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Surveyed women attributed unintended pregnancies to COVID-19 and reported constraints to contraceptive access and use in Malawi, Nepal, Niger, and Uganda.

Human HtrA3 (high-temperature requirement protease A3) is a trimeric multitasking propapoptotic serine protease associated with critical cellular functions and pathogenicity. Implicated in diseases including cancer and pre-eclampsia, its role as a tumor suppressor and potential therapeutic target cannot be ignored. Therefore, elucidating its mode of activation and regulatory switch becomes indispensable towards modulating its functions with desired effects for disease intervention. Using computational, biochemical and biophysical tools, we delineated the role of all domains, their combinations and the critical phenylalanine residues in regulating HtrA3 activity, oligomerization and specificity. Our findings underline the crucial roles of the N-terminus as well as the PDZ domain in oligomerization and formation of a catalytically competent enzyme, thus providing new insights into its structure–function coordination. Our study also reports an intricate ligand-induced allosteric switch, which redefines the existing hypothesis of HtrA3 activation besides opening up avenues for modulating protease activity favorably through suitable effector molecules.

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During cellular electroporation, the formation of transient pores allow for the diffusion of innately impermeable molecules. The diversity of cell and membrane structure results in unique properties with respect to sensitivity to electric fields. The growing use of human cell lines in biomedical research and technology has led to a demand for protocols that can effectively and economically perform electroporation. We electroporated six human cell lines using a fluorescent reporter to investigate the effects of pulse electric field strength, pulse duration, and DNA concentration during electroporation. It was found that the cell lines all responded to electric field strengths within 400-950V/cm with viability decreasing with increasing voltage. It was also observed that the concentration of DNA used directly impacts transfection efficiency and cell viability as well. To better characterize square wave electroporation, we adopted a model where the pulse is described by its energy density (J/L) with respect to the sample buffer volume. It was determined that the key electrical characteristics of electroporation can be generalized with this value to provide a simplified measure of pulse intensity. The resulting analysis was consistent with other models, indicating cell type specific optimal electrical and DNA concentrations.

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