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Benzene degradation under anoxic conditions was first reported more than 25 years ago; however, the activation mechanism in the absence of oxygen remains elusive. Progress has been hindered by the difficulty in cultivating anaerobic benzene-degrading enrichment cultures. Our laboratory has sustained a methanogenic enrichment culture harboring Desulfobacterota ORM2, a benzene fermenter distinct from any known genus but related to other known or predicted benzene degraders. ORM2's slow doubling time (∼30 days) and extended lag phase after inoculation complicate its study. We developed a fluorescent in situ hybridization (FISH) probe for ORM2, revealing rod-shaped cells of variable length that tend to cluster with other organisms, particularly methanogens. Microscopy and genomic evidence suggest that ORM2 may produce extracellular polymeric substances, facilitating cell aggregation and possibly consuming energy that contributes to the lag phase. Interestingly, higher benzene concentrations (90-120 mg/L) appeared to reduce cell aggregation. This study visualized the cells of Desulfobacterota ORM2 within a methanogenic community, offering insights into spatial organization and potential strategies to enhance its growth rate.

Chloroform and trichloroethanes are pervasive groundwater contaminants for which bioremediation has been an effective treatment strategy. Reductive dehalogenase (RDase) enzymes from organohalide-respiring bacteria are essential for their remediation under anaerobic conditions. RDases are responsible for dehalogenating these chlorinated solvents, leading to their removal. This work explores the kinetic characteristics of three closely related Dehalobacter chloroalkane-reductases─TmrA, CfrA, and AcdA─and identifies differences between their activity on chloroform (CF), 1,1,1-trichloroethane (TCA), and 1,1,2-TCA. The side-by-side comparison of these enzymes has emphasized that TmrA and AcdA are specialized toward CF with both having a 4-fold higher maximum specific activity ( V max ) on CF than 1,1,1-TCA, whereas CfrA has very similar rates on both CF and 1,1,1-TCA. AcdA is the most sensitive to substrate inhibition by CF and 1,1,2-TCA and inhibition by a common cocontaminant trichloroethene. Finally, the reduction of 1,1,2-TCA, which can produce both 1,2-dichloroethane and vinyl chloride, was assessed for each enzyme. Interestingly, each enzyme has a distinct preference for the major product it produces, indicating a favored reaction pathway. Despite over 95% sequence identity, TmrA, CfrA, and AcdA exhibit substantial differences in kinetic behavior, highlighting the importance of understanding such nuances for informed bioremediation strategies.

The archaeal class Bathyarchaeia is widely and abundantly distributed in anoxic habitats. Metagenomic studies have suggested that they are mixotrophic, capable of CO 2 fixation and heterotrophic growth, and involved in acetogenesis and lignin degradation. We analyzed 35 Bathyarchaeia metagenome-assembled genomes (MAGs), including the first complete circularized MAG (cMAG) of the Bathy-6 subgroup, from the metagenomes of three full-scale pulp and paper mill anaerobic digesters and three laboratory methanogenic enrichment cultures maintained on pre-treated poplar. Thirty-three MAGs belong to the Bathy-6, lineage while two are from the Bathy-8 lineage. In our previous analysis of the microbial community in the pulp mill digesters, Bathyarchaeia were abundant and positively correlated to hydrogenotrophic and methylotrophic methanogenesis. Several factors likely contribute to the success of the Bathy-6 lineage compared to Bathy-8 in the reactors. The Bathy-6 genomes are larger than those of Bathy-8 and have more genes involved in lignocellulose degradation, including carbohydrate-active enzymes not present in the Bathy-8. Bathy-6 also shares the Bathyarchaeal O -demethylase system recently identified in Bathy-8. All the Bathy-6 MAGs had numerous membrane-associated pyrroloquinoline quinone-domain proteins that we suggest are involved in lignin modification or degradation, together with Radical-S-adenosylmethionine (SAM) and Rieske domain proteins, and AA2, AA3, and AA6-family oxidoreductases. We also identified a complete B12 synthesis pathway and a complete nitrogenase gene locus. Finally, comparative genomic analyses revealed that Bathyarchaeia genomes are dynamic and have interacted with other organisms in their environments through gene transfer to expand their gene repertoire., Competing Interests: The authors declare no competing financial interests., (© The Author(s) 2024. Published by Oxford University Press on behalf of the International Society for Microbial Ecology.)

Purpose: Physical activity (PA) guidelines suggest a link between PA, sedentary time, and sleep quality (SQ). Step cadence is an emerging proxy for PA intensity, with zero cadence (ZC) indicating sedentary time. No research has explored the relationship between ZC and SQ. This study examines the relationship between ZC and SC, compared to traditional PA and sedentary metrics, all measured via accelerometry., Methods: Sleep and PA was assessed in 138 adults (53 male, 85 female, age = 23.5 ± 8.0 year.) via accelerometry. Sedentary, light, moderate, vigorous intensity PA, and ZC minutes per day were measured. SQ variables included sleep efficiency (SE), total sleep time (TST), and minutes of awake time after sleep onset (WASO). Correlation analysis and analysis of covariance was used to assess relationships between study variables and to assess differences in PA and sedentary behavior between normal and poor sleepers., Results: Sedentary time was negatively associated with SE (r = -0.24, P < 0.05) and TST (r = -0.51, P < 0.001). ZC was negatively associated with SE (r = -0.25, P < 0.05), TST (r = -0.39, P < 0.001) and positively associated with WASO (r = 0.17, P < 0.001). Poor sleepers accumulated greater ZC minutes than normal sleepers when categorized by SE (555.9 ± 69.1 vs. 521.6 ± 83.7, P = 0.02)) or TST (570.4 ± 77.7 vs. 524.5 ± 76.7, P < 0.01)., Conclusion: Results suggest that zero cadence minutes is a viable metric for assessing sedentary time and may be of greater utility to more traditional measure of sedentary time., Competing Interests: Declarations. Ethical approval: All procedures performed in studies involving human participants were in accordance with the ethical standards of the institutional and/or national research committee and with the 1964 Helsinki declaration and its later amendments or comparable ethical standards. Informed consent: Informed consent was obtained from all individual participants included in the study. Conflict of interest: All authors certify that they have no affiliations with or involvement in any organization or entity with any financial interest (such as honoraria; educational grants; participation in speakers’ bureaus; membership, employment, consultancies, stock ownership, or other equity interest; and expert testimony or patent-licensing arrangements), or non-financial interest (such as personal or professional relationships, affiliations, knowledge or beliefs) in the subject matter or materials discussed in this manuscript., (© 2024. The Author(s).)

We present a dataset of six metagenomes and 323 metagenome assembled genomes (MAGs) describing the microbial community of anaerobic digesters at three Canadian pulp and paper mills. Our objective was to assess the coding potential of the microbial community and obtain draft genomes of key organisms in the digesters., Competing Interests: The authors declare no conflict of interest.

Here we present two metagenomes and two Dehalobacter metagenome-assembled genomes from subcultures of an anaerobic chloroform and dichloromethane degrading microbial community used for bioremediation. Our objective was to assemble and curate the genome(s) of Dehalobacter , key biodegraders in the culture, through repeated sequencing and joint assembly with previous datasets., Competing Interests: The authors declare no conflict of interest.

