Alternative classification rules for two inverse gaussian populations with a common mean and order restricted scale-like parameters.

The problem of classification into two inverse Gaussian populations with a common mean and ordered scale-like parameters is considered. Surprisingly, the maximum likelihood estimators (MLEs) of the associated model parameters have not been utilized for classification purposes. Note that the MLEs of the model parameters, including the MLE of the common mean, do not have closed-form expressions. In this paper, several classification rules are proposed that use the MLEs and some plug-in type estimators under order restricted scale-like parameters. In the sequel, the risk values of all the proposed estimators are compared numerically, which shows that the proposed plug-in type restricted MLE performs better than others, including the Graybill-Deal type estimator of the common mean. Further, the proposed classification rules are compared in terms of the expected probability of correct classification (EPC) numerically. It is seen that some of our proposed rules have better performance than the existing ones in most of the parameter space. Two real-life examples are considered for application purposes.

Evaluation of Ayush-64 (a Polyherbal Formulation) and Its Ingredients in the Syrian Hamster Model for SARS-CoV-2 Infection Reveals the Preventative Potential of Alstonia scholaris.

In the current study, we evaluated the efficacy of Ayush-64 (A64), a polyherbal formulation containing Alstonia scholaris (L.) R. Br. (A. scholaris), Caesalpinia crista L. (C. crista), Picrorhiza kurroa Royle ex Benth (P. kurroa), and Swertia chirata (Roxb.) H. Karst. (S. chirata) against COVID-19 in a Syrian hamster infection model. Preventative use of A64 resulted in the late-phase recovery of body weight loss in severe acquired respiratory syndrome coronavirus-2 (SARS-CoV-2)-infected hamsters, suppression of pro-inflammatory cytokines, and blunted pulmonary pathology. In addition, we also investigated the efficacy of individual ingredients of A64, viz., A. scholaris, C. crista, P. kurroa, and S. chirata, in the hamster model. The hamster challenge data showed robust anti-viral and immunomodulatory potential in A. scholaris, followed by P. kurroa. However, C. crista and S. chirata of A64 showed prominent immunomodulatory potential without limiting the lung viral load. In order to better understand the immunomodulatory potential of these herbal extracts, we used an in vitro assay of helper T cell differentiation and found that A. scholaris mediated a more profound suppression of Th1, Th2, and Th17 cell differentiation as compared to A64 and other ingredients. Taken together, our animal study data identifies the ameliorative potential of A64 in mitigating coronavirus disease-19 (COVID-19) pulmonary pathology. A. scholaris, a constituent extract of A64, showed relatively higher anti-viral and immunomodulatory potential against COVID-19. The present study warrants further investigations to identify the active pharmaceutical ingredients of A. scholaris for further studies.

IL-9 aggravates SARS-CoV-2 infection and exacerbates associated airway inflammation.

SARS-CoV-2 infection is known for causing broncho-alveolar inflammation. Interleukin 9 (IL-9) induces airway inflammation and bronchial hyper responsiveness in respiratory viral illnesses and allergic inflammation, however, IL-9 has not been assigned a pathologic role in COVID-19. Here we show, in a K18-hACE2 transgenic (ACE2.Tg) mouse model, that IL-9 contributes to and exacerbates viral spread and airway inflammation caused by SARS-CoV-2 infection. ACE2.Tg mice with CD4+ T cell-specific deficiency of the transcription factor Forkhead Box Protein O1 (Foxo1) produce significantly less IL-9 upon SARS-CoV-2 infection than the wild type controls and they are resistant to the severe inflammatory disease that characterises the control mice. Exogenous IL-9 increases airway inflammation in Foxo1-deficient mice, while IL-9 blockade reduces and suppresses airway inflammation in SARS-CoV-2 infection, providing further evidence for a Foxo1-Il-9 mediated Th cell-specific pathway playing a role in COVID-19. Collectively, our study provides mechanistic insight into an important inflammatory pathway in SARS-CoV-2 infection, and thus represents proof of principle for the development of host-directed therapeutics to mitigate disease severity.

