCs that are used in various work environments based on the need. It is imperative to use appropriate levels of biosafety and types of BSCs in laboratories based on the risk assessment of the pathogen used. During the development of COVID-19 laboratories and training of laboratory staff, we came across several queries about the functions and selection of BSCs and realized that the knowledge about the detailed information on selections and applications of BSCs is scanty. There are several guidelines regarding the biosafety aspects for diagnostic and research laboratories handling infectious pathogens from national and international agencies. However, there is no detailed information on the use of appropriate types of BSCs and their functions in the context of Severe Acute Respiratory Syndrome-Coronavirus 2 (SARS-CoV-2). In view of this, the present paper describes in detail the selection and applications of BSCs, which could be useful for laboratories handling or planning to handle SARS-CoV-2 and suspected samples.

Morphological and Biochemical Characteristics of Avian Faecal Droppings and Their Impact on Survival of Avian Influenza Virus.

Environmental specimens such as faecal droppings are considered important for the detection of avian influenza viruses (AIV). In view of lower rates of AIV isolation from avian faecal droppings, characterization of droppings is imperative to elucidate contributing factors. However, there are no reports on morphological and biochemical characteristics of droppings. The objective of the present study was the characterization of droppings from different avian species and their impact on the AIV detection and isolation. A total of 373 droppings belonging to 61 avian species from 22 families of apparently healthy wild migratory, resident, domestic birds and poultry were studied during five winter migratory bird seasons between 2007 to 2012 and 2016-2017. The colour, morphology and size of the droppings varied from species-to-species. These data could be useful for the identification of avian species. Droppings from 67% of the avian species showed acidic pH (4.5-6.5); nine species showed neutral pH (7.0), and 11 species showed alkaline pH (7.5). The infectious titers of AIV in droppings with acidic pH were significantly lower (p < 0.05) than those of the droppings with neutral and alkaline pH. However, acidic pH did not hamper AIV detection by real-time RT-PCR. In order to avoid the impact of acidic pH, collecting fresh droppings into viral transport medium (pH 7.0-7.5) would help improve the rate of AIV isolation.

Oncogenes Activate an Autonomous Transcriptional Regulatory Circuit That Drives Glioblastoma.

Efforts to identify and target glioblastoma (GBM) drivers have primarily focused on receptor tyrosine kinases (RTKs). Clinical benefits, however, have been elusive. Here, we identify an SRY-related box 2 (SOX2) transcriptional regulatory network that is independent of upstream RTKs and capable of driving glioma-initiating cells. We identified oligodendrocyte lineage transcription factor 2 (OLIG2) and zinc-finger E-box binding homeobox 1 (ZEB1), which are frequently co-expressed irrespective of driver mutations, as potential SOX2 targets. In murine glioma models, we show that different combinations of tumor suppressor and oncogene mutations can activate Sox2, Olig2, and Zeb1 expression. We demonstrate that ectopic co-expression of the three transcription factors can transform tumor-suppressor-deficient astrocytes into glioma-initiating cells in the absence of an upstream RTK oncogene. Finally, we demonstrate that the transcriptional inhibitor mithramycin downregulates SOX2 and its target genes, resulting in markedly reduced proliferation of GBM cells in vivo.

Mycobacterium tuberculosis secreted antigen (MTSA-10) modulates macrophage function by redox regulation of phosphatases.

Macrophages are the primary host cells for Mycobacterium tuberculosis (Mtb). Although macrophages can mount a strong inflammatory response to dispose of invading microbial pathogens, the immune dysfunction of the Mtb-infected macrophage constitutes the hallmark of mycobacterial pathogenesis. A 10-kDa, Mtb secretory antigen (MTSA-10), encoded by ORF Rv3874, is one of the predominant members of the 'region of difference 1' locus of Mtb genome that has been strongly implicated in mycobacterial virulence. In this study, we investigated the possible role of MTSA-10 in modulating the macrophage dysfunction in a mouse macrophage cell line J774.1. We found that recombinant MTSA-10 caused extensive protein dephosphorylation in J774.1 cells as revealed by two-dimensional gel electrophoresis analysis. We also observed that MTSA-10 treatment downregulated the reactive oxygen species levels in the cells leading to activation of cellular protein phosphatases putatively responsible for the dephosphorylation phenomenon. This implied a direct role of MTSA-10 in the disruption of host cell signaling, resulting in downregulation of transcription of several genes essential for macrophage function.

Effect of binary combination of some plant-derived molluscicides with MGK-264 or piperonyl butoxide on the reproduction of the snail Lymnaea acuminata.

The effects of sub-lethal treatments (20 and 60% of 24-h LC(50)) with plant-derived molluscicides Annona squamosa, acetogenins, Argemone mexicana seed and protopine, in combination (1 + 5) with MGK-264 (ENT 8184) or piperonyl butoxide on the reproduction of Lymnaea acuminata has been studied. The plant-derived molluscicides and their active molluscicidal components, protopine and acetogenins, in combination with ENT 8184 or piperonyl butoxide caused a significant reduction in the fecundity, hatchability and survival of young snails. Combination of A squamosa seed powder with piperonyl butoxide was very effective as it caused a complete arrest of snail fecundity within 24 h of treatment. Removal of the snails to fresh water after the 96-h treatments caused a significant recovery in the fecundity of L acuminata.

Vancomycin-resistance Enterococcal Colonization in Hospitalized Patients in Relation to Antibiotic Usage in a Tertiary Care Hospital of North India.