Dehalobacter is a genus of organohalide-respiring bacteria that is recognized for its fastidious growth using reductive dehalogenases (RDases). In the SC05 culture, however, a Dehalobacter population also mineralizes dichloromethane (DCM) produced by chloroform dechlorination using the mec cassette, just downstream of its active RDase. A closed genome of this DCM-mineralizing lineage has previously evaded assembly. Here, we present the genomes of two novel Dehalobacter strains, each of which was assembled from the metagenome of a distinct subculture from SC05. A pangenomic analysis of the Dehalobacter genus, including RDase synteny and phylogenomics, reveals at least five species of Dehalobacter based on average nucleotide identity, RDase and core gene synteny, as well as differential functional genes. An integration hotspot is also pinpointed in the Dehalobacter genome, in which many recombinase islands have accumulated. This nested recombinase island encodes the active RDase and mec cassette in both SC05 Dehalobacter genomes, indicating the transfer of key functional genes between species of Dehalobacter . Horizontal gene transfer between these two novel Dehalobacter strains has implications for the evolutionary history within the SC05 subcultures and of the Dehalobacter genus as a whole, especially regarding adaptation to anthropogenic chemicals.

Here, we present the metagenome of an anaerobic chloroform respiring mixed microbial community used for bioremediation. Our objective was to obtain draft genomes of key microorganisms in the culture., Competing Interests: The authors declare no conflict of interest.

Metagenome-assembled genomes (MAGs) were recovered from metagenomic assemblies from a nitrate-reducing benzene-degrading enrichment culture. Ten MAGs of high quality or functional interest to benzene degradation are reported, seven of which are single contig genomes., Competing Interests: The authors declare no conflict of interest.

We present a metagenome assembled genome (MAG) of an anaerobic bacterium from a nitrate-reducing, benzene-degrading enrichment culture (NRBC). The draft Thermincolales genome consists of 20 contigs with a total length of 4.09 Mbp and includes putative carboxylase genes likely involved in benzene activation., Competing Interests: The authors declare no conflict of interest.

Biological fluidized bed reactor (FBR) is a promising treatment option for removing selenium oxyanions from wastewater by converting them into elemental selenium. The process can achieve high rates and be efficiently operated at low hydraulic retention times (HRT). However, the effects of HRT on the changes in microbial community in the FBR process have not been previously explored. In this study, dynamic changes of microbial communities both on biofilm carrier and in suspension of a selenate-reducing FBR were explored at various HRTs (0.3-120 h). Based on partial 16S rRNA gene sequencing of the microbial communities, alpha diversity of microbial communities in suspension rather than in the biofilm were impacted by low HRTs (0.3 h-3 h). Members from genera Geobacter, Geoalkalibacter, and Geovibrio were the main selenate-reducing bacteria on carrier throughout the FBR process. Genus Geobacter was dominant in FBR carrier at HRT of 24 h-120 h, whereas Geoalkalibacter and Geovibrio dominated at low HRT of 0.3 h-6 h. Suspended microbial communities detected in the FBR effluent were more sensitive to HRT changes than that in biofilm. "Shock loading" at HRT of 0.3 h had a great impact on microbial community compositions both in the biofilm and effluent. Reactor operation in batch mode and long HRT of 24 h helped recover the community from "shock loading" and improved selenite reduction and ethanol oxidation. Redundancy analysis revealed that HRT, influent pH and selenate loading were key operational parameters impacting both the FBR performance and the composition of microbial communities associated with both the FBR carrier and effluent. Overall, the microbial communities in FBR biofilm flexibly responded to the changes of HRT and showed resilience to the temporary shock loading, enabling efficient selenate removal., Competing Interests: Declaration of competing interest The authors declare the following financial interests/personal relationships which may be considered as potential competing interests: The authors report financial support was provided by Ontario Research Fund and China Scholarship Council., (Copyright © 2024 The Authors. Published by Elsevier Ltd.. All rights reserved.)

Carbon and chlorine isotope effects for biotransformation of chloroform by different microbes show significant variability. Reductive dehalogenases (RDase) enzymes contain different cobamides, affecting substrate preferences, growth yields, and dechlorination rates and extent. We investigate the role of cobamide type on carbon and chlorine isotopic signals observed during reductive dechlorination of chloroform by the RDase CfrA. Microcosm experiments with two subcultures of a Dehalobacter-containing culture expressing CfrA-one with exogenous cobamide (Vitamin B 12 , B12 + ) and one without (to drive native cobamide production)-resulted in a markedly smaller carbon isotope enrichment factor (ε C, bulk ) for B12 - (-22.1 ± 1.9‰) compared to B12 + (-26.8 ± 3.2‰). Both cultures exhibited significant chlorine isotope fractionation, and although a lower ε Cl, bulk was observed for B12 - (-6.17 ± 0.72‰) compared to B12 + (-6.86 ± 0.77‰) cultures, these values are not statistically different. Importantly, dual-isotope plots produced identical slopes of Λ Cl/C (Λ Cl/C, B12+  = 3.41 ± 0.15, Λ Cl/C, B12 - = 3.39 ± 0.15), suggesting the same reaction mechanism is involved in both experiments, independent of the lower cobamide bases. A nonisotopically fractionating masking effect may explain the smaller fractionations observed for the B12 - containing culture., (© 2024 The Author(s). MicrobiologyOpen published by John Wiley & Sons Ltd.)

Here, we present metagenomes from two cultures derived from an anaerobic microbial consortium used for bioremediation. One culture dechlorinates chloroform to dichloromethane, which is further mineralized to CO 2 . A second subculture was amended with only dichloromethane. We sought draft genomes of key microorganisms to identify metabolic potential in these consortia., Competing Interests: The authors declare no conflict of interest.

Compound-specific isotope analysis (CSIA) is an established tool to track the in situ transformation of organic chemicals at contaminated sites. In this work, we evaluated the potential of multi-element CSIA to assess biodegradation of 2,3-dichloroaniline (2,3-DCA), which is a major industrial feedstock. Using controlled laboratory experiments, we determined, for the first time, negligible carbon (<0.5‰) and hydrogen (<10‰) isotope fractionation and a significant inverse nitrogen isotope fractionation (>10‰) during aerobic 2,3-DCA biodegradation by a mixed enrichment culture. The tentative identification of a glutamate conjugate of 2,3-DCA as a reaction intermediate indicates that the initial multistep enzymatic reaction may be rate-limiting. The formation of the glutamate adduct would increase the bond energy at the N atom, thus likely explaining the observed inverse N isotope fractionation. The corresponding nitrogen enrichment factor was +6.8 ± 0.6‰. This value was applied to investigate the in situ 2,3-DCA biodegradation at a contaminated site where the carbon and nitrogen isotope signatures from field samples suggested similar aerobic processes by native microorganisms. Under the assumption of the applicability of the Rayleigh model in a pilot wetland treating contaminated groundwater, the extent of biodegradation was estimated to be up to 80-90%. This study proposes multi-element CSIA as a novel application to study 2,3-DCA fate in groundwater and surface water and provides insights into biodegradation pathways.

PMMoV has been widely used to normalize the concentration of severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) RNA, influenza, and respiratory syncytial virus (RSV) to account for variations in the fecal content of wastewater. PMMoV is also used as an internal RNA recovery control for wastewater-based epidemiology (WBE) tests. While potentially useful for the interpretation of WBE data, previous studies have suggested that PMMoV concentration can be affected by various physico-chemical characteristics of wastewater. There is also the possibility that laboratory methods, particularly the variability in centrifugation steps to remove supernatant from pellets can cause PMMoV variability. The goal of this study is to improve our understanding of the main drivers of PMMoV variability by assessing the relationship between PMMoV concentration, the physico-chemical characteristics of wastewater, and the methodological approach for concentrating wastewater samples. We analyzed 24-hour composite wastewater samples collected from the influent stream of three wastewater treatment plants (WWTPs) located in the City of Toronto, Ontario, Canada. Samples were collected 3 to 5 times per week starting from the beginning of March 2021 to mid-July 2023. The influent flow rate was used to partition the data into wet and dry weather conditions. Physico-chemical characteristics (e.g., total suspended solids (TSS), biological oxygen demand (BOD), alkalinity, electrical conductivity (EC), and ammonia (NH 3 )) of the raw wastewater were measured, and PMMoV was quantified. Spatial and temporal variability of PMMoV was observed throughout the study period. PMMoV concentration was significantly higher during dry weather conditions. Multiple linear regression analysis demonstrates that the number and type of physico-chemical parameters that drive PMMoV variability are site-specific, but overall BOD and alkalinity were the most important predictors. Differences in PMMoV concentration for a single WWTP between two different laboratory methods, along with a weak correlation between pellet mass and TSS using one method may indicate that differences in sample concentration and subjective subsampling bias could alter viral recovery and introduce variability to the data., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2024 The Authors. Published by Elsevier B.V. All rights reserved.)

