Discovery of active mouse, plant and fungal cytochrome P450s in endogenous proteomes and upon expression in planta.

Eukaryotes produce a large number of cytochrome P450s that mediate the synthesis and degradation of diverse endogenous and exogenous metabolites. Yet, most of these P450s are uncharacterized and global tools to study these challenging, membrane-resident enzymes remain to be exploited. Here, we applied activity profiling of plant, mouse and fungal P450s with chemical probes that become reactive when oxidized by P450 enzymes. Identification by mass spectrometry revealed labeling of a wide range of active P450s, including six plant P450s, 40 mouse P450s and 13 P450s of the fungal wheat pathogen Zymoseptoria tritici. We next used transient expression of GFP-tagged P450s by agroinfiltration to show ER-targeting and NADPH-dependent, activity-based labeling of plant, mouse and fungal P450s. Both global profiling and transient expression can be used to detect a broad range of active P450s to study e.g. their regulation and discover selective inhibitors.

Cancer-cell derived S100A11 promotes macrophage recruitment in ER+ breast cancer.

Macrophages are pivotal in driving breast tumor development, progression, and resistance to treatment, particularly in estrogen receptor-positive (ER+) tumors, where they infiltrate the tumor microenvironment (TME) influenced by cancer cell-secreted factors. By analyzing single-cell RNA-sequencing data from 25 ER+ tumors, we elucidated interactions between cancer cells and macrophages, correlating macrophage density with epithelial cancer cell density. We identified that S100A11, a previously unexplored factor in macrophage-cancer crosstalk, predicts high macrophage density and poor outcomes in ER+ tumors. We found that recombinant S100A11 enhances macrophage infiltration and migration in a dose-dependent manner. Additionally, in 3D models, we showed that S100A11 expression levels in ER+ cancer cells predict macrophage infiltration patterns. Neutralizing S100A11 decreased macrophage recruitment, both in cancer cell lines and in a clinically relevant patient-derived organoid model, underscoring its role as a paracrine regulator of cancer-macrophage interactions in the protumorigenic TME. This study offers novel insights into the interplay between macrophages and cancer cells in ER+ breast tumors, highlighting S100A11 as a potential therapeutic target to modulate the macrophage-rich tumor microenvironment.

The Effectiveness and Perceptions of Three Moderate Intensity Walking Cadence Aids and their Effects on Affective States: A Mixed Methods Study.

Substantial health benefits can be derived from walking at a moderate intensity cadence. To help regulate this cadence, three distinct aids exist 1) self-perception; 2) cadence prescription; 3) auditory cues. This study aimed to compare the effectiveness and explore perceptions of these aids to promote moderate intensity walking and effects on affective states, thereby addressing an important research gap. Individualised moderate relative intensity waking cadence was determined for participants (n = 23, Mage = 26.35, SD = 10.11). A convergent mixed-methods design was employed. A within-persons repeated measures design was used to explore the effectiveness of three aids (general guidelines, cadence prescription, and music) on promoting moderate intensity physical activity and positive affective states. Perceptions of these aids were elicited through qualitative interviews and thematic content analysis. Main effects for condition on relative physical activity intensity (η2 = .72) and positive affect (η2 = .25) were observed. Music evoked significantly higher relative physical activity intensity than other conditions (p values < .01), and higher positive affect compared to the general guidelines condition (p = .038). A significantly greater proportion of participants achieved moderate relative intensity physical activity during the music compared to general guidelines condition (p = .03). Congruently, qualitative findings suggested that participants predominantly perceived music as most effective for promoting a moderate intensity cadence and positive affect. However, individual variability existed in ability to utilise this aid. Implications of the findings for practitioners seeking to promote a moderate intensity cadence and positive affect during walking are discussed.

Impact of Rank, Provider Specialty, and Unit Sustainment Training Frequency on Military Critical Care Air Transport Team Readiness.

