Predictive discriminative accuracy of walking abilities at discharge for community ambulation levels at 6 months post-discharge among inpatients with subacute stroke.

[Purpose] This study aimed to compare the predictive accuracy of walking ability at discharge among subacute stroke inpatients at 6 months post-discharge in terms of community ambulation level and establish optimal cut-off values. [Participants and Methods] This prospective observational study included 78 patients who completed follow-up assessments. Patients were classified into three groups based on the Modified Functional Walking Category (household/most limited community walkers, least limited community walkers, and unlimited community walkers) obtained by telephone survey at 6 months post-discharge. Predictive accuracy and cut-off values for discriminating among groups were calculated from 6-minute walking distance and comfortable walking speed measured at the time of discharge using receiver operating characteristic curves. [Results] Between household/most limited and least limited community walkers, 6-minute walking distance and comfortable walking speed offered similar predictive accuracy (area under the curve, 0.6-0.7), with cut-off values of 195 m and 0.56 m/s, respectively. Between least limited and unlimited community walkers, the areas under the curve were 0.896 for 6-minute walking distance and 0.844 for comfortable walking speed, with cut-off values of 299 m and 0.94 m/s, respectively. [Conclusion] Walking endurance and walking speed among inpatients with subacute stroke provided superior predictive accuracy for unlimited community walkers at 6 months post-discharge.

Inactivation of SARS-CoV-2 by Commercially Available Disinfectants and Cleaners.

The recent emergence of severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) is a major burden for health care systems worldwide, and is a threat to our daily lives. Various effective ingredients against SARS-CoV-2 were already reported, however, since products contain various ingredients, it is also important to evaluate the effectiveness of commercially available disinfectants per se. In this study, the virucidal efficacy of forty-eight commercially available products were evaluated according to the standardized suspension method EN 14476 and the following results were obtained: Alcohol-based disinfectants, hand soaps, wet wipes, alkaline cleaners, quaternary ammonium compound sanitizers and oxygen bleach had great virucidal efficacy against SARS-CoV-2. Enveloped viruses such as SARS-CoV-2 are among the most susceptible of pathogens to formulated microbicidal actives and detergents, but as the results of this study showed, it is also necessary to pay attention to the concentration at the time of use and the required contact time.

Efficient enzymatic production of benzaldehyde from l-phenylalanine with a mutant form of 4-hydroxymandelate synthase.

Benzaldehyde is an organic compound with an almond-like aroma and one of the most important and widely used flavorings in the food industry. To develop an enzymatic process for the production of benzaldehyde from l-phenylalanine, four enzymes were expressed in Escherichia coli; l-amino acid deaminase, 4-hydroxymandelate synthase, (S)-mandelate dehydrogenase, and benzoylformate decarboxylase. Although each E. coli strain could be used to synthesize benzaldehyde from l-phenylalanine, the yield was low due to the accumulation of an intermediate, phenylpyruvic acid. We developed a second reaction step by engineering 4-hydroxymandelate synthase of Actinoplanes teichomyceticus. A quadruple mutant of 4-hydroxymandelate synthase (A199V/Q206R/I217V/K337Q) obtained by random and site-directed mutagenesis demonstrated 2.4-fold higher activity than wild type. Furthermore, the mutant-expressing strain was able to produce benzaldehyde from 100 mm l-phenylalanine at a conversion rate of 84% (wild type, 37%). We report the development of an efficient process for benzaldehyde production using l-phenylalanine as a substrate.

Acyltransferase that catalyses the condensation of polyketide and peptide moieties of goadvionin hybrid lipopeptides.

Fusions of fatty acids and peptides expand the structural diversity of natural products; however, polyketide/ribosomally synthesized and post-translationally modified peptides (PK/RiPPs) hybrid lipopeptides are relatively rare. Here we report a family of PK/RiPPs called goadvionins, which inhibit the growth of Gram-positive bacteria, and an acyltransferase, GdvG, which catalyses the condensation of the PK and RiPP moieties. Goadvionin comprises a trimethylammonio 32-carbon acyl chain and an eight-residue RiPP with an avionin structure. The positions of six hydroxyl groups and one double bond in the very-long acyl chain were determined by radical-induced dissociation tandem mass spectrometry, which collides radical ion species to generate C-C bond cleavage fragments. GdvG belongs to the Gcn5-related N-acetyltransferase superfamily. Unlike conventional acyltransferases, GdvG transfers a very long acyl chain that is tethered to an acyl carrier protein to the N-terminal amino group of the RiPP moiety. gdvG homologues flanked by PK/fatty acid and RiPP biosynthesis genes are widely distributed in microbial species, suggesting that acyltransferase-catalysed condensation of PKs and RiPPs is a general strategy in biosynthesis of similar lipopeptides.

Insulin sensor cells for the analysis of insulin secretion responses in single living pancreatic ß cells.

Investigation of the functions of insulin-secreting cells in response to glucose in single-living cells is essential for improving our knowledge on the pathogenesis of diabetes. Therefore, it is desired to develop a new convenient method that enables the direct detection of insulin secreted from single-living cells. Here, insulin-sensor-cells expressing a protein-based insulin-detecting probe immobilized on the extracellular membrane were developed to evaluate the insulin-secretion response in single-living pancreatic ß cells. The protein-based insulin-detecting probe (NαLY) was composed of a bioluminescent protein (nano-luc), the αCT segment of the insulin receptor, L1 and CR domains of the insulin receptor, and a fluorescent protein (YPet). NαLY exhibited a bioluminescence resonance energy transfer (BRET) signal in response to insulin; thus, cells of Hepa1-6 line were genetically engineered to express NαLY on the extracellular membrane. The cells were found to act as insulin-sensor-cells, exhibiting a BRET signal in response to insulin. When the insulin-sensor-cells and pancreatic ß cells (MIN6 cell line) were cocultured and stimulated with glucose, insulin-sensor-cells nearby pancreatic ß cells showed the spike-shaped BRET signal response, whereas the insulin-sensor-cells close to one pancreatic ß cell did not exhibit such signal response. However, all the insulin-sensor-cells showed a gradual increase in BRET signals, which were presumably attributed to the increase in insulin concentrations in the culture dish, confirming the function of these insulin-sensor-cells. Therefore, we demonstrated that heterogenetic insulin secretion in single-living pancreatic ß cells could be measured directly using the insulin sensor cells.