During the COVID-19 pandemic, the Province of Ontario, Canada, launched a wastewater surveillance program to monitor SARS-CoV-2, inspired by the early work and successful forecasts of COVID-19 waves in the city of Ottawa, Ontario. This manuscript presents a dataset from January 1, 2021, to March 31, 2023, with RT-qPCR results for SARS-CoV-2 genes and PMMoV from 107 sites across all 34 public health units in Ontario, covering 72% of the province's and 26.2% of Canada's population. Sampling occurred 2-7 times weekly, including geographical coordinates, serviced populations, physico-chemical water characteristics, and flowrates. In doing so, this manuscript ensures data availability and metadata preservation to support future research and epidemic preparedness through detailed analyses and modeling. The dataset has been crucial for public health in tracking disease locally, especially with the rise of the Omicron variant and the decline in clinical testing, highlighting wastewater-based surveillance's role in estimating disease incidence in Ontario., (© 2024. The Author(s).)

Chloroform (CF) and dichloromethane (DCM) are groundwater contaminants of concern due to their high toxicity and inhibition of important biogeochemical processes such as methanogenesis. Anaerobic biotransformation of CF and DCM has been well documented but typically independently of one another. CF is the electron acceptor for certain organohalide-respiring bacteria that use reductive dehalogenases (RDases) to dechlorinate CF to DCM. In contrast, known DCM degraders use DCM as their electron donor, which is oxidized using a series of methyltransferases and associated proteins encoded by the mec cassette to facilitate the entry of DCM to the Wood-Ljungdahl pathway. The SC05 culture is an enrichment culture sold commercially for bioaugmentation, which transforms CF via DCM to CO 2 . This culture has the unique ability to dechlorinate CF to DCM using electron equivalents provided by the oxidation of DCM to CO 2 . Here, we use metagenomic and metaproteomic analyses to identify the functional genes involved in each of these transformations. Though 91 metagenome-assembled genomes were assembled, the genes for an RDase-named acdA -and a complete mec cassette were found to be encoded on a single contig belonging to Dehalobacter . AcdA and critical Mec proteins were also highly expressed by the culture. Heterologously expressed AcdA dechlorinated CF and other chloroalkanes but had 100-fold lower activity on DCM. Overall, the high expression of Mec proteins and the activity of AcdA suggest a Dehalobacter capable of dechlorination of CF to DCM and subsequent mineralization of DCM using the mec cassette., Importance: Chloroform (CF) and dichloromethane (DCM) are regulated groundwater contaminants. A cost-effective approach to remove these pollutants from contaminated groundwater is to employ microbes that transform CF and DCM as part of their metabolism, thus depleting the contamination as the microbes continue to grow. In this work, we investigate bioaugmentation culture SC05, a mixed microbial consortium that effectively and simultaneously degrades both CF and DCM coupled to the growth of Dehalobacter . We identified the functional genes responsible for the transformation of CF and DCM in SC05. These genetic biomarkers provide a means to monitor the remediation process in the field., Competing Interests: The authors declare no conflict of interest.

Background: Human adenoviruses (HAdVs) can cause outbreaks of flu-like illness in university settings. Most infections in healthy young adults are mild; severe illnesses rarely occur. In Fall 2022, an adenovirus outbreak was identified in university students., Methods: HAdV cases were defined as university students 17-26 years old who presented to the University Health Service or nearby emergency department with flu-like symptoms (eg, fever, cough, headache, myalgia, nausea) and had confirmed adenovirus infections by polymerase chain reaction (PCR). Demographic and clinical characteristics were abstracted from electronic medical records; clinical severity was categorized as mild, moderate, severe, or critical. We performed contact investigations among critical cases. A subset of specimens was sequenced to confirm the HAdV type., Results: From 28 September 2022 to 30 January 2023, 90 PCR-confirmed cases were identified (51% female; mean age, 19.6 years). Most cases (88.9%) had mild illness. Seven cases required hospitalization, including 2 critical cases that required intensive care. Contact investigation identified 44 close contacts; 6 (14%) were confirmed HAdV cases and 8 (18%) reported symptoms but never sought care. All typed HAdV-positive specimens (n = 36) were type 4., Conclusions: While most students with confirmed HAdV had mild illness, 7 otherwise healthy students had severe or critical illness. Between the relatively high number of hospitalizations and proportion of close contacts with symptoms who did not seek care, the true number of HAdV cases was likely higher. Our findings illustrate the need to consider a wide range of pathogens, even when other viruses are known to be circulating., Competing Interests: Potential conflicts of interest. All authors: No reported conflicts., (Published by Oxford University Press on behalf of Infectious Diseases Society of America 2024.)

Wastewater surveillance using RT-qPCR has now been widely adopted to track circulating levels of SARS-CoV-2 virus in many sewersheds. The CDC qPCR assays targeting two regions (N1 and N2) within the N gene are commonly used, but a discrepancy between the two biomarkers has been noticed by independent studies using these methods since late 2021. The reason is presumed to be due to mutations in regions targeted by the N1 qPCR probe. In this study, we systematically investigated and unequivocally confirmed that the underlying reason for this discrepancy was mutations in the N1 probe target, and that a single mutation could cause a significant drop in signal. We first confirmed the proportion of related mutations in wastewater samples (Jan 2021-Dec 2022) using nested PCR and LC-MS. Based on relative proportions of N1 alleles, we separated the wastewater data into four time periods corresponding to different variant waves: Period I (Alpha and Delta waves with 0 mutation), Period II (BA.1/BA.2 waves with a single mutation found in all Omicron strains), Period III (BA.5.2* wave with two mutations), and Period IV (BQ.1* wave with two mutations). Significantly lower N1 copies relative to N2 copies in samples from Periods II-IV compared to those from Period I was observed in wastewater. To further pinpoint the extent to which each mutation impacted N1 quantification, we compared the qPCR response among different synthetic oligomers with corresponding mutations. This study highlighted the impact of even just one or two mutations on qPCR-based wastewater surveillance of SARS-CoV-2., Competing Interests: The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (© 2024 The Author(s).)

Background: Cognitive control training (CCT) has shown potential to reduce emotional vulnerability in adults and adolescents. However, there is scant literature testing the efficacy of CCT for the reduction of anxiety and transferring the effects to educational outcomes in children. Building on the evidence that a greater ability to suppress a prepotent response (inhibitory control) is associated with higher math achievement in children, it is plausible that training inhibitory processes using a CCT paradigm may be beneficial for reducing anxiety, improving inhibitory control, and in turn increasing math achievement., Objective: This proof-of-concept study aims to investigate the efficacy of 15 sessions of inhibitory control training for reduction in anxiety and improvement in math achievement in primary school children., Methods: We will use a 2 (group: CCT, adaptive Go/No-Go vs active control, low-load task) multiplied by 4 (time: pre- vs posttraining vs 1-month vs 3-month follow-up) randomized design in a nonselected sample of 100 children aged 8-10 years. Both groups will complete 10 minutes of daily training for 3 weeks at school. The dependent variables will be anxiety and correlates (Spence Children's Anxiety Scale, Penn State Worry Questionnaire for Children, Revised Children's Anxiety and Depression Scale, Child Response Style Questionnaire, and Modified Abbreviated Math Anxiety Scale), inhibitory control (Go/No-Go task), shifting (color-shape shifting task), updating (n-back task), and math achievement (Applied Problems, Calculation, and Math Facts Fluency subtests from the Woodcock-Johnson IV Tests of Achievement)., Results: We opened enrollment in September 2023. The initial results are expected to be published in late 2024. We predict that children in the CCT group will show a reduction in emotional symptoms; improvements in inhibition, shifting, and updating performance; and advances in math achievement from pre- to posttraining, and that these effects will be maintained at 1- and 3-month follow-ups, compared to children in the active control group., Conclusions: The CCT paradigm used in our study will provide a greater understanding of the emotional and cognitive transfer effects on children and inform future work. Specifically, the findings will advance the knowledge of deploying inhibitory control training with children and provide valuable insights into its use for reducing anxiety and advancing math achievement., Trial Registration: Open Science Framework ofs.io/de2qa; https://doi.org/10.17605/OSF.IO/DE2QA., International Registered Report Identifier (irrid): PRR1-10.2196/52929., (©Elizabeth Edwards, Khanh Linh Chu, Annemaree Carroll. Originally published in JMIR Research Protocols (https://www.researchprotocols.org), 13.03.2024.)

