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Recent research on antimicrobial resistance in the environment has emphasized targeted methods. Increased use of non-targeted methods is needed to determine how frequently novel resistance genes arise.

The design of high-intensity functional training (HIFT; e. g., CrossFit®) workouts and targeted physiological trait(s) vary on any given training day, week, or cycle. Daily workouts are typically comprised of different modality and exercise combinations that are prescribed across a wide range of intensities and durations. The only consistent aspect appears to be the common instruction to maximize effort and workout density by either completing “as many repetitions as possible” within a time limit (e.g., AMRAP, Tabata) or a list of exercises as quickly as possible. However, because effort can vary within and across workouts, the impact on an athlete's physiology may also vary daily. Programming that fails to account for this variation or consider how targeted physiological systems interrelate may lead to overuse, maladaptation, or injury. Athletes may proactively monitor for negative training responses, but any observed response must be tied to a quantifiable workload before meaningful changes (to programming) are possible. Though traditional methods exist for quantifying the resistance training loads, gymnastic movements, and cardiorespiratory modalities (e.g., cycling running) that might appear in a typical HIFT workout, those methods are not uniform, and their meaning will vary based on a specific exercise's placement within a HIFT workout. To objectively quantify HIFT workloads, the calculation must overcome differences in measurement standards used for each modality, be able to account for a component's placement within the workout and be useful regardless of how a workout is commonly scored (e.g., repetitions completed vs. time-to-completion) so that comparisons between workouts are possible. This review paper discusses necessary considerations for quantifying various HIFT workout components and structures, and then details the advantages and shortcomings of different methods used in practice and the scientific literature. Methods typically used in practice range from being excessively tedious and not conducive for making comparisons within or across workouts, to being overly simplistic, based on faulty assumptions, and inaccurate. Meanwhile, only a few HIFT-related studies have attempted to report relevant workloads and have predominantly relied on converting component and workout performance into a rate (i.e., repetitions per minute or second). Repetition completion rate may be easily and accurately tracked and allows for intra- and inter-workout comparisons. Athletes, coaches, and sports scientists are encouraged to adopt this method and potentially pair it with technology (e.g., linear position transducers) to quantify HIFT workloads. Consistent adoption of such methods would enable more precise programming alterations, and it would allow fair comparisons to be made between existing and future research.

Antimicrobial resistance is a growing public health problem that requires an integrated approach among human, agricultural, and environmental sectors. However, few studies address all three components simultaneously. We investigated the occurrence of five antibiotic resistance genes (ARGs) and the class 1 integron gene (intI1) in private wells drawing water from a vulnerable aquifer influenced by residential septic systems and land-applied dairy manure. Samples (n = 138) were collected across four seasons from a randomized sample of private wells in Kewaunee County, Wisconsin. Measurements of ARGs and intI1 were related to microbial source tracking (MST) markers specific to human and bovine feces; they were also related to 54 risk factors for contamination representing land use, rainfall, hydrogeology, and well construction. ARGs and intI1 occurred in 5-40% of samples depending on target. Detection frequencies for ARGs and intI1 were lowest in the absence of human and bovine MST markers (1-30%), highest when co-occurring with human and bovine markers together (11-78%), and intermediate when co-occurring with just one type of MST marker (4-46%). Gene targets were associated with septic system density more often than agricultural land, potentially because of the variable presence of manure on the landscape. Determining ARG prevalence in a rural setting with mixed land use allowed an assessment of the relative contribution of human and bovine fecal sources. Because fecal sources co-occurred with ARGs at similar rates, interventions intended to reduce ARG occurrence may be most effective if both sources are considered. This article is protected by copyright. All rights reserved.Investigated antibiotic resistance genes (ARGs) in groundwater influenced by septic effluent and dairy manure. Associated ARGs and intI1 with microbial source tracking (MST) markers specific to humans and bovines. Observed gradient in level of ARG contamination: no MST markers presentbovine ≈ humanboth. Land use, hydrogeology, and rainfall were positively associated with ARG contamination. Both fecal sources contribute to ARG contamination of groundwater in this setting; neither dominates.

