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The Cas9 nuclease from Staphylococcus aureus (SaCas9) holds great potential for use in gene therapy, and variants with increased fidelity have been engineered. However, we find that existing variants have not reached the greatest accuracy to discriminate base mismatches and exhibited much reduced activity when their mutations were grafted onto the KKH mutant of SaCas9 for editing an expanded set of DNA targets. We performed structure-guided combinatorial mutagenesis to re-engineer KKH-SaCas9 with enhanced accuracy. We uncover that introducing a Y239H mutation on KKH-SaCas9's REC domain substantially reduces off-target edits while retaining high on-target activity when added to a set of mutations on REC and RuvC domains that lessen its interactions with the target DNA strand. The Y239H mutation is modelled to have removed an interaction from the REC domain with the guide RNA backbone in the guide RNA-DNA heteroduplex structure. We further confirmed the greatly improved genome-wide editing accuracy and single-base mismatch discrimination of our engineered variants, named KKH-SaCas9-SAV1 and SAV2, in human cells. In addition to generating broadly useful KKH-SaCas9 variants with unprecedented accuracy, our findings demonstrate the feasibility for multi-domain combinatorial mutagenesis on SaCas9's DNA- and guide RNA- interacting residues to optimize its editing fidelity.Accelerated-phase myelofibrosis, currently defined by circulating blasts 10% to 19%, usually confers very high risk for progression and poor outcome. The outcome of hematopoietic stem cell transplantation for accelerated-phase myelofibrosis has not been evaluated yet. We analyzed the outcome of 349 clinically and genetically annotated patients with primary or secondary myelofibrosis undergoing reduced intensity transplantation, of whom 35 had accelerated-phase myelofibrosis. In comparison with chronic-phase ( less then 10% blasts) myelofibrosis, median leukocyte counts were higher, more patients had constitutional symptoms, and RAS mutations were detected more frequently in the accelerated-phase group. After a median follow-up of 5.9 years, estimated 5-year overall survival was 65% (95% confidence interval [CI], 49% to 81%) vs 64% (95% CI, 59% to 69%) for the chronic-phase group (P = .91), and median overall survival was not reached. In terms of relapse-free survival, estimated 5-year outcome for the accelerated-phase group was 49% (95% CI, 32% to 67%) vs 55% (95% CI, 50% to 61%) for the chronic-phase group (P = .65). Estimated 5-year nonrelapse mortality was 20% (95% CI, 8% to 33%) for the accelerated-phase group vs 30% (95% CI, 24% to 35%; P = .25) for the chronic-phase group. In terms of relapse, 5-year incidence was 30% (95% CI, 14% to 46%) for the accelerated-phase group vs 15% (95% CI, 11% to 19%) for the chronic-phase group (P = .02). Results were confirmed in multivariable analysis and propensity score matching. read more In conclusion, reduced intensity transplantation showed excellent survival but higher relapse for accelerated-phase myelofibrosis.

The COLCHIC database contains workplace exposure results of chemical samples collected by the French prevention network since 1987. We aimed to investigate potential associations between exposure levels and ancillary variables in COLCHIC across a broad range of chemical agents in order to provide insight into how to best interpret and exploit the information in this national database.

We selected personal and area measurements in COLCHIC and collected outside respiratory personal protective equipment (PPE), restricted to chemical agents that had at least 1000 samples available. We used Tobit models to estimate associations between exposure concentrations and sample year, sampling duration, PPE, workforce size, collective protective equipment, origin of request, and reason for request for each chemical agent for the period 1987-2019. Models for period 2002-2019 also included type of process (open/closed) and exposure frequency. We used separate models for each combination of agent, period, and personal or -agent heterogeneity of estimated effects, based on 80% prediction intervals, was lowest for sampling duration, time trends, and the presence of PPE.

Sampling duration, time trends, and the presence of PPE are important factors to take into account when analyzing COLCHIC and had similar associations with exposure levels across agents. Other variables generally showed weaker associations or variable effects. These results will be used to adjust exposure estimates for the French working population from measurements stored in COLCHIC.

Sampling duration, time trends, and the presence of PPE are important factors to take into account when analyzing COLCHIC and had similar associations with exposure levels across agents. Other variables generally showed weaker associations or variable effects. These results will be used to adjust exposure estimates for the French working population from measurements stored in COLCHIC.Lead (Pb) exposure continues to be a significant public health issue in both occupational and non-occupational settings. The vast majority of exposure and toxicological studies have focused on effects related to inhalation and gastrointestinal exposure routes. Exposure to inorganic Pb compounds through dermal absorption has been less well studied, perhaps due to the assumption that the dermal pathway is a minor contributor to aggregate exposures to Pb compounds. The aim of this rapid review was to identify and evaluate published literature on dermal exposures to support the estimation of key percutaneous absorption parameters (Kp, flux, diffusion rate) for use in occupational risk assessment. Eleven articles were identified containing information from both in vitro and in vivo systems relevant to percutaneous absorption kinetics. These articles provided 24 individual study summaries and information for seven inorganic Pb compounds. The vast majority of study summaries evaluated (n = 22, 92%) reported detectabnot high. The literature, while of limited quality, overall strongly suggests inorganic Pb has the potential for dermal uptake in meaningful amounts associated with negative health outcomes based on upper bound diffusion rate estimates. Future standard test guideline-compliant studies are needed to provide high-confidence estimates of dermal uptake. Such data are needed to allow for improved evaluation of Pb exposures in an occupational risk assessment context.