Forecasting road accidental deaths in India: an explicit comparison between ARIMA and exponential smoothing method.

The number of deaths due to road accident is increasing day by day and has become an alarming global problem over the decades. India, with her rising motorization is no stranger to this global catastrophe. In this paper two relatively simple yet powerful and versatile techniques for forecasting time series data, autoregressive integrated moving average method (ARIMA) and exponential smoothing method are used to forecast the number of deaths due to road accidents in India from the year 2022-2031. The results based on the two methods are compared and it is found that they are in sync with each other and pre-existing literature. Furthermore, this is a unique attempt to use two time series analysis techniques on the same data and carry out a comparative analysis. The data was collected from the annual report of Ministry of Road Transport and Highways, India (2020) and Accidental Deaths & Suicides in India (ADSI) Report of National Crime Record Bureau (2021). After examining all the probable models, it is observed that ARIMA (2, 2, 2) model and exponential smoothing (M, A, N) model are suitable for the given data. Amongst the two, ARIMA (2, 2, 2) model has a lower AIC and BIC value. Thus, this comes out to be the best model as per our model selection criterion. Further, the study also reveals an upward trend of number of road accidental deaths for the upcoming 10 years in India.

G4-binding drugs, chlorpromazine and prochlorperazine, repurposed against COVID-19 infection in hamsters.

The COVID-19 pandemic caused by SARS-CoV-2 has caused millions of infections and deaths worldwide. Limited treatment options and the threat from emerging variants underline the need for novel and widely accessible therapeutics. G-quadruplexes (G4s) are nucleic acid secondary structures known to affect many cellular processes including viral replication and transcription. We identified heretofore not reported G4s with remarkably low mutation frequency across >5 million SARS-CoV-2 genomes. The G4 structure was targeted using FDA-approved drugs that can bind G4s - Chlorpromazine (CPZ) and Prochlorperazine (PCZ). We found significant inhibition in lung pathology and lung viral load of SARS-CoV-2 challenged hamsters when treated with CPZ or PCZ that was comparable to the widely used antiviral drug Remdesivir. In support, in vitro G4 binding, inhibition of reverse transcription from RNA isolated from COVID-infected humans, and attenuated viral replication and infectivity in Vero cell cultures were clear in case of both CPZ and PCZ. Apart from the wide accessibility of CPZ/PCZ, targeting relatively invariant nucleic acid structures poses an attractive strategy against viruses like SARS-CoV-2, which spread fast and accumulate mutations quickly.

Pharmacological potential of Withania somnifera (L.) Dunal and Tinospora cordifolia (Willd.) Miers on the experimental models of COVID-19, T cell differentiation, and neutrophil functions.

Cytokine release syndrome (CRS) due to severe acute respiratory coronavirus-2 (SARS-CoV-2) infection leads to life-threatening pneumonia which has been associated with coronavirus disease (COVID-19) pathologies. Centuries-old Asian traditional medicines such as Withania somnifera (L.) Dunal (WS) and Tinospora cordifolia (Willd.) Miers (TC) possess potent immunomodulatory effects and were used by the AYUSH ministry, in India during the COVID-19 pandemic. In the present study, we investigated WS and TC's anti-viral and immunomodulatory efficacy at the human equivalent doses using suitable in vitro and in vivo models. While both WS and TC showed immuno-modulatory potential, WS showed robust protection against loss in body weight, viral load, and pulmonary pathology in the hamster model of SARS-CoV2. In vitro pretreatment of mice and human neutrophils with WS and TC had no adverse effect on PMA, calcium ionophore, and TRLM-induced ROS generation, phagocytosis, bactericidal activity, and NETs formation. Interestingly, WS significantly suppressed the pro-inflammatory cytokines-induced Th1, Th2, and Th17 differentiation. We also used hACE2 transgenic mice to further investigate the efficacy of WS against acute SARS-CoV2 infection. Prophylactic treatment of WS in the hACE2 mice model showed significant protection against body weight loss, inflammation, and the lung viral load. The results obtained indicate that WS promoted the immunosuppressive environment in the hamster and hACE2 transgenic mice models and limited the worsening of the disease by reducing inflammation, suggesting that WS might be useful against other acute viral infections. The present study thus provides pre-clinical efficacy data to demonstrate a robust protective effect of WS against COVID-19 through its broader immunomodulatory activity.

