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Background: The rapid emergence of the SARS-CoV-2 Omicron variant that evades many therapies illustrates the need for antiviral treatments with high genetic barriers to resistance. The small molecule PAV-104, identified through a moderate-throughput screen involving cell-free protein synthesis, was recently shown to target a subset of host protein assembly machinery in a manner specific to viral assembly with minimal host toxicity. The chemotype shows broad activity against respiratory viral pathogens, including Orthomyxoviridae, Paramyxoviridae, Adenoviridae, Herpesviridae, and Picornaviridae, with low susceptibility to evolutionary escape. Here, we investigated the capacity of PAV-104 to inhibit SARS-CoV-2 replication in human airway epithelial cells (AECs). Method(s): Dose-dependent cytotoxicity of PAV-104 in Calu-3 cells was determined by MTT assay. Calu-3 cells were infected with SARS-CoV-2 isolate USA-WA1/2020 (MOI=0.01). Primary AECs were isolated from healthy donor lung transplant tissue, cultured at air liquid interface (ALI), and infected with SARS-CoV-2 Gamma, Delta, and Omicron variants (MOI=0.1). SARS-CoV-2 replication was assessed by RT-PCR quantitation of the N gene, immunofluorescence assay (IFA) of nucleocapsid (N) protein, and titration of supernatant (TCID50). Transient co-expression of four SARS-CoV-2 structural proteins (N, M, S, E) to produce virus-like particles (VLPs) was used to study the effect of PAV-104 on viral assembly. Drug resin affinity chromatography was performed to study the interaction between PAV-104 and N. Glycerol gradient sedimentation was used to assess N oligomerization. Total RNA-seq and the REACTOME database were used to evaluate PAV-104 effects on the host transcriptome. Result(s): PAV-104 reached 50% cytotoxicity in Calu-3 cells at 3732 nM (Fig.1A). 50 nM PAV-104 inhibited >99% of SARS-CoV-2 infection in Calu-3 cells (p< 0.01) and in primary AECs (p< 0.01) (Fig.1B-E). PAV-104 specifically inhibited SARS-CoV-2 post entry, and suppressed production of SARS-CoV-2 VLPs without affecting viral protein synthesis. PAV-104 interacted with SARS-CoV-2 N and interfered with N oligomerization. Transcriptome analysis revealed that PAV-104 treatment reversed SARS-CoV-2 induction of the interferon and maturation of nucleoprotein signaling pathways. Conclusion(s): PAV-104 is a pan-respiratory virus small molecule inhibitor with promising activity against SARS-CoV-2 in human airway epithelial cells that should be explored in animal models and clinical studies.
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Introduction: The prevalence of obesity among school children in Kerala is on a steady rise. Consumption of food with high glycaemic index, change in sleep patterns, reduced physical activity and the use of screen has been linked to obesity in children. Published literature on this association is scarce from urban Thiruvananthapuram, hence, the present study. Aim(s): To identify the association of various risk factors such as frequency of junk food consumption, dietary preferences, physical activity and daily screen time and weight related disorders among school going children (8-10 years) in Thiruvananthapuram. Material(s) and Method(s): The present cross-sectional case-control study was conducted in one Rural Government School (Venjaramoodu Government Upper Primary School) and one Urban Private School (S.N. Public School, Chenkottukonam) of Thiruvananthapuram, Kerala, India, and enrolled school going children aged 8-10 years with higher than recommended Body Mass Index (BMI) for age as cases, age and gender-matched children with normal BMI as controls. Participants with BMI above 23rd and below 27th adult equivalent for age and gender were considered overweight and those above 27th adult equivalent for age and gender were considered as obese. A structured questionnaire was sent home with the children, and the parents were requested to answer the questions along with written informed consent. Socio-demographic parameters, anthropometric measurements were obtained by trained staff, dietary habits, and details regarding physical activity and screen usage were collected. Variables were categorised according to the standard recommendations by World Health Organisation (WHO) and Indian Association of Paediatrics (IAP). Variables were expressed as frequencies and the tests of significance used were Chi-square test and Odds ratio, to express the strength of association between parameters. A p-value <0.05 was considered statistically significant. Result(s): The mean age of cases and controls was nine years. A total of 708 school children were screened and 352 participants (175 cases and 177 controls) were enrolled in the present study. The BMI of cases was 29.3 kg/m2 and of controls was 20.2 kg/m2. Higher than recommended screen time (p<0.001), more frequent junk food consumption (p<0.001) and lack of physical activity (p<0.001) were found to be significantly associated with obesity and overweight. Dietary preference was not associated with obesity or overweight and obesity and overweight was more common in children studying in private schools (p<0.001). Conclusion(s): Reducing screen time, reducing junk food consumption and increasing physical activity will help in reducing the prevalence of life style diseases among school children. Further evaluation is necessary to determine the factors contributing to the increased prevalence of these disorders in private schools.Copyright © 2023 Journal of Clinical and Diagnostic Research. All rights reserved.
