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Whilst research has assessed the multidimensionality of organizational stressors, severity has been overlooked. This study aimed to develop and validate a severity response scale, before examining the relationship between organizational stressor severity, physical health, and well-being, and if severity mediated the relationship between stressor occurrence and outcomes. 403 sport performers (186 male; Mage = 21.43) completed a questionnaire assessing study variables and data were analysed using structural equation modelling. Organizational stressor severity had a significant, negative relationship with physical health (r = -.374, p < .001), but notwith hedonic or eudaimonic well-being (p = .624 and .203). Direct effects were found for neuroticism (ß = -0.632, p < .001) and stressor frequency (ß = -0.226, p = .016) on physical health; however, when adding severity as a mediator, stressor frequency positively predicted stressor severity (ß = 1.025, p < .001); however, the indirect effect of stressor frequency on physical health via stressor severity (ß = -0.018, p = .885) and the direct effect between stressor frequency and physical health (ß = -0.191, p = .204) were non-significant. These novel findings highlight that higher stressor severity is associated with higher physical health complaints, and that repeated exposure to stressors may heighten an vulnerability to future stressors.
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Estado de Salud , Estrés Psicológico , Humanos , Masculino , Adulto Joven , Femenino , Encuestas y Cuestionarios , Adulto , Atletas/psicología , Neuroticismo , AdolescenteRESUMEN
BACKGROUND: Mixed martial arts (MMA) is experiencing a surge in popularity in Australia. Previous research has suggested knockout (KO) and technical knockout (TKO) are frequent outcomes during competition, raising concern about the brain health of athletes. This study aims to describe fight outcomes in Australian MMA and to explore differences in fight-ending outcomes between male and female athletes, amateur and professional competition, and different weight classes. HYPOTHESIS: There is no difference in the incidence of KO/TKO between level of competition, sex, and weight class. STUDY DESIGN: Descriptive epidemiology study. LEVEL OF EVIDENCE: Level 3. METHODS: Retrospective analysis of 143 Australian MMA events from 2020 to 2023 was conducted using video replay to assess fight outcomes between sex and level of competition. Binary logistic regression analysis was used to determine relationships between weight class and KO/TKO fight outcomes. RESULTS: Male competition (34%) had a significantly greater number of KO/TKO secondary to head strikes fight outcomes compared with female competition (23%) (P = 0.01). The KO/TKO rate secondary to head strikes for amateur and professional male competition was 16.6 and 18.7 per 100 athlete-exposures (AEs), respectively. The amateur and professional female rate was 12.6 and 7.4 per 100 AEs, respectively. Amateur male light heavyweight and heavyweight, and professional male heavyweight were at greater odds of a KO or TKO compared with other weight classes in their equivalent level of competition. CONCLUSION: There is a sex and professional level disparity in the incidence of fight-ending head trauma in Australian MMA. The study findings highlight the urgent need for targeted safety protocols and medical oversight, particularly for men in heavier weight classes. CLINICAL RELEVANCE: This study highlights the need for enhanced safety protocols and medical oversight in Australian MMA, particularly for male athletes in heavier weight divisions.