Statistical modeling and estimating number of healthy life years lost and healthy life expectancy in India, 2000-2019.

Objective: In this study, our objective is to propose various models to estimate healthy life year lost (HLYL) and healthy life expectancy (HLE) in India. Methods: The HLYL and HLE were estimated and further these estimates were compared with the direct life table method and the World Health Organization (WHO) method. From the mortality perspective, we have developed a log-logistic model for estimating the parameter (bx), which is characterized by HLYL. The results were compared with other models, such as the Gompertz and Weibull model. Here, we have also obtained the HLE by subtracting HLYL from the total life expectancy. Results: The result shows an increasing trend of HLYL among the male, female, and the total population in India. Conclusion: From the log-logistic model, the HLYL was estimated as 8.79 years, 8.36 years, and 9.38 years for the total, male, and female populations, respectively, in India during 2019.

A broadly neutralizing monoclonal antibody overcomes the mutational landscape of emerging SARS-CoV-2 variants of concern.

The emergence of new variants of SARS-CoV-2 necessitates unremitting efforts to discover novel therapeutic monoclonal antibodies (mAbs). Here, we report an extremely potent mAb named P4A2 that can neutralize all the circulating variants of concern (VOCs) with high efficiency, including the highly transmissible Omicron. The crystal structure of the P4A2 Fab:RBD complex revealed that the residues of the RBD that interact with P4A2 are a part of the ACE2-receptor-binding motif and are not mutated in any of the VOCs. The pan coronavirus pseudotyped neutralization assay confirmed that the P4A2 mAb is specific for SARS-CoV-2 and its VOCs. Passive administration of P4A2 to K18-hACE2 transgenic mice conferred protection, both prophylactically and therapeutically, against challenge with VOCs. Overall, our data shows that, the P4A2 mAb has immense therapeutic potential to neutralize the current circulating VOCs. Due to the overlap between the P4A2 epitope and ACE2 binding site on spike-RBD, P4A2 may also be highly effective against a number of future variants.

Prophylactic treatment of Glycyrrhiza glabra mitigates COVID-19 pathology through inhibition of pro-inflammatory cytokines in the hamster model and NETosis.

Severe coronavirus disease (COVID-19) is accompanied by acute respiratory distress syndrome and pulmonary pathology, and is presented mostly with an inflammatory cytokine release, a dysregulated immune response, a skewed neutrophil/lymphocyte ratio, and a hypercoagulable state. Though vaccinations have proved effective in reducing the COVID-19-related mortality, the limitation of the use of vaccine against immunocompromised individuals, those with comorbidity, and emerging variants remains a concern. In the current study, we investigate for the first time the efficacy of the Glycyrrhiza glabra (GG) extract, a potent immunomodulator, against SARS-CoV-2 infection in hamsters. Prophylactic treatment with GG showed protection against loss in body weight and a 35%-40% decrease in lung viral load along with reduced lung pathology in the hamster model. Remarkably, GG reduced the mRNA expression of pro-inflammatory cytokines and plasminogen activator inhibitor-1 (PAI-1). In vitro, GG acted as a potent immunomodulator by reducing Th2 and Th17 differentiation and IL-4 and IL-17A cytokine production. In addition, GG also showed robust potential to suppress ROS, mtROS, and NET generation in a concentration-dependent manner in both human polymorphonuclear neutrophils (PMNs) and murine bone marrow-derived neutrophils (BMDNs). Taken together, we provide evidence for the protective efficacy of GG against COVID-19 and its putative mechanistic insight through its immunomodulatory properties. Our study provides the proof of concept for GG efficacy against SARS-CoV-2 using a hamster model and opens the path for further studies aimed at identifying the active ingredients of GG and its efficacy in COVID-19 clinical cases.

Proinflammatory Innate Cytokines and Distinct Metabolomic Signatures Shape the T Cell Response in Active COVID-19.