Objective: To examine the effect of varying fatty acid composition in a HFM on eNO, pulmonary function, and airway resistance., Methods: Fifteen individuals [6 M/9 F; 21.9 ± 1.5 years old] each completed three HFM conditions {SF, O6FA, and O3FA; 12 kcal/kg body weight, 63% total fat, and 0.72 g/kg sugar smoothies} in random order separated by at least 48 h. Airway inflammation assessed via eNO, pulmonary function measured using the maximum flow volume loop (MFVL) and airway resistance measured using impulse oscillometry (iOS) were taken at baseline, 2h and 4h postprandially., Results: There was no difference in eNO or iOS across time in any condition or between conditions ( p  > 0.05). There was a significant time by condition effect for FEV 1 post-HFM in the SF and O6FA conditions ( p  < 0.05)., Conclusion: Different fatty acid compositions do not increase eNO or iOS in healthy, college-aged participants after consumption of a HFM, though the minimally processed meals with fruit added may contribute to these findings.

Laccase-like multicopper oxidases are recognized for their potential to alter the reactivity of lignins for application in value-added products. Typically, model compounds are employed to discover such enzymes; however, they do not represent the complexity of industrial lignin substrates. In this work, a screening pipeline was developed to test enzymes simultaneously on model compounds and industrial lignins. A total of 12 lignin-active fungal multicopper oxidases were discovered, including 9 enzymes active under alkaline conditions (pH 11.0). Principal component analysis revealed the poor ability of model compounds to predict enzyme performance on industrial lignins. Additionally, sequence similarity analyses grouped these enzymes with Auxiliary Activity-1 sub-families with few previously characterized members, underscoring their taxonomic novelty. Correlation between the lignin-activity of these enzymes and their taxonomic origin, however, was not observed. These are critical insights to bridge the gap between enzyme discovery and application for industrial lignin valorization., Competing Interests: Declaration of competing interest The authors have no competing interests to declare., (Copyright © 2023 The Authors. Published by Elsevier Ltd.. All rights reserved.)

We present a data set of four metagenomes and 281 metagenome-assembled genomes describing the microbial community of a laboratory-scale high solids anaerobic digester. Our objective was to obtain information on the coding potential of the microbial community and draft genomes of the most abundant organisms in the digester., Competing Interests: The authors declare no conflict of interest.

Microbial carboxylases and decarboxylases play important roles in the global carbon cycle and have many potential applications in biocatalysis and synthetic biology. The widespread family of reversible UbiD-like (de)carboxylases are of particular interest because these enzymes are active against a diverse range of substrates. Several characterized UbiD enzymes have been shown to catalyze reversible (de)carboxylation of aromatic and aliphatic substrates using the recently discovered prenylated FMN (prFMN) cofactor, which is produced by the associated family of UbiX FMN prenyltransferases. However, discovery and investigation of novel UbiD (de)carboxylases are delayed by our limited knowledge and the experimental complexities associated with FMN prenylation and UbiD activation resulting in the production of inactive recombinant UbiD enzymes. Therefore, there is a need for developing robust methods for efficient in vitro and in vivo FMN prenylation and UbiD activation for heterologous production of active UbiD enzymes. In this chapter, we present two protocols for in vitro and in vivo FMN prenylation and UbiD activation under aerobic conditions using enzyme cascades with regenerating systems and recombinant E. coli cells., (Copyright © 2024. Published by Elsevier Inc.)

Compound specific isotope analysis (CSIA) is a powerful technique to demonstrate in situ degradation of traditional groundwater contaminants when concentrations are typically in the mg/L range. Currently, an efficient preconcentration method is lacking to expand CSIA to low aqueous concentration environmental samples. Specially for the H- and N-CSIA of heteroatom-bearing non-traditional compounds, the CSIA analytical detection limits are significantly higher than that of the C-CSIA. This work demonstrates the compatibility of polar organic chemical integrative sampler (POCIS) with C-, H-, and N-CSIA using four nitro- and amino-substituted chlorobenzenes that are common industrial feedstocks for numerous applications and are commonly detected in the environment at mg/L to μg/L range. Using lab experiments, we showed isotopic equilibrium in POCIS was achieved after 30 days with either a negligible (<0.5 ‰) or a constant shift for C (<1 ‰) and N (<2 ‰). Similar negligible (<5 ‰) or constant shift (<20 ‰) was evident for H isotope except for 3,4-dichloroaniline. The method quantification limits for the combined sorbent and membrane of one POCIS were comparable to that of the solid phase extraction (SPE) using 10 L water. Next, we demonstrated the field applicability of POCIS for C- and N-CSIA after a 60-day deployment in a pilot constructed wetland by showing <1 ‰ difference between the δ 13 C and δ 15 N obtained from POCIS and SPE. Finally, we evaluated whether the biofilm development on POCIS membrane could affect the isotope signature of the sampled compounds during field deployment. Although a diverse microbial community was identified on the membrane after a 60-day deployment, we did not observe significant isotope fractionation. This was likely due to either slower diffusion in the biofilm or microbial degradation of the sampled compounds. This work demonstrates the potential of using POCIS-CSIA as a simple, fast, and sensitive method for low-concentration contaminants, such as pesticides, pharmaceuticals, and flame-retardants., Competing Interests: Declaration of competing interest The authors declare the following financial interests/personal relationships which may be considered as potential competing interests: Elodie Passeport reports financial support was provided by Natural Sciences and Engineering Research Council of Canada (NSERC) and a MITACS grant., (Copyright © 2023 Elsevier B.V. All rights reserved.)

Objective: To assess changes in physical activity (PA) after a COVID-19 shutdown on a primarily residential university campus., Methods: Eighty students, faculty, and staff (FS) of a university (age: 32.2 ± 13.6 yr) who wore a consumer wearable technology (CWT) device completed an anonymous survey by inputting data for 30 days prior to- and 30 days following an academic break in 2020, in which the university transitioned to remote learning., Results: Steps decreased after spring break in all subjects ( p  < .001), but steps were impacted to a greater extent in students. 30-day, weekday, and weekend step averages all decreased in students ( p < .001). FS were able to maintain their weekend step averages., Conclusions: PA decreased in a university community after the COVID-19 shutdown. Students, no longer active transport for campus life, saw a greater impact on their PA. These changes could have an impact on health status.