The design of high-intensity functional training (HIFT; e. g., CrossFit

Infection risk from waterborne pathogens can be estimated via quantitative microbial risk assessment (QMRA) and forms an important consideration in the management of public groundwater systems. However, few groundwater QMRAs use site-specific hazard identification and exposure assessment, so prevailing risks in these systems remain poorly defined. We estimated the infection risk for 9 waterborne pathogens based on a 2-year pathogen occurrence study in which 964 water samples were collected from 145 public wells throughout Minnesota, USA. Annual risk across all nine pathogens combined was 3.3 × 10

The spike protein of severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) is critical for virus infection through the engagement of the human ACE2 protein1 and is a major antibody target. Here we show that chronic infection with SARS-CoV-2 leads to viral evolution and reduced sensitivity to neutralizing antibodies in an immunosuppressed individual treated with convalescent plasma, by generating whole-genome ultra-deep sequences for 23 time points that span 101 days and using in vitro techniques to characterize the mutations revealed by sequencing. There was little change in the overall structure of the viral population after two courses of remdesivir during the first 57 days. However, after convalescent plasma therapy, we observed large, dynamic shifts in the viral population, with the emergence of a dominant viral strain that contained a substitution (D796H) in the S2 subunit and a deletion (ΔH69/ΔV70) in the S1 N-terminal domain of the spike protein. As passively transferred serum antibodies diminished, viruses with the escape genotype were reduced in frequency, before returning during a final, unsuccessful course of convalescent plasma treatment. In vitro, the spike double mutant bearing both ΔH69/ΔV70 and D796H conferred modestly decreased sensitivity to convalescent plasma, while maintaining infectivity levels that were similar to the wild-type virus.The spike substitution mutant D796H appeared to be the main contributor to the decreased susceptibility to neutralizing antibodies, but this mutation resulted in an infectivity defect. The spike deletion mutant ΔH69/ΔV70 had a twofold higher level of infectivity than wild-type SARS-CoV-2, possibly compensating for the reduced infectivity of the D796H mutation. These data reveal strong selection on SARS-CoV-2 during convalescent plasma therapy, which is associated with the emergence of viral variants that show evidence of reduced susceptibility to neutralizing antibodies in immunosuppressed individuals.

Anaerobic digestion has been suggested as an intervention to attenuate antibiotic resistance genes (ARGs) in livestock manure but supporting data have typically been collected at laboratory scale. Few studies have quantified ARG fate during full-scale digestion of livestock manure. We sampled untreated manure and digestate from seven full-scale mesophilic dairy manure digesters to assess ARG fate through each system. Samples were collected biweekly from December through August (i.e., winter, spring, and summer; n = 235 total) and analyzed by quantitative polymerase chain reaction for intI1, erm(B), sul1, tet(A), and tet(W). Concentrations of intI1, sul1, and tet(A) decreased during anaerobic digestion, but their removal was less extensive than expected based on previous laboratory studies. Removal for intI1 during anaerobic digestion equaled 0.28 ± 0.03 log

This practical theological multidisciplinary research investigates the leadership stances in a Christian developmental organisation operating in gangster-ridden communities in the Cape Flats within the Cape Town conurbation. In this article “stance” is used as a technical term referring to certain well-defined aspects of a leader’s outlook and approach. The organisation is evaluated using a Christian transcendental leadership stance developed by integrating the transactional, transformational and transcendental business leadership stances with Christian spirituality. The analysis reveals that, within the overall transcendental leadership stance of the researched organisation, the primary purpose and motivational incentivisation is transformational. It is suggested that the research may provide leaders of other and African Christian developmental organisations with useful and productive insights into their own organisations.