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PURPOSE: To characterize the response and survival outcomes of yttrium-90 transarterial radioembolization (90Y-TARE) for unresectable, liver-dominant metastases from primary neoplasms other than colorectal carcinoma. MATERIALS AND METHODS: This study included 1474 patients enrolled in the RESiN registry who received resin 90Y-TARE as part of their oncologic management for unresectable primary or secondary liver tumors (NCT02685631). 33% (481/1474) were treated for liver metastases of non-colorectal origin (m-nonCRC), compared to 34% (497/1474) treated for colorectal liver metastases (mCRC) and 34% (496/1474) treated for hepatocellular carcinoma (HCC). Treatment response and cancer survival probabilities were computed and compared for each primary cancer type. The Kaplan-Meier method and log-rank test were used to compare survival outcomes. RESULTS: Radiological responses were observed in 12 unique cancer types, mostly heavily pre-treated malignancies refractory to multiple lines of systemic therapies. The overall use of resin 90Y-TARE in m-nonCRC resulted in better treatment outcomes in terms of duration of response, progression free survival, time to progression and overall survival (P = 0.04, P = 0.02, P = 0.01, P = 0.04). Analyses of cancer cell types revealed that metastatic neuroendocrine tumor, sarcoma, and ovarian, renal, prostate, and breast cancers were associated with superior treatment outcomes, whereas worse treatment outcomes were observed in metastatic lung, gastric, pancreatic and esophageal cancers. CONCLUSION: Real-world data demonstrate the use of resin 90Y-TARE in m-nonCRC refractory to standard chemotherapy. For some cell types, this expanded use achieved superior treatment outcomes relative to the reference standard of mCRC, suggesting the need for inquiry into broadened indications for 90Y-TARE.
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PURPOSE: To provide guidance, via multidisciplinary consensus statements, on the safety interactions between systemic anticancer agents (such as radiosensitizing chemotherapy, immunotherapy, targeted therapy, and peptide receptor radionuclide therapy) and transarterial radioembolization (TARE) with yttrium-90 (90Y)-labeled microspheres in the treatment of primary and metastatic liver malignancies. MATERIALS AND METHODS: A literature search identified 59 references that informed 26 statements on the safety of 90Y TARE combined with systemic therapies. Modified Delphi method was used to develop consensus on statements through online anonymous surveys of the 12 panel members representing the fields of interventional radiology, medical oncology, surgical oncology, hepatology, and pharmacy, focusing on hepatocellular carcinoma (HCC), metastatic colorectal cancer (mCRC), neuroendocrine tumors, metastatic breast cancer, and intrahepatic cholangiocarcinoma. RESULTS: High-level evidence was limited. Level 1 data in patients with mCRC suggest that some radiosensitizing chemotherapies (eg, oxaliplatin) require temporary dose reduction when used concomitantly with 90Y TARE, and some targeted therapies (eg, vascular endothelial growth factor inhibitors and antiangiogenic tyrosine kinase inhibitors) should be avoided for at least 4 weeks before 90Y TARE. In patients with HCC, the feasibility of 90Y TARE and immunotherapy has been demonstrated with Level 4 evidence. Data are more limited for other primary and secondary liver malignancies, and consensus statements were driven by expert opinion (Level 5). CONCLUSIONS: Given the absence of evidence-based guidelines on the safety of 90Y TARE in combination with systemic anticancer therapy, these consensus statements provide expert guidance on the potential risks when considering specific combinations.
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INTRODUCTION: During high-fidelity simulations in the Critical Care Air Transport (CCAT) Advanced course, we identified a high frequency of insulin medication errors and sought strategies to reduce them using a human factors approach. MATERIALS AND METHODS: Of 169 eligible CCAT simulations, 22 were randomly selected for retrospective audio-video review to establish a baseline frequency of insulin medication errors. Using the Human Factors Analysis Classification System, dosing errors, defined as a physician ordering an inappropriate dose, were categorized as decision-based; administration errors, defined as a clinician preparing and administering a dose different than ordered, were categorized as skill-based. Next, 3 a priori interventions were developed to decrease the frequency of insulin medication errors, and these were grouped into 2 study arms. Arm 1 included a didactic session reviewing a sliding-scale insulin (SSI) dosing protocol and a hands-on exercise requiring all CCAT teams to practice preparing 10 units of insulin including a 2-person check. Arm 2 contained arm 1 interventions