We sequenced and characterized the inhibitor of apoptosis (iap) 1 gene from Aedes albopictus, designated as Aaiap1. The Aaiap1 gene rescued Spodoptera frugiperda (Sf9) cells from apoptosis when cotransfected with the Drosophila pro-apoptotic hid gene. The antiapoptotic function of the Aaiap1 gene was evaluated in the bluetongue virus (BTV)-induced apoptosis system. BTV infection induced apoptosis in vertebrate cells via the intrinsic apoptotic pathway. This was shown by the translocation of cytochrome C and the second mitochondria-derived activator of caspase (Smac, also known as DIABLO) from the mitochondria and the subsequent activation of caspase-9 and -3. Stable expression of the Aaiap1 gene in derivative baby hamster kidney cells delayed BTV-induced apoptosis by 24 h and reduced the BTV progeny yield by 10-fold. This study provides the first evidence that the mosquito AaIAP1 protein possesses antiapoptotic activity.

A case-control study of maternal mortality was conducted in selected rural areas of two provinces in China: Henan province, which has a relatively lower socio-economic status and higher maternal mortality rates, and Jiangsu province with higher socio-economic status and lower maternal mortality rates. The major cause of maternal mortality in the two provinces was postpartum hemorrhage and the largest proportion of deaths occurred on the road between the women's home and the health care facility. Results indicate that the expectant mother's socio-economic status, knowledge of maternal care, and the nature and level of maternal care provided all influence rural maternal death rates. However, socio-economic factors were only significant predictors of mortality in the poorer province. Implications for health policy and future research are discussed.

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Cigarette smoking is associated with COVID-19 prevalence and severity, but the mechanistic basis for how smoking alters SARS-CoV-2 pathogenesis is unknown. A potential explanation is that smoking could alter expression of angiotensin converting enzyme-2 (ACE2), which functions as the cellular receptor and point of entry. Here we investigated the severity of SARS CoV-2 infection ex vivo and in vitro and using tissue samples and primary ferret and human airway epithelial cells with and without antecedent exposure to cigarette smoke. Methods: ACE2 expression measured by Quantitative PCR (Q-PCR) of ferret lungs exposed to 6 months of cigarette smoke, and findings validated by immunofluorescence (IF). Primary airway cells isolated from airways of ferrets or human non-smokers or COPD subjects were grown until terminally differentiated at air liquid interface. Cells were then exposed to cigarette smoke extract (CSE) or vehicle control, then infected with SARS CoV-2 (3 MOI) or mock control. Viral copy was measured by Q-PCR. Viral infection was quantified by foci forming assay (FFU/mL) using VeroE6 cells. Results: Ferret lungs following 6 months of smoke exposure had increased ACE2 as compared to air controls by Q-PCR (>1.5 fold, P<0.05, N=6), and IF staining. Higher ACE2 expression was also observed in normal ferret airway cells exposed to CSE (>1.5 fold, P<0.05, N=3), normal Human Bronchial Epithelial (HBE) cells exposed to CSE (>2 fold, P<0.001, N=4), and HBE cells from COPD donors as compared to healthy controls (>2 fold, P<0.001, N=4). When ferret airway cells were inoculated with SARS-CoV-2, intracellular viral load of SARS-CoV-2 was increased in CSE exposed cells as compared to vehicle controls (10 3 -10 4 vehicle Vs CSE 10 5 10 6 copy/μL). Viral infection was also increased >2 fold (P<0.01, N=5). Likewise, CSE exposed (10 5 -10 7 copy/μL, P<0.0001, N=4) and COPD (10 6 -10 8 copy/μL, P<0.0001, N=4) HBE had increased viral load as compared to controls (10 3 -10 4 copy/μL, P <0.001, N=4), and 2 to 3-fold increase in viral infection, respectively. TUNEL staining was increased in infected cells, indicating apoptosis. Transcript analysis of HBE cells with and without SARS-CoV-2 by RNASeq to identify differentially expressed genes in CSE exposed cells as compared to controls is in progress. Conclusion: Cigarette smoke and CSE increased ACE2 expression in ferrets, and ferret and human cells respectively. CSE-induction increased viral replication and infection severity, resulting in increased apoptosis. Cigarette smoking likely influences the severity of SARS-CoV-2 infection by altering expression of ACE2, inducing airway cell apoptosis upon infection.

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