Background: Groundwater quality in the Silurian dolomite aquifer in northeastern Wisconsin, USA, has become contentious as dairy farms and exurban development expand. Objectives: We investigated private household wells in the region, determining the extent, sources, and risk factors of nitrate and microbial contamination. Methods: Total coliforms, Escherichia coli, and nitrate were evaluated by synoptic sampling during groundwater recharge and no-recharge periods. Additional seasonal sampling measured genetic markers of human and bovine fecal-associated microbes and enteric zoonotic pathogens. We constructed multivariable regression models of detection probability (log-binomial) and concentration (gamma) for each contaminant to identify risk factors related to land use, precipitation, hydrogeology, and well construction. Results: Total coliforms and nitrate were strongly associated with depth-to-bedrock at well sites and nearby agricultural land use, but not septic systems. Both human wastewater and cattle manure contributed to well contamination. Rotavirus group A, Cryptosporidium, and Salmonella were the most frequently detected pathogens. Wells positive for human fecal markers were associated with depth-to-groundwater and number of septic system drainfield within 229m. Manure-contaminated wells were associated with groundwater recharge and the area size of nearby agricultural land. Wells positive for any fecal-associated microbe, regardless of source, were associated with septic system density and manure storage proximity modified by bedrock depth. Well construction was generally not related to contamination, indicating land use, groundwater recharge, and bedrock depth were the most important risk factors. Discussion: These findings may inform policies to minimize contamination of the Silurian dolomite aquifer, a major water supply for the U.S. and Canadian Great Lakes region. https://doi.org/10.1289/EHP7813

Pd0 -catalyzed Mizoroki-Heck reactions traditionally exhibit poor reactivity with polysubstituted, unbiased alkenes. Intermolecular reactions with simple, all-carbon tetrasubstituted alkenes are unprecedented. Herein we report that pendant carboxylic acids, combined with bulky monophospine ligands on palladium, can direct the arylation of tri- and tetrasubstituted olefins. Quaternary carbons are established at high Fsp3 attached-ring junctures and the carboxylate directing group can be removed after coupling. Carboxylate directivity prevents over-arylation of the new, less substituted alkene, which can be diversified in subsequent reactions.

Objectives\ud \ud Rugby Union has adapted the Sports Concussion Assessment Tool (SCAT) into an abridged off-field concussion screen and the complete SCAT is used during diagnostic screens performed after head impact events. No firm guidelines exist as to what should be considered “abnormal” and warrant further evaluation. This study evaluates SCAT performances in 13,479 baseline SCAT assessments, and proposes clear reference limits for each sub-component of the SCAT5. Baseline reference limits are proposed to guide management of baseline testing by identifying abnormal sub-tests, enhancing the clinical validity of baseline screens, while clinical reference limits are identified to support concussion diagnosis when no baseline is available.\ud \ud \ud \ud Design\ud \ud Cross sectional census sample.\ud \ud \ud \ud Methods\ud \ud 13,479 baseline SCATs from 7565 elite male rugby players were evaluated. Baseline reference limits were identified for each sub-test as the sub-test result achieved by approximately 5% of the population, while clinical references limits corresponded to the sub-test score achieved by as close as possible to 50% of the cohort.\ud \ud \ud \ud Results\ud \ud Players reported symptoms 35% (95% CI 1.29–1.42) more frequently during SCAT5 than SCAT3 baseline assessments (mean 1.4 ± 2.7 vs 1.0 ± 2.4). Ceiling effects were identified for many cognitive sub-tests within the SCAT. Baseline and Clinical reference limits corresponding to the worst performing 5th percentile and 50th percentile were described.\ud \ud \ud \ud Conclusions\ud \ud Targeted baseline re-testing should be repeated when abnormal sub-tests are identified according to proposed baseline reference limits, while a more conservative clinical reference limit supports concussion diagnosis during screens in diagnostic settings.