Chloroform (CF) and dichloromethane (DCM) contaminate groundwater sites around the world but can be cleaned up through bioremediation. Although several strains of Dehalobacter restrictus can reduce CF to DCM and multiple Peptococcaceae can ferment DCM, these processes cannot typically happen simultaneously due to CF sensitivity in the known DCM-degraders or electron donor competition. Here, we present a mixed microbial culture that can simultaneously metabolize CF and DCM and create an additional enrichment culture fed only DCM. Through genus-specific quantitative polymerase chain reaction, we find that Dehalobacter grows while either CF alone or DCM alone is converted, indicating its involvement in both metabolic steps. Additionally, the culture was maintained for over 1400 days without the addition of an exogenous electron donor, and through electron balance calculations, we show that DCM metabolism would produce sufficient reducing equivalents (likely hydrogen) for CF respiration. Together, these results suggest intraspecies electron transfer could occur to continually reduce CF in the culture. Minimizing the addition of electron donor reduces the cost of bioremediation, and "self-feeding" could prolong bioremediation activity long after donor addition ends. Overall, understanding this mechanism informs strategies for culture maintenance and scale-up and benefits contaminated sites where the culture is employed for remediation worldwide.

Cancer survivors commonly contend with concurrent cognitive difficulties such as problems with attention and concentration, and psychological distress, including anxiety and depression. However, the associations between attentional and emotional difficulties within the specific context of melanoma survivors remain relatively unexplored. Premised on attentional control theory, the current study employed a cross-sectional design to explore the interplay among trait anxiety (dispositional) and situational anxiety (cancer-related worry), depression and attentional control (ability to inhibit distractors and flexibly shift within and between tasks) in a sample of 187 melanoma survivors aged 18 to 58 years ( M age = 36.83 years, SD age = 5.44 years; 93% female). Data were analyzed using a moderated multiple regression, with anxiety, cancer worry and depression as predictors, and attentional control as the criterion variable. After statistically controlling for the variance of chemotherapy, we found that individuals with higher trait anxiety and higher cancer-related worry reported greater attentional control at low levels of depression, yet poorer attentional control at high depression, relative to individuals with low anxiety. Our findings suggest that anxiety and depression are differentially related to attentional control in melanoma survivors. The results provide a marker for clinicians addressing anxiety and depression in this population. Implications for primary healthcare are discussed.

Background: Performing an episiotomy where clinically indicated is a key intervention in the Obstetric Anal Sphincter Injury Care Bundle (OASI-CB) implemented across England and Wales to reduce the risk and increase the detection of severe perineal trauma after birth. Standards of consent provided to people in maternity care generally and for episiotomy specifically have been reported as suboptimal. Compromising birthing people's personal autonomy or sense of control has been linked to a dissatisfying birth experience, negative psychological sequelae, and litigation., Methods: This study explored experienced midwives' practice of informed consent for episiotomy during a midwife-led birth. We sampled 43 midwives across eight NHS Trusts in England and Wales using online focus groups and telephone interviews about their experience of consent in episiotomy. Using qualitative content analysis and art-based co-analysis methods with eight midwives from across the research sites, we co-analyzed and co-constructed three themes and four practice recommendations from the data., Results: Three themes were constructed from the data: Assent rather than consent, Change in culture to support best practice, and Standardized information. These themes informed the shaping of four recommendations for best practice in episiotomy informed consent., Conclusion: This study has shown how variations in midwifery practice and culture may impact birthing people's experience of informed consent in episiotomy. Midwives may not have the knowledge or skills to conduct a detailed consent conversation, leading to variation in practice and messages for birthing people. The use of antenatal discussion aids can offer women the opportunity to become informed and fully participate in the decision-making process., (© 2023 The Authors. Birth published by Wiley Periodicals LLC.)

Background: Acute fluid ingestion increases estimated body fat percentage (BF%) measurements by single frequency (SF-BIA) and multi-frequency bioelectrical impedance (MF-BIA). It is unknown if MF-BIA accurately measures total BF% and total body water (TBW) after creatine supplementation, which causes fluid retention, and resultant increases in fat-free mass and TBW. The purpose of this study was to analyze the effect of creatine supplementation on body composition and TBW measured through a popular MF-BIA device (InBody 770)., Methods: Thirteen male and 14 female subjects (18-22 years) completed one week of creatine monohydrate (0.3 g/kg body weight) or maltodextrin. Pre- and post-supplementation body composition measurements included dual-energy X-ray absorptiometry (DEXA), SF-BIA measured by an Omron HBF-306C device, and MF-BIA measured by an InBody 770 device to measure BF%, fat free mass (FFM), and fat mass (FM). Additionally, intracellular water (ICW), extracellular water (ECW), and TBW were estimated by MF- BIA., Results: FFM increased more in the creatine group than the placebo group measured by all body composition modes (1.2 kg, 1.9 kg, and 1.1 kg increase for SF-BIA, MF-BIA, and DEXA respectively, P<0.05). Creatine supplementation resulted in a 2% increase (P<0.05) in TBW measured by MF-BIA (40.4±9.5 to 41.2±9.6 kg)., Conclusions: One week of creatine supplementation increased TBW as detected by the InBody 770 device. Changes in body composition that occurred due to the increase in TBW were detected as an increase in FFM measured by SF-BIA, MF-BIA, and DEXA.

Problematic smartphone use has been associated with poorer mental health in different population groups; however, little is known about how levels of smartphone use were associated with mental health outcomes of adults in Australia. Using data from a cross-sectional survey among Australian adults aged 18-59 years (n = 655, Mean = 24.55 [SD = 5.59] years; 66% female), the current study aimed to examine association between problematic smartphone use and different psychological outcomes. Participants completed measures of problematic smartphone use with Mobile Phone Problem Use Scale (MPPUS), mental health outcomes with Depression, Anxiety, and Stress Scale (DASS-21), and Pittsburgh Sleep Quality Index (PSQI), in addition to some socio-demographics. Smartphone use was categorised into three groups: low-moderate, moderate-high, and high-severe. A total of 160 adults (24.4%) reported high-severe smartphone use. Multivariable linear regression analyses showed that smartphone use was inversely associated with psychological outcomes in a dose-dependent manner with high-severe smartphone uses having the most adverse effects. Compared to low-moderate use, average depression score was 3.5 points higher for moderate-high smartphone use (β = 3.51, 95% CI: 1.63-5.40) and 6.9 points higher for high-severe smartphone use (β = 6.91, 95% CI: 4.74-9.07). Similarly, average stress score was 3.4 points higher for moderate-high smartphone use (β = 3.40, 95% CI: 1.75-5.06) and 7.0 points higher for high-severe smartphone use (β = 7.02, 95% CI: 5.11-8.93). Similar association estimates were found for anxiety and sleep quality. Reducing smartphone use has the potential to optimise depression, anxiety, stress, and sleep quality; however, longitudinal research is warranted to establish directionality of the association., (© 2023. The Author(s).)