BACKGROUND: The Critical Care Air Transport (CCAT) Advanced Course utilizes fully immersive high-fidelity simulations to assess personnel readiness for deployment. This study aims to determine whether simple well-defined demographic identifiers can be used to predict CCAT students' performance at CCAT Advanced. MATERIALS AND METHODS: CCAT Advanced student survey data and course status (pass/fail) between March 2006 and April 2020 were analyzed. The data included students' Air Force Specialty Code (AFSC), military status (active duty and reserve/guard), CCAT deployment experience (yes/no), prior CCAT Advanced training (yes/no), medical specialty, rank, and unit sustainment training frequency (never, frequency less often than monthly, and frequency at least monthly). Following descriptive analysis and comparative tests, multivariable regression was used to identify the predictors of passing the CCAT Advanced course for each provider type. RESULTS: A total of 2,576 student surveys were analyzed: 694 (27%) physicians (MDs), 1,051 (40%) registered nurses (RNs), and 842 (33%) respiratory therapists (RTs). The overall passing rates were 92.2%, 90.3%, and 85.4% for the MDs, RNs, and RTs, respectively. The students were composed of 579 (22.5%) reserve/guard personnel, 636 (24.7%) with CCAT deployment experience, and 616 (23.9%) with prior CCAT Advanced training. Regression analysis identified groups with lower odds of passing; these included (1) RNs who promoted from Captain to Major (post-hoc analysis, P = .03), (2) RTs with rank Senior Airman, as compared to Master Sergeants (post-hoc analysis, P = .04), and (3) MDs with a nontraditional AFSC (P = .0004). Predictors of passing included MDs and RNs with CCAT deployment experience, odds ratio 2.97 (P = .02) and 2.65 (P = .002), respectively; and RTs who engaged in unit CCAT sustainment at least monthly (P = .02). The identifiers prior CCAT Advanced training or reserve/guard military status did not confer a passing advantage. CONCLUSION: Our main result is that simple readily available metrics available to unit commanders can identify those members at risk for poor performance at CCAT Advanced readiness training; these include RNs with rank Major or above, RTs with rank Senior Airman, and RTs who engage in unit sustainment training less often than monthly. Finally, MD specialties which are nontraditional for CCAT have significantly lower CCAT Advanced passing rates, reserve/guard students did not outperform active duty students, there was no difference in the performance between different RN specialties, and for MD and RN students' previous deployment experience was a strong predictor of passing.

The influence of habitual physical activity and sedentary behavior on objective and subjective hot flashes at midlife.

OBJECTIVE: The purpose of this study was to evaluate relationships between physical activity, sedentary time, and hot flashes during both waking and sleeping periods using concurrent objective and subjective measures of hot flashes in midlife women. METHODS: Women aged 45 to 55 years (n = 196) provided self-reported data on physical activity and underwent 24 hours of hot flash monitoring using sternal skin conductance. Participants used event marking and logs to indicate when hot flashes were perceived. Wake and sleep periods were defined by actigraphy. Mean ambient temperature and humidity were recorded during the study period. Generalized linear regression modeling was used to evaluate the effect of physical activity types and sedentary time on hot flash outcomes. Isotemporal substitution modeling was used to study the effect of replacing sedentary time with activity variables on hot flash frequency. RESULTS: Modeled results indicated that increasing sitting by 1 hour was associated with a 7% increase in the rate of objectively measured but not subjectively reported hot flashes during sleep. Replacing 1 hour of sitting with 1 hour of vigorous activity was associated with a 100% increase in subjectively reported but not objectively measured waking hot flashes. There was little evidence for an effect of temperature or humidity on any hot flash outcome. CONCLUSIONS: These data provide support for relations between sedentary time, physical activity, and hot flashes and highlight the importance of using objective and subjective assessments to better understand the 24-hour hot flash experience.

Simulation of Image-Guided Microwave Ablation Therapy Using a Digital Twin Computational Model.