The underlying factors contributing to the evolution of SARS-CoV-2-specific T cell responses during COVID-19 infection remain unidentified. To address this, we characterized innate and adaptive immune responses with metabolomic profiling longitudinally at three different time points (0-3, 7-9, and 14-16 days post-COVID-19 positivity) from young, mildly symptomatic, active COVID-19 patients infected during the first wave in mid-2020. We observed that anti-RBD IgG and viral neutralization are significantly reduced against the delta variant, compared to the ancestral strain. In contrast, compared to the ancestral strain, T cell responses remain preserved against the delta and omicron variants. We determined innate immune responses during the early stage of active infection, in response to TLR 3/7/8-mediated activation in PBMCs and serum metabolomic profiling. Correlation analysis indicated PBMCs-derived proinflammatory cytokines, IL-18, IL-1ß, and IL-23, and the abundance of plasma metabolites involved in arginine biosynthesis were predictive of a robust SARS-CoV-2-specific Th1 response at a later stage (two weeks after PCR positivity). These observations may contribute to designing effective vaccines and adjuvants that promote innate immune responses and metabolites to induce a long-lasting anti-SARS-CoV-2-specific T cell response.

Golden Syrian hamster as a model to study cardiovascular complications associated with SARS-CoV-2 infection.

Severe acute respiratory syndrome coronavirus-2 (SARS-CoV-2) infection in the Golden Syrian hamster causes lung pathology that resembles human coronavirus disease (COVID-19). However, extrapulmonary pathologies associated with SARS-CoV-2 infection and post-COVID sequelae remain to be understood. Here, we show, using a hamster model, that the early phase of SARS-CoV-2 infection leads to an acute inflammatory response and lung pathologies, while the late phase of infection causes cardiovascular complications (CVCs) characterized by ventricular wall thickening associated with increased ventricular mass/body mass ratio and interstitial coronary fibrosis. Molecular profiling further substantiated our findings of CVC as SARS-CoV-2-infected hamsters showed elevated levels of serum cardiac troponin I, cholesterol, low-density lipoprotein, and long-chain fatty acid triglycerides. Serum metabolomics profiling of SARS-CoV-2-infected hamsters identified N-acetylneuraminate, a functional metabolite found to be associated with CVC, as a metabolic marker was found to be common between SARS-CoV-2-infected hamsters and COVID-19 patients. Together, we propose hamsters as a suitable animal model to study post-COVID sequelae associated with CVC, which could be extended to therapeutic interventions.

Effect of Prophylactic Use of Intranasal Oil Formulations in the Hamster Model of COVID-19.

Severe acute respiratory syndrome coronavirus 2 (SARS-CoV2) infection initiates with viral entry in the upper respiratory tract, leading to coronavirus disease 2019 (COVID-19). Severe COVID-19 is characterized by pulmonary pathologies associated with respiratory failure. Thus, therapeutics aimed at inhibiting the entry of the virus or its internalization in the upper respiratory tract are of interest. Herein, we report the prophylactic application of two intranasal formulations provided by the National Medicinal Plant Board (NMPB), Anu oil and til tailya, in the hamster model of SARS-CoV-2 infection. Prophylactic intra-nasal instillation of these oil formulations exhibited reduced viral load in lungs and resulted in reduced body weight loss and lung-pneumonitis. In line with reduced viral load, histopathological analysis revealed a reduction in lung pathology in the Anu oil group as compared to the control infected group. However, the til tailya group did not show a significant reduction in lung pathology. Furthermore, molecular analysis using mRNA expression profiling indicated reduced expression of pro-inflammatory cytokine genes, including Th1 and Th17 cytokines for both the intranasal formulations as a result of decreased viral load. Together, the prophylactic intranasal application of Anu oil seems to be useful in limiting both viral load and severity in SARS-CoV2 infection in the hamster model.

High-salt diet mediates interplay between NK cells and gut microbiota to induce potent tumor immunity.