and added an SSI cognitive aid available to students during simulation. Frequency and type of insulin medication errors were collected for both arms with 93 simulations for arm 1 (January-August 2021) and 139 for arm 2 (August 2021-July 2022). The frequency of decision-based and skill-based errors was compared across control and intervention arms. RESULTS: Baseline insulin medication error rates were as follows: decision-based error occurred in 6/22 (27.3%) simulations and skill-based error occurred in 6/22 (27.3%). Five of the 6 skill-based errors resulted in administration of a 10-fold higher dose than ordered. The post-intervention decision-based error rates were 9/93 (9.7%) and 23/139 (2.2%), respectively, for arms 1 and 2. Compared to baseline error rates, both arm 1 (P = .04) and arm 2 (P < .001) had a significantly lower rate of decision-based errors. Additionally, arm 2 had a significantly lower decision-based error rate compared to arm 1 (P = .015). For skill-based preparation errors, 1/93 (1.1%) occurred in arm 1 and 4/139 (2.9%) occurred in arm 2. Compared to baseline, this represents a significant decrease in skill-based error in both arm 1 (P < .001) and arm 2 (P < .001). There were no significant differences in skill-based error between arms 1 and 2. CONCLUSIONS: This study demonstrates the value of descriptive error analysis during high-fidelity simulation using audio-video review and effective risk mitigation using training and cognitive aids to reduce medication errors in CCAT. As demonstrated by post-intervention observations, a human factors approach successfully reduced decision-based error by using didactic training and cognitive aids and reduced skill-based error using hands-on training. We recommend the development of a Clinical Practice Guideline including an SSI protocol, guidelines for a 2-person check, and a cognitive aid for implementation with deployed CCAT teams. Furthermore, hands-on training for insulin preparation and administration should be incorporated into home station sustainment training to reduced medication errors in the operational environment.
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Total wrist arthroplasty (TWA) aims to restore pain-free motion to diseased joints. One such TWA, the Motec, has demonstrated good results with acceptable complication rates. It has recently been suggested that the metal-on-carbon fibre reinforced poly ether ether ketone (Mo-CFR-PEEK) version of the Motec TWA be implanted instead of the metal-on-metal version. An explant analysis was undertaken on seven Motec Mo-CFR-PEEK TWAs, revised for a variety of reasons, after a mean time of 2 years in vivo. Compared to a new Motec implant, five of the explanted metal heads and three of the CFR-PEEK cups became smoother in vivo, suggesting self-polishing and negative skewness, indicating some material loss in vivo. Two explanted cups showed indentation marks on their rims and one of these was from component impingement with embedded metallic debris. In the short-term, the articulating surfaces of Motec Mo-CFR-PEEK TWAs did not show major damage.Level of evidence: IV.
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Surgical intervention is a common option for the treatment of wrist joint arthritis and traumatic wrist injury. Whether this surgery is arthrodesis or a motion preserving procedure such as arthroplasty, wrist joint biomechanics are inevitably altered. To evaluate effects of surgery on parameters such as range of motion, efficiency and carpal kinematics, repeatable and controlled motion of cadaveric specimens is required. This study describes the development of a device that enables cadaveric wrist motion to be simulated before and after motion preserving surgery in a highly controlled manner. The simulator achieves joint motion through the application of predetermined displacements to the five major tendons of the wrist, and records tendon forces. A pilot experiment using six wrists aimed to evaluate its accuracy and reproducibility. Biplanar X-ray videoradiography (BPVR) and X-Ray Reconstruction of Moving Morphology (XROMM) were used to measure overall wrist angles before and after total wrist arthroplasty. The simulator was able to produce flexion, extension, radioulnar deviation, dart thrower's motion and circumduction within previously reported functional ranges of motion. Pre- and post-surgical wrist angles did not significantly differ. Intra-specimen motion trials were repeatable; root mean square errors between individual trials and average wrist angle and tendon force profiles were below 1° and 2 N respectively. Inter-specimen variation was higher, likely due to anatomical variation and lack of wrist position feedback. In conclusion, combining repeatable intra-specimen cadaveric motion simulation with BPVR and XROMM can be used to determine potential effects of motion preserving surgeries on wrist range of motion and biomechanics.