BACKGROUND: Alarming rise of vancomycin-resistant enterococci (VRE) is a global cause of concern. Several factors have been held responsible for such rise, of which antibiotic usage is a prominent one. OBJECTIVE: This study was undertaken to determine the intestinal VRE colonization rate amongst hospitalized patients in relation to use of various antibiotics in the Intensive Care Unit (ICU) of a tertiary care university hospital, India. MATERIALS AND METHODS: Stool samples were collected weekly from all the patients in the adult ICU for a period of 6 months and processed for isolation and phenotypic and genotypic characterization of VRE isolates. Patient and treatment details were noted and cases (those with VRE in stool) and controls (those without VRE in stool) were compared statistically. Further, a multivariate analysis was done to identify those antibiotics as independent risk factors for VRE colonization. RESULTS: VRE colonization was found in 34.56% (28/81) of the patients studied, with the majority 75% (21/28) carrying the vanA gene. The cases had significantly more (P < 0.05) duration of hospital stay and antibiotic exposure. Intake of metronidazole, vancomycin, and piperacillin-tazobactam were identified as significant risk factors both in univariate and multivariate analysis. CONCLUSION: A potential reservoir of VRE was thus revealed even in low VRE prevalence setting. Based on this high colonization status, restriction of empirical antibiotic use, reviewing of the ongoing antibiotic policy, and active VRE surveillance as an integral part of infection control strategy were suggested.

The primary phase of infection by pathogenic simian-human immunodeficiency virus results in disruption of the blood-brain barrier.

Using the simian-human immunodeficiency virus (SHIV), we have investigated whether the blood-brain barrier (BBB) is compromised during the early stages of infection. Five macaques were inoculated with pathogenic SHIV(50OLNV) for 2 weeks at which time macaques were anesthetized, perfused with saline, and sacrificed. The brains were removed and examined for the disruption of the blood-brain barrier by immunohistochemical staining for the plasma protein fibrinogen in the neural parenchyma. Our results indicate a disruption of the BBB in the five of five macaques inoculated with SHIV(50OLNV) for 2 weeks. Zonula occludens 1 (ZO-1), which is a marker for the tight junctions formed by brain vascular endothelial cells, was largely absent in areas that showed fibrinogen deposition in all five macaques. To determine if the BBB integrity correlated with the initial stages of infection, the brains from two macaques were analyzed that had progressed to end-stage disease following inoculation with pathogenic SHIV(50OLNV) but developed no neuropathology and from two macaques that were inoculated with a gene-deleted, nonpathogenic virus (novpuSHIV(KU-1bMC33)) for over 1 year. Our results indicate that unlike the macaques sacrificed during the acute phase of infection, immunohistochemical staining for fibrinogen in the neural parenchyma was negative and ZO-1 staining was readily detected in the endothelial cells of the blood vessels. The results of this study indicate that the transient loss of BBB integrity is a function of the high level of virus replication that occurs during the acute phase of infection and provides important information on the early stages of lentivirus neuroinvasion.

Infection of human astrocytoma cells with simian-human immunodeficiency virus results in up-regulation of gene expression and altered growth properties.

Recent reports of human immunodeficiency virus-1 (HIV-1) infection of astrocytes suggest a role for astrocytes in HIV encephalitis. In this study, we infected a human astrocytoma cell line with a pathogenic simian HIV (SHIV(50OLNV)) and examined growth patterns and immunomodulatory genes. Approximately 1% of uninfected cells in culture expressed glial fibrillary acid protein (GFAP) whereas 40% of the cells expressed GFAP at 7 days post-inoculation along altered growth patterns. Using targeted cytokine cDNA arrays, we found that SHIV(50OLNV) infection resulted in the up-regulation of several genes including metalloproteinase bone morphogenic protein 1 and chemokines monocyte chemoattractant protein 1 and stromal cell derived factor 1alpha. These data suggest that astrocytic activation, altered morphology and up-regulation of immunomodulatory genes in response to SHIV infection may participate in initiation of inflammation and trafficking of infected monocytes/macrophages into the central nervous system, potentiating the development of HIV encephalitis.

Evaluating the role of flupirtine for postcraniotomy pain and compare it with diclofenac sodium: a prospective, randomized, double blind, placebo-controlled study.

BACKGROUND: Patients undergoing craniotomy, experience moderate to severe pain in postoperative period. Flupirtine does not have side effects like sedation and increase postoperative bleeding, so it may be a useful analgesic in neurosurgical patients. We designed this prospective, randomized, double blind, placebo-controlled study to evaluate the role of flupirtine for postcraniotomy pain and compare it with diclofenac sodium. MATERIALS AND METHODS: A total of 390 adults (18 to 70 y), American Society of Anaesthesiologists I and II, of either sex, undergoing elective craniotomy, were randomly divided into 3 equal groups of 130 each. Group 1 (control) received placebo, group 2 (diclofenac) received tablet diclofenac 50 mg, and group 3 (flupirtine) received capsule flupirtine 100 mg. All medications were given 8 hourly on second postoperative day for 48 hours. Visual Analogue Scale score, level of sedation and incidence of side effects were observed. RESULTS: Nineteen patients were dropped from the study and therefore subsequent analysis was carried out for 371 patients only. There was significant reduction of Visual Analogue Scale score in flupirtine and diclofenac group when compared to control (P<0.0001). Pain relief observed in control, flupirtine, and diclofenac group was 69.8%, 90.2%, and 90.5%, respectively. Need of rescue analgesia was significantly less in flupirtine and diclofenac group as compared to control (P<0.0001). No significant difference was observed among the groups in regards to adverse effects. CONCLUSION: We conclude that oral flupirtine 100 mg is safe and as effective as oral diclofenac sodium 50 mg in reducing postcraniotomy pain.