The acute effect of exercise on β-cell function during a high-fat meal (HFM) in young adults (YA) versus old adults (OA) is unclear. In this randomized crossover trial, YA ( n = 5 M/7 F, 23.3 ± 3.9 yr) and OA ( n = 8 M/4 F, 67.7 ± 6.0 yr) underwent a 180-min HFM (12 kcal/kg body wt; 57% fat, 37% CHO) after a rest or exercise [∼65% heart rate peak (HR peak )] condition ∼12 h earlier. After an overnight fast, plasma lipids, glucose, insulin, and free fatty acid (FFA) were determined to estimate peripheral, or skeletal muscle, insulin sensitivity (Matsuda index) as well as hepatic [homeostatic model assessment of insulin resistance (HOMA-IR)] and adipose insulin resistance (adipose-IR). β-Cell function was derived from C-peptide and defined as early-phase (0-30 min) and total-phase (0-180 min) disposition index [DI, glucose-stimulated insulin secretion (GSIS) adjusted for insulin sensitivity/resistance]. Hepatic insulin extraction (HIE), body composition [dual-energy X-ray absorptiometry (DXA)], and peak oxygen consumption (V̇o 2peak ) were also assessed. OA had higher total cholesterol (TC), LDL, HIE, and DI across organs as well as lower adipose-IR (all, P < 0.05) and V̇o 2peak ( P = 0.056) despite similar body composition and glucose tolerance. Exercise lowered early-phase TC and LDL in OA versus YA ( P < 0.05). However, C-peptide area under the curve (AUC), total phase GSIS, and adipose-IR were reduced postexercise in YA versus OA ( P < 0.05). Skeletal muscle DI increased in YA and OA after exercise ( P < 0.05), whereas adipose DI tended to decline in OA ( P = 0.06 and P = 0.08). Exercise-induced skeletal muscle insulin sensitivity ( r = -0.44, P = 0.02) and total-phase DI ( r = -0.65, P = 0.005) correlated with reduced glucose AUC 180min . Together, exercise improved skeletal muscle insulin sensitivity/DI in relation to glucose tolerance in YA and OA, but only raised adipose-IR and reduced adipose-DI in OA. NEW & NOTEWORTHY High-fat diets may induce β-cell dysfunction. This study compared how young and older adults responded to a high-fat meal with regard to β-cell function and whether exercise comparably impacted glucose regulation. Older adults secreted more insulin during the high-fat meal than younger adults. Although exercise increased β-cell function adjusted for skeletal muscle insulin sensitivity in relation to glucose tolerance, it raised adipose insulin resistance and reduced pancreatic β-cell function relative to adipose tissue in older adults. Additional work is needed to discern nutrient-exercise interactions across age to mitigate chronic disease risk.

Wastewater-based surveillance has become an effective tool around the globe for indirect monitoring of COVID-19 in communities. Variants of Concern (VOCs) have been detected in wastewater by use of reverse transcription polymerase chain reaction (RT-PCR) or whole genome sequencing (WGS). Rapid, reliable RT-PCR assays continue to be needed to determine the relative frequencies of VOCs and sub-lineages in wastewater-based surveillance programs. The presence of multiple mutations in a single region of the N-gene allowed for the design of a single amplicon, multiple probe assay, that can distinguish among several VOCs in wastewater RNA extracts. This approach which multiplexes probes designed to target mutations associated with specific VOC's along with an intra-amplicon universal probe (non-mutated region) was validated in singleplex and multiplex. The prevalence of each mutation (i.e. VOC) is estimated by comparing the abundance of the targeted mutation with a non-mutated and highly conserved region within the same amplicon. This is advantageous for the accurate and rapid estimation of variant frequencies in wastewater. The N200 assay was applied to monitor frequencies of VOCs in wastewater extracts from several communities in Ontario, Canada in near real time from November 28, 2021 to January 4, 2022. This includes the period of the rapid replacement of the Delta variant with the introduction of the Omicron variant in these Ontario communities in early December 2021. The frequency estimates using this assay were highly reflective of clinical WGS estimates for the same communities. This style of qPCR assay, which simultaneously measures signal from a non-mutated comparator probe and multiple mutation-specific probes contained within a single qPCR amplicon, can be applied to future assay development for rapid and accurate estimations of variant frequencies., Competing Interests: Declaration of competing interest The authors declare the following financial interests/personal relationships which may be considered as potential competing interests: Mark Servos reports financial support was provided by Ontario Ministry of the Environment Conservation and Parks. Mark Servos reports financial support was provided by Natural Sciences and Engineering Research Council of Canada. Mark Servos reports financial support was provided by Global Water Futures., (Copyright © 2023 Elsevier B.V. All rights reserved.)

The Candidate Phyla Radiation (CPR), also referred to as superphylum Patescibacteria , is a very large group of bacteria with no pure culture representatives discovered by 16S rRNA sequencing or genome-resolved metagenomic analyses of environmental samples. Within the CPR, candidate phylum Parcubacteria , previously referred to as OD1, is prevalent in anoxic sediments and groundwater. Previously, we had identified a specific member of the Parcubacteria (referred to as DGGOD1a) as an important member of a methanogenic benzene-degrading consortium. Phylogenetic analyses herein place DGGOD1a within the clade " Candidatus Nealsonbacteria." Because of its persistence over many years, we hypothesized that " Ca. Nealsonbacteria" DGGOD1a must play an important role in sustaining anaerobic benzene metabolism in the consortium. To try to identify its growth substrate, we amended the culture with a variety of defined compounds (pyruvate, acetate, hydrogen, DNA, and phospholipid), as well as crude culture lysate and three subfractions thereof. We observed the greatest (10-fold) increase in the absolute abundance of " Ca. Nealsonbacteria" DGGOD1a only when the consortium was amended with crude cell lysate. These results implicate " Ca. Nealsonbacteria" in biomass recycling. Fluorescence in situ hybridization and cryogenic transmission electron microscope images revealed that " Ca. Nealsonbacteria" DGGOD1a cells were attached to larger archaeal Methanothrix cells. This apparent epibiont lifestyle was supported by metabolic predictions from a manually curated complete genome. This is one of the first examples of bacterial-archaeal episymbiosis and may be a feature of other " Ca. Nealsonbacteria" found in anoxic environments. IMPORTANCE An anaerobic microbial enrichment culture was used to study members of candidate phyla that are difficult to grow in the lab. We were able to visualize tiny " Candidatus Nealsonbacteria" cells attached to a large Methanothrix cell, revealing a novel episymbiosis., Competing Interests: The authors declare no conflict of interest.

Draft and complete metagenome assembled genomes (MAGs) were created from multiple metagenomic assemblies of DGG-B, a strictly anaerobic, stable mixed microbial consortium that degrades benzene completely to methane and CO 2 . Our objective was to obtain closed genome sequences of benzene-fermenting bacteria to enable the elucidation of their elusive anaerobic benzene degradation pathway., Competing Interests: The authors declare no conflict of interest.

Background: Knowledge of the specific dynamics of influenza introduction and spread in university settings is limited., Methods: Persons with acute respiratory illness symptoms received influenza testing by molecular assay during October 6-November 23, 2022. Viral sequencing and phylogenetic analysis were conducted on nasal swab samples from case-patients. Case-control analysis of a voluntary survey of persons tested was used to identify factors associated with influenza; logistic regression was conducted to calculate odds ratios and 95% CIs. A subset of case-patients tested during the first month of the outbreak was interviewed to identify sources of introduction and early spread., Results: Among 3268 persons tested, 788 (24.1%) tested positive for influenza; 744 (22.8%) were included in the survey analysis. All 380 sequenced specimens were influenza A (H3N2) virus clade 3C.2a1b.2a.2, suggesting rapid transmission. Influenza (OR [95% CI]) was associated with indoor congregate dining (1.43 [1.002-2.03]), attending large gatherings indoors (1.83 [1.26-2.66]) or outdoors (2.33 [1.64-3.31]), and varied by residence type (apartment with ≥1 roommate: 2.93 [1.21-7.11], residence hall room alone: 4.18 [1.31-13.31], or with roommate: 6.09 [2.46-15.06], or fraternity/sorority house: 15.13 [4.30-53.21], all compared with single-dwelling apartment). Odds of influenza were lower among persons who left campus for ≥1 day during the week before their influenza test (0.49 [0.32-0.75]). Almost all early cases reported attending large events., Conclusions: Congregate living and activity settings on university campuses can lead to rapid spread of influenza following introduction. Isolating following a positive influenza test or administering antiviral medications to exposed persons may help mitigate outbreaks., (© 2023 The Authors. Influenza and Other Respiratory Viruses published by John Wiley & Sons Ltd. This article has been contributed to by U.S. Government employees and their work is in the public domain in the USA.)