Transmission of SARS-CoV-2 is uncontrolled in many parts of the world; control is compounded in some areas by the higher transmission potential of the B.1.1.7 variant1, which has now been reported in 94 countries. It is unclear whether the response of the virus to vaccines against SARS-CoV-2 on the basis of the prototypic strain will be affected by the mutations found in B.1.1.7. Here we assess the immune responses of individuals after vaccination with the mRNA-based vaccine BNT162b22. We measured neutralizing antibody responses after the first and second immunizations using pseudoviruses that expressed the wild-type spike protein or a mutated spike protein that contained the eight amino acid changes found in the B.1.1.7 variant. The sera from individuals who received the vaccine exhibited a broad range of neutralizing titres against the wild-type pseudoviruses that were modestly reduced against the B.1.1.7 variant. This reduction was also evident in sera from some patients who had recovered from COVID-19. Decreased neutralization of the B.1.1.7 variant was also observed for monoclonal antibodies that target the N-terminal domain (9 out of 10) and the receptor-binding motif (5 out of 31), but not for monoclonal antibodies that recognize the receptor-binding domain that bind outside the receptor-binding motif. Introduction of the mutation that encodes the E484K substitution in the B.1.1.7 background to reflect a newly emerged variant of concern (VOC 202102/02) led to a more-substantial loss of neutralizing activity by vaccine-elicited antibodies and monoclonal antibodies (19 out of 31) compared with the loss of neutralizing activity conferred by the mutations in B.1.1.7 alone. The emergence of the E484K substitution in a B.1.1.7 background represents a threat to the efficacy of the BNT162b2 vaccine.

The unprecedented widespread closing of buildings due to the COVID-19 pandemic has allowed water to stagnate in premise plumbing systems, creating conditions that may facilitate the growth of opportunistic pathogens. In this study, we flushed and collected samples from showers in buildings that had been unoccupied for approximately two months and quantified Legionella pneumophila using a commercial cultivation-based assay. In addition, all bacteria, Legionella spp., L. pneumophila, L. pneumophila serogroup 1, non-tuberculous mycobacteria (NTM), and Mycobacterium avium complex (MAC) were analyzed using quantitative PCR (qPCR). Despite low or negligible total chlorine in the stagnant pre-flush water samples, L. pneumophila were not detected by either method; Legionella spp., NTM, and MAC, however, were widespread. Using quantitative microbial risk assessment (QMRA), estimated risks of clinical illness from exposure to legionella and MAC via showering were generally low, but the risk of subclinical infection via Legionella spp. could exceed a 10-7 daily risk threshold if just a small fraction (≥0.1 %) of those legionellae detected by qPCR are highly infectious. Flushing cold and hot water lines rapidly restored a total chlorine (as chloramine) residual and decreased all bacterial gene targets to building inlet water levels within 30 min. Following flushing, the chlorine residual rapidly dissipated and bacterial gene targets rebounded, approaching pre-flush concentrations after 6 to 7 days of stagnation. These results suggest that stagnant water in premise plumbing may contain elevated levels of opportunistic pathogens; flushing, however, can rapidly improve water quality and reduce the health risk but the improvement will be short-lived if building disuse persists.

Combined sewer overflows (CSOs) are known contributors of human fecal pollution in urban waterways. Exposure to these waterways occurs during recreational activities, including swimming, wading, and fishing. This study used quantitative microbial risk assessment (QMRA) to estimate the risk of acute gastrointestinal illness (AGI) due to recreation during CSO-impacted ( 24 hours after a CSO) conditions. Water samples (n = 69) were collected from two creeks and one river in Philadelphia from June-August 2017-2019. HF183 concentrations were measured to estimate concentrations of five reference pathogens: Cryptosporidium, Giardia, norovirus, E. coli O157:H7, and Salmonella. Observational data on the types and frequency of recreational exposures were also collected. Results found that recreating 24 hours after a CSO. However, estimated health risks were still high for some exposure scenarios that occurred >24 hours after a CSO. Crudes estimates determined that recreational activities along known CSO-impacted sites may account for 1-8% of all cases of salmonellosis, cryptosporidiosis, and giardiasis in the city of Philadelphia. Findings support risk reduction strategies that aim to reduce the frequency of CSOs in urban settings and may help target risk mitigation strategies.