Agriculture is one of the most hazardous occupations in the USA. Especially, tractor rollover incidents are the leading cause of farming-related injuries or deaths. This study examines the effect of a VR intervention (Virtual Reality Intervention for Safety Education; VRISE) on behavioral intentions for occupational safety and identifies a psychological mechanism that shows how the immersive technology works.

VRISE was developed by a multidisciplinary team of agricultural educators, computer scientists and communication specialists. It was designed to provide a virtual environment where users practice tractor operation and try to avoid several rollover hazards. The participants (291 high school students) were recruited at the 2019 National Future Farmers Association Convention & Expo and randomly assigned to one of three conditions two different types of control groups (Control1 No treatment group and Control2 2D Screen group) and the treatment group.

Findings show that, through the immersive VR experience, the VR intervention enhanced perceived threat of tractor-related accidents which in turn, led to improved behavioral intentions for tractor safety.

Findings shed light on the effectiveness of a VR intervention to improve public health outcomes, especially in occupational safety education, where unsafe practices often result in injury and fatality.

Findings shed light on the effectiveness of a VR intervention to improve public health outcomes, especially in occupational safety education, where unsafe practices often result in injury and fatality.Asbestos fibre counting by phase-contrast microscope is subject to many sources of variation, including those dependent on the analyst. In this study, asbestos sample slides prepared with relocatable coverslips have been used for fibre counting among voluntary analysts to evaluate their proficiency. One slide of amosite and one of chrysotile were distributed to all the analysts, and three proficiency testing rounds were conducted for amosite and four for chrysotile. Each relocatable coverslip has a report in which are reported for each viewing field both the number of certified fibres (Verified Fibres) and a drawing representing the shape and position of the individual fibres. In the first round, the analysts were asked to report only the number of fibres counted in each of the predesignated fields of view. In the other rounds, subsequently developed, the analysts had to report the number and the position of the fibres for each field. The reported number of fibres and their position in each of the designed fito consider an analyst as proficient. The results of chrysotile fibres showed that in the first round all analysts counted less than fifty per cent of the verified fibres. In the second round 10 analysts out of 13 reached a score of ≥50, 8 of 16 in the third and 10 of 12 in the fourth. For chrysotile fibres, the error relating to the counting of fibres shorter than 5 µm was of 56% of extra fibres, but the error that most influenced the results was the number of oversight-missing fibres. This type of error accounted for 97% of the missing fibres and for the 29% under-estimation of the chrysotile fibres. For amosite fibre counting, results of this study show an improvement of the analyst's performance. For the chrysotile fibre count, although there is a significant improvement in the comparison between some rounds, this is not continuous over time.α-Diketones such as diacetyl (2,3-butanedione) and 2,3-pentanedione are generated during the roasting and fermentation of foods and are also used as flavoring compounds. Exposure to these compounds has been associated with obliterative bronchiolitis in workers. We report indoor air concentrations of diacetyl and 2,3-pentanedione, as well as acetoin (3-hydroxy-2-butanone), in several small coffee roasteries and breweries using standard integrated air sampling sorbent tubes followed by gas chromatography tandem mass spectrometry as well as the first use of on-site continuous real-time proton-transfer reaction time-of-flight mass spectrometry (PTR-ToF-MS). Diacetyl and 2,3-pentanedione were detected in most of the sorbent samples at concentrations between 0.02 and 8 ppbv, and in general were higher in coffee roasteries compared with breweries. Three integrated air samples, all from the barista area at one facility, exceeded the NIOSH recommended exposure limit (REL) of 5 ppbv for diacetyl. 2,3-Pentanedione concepbv higher and the post-shift FENO was 7.1 (-1.9, 16.1) ppbv higher for workers at coffee roasteries compared with breweries. The cross-shift change in FENO was 3.4 (-2.8, 9.6) ppbv higher for workers at coffee roasteries compared with breweries. However, none of these differences were statistically significant, and the cross-shift change in FENO was not statistically different from zero for either group of workers. The findings from this pilot study demonstrate that α-diketones and related compounds are present in the indoor air of both breweries and coffee roasteries and may exceed health protective guidelines in coffee roasteries. Additional studies are required to fully characterize worker exposures in these settings and to identify specific work activities and processes associated with high exposures. Engineering controls, including targeted exhaust ventilation and the use of low-cost sensors, are recommended as an approach to protect workers from exposure to hazardous levels of α-diketones.