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Cadáver , Rango del Movimiento Articular , Articulación de la Muñeca , Humanos , Articulación de la Muñeca/cirugía , Articulación de la Muñeca/diagnóstico por imagen , Articulación de la Muñeca/fisiología , Articulación de la Muñeca/anatomía & histología , Fenómenos Biomecánicos , Radiografía/métodos , Masculino , Anciano , Reproducibilidad de los Resultados , Tendones/cirugía , Tendones/diagnóstico por imagen , Tendones/fisiología , Tendones/anatomía & histología , FemeninoRESUMEN
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.
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Sistema Enzimático del Citocromo P-450 , Proteínas Fúngicas , Proteoma , Animales , Sistema Enzimático del Citocromo P-450/metabolismo , Sistema Enzimático del Citocromo P-450/genética , Ratones , Proteoma/metabolismo , Proteínas Fúngicas/metabolismo , Proteínas Fúngicas/genética , Ascomicetos/metabolismo , Proteínas de Plantas/metabolismo , Proteínas de Plantas/genéticaRESUMEN
OBJECTIVE: This study determined the association between acute changes in physical activity, temperature, and humidity and 24-hour subjective and objective hot flash experience. METHODS: Data collection occurred during the cooler months of the year in Western Massachusetts (October-April). Women aged 45-55 across three menopause stages (n = 270) were instrumented with ambulatory monitors to continuously measure hot flashes, physical activity, temperature, and humidity for 24 hours. Objective hot flashes were assessed via sternal skin conductance, and subjective hot flashes were recorded by pressing an event marker and data logging. Physical activity was measured with wrist-worn accelerometers and used to define sleep and wake periods. Logistic multilevel modeling was used to examine the differences in physical activity, humidity, and temperature in the 10 minutes preceding a hot flash versus control windows when no hot flashes occurred. The odds of hot flashes were considered separately for objective and subjective hot flashes as well as for wake and sleep periods. RESULTS: Data from 188 participants were included in the analyses. There was a significantly greater odds of a hot flash following acute increases in physical activity for objective waking hot flashes (odds ratio [OR], 1.31; 95% confidence interval [CI], 1.17-1.47; P < 0.001) and subjective waking hot flashes (OR, 1.16; 95% CI, 1.0-1.33; P = 0.03). Acute increases in the actigraphy signal were associated with significantly higher odds of having an objective (OR, 1.17; 95% CI, 1.03-1.35; P < 0.01) or subjective (OR, 1.72; 95% CI, 1.52-2.01; P < 0.001) sleeping hot flash. Increases in temperature were significantly related to the odds of subjective sleeping hot flashes only (OR, 1.38; 95% CI, 1.15-1.62; P < 0.001). There was no evidence for a relationship between humidity and odds of experiencing any hot flashes. CONCLUSION: These results indicate that acute increases in physical activity increase the odds of hot flashes that are objectively measured and subjectively reported during waking and sleeping periods. Temperature increases were only related to subjectively reported nighttime hot flashes.
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Ejercicio Físico , Sofocos , Menopausia , Sueño , Humanos , Femenino , Sofocos/fisiopatología , Persona de Mediana Edad , Ejercicio Físico/fisiología , Menopausia/fisiología , Sueño/fisiología , Humedad , Temperatura , Massachusetts/epidemiologíaRESUMEN
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.
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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.
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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.
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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.
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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.
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Ejercicio Físico , Sofocos , Menopausia , Conducta Sedentaria , Sueño , Humanos , Femenino , Sofocos/fisiopatología , Persona de Mediana Edad , Ejercicio Físico/fisiología , Sueño/fisiología , Menopausia/fisiología , Autoinforme , ActigrafíaRESUMEN
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.
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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.
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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.
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Leucocitos Mononucleares , Análisis de la Célula Individual , Humanos , Animales , Ratones , RNA-Seq , Análisis de Secuencia de ARN/métodos , Análisis de la Célula Individual/métodos , Algoritmos , Perfilación de la Expresión Génica/métodosRESUMEN
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, VÌO2peak 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 â¼1.5 % 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.