Emerging computational tools such as healthcare digital twin modeling are enabling the creation of patient-specific surgical planning, including microwave ablation to treat primary and secondary liver cancers. Healthcare digital twins (DTs) are anatomically one-to-one biophysical models constructed from structural, functional, and biomarker-based imaging data to simulate patient-specific therapies and guide clinical decision-making. In microwave ablation (MWA), tissue-specific factors including tissue perfusion, hepatic steatosis, and fibrosis affect therapeutic extent, but current thermal dosing guidelines do not account for these parameters. This study establishes an MR imaging framework to construct three-dimensional biophysical digital twins to predict ablation delivery in livers with 5 levels of fat content in the presence of a tumor. Four microwave antenna placement strategies were considered, and simulated microwave ablations were then performed using 915 MHz and 2450 MHz antennae in Tumor Naïve DTs (control), and Tumor Informed DTs at five grades of steatosis. Across the range of fatty liver steatosis grades, fat content was found to significantly increase ablation volumes by approximately 29-l42% in the Tumor Naïve and 55-60% in the Tumor Informed DTs in 915 MHz and 2450 MHz antenna simulations. The presence of tumor did not significantly affect ablation volumes within the same steatosis grade in 915 MHz simulations, but did significantly increase ablation volumes within mild-, moderate-, and high-fat steatosis grades in 2450 MHz simulations. An analysis of signed distance to agreement for placement strategies suggests that accounting for patient-specific tumor tissue properties significantly impacts ablation forecasting for the preoperative evaluation of ablation zone coverage.

Combining range-general and range-specific techniques for late-time focused image reconstructiona).

Synthetic aperture sonar (SAS) is an acoustic method for detecting objects in an environment. Conventional SAS image reconstruction techniques invert a forward model based on geometric scattering and straight-line propagation. Acoustic features that do not fit this model, such as multiple scattering and late-time returns, appear out of focus. This paper describes an image reconstruction technique that selectively applies range-general and range-specific methods to improve the focus of late-time returns while maintaining image quality away from the focal plane. The technique is demonstrated on experimental data and compared with a range-specific algorithm.

Can the Cans: Determinants of Container Deposit Behavior before and after Introduction of a Container Refund Scheme.

OBJECTIVE: Container deposit schemes are often hailed as a useful avenue to increase consumer recycling rates. Yet, there is little research investigating within-person changes in people's beliefs and behavior following the implementation of these schemes, or tests of the mechanisms by which such change has occurred. METHODS: The current study fills this knowledge gap and assessed container recycling behavior and habits as well as the social cognition factors of attitudes, subjective norms, perceived behavioral control, and intentions in a sample of 90 Queenslanders before the implementation of the container deposit scheme and one and three months post-implementation. RESULTS: Analysis of variance indicated more frequent recycling behavior following the implementation of the scheme, as well as stronger habits, intentions, and perceived behavioral control. CONCLUSIONS: Such a concomitant change in behavior, beliefs, and habits provides support for behavior change theory, while also flagging potential targets for strategies that can be paired with container deposit schemes to enhance their efficacy and uptake.

Predictive Dosimetry and Outcomes of Hepatocellular Carcinoma Treated by Yttrium-90 Resin Microsphere Radioembolization: A Retrospective Analysis Using Technetium-99m Macroaggregated Albumin SPECT/CT and Planning Software.

PURPOSE: To characterize estimated mean absorbed tumor dose (ADT), objective response (OR), and estimated target dose of hepatocellular carcinoma (HCC) after resin microsphere yttrium-90 (90Y) radioembolization using partition dosimetry. MATERIALS AND METHODS: In this retrospective, single-center study, multicompartment dosimetry of index tumors receiving 90Y radioembolization between October 2015 and June 2022 was performed using a commercial software package and pretreatment technetium-99m macroaggregated albumin single photon emission computed tomography (SPECT)/computed tomography (CT). In total, 101 patients with HCC underwent 102 treatments of 127 index tumors. Patients underwent imaging every 2-3 months after treatment to determine best response per modified Response Evaluation Criteria in Solid Tumors (mRECIST). Best response was defined as the greatest response category per mRECIST and categorized as OR or nonresponse (NR). A Cox proportional hazards model evaluated the probability of tumor OR and progression-free survival using ADT. RESULTS: The median follow-up period was 148 days (interquartile range [IQR], 92-273 days). The median ADT of OR was 141.9 Gy (IQR, 89.4-215.8 Gy) compared with the median ADT of NR treatments of 70.8 Gy (IQR, 42.0-135.3 Gy; P < .001). Only ADT was predictive of response (hazard ratio = 2.79 [95% confidence interval {CI}: 1.44-5.40]; P = .003). At 6 months, an ADT of 157 Gy predicted 90.0% (95% CI: 41.3%-98.3%) probability of OR. At 1 year, an ADT of 157 Gy predicted 91.6% (95% CI: 78.3%-100%) probability of progression-free survival. Partition modeling and delivered activity were predictive of progression (P = .021 and P = .003, respectively). CONCLUSIONS: For HCC treated with resin microspheres, tumors receiving higher ADT exhibited higher rates of OR. An ADT of 157 Gy predicted 90.0% OR at 6 months.

