Substrate specificity mapping of fungal CAZy AA3_2 oxidoreductases.

BACKGROUND: Oxidative enzymes targeting lignocellulosic substrates are presently classified into various auxiliary activity (AA) families within the carbohydrate-active enzyme (CAZy) database. Among these, the fungal AA3 glucose-methanol-choline (GMC) oxidoreductases with varying auxiliary activities are attractive sustainable biocatalysts and important for biological function. CAZy AA3 enzymes are further subdivided into four subfamilies, with the large AA3_2 subfamily displaying diverse substrate specificities. However, limited numbers of enzymes in the AA3_2 subfamily are currently biochemically characterized, which limits the homology-based mining of new AA3_2 oxidoreductases. Importantly, novel enzyme activities may be discovered from the uncharacterized parts of this large subfamily. RESULTS: In this study, phylogenetic analyses employing a sequence similarity network (SSN) and maximum likelihood trees were used to cluster AA3_2 sequences. A total of 27 AA3_2 proteins representing different clusters were selected for recombinant production. Among them, seven new AA3_2 oxidoreductases were successfully produced, purified, and characterized. These enzymes included two glucose dehydrogenases (TaGdhA and McGdhA), one glucose oxidase (ApGoxA), one aryl alcohol oxidase (PsAaoA), two aryl alcohol dehydrogenases (AsAadhA and AsAadhB), and one novel oligosaccharide (gentiobiose) dehydrogenase (KiOdhA). Notably, two dehydrogenases (TaGdhA and KiOdhA) were found with the ability to utilize phenoxy radicals as an electron acceptor. Interestingly, phenoxy radicals were found to compete with molecular oxygen in aerobic environments when serving as an electron acceptor for two oxidases (ApGoxA and PsAaoA), which sheds light on their versatility. Furthermore, the molecular determinants governing their diverse enzymatic functions were discussed based on the homology model generated by AlphaFold. CONCLUSIONS: The phylogenetic analyses and biochemical characterization of AA3_2s provide valuable guidance for future investigation of AA3_2 sequences and proteins. A clear correlation between enzymatic function and SSN clustering was observed. The discovery and biochemical characterization of these new AA3_2 oxidoreductases brings exciting prospects for biotechnological applications and broadens our understanding of their biological functions.

A Novel Metric "Exercise Cardiac Load" Proposed to Track and Predict the Deterioration of the Autonomic Nervous System in Division I Football Athletes.

Current metrics like baseline heart rate (HR) and HR recovery fail in predicting overtraining (OT), a syndrome manifesting from a deteriorating autonomic nervous system (ANS). Preventing OT requires tracking the influence of internal physiological loads induced by exercise training programs on the ANS. Therefore, this study evaluated the predictability of a novel, exercise cardiac load metric on the deterioration of the ANS. Twenty male American football players, with an average age of 21.3 years and body mass indices ranging from 23.7 to 39.2 kg/m2 were included in this study. Subjects participated in 40 strength- and power-focused exercise sessions over 8 weeks and wore armband monitors (Warfighter Monitor, Tiger Tech Solutions) equipped with electrocardiography capabilities. Exercise cardiac load was the product of average training HR and duration. Baseline HR, HR variability (HRV), average HR, and peak HR were also measured. HR recovery was measured on the following day. HRV indices assessed included the standard deviation of NN intervals (SDNN) and root mean square of successive RR interval differences (rMSSD) Linear regression models assessed the relationships between each cardiac metric and HR recovery, with statistical significance set at α < 0.05. Subjects were predominantly non-Hispanic black (70%) and aged 21.3 (±1.4) years. Adjusted models showed that exercise cardiac load elicited the strongest negative association with HR recovery for previous day (ß = -0.18 ± 0.03; p < 0.0000), one-week (ß = -0.20 ± 0.03; p < 0.0000) and two-week (ß = -0.26 ± 0.03; p < 0.0000) training periods compared to average HR (ßetas: -0.09 to -0.02; p < 0.0000) and peak HR (ßetas: -0.13 to -0.23; p < 0.0000). Statistically significant relationships were also found for baseline HR (p < 0.0000), SDNN (p < 0.0000) and rMSSD (p < 0.0000). Exercise cardiac load appears to best predict ANS deterioration across one- to two-week training periods, showing a capability for tracking an athlete's physiological tolerance and ANS response. Importantly, this information may increase the effectiveness of exercise training programs, enhance performance, and prevent OT.

Exercise Cardiac Load and Autonomic Nervous System Recovery during In-Season Training: The Impact on Speed Deterioration in American Football Athletes.

Fully restoring autonomic nervous system (ANS) function is paramount for peak sports performance. Training programs failing to provide sufficient recovery, especially during the in-season, may negatively affect performance. This study aimed to evaluate the influence of the physiological workload of collegiate football training on ANS recovery and function during the in-season. Football athletes recruited from a D1 college in the southeastern US were prospectively followed during their 13-week "in-season". Athletes wore armband monitors equipped with ECG and inertial movement capabilities that measured exercise cardiac load (ECL; total heartbeats) and maximum running speed during and baseline heart rate (HR), HR variability (HRV) 24 h post-training. These metrics represented physiological load (ECL = HR·Duration), ANS function, and recovery, respectively. Linear regression models evaluated the associations between ECL, baseline HR, HRV, and maximum running speed. Athletes (n = 30) were 20.2 ± 1.5 years, mostly non-Hispanic Black (80.0%). Negative associations were observed between acute and cumulative exposures of ECLs and running speed (ß = -0.11 ± 0.00, p < 0.0000 and ß = -0.15 ± 0.04, p < 0.0000, respectively). Similarly, negative associations were found between baseline HR and running speed (ß = -0.45 ± 0.12, 95% CI: -0.70, -0.19; p = 0.001). HRV metrics were positively associated with running speed: (SDNN: ß = 0.32 ± 0.09, p < 0.03 and rMSSD: ß = 0.35 ± 0.11, p < 0.02). Our study demonstrated that exposure to high ECLs, both acutely and cumulatively, may negatively influence maximum running speed, which may manifest in a deteriorating ANS. Further research should continue identifying optimal training: recovery ratios during off-, pre-, and in-season phases.