We review a technique for solving a class of classical linear partial differential systems of relevance to physics in Minkowski spacetime. All the equations are amenable to analysis in terms of complex solutions in the kernel of the scalar Laplacian and a complexified Hertz potential. The complexification prescription ensures the existence of regular physical solutions with chirality and propagating, non-singular, pulse-like characteristics that are bounded in all three spatial dimensions. The technique is applied to the source-free Maxwell, Bopp-Land\'e-Podolsky and linearised Einstein field systems, and particular solutions are used for constructing classical models describing single-cycle laser pulses and a mechanism is discussed for initiating astrophysical jets. Our article concludes with a brief introduction to spacetime Clifford algebra ideals that we use to represent spinor fields. We employ these to demonstrate how the same technique used for tensor fields enables one to construct new propagating, chiral, non-singular, pulse-like spinor solutions to the massless Dirac equation in Minkowski spacetime., Comment: 31 pages, 6 figures, written for Tekin Dereli's 70th birthday Festschrift celebration. arXiv admin note: text overlap with arXiv:1805.08825

Neural implicit shape representations are an emerging paradigm that offers many potential benefits over conventional discrete representations, including memory efficiency at a high spatial resolution. Generalizing across shapes with such neural implicit representations amounts to learning priors over the respective function space and enables geometry reconstruction from partial or noisy observations. Existing generalization methods rely on conditioning a neural network on a low-dimensional latent code that is either regressed by an encoder or jointly optimized in the auto-decoder framework. Here, we formalize learning of a shape space as a meta-learning problem and leverage gradient-based meta-learning algorithms to solve this task. We demonstrate that this approach performs on par with auto-decoder based approaches while being an order of magnitude faster at test-time inference. We further demonstrate that the proposed gradient-based method outperforms encoder-decoder based methods that leverage pooling-based set encoders., Project website: https://vsitzmann.github.io/metasdf/

Objectives\ud \ud The vast majority of rugby union (‘rugby’) participants are community-based players; however, the majority of injury surveillance studies reported relate to the elite, professional game. A potential reason for this dearth of studies could be the perceived difficulty of using the consensus statement for injury recording at the community level. The aim of this study was to identify areas where the consensus statement could be adapted for easier and more appropriate implementation within the community setting.\ud \ud Design\ud \ud Round-table discussion\ud \ud Methods\ud \ud All community-based injury surveillance issues were discussed during a 2-day facilitated round-table meeting, by an 11-person working group consisting of researchers currently active in rugby-related injury surveillance, sports medicine and sports science issues. The outcomes from the meeting were summarised in a draft guidance document that was then subjected to an extensive iterative review prior to producing methodological recommendations.\ud \ud Results\ud \ud Each aspect of the rugby-specific consensus statement was reviewed to determine whether it was feasible to implement the standards required in the context of non-elite rugby and the resources available within in a community setting. Final recommendations are presented within a community-based injury report form.\ud \ud Conclusions\ud \ud It is recommended that whenever possible the rugby-specific consensus statement for injury surveillance studies be used: this paper presents an adapted report form that can be used to record injury surveillance information in community rugby if suitable medical support is not available.

Anaerobic digestion can inactivate zoonotic pathogens present in cattle manure, which reduces transmission of these pathogens from farms to humans through the environment. However, the variability of inactivation across farms and over time is unknown because most studies have examined pathogen inactivation under ideal laboratory conditions or have focused on only one or two full‐scale digesters at a time. In contrast, we sampled seven full‐scale digesters treating cattle manure in Wisconsin for 9 mo on a biweekly basis (n = 118 pairs of influent and effluent samples) and used real‐time quantitative polymerase chain reaction to analyze these samples for 19 different microbial genetic markers. Overall, inactivation of pathogens and fecal indicators was highly variable. When aggregated across digester and season, log‐removal values for several representative microorganisms—bovine Bacteroides, Bacteroidales‐like CowM3, and bovine polyomavirus—were 0.78 ± 0.34, 0.70 ± 0.50, and 0.53 ± 0.58, respectively (mean ± SD). These log‐removal values were up to two times lower than expected based on the scientific literature. Thus, our study indicates that full‐scale anaerobic digestion of cattle manure requires optimization with regard to pathogen inactivation. Future studies should focus on identifying the potential causes of this suboptimal performance (e.g., overloading, poor mixing, poor temperature control). Our study also examined the fate of pathogens during manure separation and found that the majority of microbes we detected ended up in the liquid fraction of separated manure. This finding has important implications for the transmission of zoonotic pathogens through the environment to humans. Core Ideas Pathogen inactivation is highly variable among full‐scale anaerobic digesters.Pathogen inactivation by full‐scale digesters on cattle farms needs optimization.Most microbes end up in the liquid fraction during solids separation of manure.