Validation of reaction norm breeding values for robustness in Australian sheep.

BACKGROUND: There can be variation between animals in how stable their genetic merit is across different environments due to genotype-by-environment (G×E) interactions. This variation could be used in breeding programs to select robust genotypes that combine high overall performance with stable genetic ranking across environments. There have been few attempts to validate breeding values for robustness in livestock, although this is a necessary step towards their implementation in selection decisions. The objective of this study was to validate breeding values for the robustness of body weight across different growth environments that were estimated using reaction norm models in sheep data. RESULTS: Using threefold cross-validation for the progeny of 337 sires, the average correlation between single-step breeding values for the reaction norm slope and the realised robustness of progeny across different growth environments was 0.21. The correlation between breeding values for the reaction slope estimated independently in two different datasets linked by common sires was close to the expected correlation based on theory. CONCLUSIONS: Slope estimated breeding values (EBV) obtained using reaction norm models were predictive of the phenotypic robustness of progeny across different environments and were consistent for sires with progeny in two different datasets. Selection based on reaction norm EBV could be used to increase the robustness of a population to environmental variation.

A risk-reward examination of sample multiplexing reagents for single cell RNA-Seq.

Single-cell RNA sequencing (scRNA-Seq) has emerged as a powerful tool for understanding cellular heterogeneity and function. However the choice of sample multiplexing reagents can impact data quality and experimental outcomes. In this study, we compared various multiplexing reagents, including MULTI-Seq, Hashtag antibody, and CellPlex, across diverse sample types such as human peripheral blood mononuclear cells (PBMCs), mouse embryonic brain and patient-derived xenografts (PDXs). We found that all multiplexing reagents worked well in cell types robust to ex vivo manipulation but suffered from signal-to-noise issues in more delicate sample types. We compared multiple demultiplexing algorithms which differed in performance depending on data quality. We find that minor improvements to laboratory workflows such as titration and rapid processing are critical to optimal performance. We also compared the performance of fixed scRNA-Seq kits and highlight the advantages of the Parse Biosciences kit for fragile samples. Highly multiplexed scRNA-Seq experiments require more sequencing resources, therefore we evaluated CRISPR-based destruction of non-informative genes to enhance sequencing value. Our comprehensive analysis provides insights into the selection of appropriate sample multiplexing reagents and protocols for scRNA-Seq experiments, facilitating more accurate and cost-effective studies.

Perceived dehydration impairs endurance cycling performance in the heat in active males.

Dehydration of >3 % body mass impairs endurance performance irrespective of the individual's knowledge of their hydration status, but whether knowledge of hydration status influences performance at lower levels of dehydration is unknown. This study examined whether perception of hydration status influenced endurance performance. After familiarisation, nine active males (age 25 ± 2 y, V̇O2peak 52.5 ± 9.1 mL kg min-1) completed two randomised trials at 34 °C. Trials involved an intermittent exercise preload (8 × 10 min cycling/5 min rest), followed by a 15 min all-out cycling performance test. During the preload in both trials, water was ingested orally every 10 min (0.3 mL kg body mass-1), with additional water infused into the stomach via gastric feeding tube to produce dehydration of ∼1.5 % body mass pre-performance test. Participants were told intra-gastric infusion was manipulated to produce euhydration (0 % dehydration; Perceived-EUH) or dehydration (2 % dehydration; Perceived-DEH) pre-performance test, which was told to them pre-preload and confirmed after body mass measurement pre-performance test. Body mass loss during the preload (Perceived-EUH 1.6 ± 0.2 %, Perceived-DEH 1.7 ± 0.2 %; P = 0.459), heart rate, gastrointestinal temperature and RPE (P ≥ 0.110) were not different between trials. Thirst was greater at the end of the preload and performance test in Perceived-DEH (P ≤ 0.040). Work completed during the performance test was 5.6 ± 6.1 % lower in Perceived-DEH (187.4 ± 37.0 kJ vs. 176.9 ± 36.0 kJ; P = 0.038). These results suggest that at lower levels of dehydration (<2 % body mass), an individual's perception of their hydration status could impair their performance, as well as their thirst perception.