Glacial changes in sea level modulated millennial-scale variability of Southeast Asian autumn monsoon rainfall.

Most paleoclimate studies of Mainland Southeast Asia hydroclimate focus on the summer monsoon, with few studies investigating rainfall in other seasons. Here, we present a multiproxy stalagmite record (45,000 to 4,000 years) from central Vietnam, a region that receives most of its annual rainfall in autumn (September-November). We find evidence of a prolonged dry period spanning the last glacial maximum that is punctuated by an abrupt shift to wetter conditions during the deglaciation at ~14 ka. Paired with climate model simulations, we show that sea-level change drives autumn monsoon rainfall variability on glacial-orbital timescales. Consistent with the dry signal in the stalagmite record, climate model simulations reveal that lower glacial sea level exposes land in the Gulf of Tonkin and along the South China Shelf, reducing convection and moisture delivery to central Vietnam. When sea level rises and these landmasses flood at ~14 ka, moisture delivery to central Vietnam increases, causing an abrupt shift from dry to wet conditions. On millennial timescales, we find signatures of well-known Heinrich Stadials (HS) (dry conditions) and Dansgaard-Oeschger Events (wet conditions). Model simulations show that during the dry HS, changes in sea surface temperature related to meltwater forcing cause the formation of an anomalous anticyclone in the Western Pacific, which advects dry air across central Vietnam, decreasing autumn rainfall. Notably, sea level modulates the magnitude of millennial-scale dry and wet phases by muting dry events and enhancing wet events during periods of low sea level, highlighting the importance of this mechanism to autumn monsoon variability.

Positive Minds for Refugees: A Qualitative Study to Inform Intervention Development.

Refugees face significantly worse mental health outcomes compared to the general population within their host country; however, few refugee-specific mental health programs exist within the United States. Utilizing Community-based Participatory Research methods, a community-based mental health intervention named Positive Minds for Refugees (PMR) is in development. In this preliminary study, we shared the in-development intervention with refugees (n = 8), holding a series of 3 focus groups to gain feedback on the intervention content and determine acceptability. Findings suggest that the intervention is generally acceptable and relevant to the refugee community. Mental health for refugees is closely linked with their ability to navigate new social, cultural, and physical environments during resettlement; therefore, to address unmet needs, additional content should be added on: American cultural norms, navigating US society, and coping with social isolation and self-care. This study highlights cultural considerations for presenting written intervention content and implementing sessions.

Exposures to Elevated Core Temperatures during Football Training: The Impact on Autonomic Nervous System Recovery and Function.

Exercising with elevated core temperatures may negatively affect autonomic nervous system (ANS) function. Additionally, longer training duration under higher core temperatures may augment these negative effects. This study evaluated the relationship between exercise training duration and 24 h ANS recovery and function at ≥37 °C, ≥38 °C and ≥39 °C core temperature thresholds in a sample of male Division I (D1) collegiate American football athletes. Fifty athletes were followed over their 25-week season. Using armband monitors (Warfighter MonitorTM, Tiger Tech Solutions, Inc., Miami, FL, USA), core temperature (°C) and 24 h post-exercise baseline heart rate (HR), HR recovery and heart rate variability (HRV) were measured. For HRV, two time-domain indices were measured: the root mean square of the standard deviation of the NN interval (rMSSD) and the standard deviation of the NN interval (SDNN). Linear regression models were performed to evaluate the associations between exercise training duration and ANS recovery (baseline HR and HRV) and function (HR recovery) at ≥37 °C, ≥38 °C and ≥39 °C core temperature thresholds. On average, the athletes were 21.3 (± 1.4) years old, weighed 103.0 (±20.2) kg and had a body fat percentage of 15.4% (±7.8%, 3.0% to 36.0%). The duration of training sessions was, on average, 161.1 (±40.6) min and they ranged from 90.1 to 339.6 min. Statistically significant associations between training duration and 24 h ANS recovery and function were observed at both the ≥38.0 °C (baseline HR: ß = 0.10 ± 0.02, R2 = 0.26, p < 0.0000; HR recovery: ß = -0.06 ± 0.02, R2 = 0.21, p = 0.0002; rMSSD: ß = -0.11 ± 0.02, R2 = 0.24, p < 0.0000; and SDNN: ß = -0.16 ± 0.04, R2 = 0.22, p < 0.0000) and ≥39.0 °C thresholds (ß = 0.39 ± 0.05, R2 = 0.62, p < 0.0000; HR recovery: ß = -0.26 ± 0.04, R2 = 0.52, p < 0.0000; rMSSD: ß = -0.37 ± 0.05, R2 = 0.58, p < 0.0000; and SDNN: ß = -0.67 ± 0.09, R2 = 0.59, p < 0.0000). With increasing core temperatures, increases in slope steepness and strengths of the associations were observed, indicating accelerated ANS deterioration. These findings demonstrate that exercise training under elevated core temperatures (≥38 °C) may negatively influence ANS recovery and function 24 h post exercise and progressively worsen.