Residual wastewater solids are a significant reservoir of antibiotic resistance genes (ARGs). While treatment technologies can reduce ARG levels in residual wastewater solids, the effects of these technologies on ARGs in soil during subsequent land-application are unknown. In this study we investigated the use of numerous treatment technologies (air drying, aerobic digestion, mesophilic anaerobic digestion, thermophilic anaerobic digestion, pasteurization, and alkaline stabilization) on the fate of ARGs and class 1 integrons in wastewater solids-amended soil microcosms. Six ARGs [erm(B), qnrA, sul1, tet(A), tet(W), and tet(X)], the integrase gene of class 1 integrons (intI1), and 16S rRNA genes were quantified using quantitative polymerase chain reaction. The quantities of ARGs and intI1 decreased in all microcosms, but thermophilic anaerobic digestion, alkaline stabilization, and pasteurization led to the most extensive decay of ARGs and intI1, often to levels similar to that of the control microcosms to which no wastewater solids had been applied. In contrast, the rates by which ARGs and intI1 declined using the other treatment technologies were generally similar, typically varying by less than 2 fold. These results demonstrate that wastewater solids treatment technologies can be used to decrease the persistence of ARGs and intI1 during their subsequent application to soil.

Chemical genetic screening of small-molecule libraries has been a promising strategy for discovering unique and novel therapeutic compounds. However, identifying the targets of lead molecules that arise from these screens has remained a major bottleneck in understanding the mechanism of action of these compounds. Here, we have coupled the screening of a cysteine-reactive fragment-based covalent ligand library with an isotopic tandem orthogonal proteolysis-enabled activity-based protein profiling (isoTOP-ABPP) chemoproteomic platform to rapidly couple the discovery of lead small molecules that impair pancreatic cancer pathogenicity with the identification of druggable hotspots for potential cancer therapy. Through this coupled approach, we have discovered a covalent ligand DKM 2-93 that impairs pancreatic cancer cell survival and in vivo tumor growth through covalently modifying the catalytic cysteine of the ubiquitin-like modifier activating enzyme 5 (UBA5), thereby inhibiting its activity as a protein that activates the ubiquitin-like protein UFM1 to UFMylate proteins. We show that UBA5 is a novel pancreatic cancer therapeutic target and show DKM 2-93 as a relatively selective lead inhibitor of UBA5. Our results underscore the utility of coupling the screening of covalent ligand libraries with isoTOP-ABPP platforms for mining the proteome for druggable hotspots for cancer therapy.

Giardia is a zoonotic gastrointestinal parasite responsible for a substantial global public health burden, and quantitative microbial risk assessment (QMRA) is often used to forecast and manage this burden. QMRA requires dose-response models to extrapolate available dose-response data, but the existing model for Giardia ignores valuable dose-response information, particularly data from several well-documented waterborne outbreaks of giardiasis. The current study updates Giardia dose-response modeling by synthesizing all available data from outbreaks and experimental studies using a Bayesian random effects dose-response model. For outbreaks, mean doses (D) and the degree of spatial and temporal aggregation among cysts were estimated using exposure assessment implemented via two-dimensional Monte Carlo simulation, while potential overreporting of outbreak cases was handled using published overreporting factors and censored binomial regression. Parameter estimation was by Markov chain Monte Carlo simulation and indicated that a typical exponential dose-response parameter for Giardia is r = 1.6 × 10-2 [3.7 × 10-3 , 6.2 × 10-2 ] (posterior median [95% credible interval]), while a typical morbidity ratio is m = 3.8 × 10-1 [2.3 × 10-1 , 5.5 × 10-1 ]. Corresponding (logistic-scale) variance components were σr = 5.2 × 10-1 [1.1 × 10-1 , 9.6 × 10-1 ] and σm = 9.3 × 10-1 [7.0 × 10-2 , 2.8 × 100 ], indicating substantial variation in the Giardia dose-response relationship. Compared to the existing Giardia dose-response model, the current study provides more representative estimation of uncertainty in r and novel quantification of its natural variability. Several options for incorporating variability in r (and m) into QMRA predictions are discussed, including incorporation via Monte Carlo simulation as well as evaluation of the current study's model using the approximate beta-Poisson.