High-salt diet (HSD) modulates effector and regulatory T cell functions and promotes tissue inflammation in autoimmune diseases. However, effects of HSD and its association with gut microbiota in tumor immunity remain undefined. Here, we report that HSD induces natural killer (NK) cell­mediated tumor immunity by inhibiting PD-1 expression while enhancing IFNγ and serum hippurate. Salt enhanced tumor immunity when combined with a suboptimal dose of anti-PD1 antibody. While HSD-induced tumor immunity was blunted upon gut microbiota depletion, fecal microbiota transplantation (FMT) from HSD mice restored the tumor immunity associated with NK cell functions. HSD increased the abundance of Bifidobacterium and caused increased gut permeability leading to intratumor localization of Bifidobacterium, which enhanced NK cell functions and tumor regression. Intratumoral injections of Bifidobacterium activated NK cells, which inhibited tumor growth. These results indicate that HSD modulates gut microbiome that induces NK cell­dependent tumor immunity with a potential translational perspective.

Forecasting suicide rates in India: An empirical exposition.

INTRODUCTION: Suicide is a major social and health issue in India. Yearly statistics show a concerning increasing pattern of suicidal deaths in India which is higher in comparison to the global trend. There is limited evidence regarding historical analysis of suicide or any forecasting for suicide in India towards predicting the possible risks of death due to suicide. METHODS: This paper examines the trend of suicide rate and characteristics of suicide victims in India, based on the longitudinal time series data over the last 50 years-collected from the National Crime Record Bureau Reports (1969 to 2018) of the Government of India. In our analysis, we have used the time series model to forecast the suicide rates in India for the next decade. ARIMA (4,1,0) model is found to be the best fit model for forecasting the data. FINDINGS: There has been an observable and rising trend of suicide rates in India over the last five decades. The forecast indicates a continuance of rising suicide cases for an upcoming couple of years in India with a limited decline in the following years. The prediction model indicates a future relatively consistent pattern of suicide in India which does not seem to be a very encouraging trend. As we have not included the period staring the year 2020 onwards affected by Covid-19 and which has several disruptions in personal and family spaces, the projected suicide trend during the period of next two to three years (2020-22) may rise far high and then it may show a declining path. Along with this, there is a shift in means of suicide in the last couple of decades. Constituting the second-highest number of cases, Illness associated suicide was visibly a serious concern. CONCLUSION: The present analysis finds that there is no visible substantial relief for suicide deaths during the coming years in India. On the other hand, more extensive exploration of sample cases may provide important information for suicide prevention. Availability of detailed and more inclusive data will be highly useful for analysis and suicide preventive policies. Investment in public health care and other welfare activities like education and employment generation will yield visible positive results in suicide control.

Author Correction: Au nanoparticles modified CuO nanowire electrode based non-enzymatic glucose detection with improved linearity.

An amendment to this paper has been published and can be accessed via a link at the top of the paper.

Au nanoparticles modified CuO nanowireelectrode based non-enzymatic glucose detection with improved linearity.

This paper explores gold nanoparticle (GNP) modified copper oxide nanowires(CuO NWs)based electrode grown on copper foil for non-enzymatic glucose detection in a wide linear ranging up to 31.06 mM, and 44.36 mM at 0.5 M NaOH and 1 M NaOH concentrations. The proposed electrode can be used to detect a very low glucose concentration of 0.3 µM with a high linearity range of 44.36mM and sensitivity of 1591.44 µA mM-1 cm-2. The electrode is fabricated by first synthesizing Cu (OH)2 NWs on a copper foil by chemical etching method and then heat treatment is performed to convert Cu (OH)2 NWs into CuO NWs. The GNPs are deposited on CuO NWs to enhance the effective surface-to-volume ratio of the electrode with improved catalytic activity. The surface morphology has been investigated by XRD, XPS, FE-SEM and HR-TEM analysis. The proposed sensor is expected to detect low-level of glucose in urine, and saliva. At the same time, it can also be used to measure extremely high sugar levels (i.e. hyperglycemia) of ~ 806.5454 mg/dl. The proposed sensor is also capable of detecting glucose after multiple bending of the GNP modified CuO NWs electrode. The proposed device is also used to detect the blood sugar level in human being and it is found that this sensor's result is highly accurate and reliable.