Combined purebred and crossbred genetic evaluation of Columbia, Suffolk, and crossbred lamb birth and weaning weights: systematic effects and heterogeneous variances.

Despite the benefits of crossbreeding on animal performance, genetic evaluation of sheep in the U.S. does not directly incorporate records from crossbred lambs. Crossbred animals may be raised in different environments as compared to purebreds. Systemic factors such as age of dam and birth and rearing type may, therefore, affect purebred and crossbred performance differently. Furthermore, crossbred performance may benefit from heterozygosity, and genetic and environmental variances may be heterogeneous in different breeds and their crosses. Such issues must be accounted for in a combined (purebred and crossbred) genetic evaluation. The objectives of this study were to i) determine the effect of dam age and birth type on birth weight, and dam age and birth-rearing type on weaning weight, in purebred and crossbred lambs, ii) test for heterogeneous genetic and environmental variances in those weights, and iii) assess the impact of including weights on crossbred progeny on sire estimated breeding values (EBV). Performance records were available on purebred Columbia and Suffolk lambs. Crossbred information was available on lambs sired by Suffolk, Columbia or Texel rams mated to Columbia, Suffolk, or crossbred ewes. A multiple-trait animal model was fitted in which weights from Columbia, Suffolk, or crossbred lambs were considered different traits. At birth, there were 4,160, 2,356, and 5,273 Columbia, Suffolk, and crossbred records, respectively, with means (SD) of 5.14 (1.04), 5.32 (1.14), and 5.43 (1.23) kg, respectively. At weaning, on average at 122 (12) d, there were 2,557, 980, and 3,876 Columbia, Suffolk, and crossbred records, respectively, with corresponding means of 39.8 (7.2), 40.3 (7.9), and 39.6 (8.0) kg. Dam age had a large positive effect on birth and weaning weight in pure and crossbred lambs. At birth, however, the predicted effect was larger in crossbred and Suffolk lambs. While an increase in a number of lambs born and reared had a strong and negative influence on birth and weaning weight, the size of the effect did not differ across-breed types. Environmental variances were similar at birth and weaning, but additive variances differed among breed types for both weights. Combining purebred and crossbred information in the evaluation not only improved predictions of genetic merit in purebred sires but also allowed for direct comparisons of sires of different breeds. Breeders thus can benefit from an additional tool for making selection decisions.

Combining multiple breeds in a genetic evaluation allows for their direct comparison. However, differences in management and other systematic effects among breeds may affect the evaluation. Estimates of genetic merit of sires may also be biased by heterosis in crossbred progeny. We examined genetic and environmental factors that affect the efficacy of a multi-breed genetic evaluation. Birth and weaning weights of Columbia, Suffolk, and their cross, were available. Depending on the breed type, the systematic effects of dam age and either birth or birth-rearing type on weights differed. Separately for birth and weaning, weights were defined as a different trait for each breed type. A multi-breed, multi-trait model was fitted that accounted for systematic effects unique to a breed type, and heterosis. Estimated direct and maternal heritabilities were moderate. Genetic correlations between breeds were moderate to high. Estimates of genetic merit of Columbia and Suffolk sires were unaffected by bias due to heterosis and environmental effects when crossbred lambs were included in a purebred or a combined Columbia, Suffolk, and crossbred evaluation. For direct across-breed comparisons, breed type-specific adjustments for systematic effects are necessary when combining weight data on pure and crossbred lambs in a joint genetic evaluation.

Modes of failure following locking plate fixation of the distal radius: a thematic analysis of 517 fractures.