Parthenolide, a natural product from the feverfew plant and member of the large family of sesquiterpene lactones, exerts multiple biological and therapeutic activities including anti-inflammatory and anti-cancer effects. Herein, we further study parthenolide mechanism of action using activity-based protein profiling (ABPP)-based chemoproteomic platforms to map additional covalent targets engaged by parthenolide in human breast cancer cells. We find that parthenolide, as well as other related exocyclic methylene lactone-containing sesquiterpenes, covalently modify cysteine 427 (C427) of focal adhesion kinase 1 (FAK1) leading to impairment of FAK1-dependent signaling pathways and breast cancer cell proliferation, survival, and motility. These studies reveal a novel functional target exploited by members of a large family of anticancer natural products.

Metal-hydride hydrogen atom transfer (MHAT) functionalizes unbiased alkenes with predictable branched (Markovnikov) selectivity. The breadth of these transformations has been confined to π-radical traps; no sp3 electrophiles have been reported. Here we describe a Mn/Ni dual catalytic system that allows for the hydroalkylation of unactivated olefins by unactivated alkyl halides, yielding aliphatic quaternary carbons.

Robert L Hill,1,* John G Wilmot,1,* Beth A Belluscio,1Kevin Cleary,2 David Lindisch,3Robin Tucker,4Emmanuel Wilson,2Rajesh B Shukla11Meridian Medical Technologies Inc., Columbia, MD, 2Children’s National Medical Center, 3Washington DC VA Medical Center, 4Georgetown University Medical Center, Washington, DC, USA *These authors have contributed equally to this work Abstract: Parenteral routes of drug administration are often selected to optimize actual dose of drug delivered, assure high bioavailability, bypass first-pass metabolism or harsh gastrointestinal environments, as well as maximize the speed of onset. Intramuscular (IM) delivery can be preferred to intravenous delivery when initiating intravenous access is difficult or impossible. Drugs can be injected intramuscularly using a syringe or an automated delivery device (autoinjector). Investigation into the IM delivery dynamics of these methods may guide further improvements in the performance of injection technologies. Two porcine model studies were conducted to compare differences in dispersion of injectate volume for different methods of IM drug administration. The first study compared the differences in the degree of dispersion and uptake of injectate following the use of a manual syringe and an autoinjector. The second study compared the spatial spread of the injected formulation, or dispersion volume, and uptake of injectate following the use of five different autoinjectors (EpiPen® [0.3 mL], EpiPen® Jr [0.3 mL], Twinject® [0.15 mL, 0.3 mL], and Anapen® 300 [0.3 mL]) with varying needle length, needle gauge, and force applied to the plunger. In the first study, the autoinjector provided higher peak volumes of injectate, indicating a greater degree of dispersion, compared with manual syringe delivery. In the second study, EpiPen autoinjectors resulted in larger dispersion volumes and higher initial dispersion ratios, which decreased rapidly over time, suggesting a greater rate of uptake of injectate than the other autoinjectors. The differences in dispersion and uptake of injectate are likely the result of different functional characteristics of the delivery systems. Both studies demonstrate that the functional characteristics of the method for delivering IM injections impact the dispersion and uptake of the material injected, which could significantly affect the pharmacokinetics and, ultimately, the effectiveness of the drug. Keywords: anaphylaxis, autoinjector device, injector pen, intramuscular drug administration, dispersion volume