Naphthoquinone (1,4-NQ) and its derivatives (NQs, juglone, plumbagin, 2-methoxy-1,4-NQ, and menadione) have a variety of therapeutic applications, many of which are attributed to redox cycling and the production of reactive oxygen species (ROS). We previously demonstrated that NQs also oxidize hydrogen sulfide (H 2 S) to reactive sulfur species (RSS), potentially conveying identical benefits. Here we use RSS-specific fluorophores, mass spectroscopy, EPR and UV-Vis spectrometry, and oxygen-sensitive optodes to examine the effects of thiols and thiol-NQ adducts on H 2 S-NQ reactions. In the presence of glutathione (GSH) and cysteine (Cys), 1,4-NQ oxidizes H 2 S to both inorganic and organic hydroper-/hydropolysulfides (R 2 S n , R=H, Cys, GSH; n = 2-4) and organic sulfoxides (GS n OH, n = 1, 2). These reactions reduce NQs and consume oxygen via a semiquinone intermediate. NQs are also reduced as they form adducts with GSH, Cys, protein thiols, and amines. Thiol, but not amine, adducts may increase or decrease H 2 S oxidation in reactions that are both NQ- and thiol-specific. Amine adducts also inhibit the formation of thiol adducts. These results suggest that NQs may react with endogenous thiols, including GSH, Cys, and protein Cys, and that these adducts may affect both thiol reactions as well as RSS production from H 2 S.

Background: One of four key points in the Obstetric Anal Sphincter Injury Care Bundle, first piloted in the UK in 2016, was the directive to perform episiotomy when clinically indicated. Midwives are the primary health care professional for straightforward births in the UK and there is very little published literature that relates to their practice in this area., Aim: The aim of the study was to explore experienced midwives' decision-making processes in their assessments for episiotomy during birth., Methods: 43 midwives self-identifying as confident in performing episiotomy were sampled across 8 NHS Trusts in England and Wales. Data collection was via online focus groups and 1:1 interviews. Primary thematic analysis was undertaken by the research team. Preliminary themes were used to structure a co-production analysis workshop where eight experienced midwives undertook a secondary analysis of the data resulting in four overall themes., Findings: Four themes were identified, 'Optimising Perineal Function', 'Red Flags to Stimulate Decision-Making', 'The Midwives' Episiotomy' and 'Infiltration as a Catalyst for Birth'., Discussion: Midwives use a number of visual, auditory and touch cues to inform their assessments for episiotomy during birth., Conclusion: This study provides valuable insight into the cues that guide experienced midwives' decision-making in relation to episiotomy and contributes evidence related to performing episiotomy when clinically indicated in spontaneous vaginal birth., Competing Interests: Conflict of interest The authors have no financial or other interest in the product or distributor of the product., (Copyright © 2022. Published by Elsevier Ltd.)

A 50-kg scale, high solids anaerobic digester (AD) comprising six sequentially fed leach beds with a leachate recirculation system was operated at 37°C for 88 weeks. The solid feedstock contained a constant fibre fraction (a mix of cardboard, boxboard, newsprint, and fine paper) and varying proportions of food waste. Previously, we reported on the stable operation of this digestion system, where significantly enhanced methane production from the fibre fraction was observed as the proportion of food waste increased. The objective of this study was to identify relationships between process parameters and the microbial community. Increasing food waste led to a large increase in the absolute microbial abundance in the circulating leachate. While 16S rRNA amplicons for Clostridium butyricum were most abundant and correlated with the amount of FW in the system and with the overall methane yield, it was more cryptic Candidatus Roizmanbacteria and Spirochaetaceae that correlated specifically with enhanced methane from the fiber fraction. A faulty batch of bulking agent led to hydraulic channeling, which was reflected in the leachate microbial profiles matching that of the incoming food waste. The system performance and microbial community re-established rapidly after reverting to better bulking agent, illustrating the robustness of the system., (© The Author(s) 2023. Published by Oxford University Press on behalf of FEMS.)

Introduction: Chronic diets high in saturated fat (SF) and omega-6-fatty acids (O6FAs) elevate fasting triglycerides (TRGs) and glucose (GLU). Postprandial TRGs, GLU, and Metabolic Load Index (MLI) are better predictors of disease risk compared to fasting levels alone. Conversely, diets high in omega-3 fatty acids (O3FAs) may be cardioprotective. Unfortunately, many existing postprandial studies are not standardized to body weight and given in an amount individuals would typically consume in their daily lives; the MLI is not calculated, and varying types of fat content are not examined. Therefore, we sought to determine whether SF, O3FAs, or O6FAs altered postprandial TRGs, GLU, and MLI from a standardized mixed meal. Methods: Fifteen individuals (6 M and 9 F) visited the laboratory three times, separated by at least 48 h, to consume HFM smoothies with varying FA composition (SF, high O6FAs, and high O3FAs). The smoothies were standardized to 12 kcal/kg body weight, 63% total fat, and 0.72 g/kg sugar. TRGs and GLU were collected at baseline and at 2 h and 4 h postprandially; the MLI was calculated by summing the TRG and GLU responses at each time point. Results: There was a significant increase in TRGs across time points ( p < 0.001). For TRGs, there was a trend toward a significant interaction between smoothie type and time ( p = 0.06) due to the increase in TRGs in the SF compared to the O3FA smoothie. There was an increase in postprandial GLU that varied across smoothie types ( p = 0.036). Taken together, the MLI was elevated in the SF smoothie compared to the O3FAs at 2 h ( p = 0.041). Conclusion: A SF smoothie in the morning elevated the metabolic load compared to an O3FA smoothie. Mechanisms of action in the competing clearance of TRGs and GLU warrant further investigation.

Background: High screen use has been adversely linked with mental wellbeing; however, little is known about how active versus passive screen time are associated with sleep-onset difficulties among adolescents., Methods: We analysed data from 38 European and North American countries that participated in the 2014 Health Behaviour in School-aged Children (HBSC) survey. Difficulties in falling sleep were assessed using a self-reported item with a 5-point Likert scale, and then dichotomised. Participants reported h/day of discretional time spent watching television, electronic gaming, and computer use., Results: Of the 195,668 participants (M age 13.59 [1.62] years; 51 % girls), about 25 % of girls reported sleep-onset difficulties, while the rate was 18 % in boys. Adolescents who played electronic games >4 h/day (≤1 h/day as reference) had 30 % higher odds in boys (OR 1.30; 95 % CI: 1.23-1.38) and 38 % higher odds in girls (OR 1.38; 95 % CI: 1.31-1.45) of reporting sleep difficulties. High computer use (>4 h/day) increased the odds of sleep difficulties by 41 % in boys (OR 1.41, 95 % CI: 1.33-1.49) and 61 % in girls (OR 1.61, 95 % CI: 1.53-1.69). Similarly, high television time (>4 h/day) had increased the odds of sleep difficulties by 15 % in boys (OR 1.15, 95 % CI: 1.08-1.22) and 19 % in girls (OR 1.19, 95 % CI: 1.12-1.25)., Limitations: Cross-sectional analyses cannot establish causality of the associations., Conclusions: Higher levels of recreational screen use of any type were associated with sleep-onset difficulties among adolescents with adverse effects being more prevalent in active than passive screen time. Prospective research with objective measures is warranted to understand causality of these relationships., Competing Interests: Conflict of interest None to declare., (Copyright © 2022 Elsevier B.V. All rights reserved.)