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Introduction: The rapid emergence of the SARS-CoV-2 Omicron variant that evades many monoclonal antibody therapies illustrates the need for anti-viral treatments with low susceptibility to evolutionary escape. The small molecule PAV-104, identified through a moderate-throughput screen involving cell-free protein synthesis, was recently shown to target a subset of host protein assembly machinery in a manner specific to viral assembly. This compound has minimal host toxicity, including once daily oral dosing in rats that achieves >200-fold of the 90% effective concentration (EC90) in blood. The chemotype shows broad activity against respiratory viral pathogens, including Orthomyxoviridae, Paramyxoviridae, Adenoviridae, Herpesviridae, and Picornaviridae, with low suceptability to evolutionary escape. We hypothesized that PAV-104 would be active against SARSCoV- 2 variants in human airway epithelial cells. Methods: Airway epithelial cells were differentiated from lung transplant tissue at air-liquid interface (ALI) for four weeks prior to challenge with Alpha (Pango lineage designation B.1.1.7), Beta (B.1.351), Gamma (P.1), and Delta (B.1.617.2) SARS-CoV-2 variants. Viral replication was determined by quantitative PCR measurement of the SARS-CoV-2 nucleocapsid (N) gene. Dose-dependent virus inhibition and cytotoxicity of PAV-104 in the Calu-3 airway epithelial cell line was determined by PCR and MTT assay. Student's t-tests were used to evaluate statistical significance. Results: Alpha, Beta, Gamma, and Delta variants of SARS-CoV-2 showed comparable infectivity in human primary airway epithelial cells at ALI (N=3 donors), 47- to 550-fold higher than the parent (USA-WA1/2020) strain. PAV-104 reached 50% cytotoxicity in Calu-3 cells at 240 nM (Fig. 1A). Dose-response studies in Calu-3 cells demonstrated PAV-104 has a 6 nM 50% inhibitory concentration (IC50) for blocking replication of SARS-CoV-2 (USA-WA1/2020) (Fig.1B). In primary cells at ALI from 3 donors tested, there was >99% inhibition of infection by SARS-CoV-2 Gamma variant (N=3, MOI 0.1, P <0.01) with 100 nM PAV-104 (Fig. 1C). Addition of 100 nM PAV-104 2-hours post-infection, but not pre-infection, resulted in >99% suppression of viral replication, indicating a post-entry drug mechanism. PAV-104 bound a small subset of the known allosteric modulator 14-3-3, itself implicated in the interactome of SARS-CoV-2. Conclusion: PAV-104 is a host-targeted, orally bioavailable, pan-viral small molecule inhibitor with promising activity against SARS-CoV-2 variants in human primary airway epithelial cells. (Figure Presented).
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BACKGROUND: With the emerging pandemic of corona virus disease 19 (Covid-19) infection around the world, the need to identify the course of this disease in pregnant women becomes the need of the hour. Its effect on pregnancy and the neonatal outcome is not well known because of lack of any reliable data. We wanted to evaluate the clinical features, laboratory manifestations and obstetric outcome of Covid-19 in the term of antenatal mothers who were either admitted in labour, induction of labour or elective caesarean section and rule out vertical transmission by doing a routine neonatal screening for Covid-19. METHODS: A descriptive cross-sectional study conducted at Sri Ramachandra Medical College & Research Institute, Chennai from March 2020 to September 2020 includes all antenatal women who were tested positive for severe acute respiratory syndrome (SARS- Cov2) reverse transcription - polymerase chain reaction (RT-PCR). 43 Covid positive patients were included in the study. RESULTS: Incidence of Covid positive antenatal women was found to be 1.7%. Only 3% were symptomatic with mild disease. Gestational diabetes mellitus accounted for (27%), Class II obesity (7%), hypertensive disorders of pregnancy (13%) of the screen positive mothers. 20% of the labouring women had meconium-stained liquor. Lymphopenia was seen in 73% of cases. Elevated d-dimer in 13%, requiring thromboprophylaxis. Negative RT-PCR for SARS-CoV-2 in the neonates ruled out vertical transmission. CONCLUSIONS: Covid-19 in pregnancy is more common in those with gestational diabetes, obesity, with development of complications like hypertension and meconium-stained liquor. The impact of Covid-19 is not as disabling as it is in the non-pregnant population.
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N95 respirators are personal protective equipment most often used to control exposures to infections transmitted via the airborne route. Supplies of N95 respirators can become depleted during pandemics or when otherwise in high demand. In this paper, we offer strategies for optimizing supplies of N95 respirators in health care settings while maximizing the level of protection offered to health care personnel when there is limited supply in the United States during the 2019 coronavirus disease pandemic. The strategies are intended for use by professionals who manage respiratory protection programs, occupational health services, and infection prevention programs in health care facilities to protect health care personnel from job-related risks of exposure to infectious respiratory illnesses. Consultation with federal, state, and local public health officials is also important. We use the framework of surge capacity and the occupational health and safety hierarchy of controls approach to discuss specific engineering control, administrative control, and personal protective equipment measures that may help in optimizing N95 respirator supplies.