Anterior locking plate fixation of the distal radius is a common procedure with reliable results. Failure of fixation is sometimes seen. The aim of the present study was to identify the reasons for failure. In total, 517 cases met the study inclusion criteria. Of them, 23 cases had failure of fixation (4.4%). Failure analysis generated qualitative data. Subsequent thematic analysis identified the primary mode of failure and contributing factors. Primary modes were identified as failure to support all key fracture fragments (n = 20), wrong choice of implant (n = 1), failure of union (n = 1) and poor bone quality (n = 1). Contributing factors were errors in plate positioning, fracture reduction, implant selection and screw configuration, as well as fracture pattern complexity and poor bone quality. Most failed fixations had a primary mode and two or three contributing factors. Overall anterior plating is reliable with a low rate of surgical failure. Knowledge of failure modes will aid operative planning and prevent failure.Level of evidence: V.

Impingement-related osteolysis in Motec total wrist arthroplasty: an explant analysis and review of the literature.

We present five cases of osteolysis in the Motec total wrist prosthesis, three around the radial implant, one around the metacarpal implant and one around both. Three of these were progressive and required revision, and biomechanical explant analyses of these revised prostheses were performed. Ex vivo testing of the contact points of the Motec implants was also performed at maximum extension. Here, impingement occurs between the metacarpal screw and the dorsal rim of the cup (non-articulating surfaces) with the short-necked prosthesis, leading to metacarpal screw damage, titanium debris formation and osteolysis. An analysis of three previously published cases suggests that this may have been the likely mode of failure in those cases. This complication is preventable by avoiding use of the short-neck prosthesis.Level of evidence: IV.

The Minnesota Medical Operations Coordination Center: A COVID-19 Statewide Response to Ensure Access to Critical Care and Medical-Surgical Beds.

BACKGROUND: COVID-19 led to unprecedented inpatient capacity challenges, particularly in ICUs, which spurred development of statewide or regional placement centers for coordinating transfer (load-balancing) of adult patients needing intensive care to hospitals with remaining capacity. RESEARCH QUESTION: Do Medical Operations Coordination Centers (MOCC) augment patient placement during times of severe capacity challenges? STUDY DESIGN AND METHODS: The Minnesota MOCC was established with a focus on transfer of adult ICU and medical-surgical patients; trauma, cardiac, stroke, burn, and extracorporeal membrane oxygenation cases were excluded. The center operated within one health care system's bed management center, using a dedicated 24/7 telephone number. Major health care systems statewide and two tertiary centers in a neighboring state participated, sharing information on system status, challenges, and strategies. Patient volumes and transfer data were tracked; client satisfaction was evaluated through an anonymous survey. RESULTS: From August 1, 2020, through March 31, 2022, a total of 5,307 requests were made, 2,008 beds identified, 1,316 requests canceled, and 1,981 requests were unable to be fulfilled. A total of 1,715 patients had COVID-19 (32.3%), and 2,473 were negative or low risk for COVID-19 (46.6%). COVID-19 status was unknown in 1,119 (21.1%). Overall, 760 were patients on ventilators (49.1% COVID-19 positive). The Minnesota Critical Care Coordination Center placed most patients during the fall 2020 surge with the Minnesota Governor's stay-at-home order during the peak. However, during the fall 2021 surge, only 30% of ICU patients and 39% of medical-surgical patients were placed. Indicators characterizing severe surge include the number of Critical Care Coordination Center requests, decreasing placements, longer placement times, and time series analysis showing significant request-acceptance differences. INTERPRETATION: Implementation of a large-scale Minnesota MOCC program was effective at placing patients during the first COVID-19 pandemic fall 2020 surge and was well regarded by hospitals and health systems. However, under worsening duress of limited resources during the fall 2021 surge, placement of ICU and medical-surgical patients was greatly decreased.

Learning curves in Motec total wrist arthroplasty: an international cohort study.

We examined the learning curve of Motec total wrist arthroplasty (TWA) of six experienced surgeons in their first 30 cases. Three times more complications/revisions were encountered in the first half of the study compared with the second half. Motec TWA surgery should be concentrated in a smaller number of centres performing higher volumes.