Clinical testing has been the cornerstone of public health monitoring and infection control efforts in communities throughout the COVID-19 pandemic. With the anticipated reduction of clinical testing as the disease moves into an endemic state, SARS-CoV-2 wastewater surveillance (WWS) will have greater value as an important diagnostic tool. An in-depth analysis and understanding of the metrics derived from WWS is required to interpret and utilize WWS-acquired data effectively (McClary-Gutierrez et al., 2021; O'Keeffe, 2021). In this study, the SARS-CoV-2 wastewater signal to clinical cases (WC) ratio was investigated across seven cities in Canada over periods ranging from 8 to 21 months. This work demonstrates that significant increases in the WC ratio occurred when clinical testing eligibility was modified to appointment-only testing, identifying a period of insufficient clinical testing (resulting in a reduction to testing access and a reduction in the number of daily tests) in these communities, despite increases in the wastewater signal. Furthermore, the WC ratio decreased significantly in 6 of the 7 studied locations, serving as a potential signal of the emergence of the Alpha variant of concern (VOC) in a relatively non-immunized community (40-60 % allelic proportion), while a more muted decrease in the WC ratio signaled the emergence of the Delta VOC in a relatively well-immunized community (40-60 % allelic proportion). Finally, a significant decrease in the WC ratio signaled the emergence of the Omicron VOC, likely because of the variant's greater effectiveness at evading immunity, leading to a significant number of new reported clinical cases, even when community immunity was high. The WC ratio, used as an additional monitoring metric, could complement clinical case counts and wastewater signals as individual metrics in its potential ability to identify important epidemiological occurrences, adding value to WWS as a diagnostic technology during the COVID-19 pandemic and likely for future pandemics., Competing Interests: Declaration of competing interest The authors declare that no known competing financial interests or personal relationships influenced the work reported in this manuscript., (Copyright © 2022 The Authors. Published by Elsevier B.V. All rights reserved.)

Sports participation has been linked to various health outcomes; however, there is scant literature exploring this relationship in developing countries. We used a mixed method approach to examine the association between sports participation and mental health of Bangladeshi adolescents ( n = 320; 13-17 years; 59% boys) and to explore sports preferences and barriers to sports participation across genders. A survey collected team and non-team sports participation, depression, and life satisfaction. Focus groups (16 boys, 16 girls) explored preferences for, and barriers to, sports participation. Regression analysis showed that higher team and non-team sports participation were associated with lower depressive symptoms in boys (β = -1.22, 95% CI:-2.55 to -0.10; β = -2.50, 95% CI:-3.83 to -1.16, respectively), while greater participation in team sports was associated with less depression in girls (β = -2.44, 95% CI:-4.63 to -0.24). Participation in team and non-team sports was positively associated with life satisfaction for boys and girls. Boys reported preferences for playing football and cricket, while girls favored skipping and running. Prolonged time on electronic devices was reported as barriers to sports participation in both genders. Furthermore, household chores, family restrictions, and unsafe environment were reported by girls. Participation in team sports may provide mental health benefits for both genders, while non-team sports may be more beneficial to boys than girls.

The present study investigated the relationship between anxiety, social support, living arrangements and cognitive performance of university students during the global pandemic. Two hundred and fifteen students participated by completing online questionnaires. Separate moderated multiple regression models were used to test whether social support (Family, Friends, Significant Other subscales of the Multidimensional Scale of Perceived Social Support) moderated the relationship between anxiety (Anxiety subscale of Depression, Anxiety Stress Scale), living arrangements (Living Alone vs Living with Friends and Family) and cognitive performance (Cognitive Failures Questionnaire), after controlling for comorbid depression. The results for each level of perceived social support suggested that anxiety was negatively associated with cognitive performance. Our most significant finding was that for students living alone, social support from a significant other offered a protective factor, whereby buffering the anxiety related cognitive deficits prevalent in those who reported lower social support. These data have important practical implications for supporting the social-emotional and academic needs of university students during the global pandemic., (© 2022 Elsevier Ltd. All rights reserved.)

We investigated the impact of oxygen on a strictly anaerobic, methanogenic benzene-degrading enrichment culture derived decades ago from oil-contaminated sediment. The culture includes a benzene fermenter from Deltaproteobacteria candidate clade Sva0485 (referred to as ORM2) and methanogenic archaea. A one-time injection of 0.1 mL air , simulating a small leak into 30 mL batch culture bottle, had no measurable impact on benzene degradation rates, although retrospectively, a tiny enrichment of aerobic taxa was detected. A subsequent 100 times larger injection of air stalled methanogenesis and caused drastic perturbation of the microbial community. A benzene-degrading Pseudomonas became highly enriched and consumed all available oxygen. Anaerobic benzene-degrading ORM2 cell numbers plummeted during this time; re-growth and associated recovery of methanogenic benzene degradation took almost 1 year. These results highlight the oxygen sensitivity of this methanogenic culture and confirm that the mechanism for anaerobic biotransformation of benzene is independent of oxygen, fundamentally different from established aerobic pathways, and is carried out by distinct microbial communities. The study also highlights the importance of including microbial decay in characterizing and modeling mixed microbial communities.

The purpose of the present study was to determine fasting and high-fat meal (HFM)-induced post-prandial systemic inflammation and airway inflammation (exhaled nitric oxide (eNO)) in older adults (OAs) compared to younger adults (YAs) before and after acute exercise. Twelve YAs (23.3 ± 3.9 y n = 5 M/7 F) and 12 OAs (67.7 ± 6 y, n = 8 M/4 F) completed two HFM challenges. After an overnight fast, participants underwent an HFM session or pre-prandial exercise (EX, 65% VO2Peak to expend 75% of the caloric content of the HFM) plus HFM (EX + HFM) in a randomized order. Systemic inflammatory cytokines were collected at 0, 3, and 6 h, while eNO was determined at 0, 2, and 4 h after the HFM (12 kcal/kg body weight: 61% fat, 35% CHO, 4% PRO). TNF-α was higher in OAs compared to YAs (p = 0.005) and decreased across time from baseline to 6 h post-HFM (p = 0.007). In response to the HFM, IL-6 decreased from 0 to 3 h but increased at 6 h regardless of age or exercise (p = 0.018). IL-8 or IL-1β did not change over the HFM by age or exercise (p > 0.05). eNO was also elevated in OAs compared to YAs (p = 0.003) but was not altered by exercise (p = 0.108). There was a trend, however, towards significance post-prandially in OAs and YAs from 0 to 2 h (p = 0.072). TNF-α and eNO are higher in OAs compared to YAs but are not elevated more in OAs post-prandially compared to YAs. Primary systemic inflammatory cytokines and eNO were not modified by acute exercise prior to an HFM.

Chlorothalonil (2,4,5,6-tetrachloroisophthalonitrile, TePN) is one of the most widely used fungicides all over the world. Its major environmental transformation product 4-hydroxy-chlorothalonil (4-hydroxy-2,5,6-trichloroisophthalonitrile, 4-OH-TPN) is more persistent, mobile, and toxic and is frequently detected at a higher concentration in various habitats compared to its parent compound TePN. Further microbial transformation of 4-OH-TPN has never been reported. In this study, we demonstrated that 4-OH-TPN underwent complete microbial reductive dehalogenation to 4-hydroxy-isophthalonitrile via 4-hydroxy-dichloroisophthalonitrile and 4-hydroxy-monochloroisophthalonitrile. 16S rRNA gene amplicon sequencing demonstrated that Dehalogenimonas species was enriched from 6% to 17-22% after reductive dechlorination of 77.24 μmol of 4-OH-TPN. Meanwhile, Dehalogenimonas copies increased by one order of magnitude and obtained a yield of 1.78 ± 1.47 × 10 8 cells per μmol Cl - released ( N = 6), indicating that 4-OH-TPN served as the terminal electron acceptor for organohalide respiration of Dehalogenimonas species. A draft genome of Dehalogenimonas species was assembled through metagenomic sequencing, which harbors 30 putative reductive dehalogenase genes. Syntrophobacter , Acetobacterium , and Methanosarcina spp. were found to be the major non-dechlorinating populations in the microbial community, who might play important roles in the reductive dechlorination of 4-OH-TPN by the Dehalogenimonas species. This study first reports that Dehalogenimonas sp. can also respire on the seemingly dead-end product of TePN, paving the way to complete biotransformation of the widely present TePN and broadening the substrate spectrum of Dehalogenimonas sp. to polychlorinated hydroxy-benzonitrile.