A Personal Respirator to Improve Protection for Healthcare Workers Treating COVID-19 (PeRSo)

Introduction: SARS-CoV-2 infection is a global pandemic. Personal Protective Equipment (PPE) to protect healthcare workers has been a recurrent challenge in terms of global stocks, supply logistics and suitability. In some settings, around 20% of healthcare workers treating COVID-19 cases have become infected, which leads to staff absence at peaks of the pandemic, and in some cases mortality.Methods: To address shortcomings in PPE, we developed a simple powered air purifying respirator, made from inexpensive and widely available components. The prototype was designed to minimize manufacturing complexity so that derivative versions could be developed in low resource settings with minor modification.Results: The “Personal Respirator – Southampton” (PeRSo) delivers High-Efficiency Particulate Air (HEPA) filtered air from a battery powered fan-filter assembly into a lightweight hood with a clear visor that can be comfortably worn for several hours. Validation testing demonstrates that the prototype removes microbes, avoids excessive CO2 build-up in normal use, and passes fit test protocols widely used to evaluate standard N95/FFP2 and N99/FFP3 face masks. Feedback from doctors and nurses indicate the PeRSo prototype was preferred to standard FFP2 and FFP3 masks, being more comfortable and reducing the time and risk of recurrently changing PPE. Patients report better communication and reassurance as the entire face is visible.Conclusion: Rapid upscale of production of cheaply produced powered air purifying respirators, designed to achieve regulatory approval in the country of production, could protect healthcare workers from infection and improve healthcare delivery during the COVID-19 pandemic

An arabidopsis cDNA encodes an apparent polyprotein of two non-identical thylakoid membrane proteins that are associated with photosystem II and homologous to algal ycf32 open reading frames

We have characterised an Arabidopsis thaliana cDNA homologous to the ycf32 open reading frames present in the Synechocystis genome and the plastid genomes of several eukaryotic algae. The predicted protein is also homologous to a novel protein reported to be associated with photosystem II. The protein is synthesised as a 23 kDa precursor with an N-terminal presequence that appears to be bipartite in structure, and the protein is targeted into the thylakoid membrane of pea chloroplasts. Although the Ycf32 presequence contains an apparent signal peptide, we find that this protein is not imported by either of the standard Sec- or ΔpH-dependent pathways. The mature protein is also unusual in two respects. First, there are two distinct, non-identical copies of typical single-span Ycf32 sequences in the Arabidopsis sequence, separated by an additional hydrophobic region. Secondly, the imported protein runs as a doublet of 6 kDa and 7 kDa polypeptides whereas the mature protein is predicted to be 14 kDa. We speculate that the protein undergoes further maturation once inserted into the thylakoid membrane to yield two separate Ycf32-like polypeptides.</p

Targeting of proteins into and across the thylakoid membrane

The assembly of the photosynthetic apparatus utilizes component proteins that are synthesized by two genomes and then targeted into and across the thylakoid membrane. The emerging picture is one of a remarkably complex system of protein trafficking, in which at least four distinct pathways operate within the chloroplast - two for lumenal proteins and two for integral membrane proteins. Some of the pathways can be traced back to the prokaryotic ancestor of the chloroplast, whereas others appear to have arisen more recently - one in response to the transfer of genes to the plant nucleus and another, possibly, in response to the acquisition of new photosynthetic proteins. Remarkably, proteins in three of these pathways are synthesized with clearable signal-type peptides that are almost identical in overall structure, yet that execute entirely different functions. Recent studies have begun to reconcile the function of these targeting signals with the nature of the protein being targeted.</p

Determination of the unfrozen water content of maximally freeze-concentrated carbohydrate solutions

The heat capacity change at T′g has been studied in freeze-concentrated carbohydrate solutions. The values obtained have been compared with those found for high concentration solutions that do not undergo freezing above Tg. The analysis has indicated that the freezing process influences the degree of stress in the glassy phase. This results in a complex power-time curve when frozen solutions are heated in a differential scanning calorimeter. The endotherm produced by the stress relaxation can cause considerable error in W′g measurement obtained by any method that relies on the integration of the power-time curve. A more reliable method for W′g determination is via the intersection of T′g with a previously prepared Tg/Wg calibration curve.</p

Acid sensitive background potassium channels K2p3.1 and K2p9.1 undergo rapid dynamin-dependent endocytosis

Acid-sensitive, two-pore domain potassium channels, K2p3.1 and K2p9.1, are implicated in cardiac and nervous tissue responses to hormones, neurotransmitters and drugs. K2p3.1 and K2p9.1 leak potassium from the cell at rest and directly impact membrane potential. Hence altering channel number on the cell surface drives changes in cellular electrical properties. The rate of K2p3.1 and K2p9.1 delivery to and recovery from the plasma membrane determines both channel number at the cell surface and potassium leak from cells. This study examines the endocytosis of K2p3.1 and K2p9.1. Plasma membrane biotinylation was used to follow the fate of internalized GFP-tagged rat K2p3.1 and K2p9.1 transiently expressed in HeLa cells. Confocal fluorescence images were analyzed using Imaris software, which revealed that both channels are endocytosed by a dynamin-dependent mechanism and over the course of 60 min, move progressively toward the nucleus. Endogenous endocytosis of human K2p3.1 and K2p9.1 was examined in the lung carcinoma cell line, A549. Endogenous channels are endocytosed over a similar time-scale to the channels expressed transiently in HeLa cells. These findings both validate the use of recombinant systems and identify an endogenous model system in which K2p3.1 and K2p9.1 trafficking can be further studied