Numerous natural and anthropogenic processes in a watershed produce the geochemical composition of a river, which can be altered over time by snowmelt and rainfall events and by built infrastructure (i.e., dams and diversions). Trace element concentrations coupled with isotopic ratios offer valuable insights to disentangle the effects of these processes on water quality. In this study, we measured a suite of 40+ trace and major elements (including As, Cd, Ce, Cr, Cs, Fe, La, Li, Mo, Pb, Rb, Sb, Se, Sr, Ti, Tl, U, and Zn), Sr isotopes (87Sr/86Sr), and stable isotopes of H and O (δD and δ18O) to investigate natural and anthropogenic processes impacting the Provo River in northern Utah, USA. The river starts as a pristine mountain stream and passes through agricultural and urban areas, with two major reservoirs and several major diversions to and from the river. We sampled the entire 120 km length of the Provo River at 13 locations from the Uinta Mountains to Utah Valley, as well as two important tributaries, across the range of hydrologic conditions from low flow to snowmelt runoff during the 2013 water year. We also sampled the furthest downstream site in the Utah Valley urban area during a major flood event. Trace element concentrations indicate that a variety of factors potentially influence Provo River chemistry, including inputs from weathering of carbonate/siliciclastic rocks (Sr) and black shales (Se and U), geothermal groundwater (As, Cs, Li, and Rb), soil erosion during snowmelt runoff (Ce, Cr, Fe, La, Pb, and Ti), legacy mining operations (Mo, Sb, and Tl), and urban runoff (Cr, Pb, and Zn). Although specific elements overlap between different groups, the combination of different elements together with isotopic measurements and streamflow observations may act as diagnostic tools to identify sources. 87Sr/86Sr ratios indicate a strong influence of siliciclastic bedrock in the headwaters with values exceeding 0.714 and carbonate bedrock in the lower reaches of the river with values approaching 0.709. δD and δ18O changed little throughout the year in the Provo River, suggesting that the river is primarily fed by snowmelt during spring runoff and snowmelt-fed groundwater during baseflow. Based on nonmetric multidimensional scaling (NMS) water chemistry was unique across the upper, middle, and lower portions of the river, with high temporal variability above the first reservoir but minimal temporal variability below the reservoir. Thus, the results show that dams alter water chemistry by allowing for settling of particle-associated elements and also by homogenizing inflows throughout the year to minimize dilution during snowmelt runoff. Taken together, trace element concentrations and isotopic measurements can be used to evaluate the complex geochemical patterns of rivers and their variability in space and time. These measurements are critical for identifying natural and anthropogenic impacts on river systems.

Pd(0)-catalyzed, non-chain-walking Mizoroki-Heck reactions exhibit poor reactivity with polysubsti- tuted, unbiased alkenes. Intermolecular reactions with simple, all-carbon tetrasubstituted alkenes are unprecedented. Here we report that pendant carboxylic acids, combined with bulky monophospine ligands on palladium, can direct the arylation of tri- and tetrasubstituted olefins. Quaternary carbons are established at high Fsp3 attached-ring junctures and the carboxylate directing group can be removed after coupling. Carboxylate directivity prevents over-arylation of the new, less hindered alkene, which can be diversified in subsequent reactions.

This study investigated the use of thermophilic anaerobic digestion for removing antibiotic resistance genes (ARGs) from residual municipal wastewater solids. Four laboratory-scale anaerobic digesters were operated in 8-day batch cycles at temperatures of 40, 56, 60, and 63 °C. Two tetracycline resistance genes (tet(W) and tet(X)), a fluoroquinolone resistance gene (qnrA), the integrase gene of class 1 integrons (intI1), 16S rRNA genes of all Bacteria, and 16S rRNA genes of methanogens were quantified using real-time quantitative PCR. ARG and intI1 quantities decreased at all temperatures and were described well by a modified form of the Collins-Selleck disinfection kinetic model. The magnitudes of Collins-Selleck kinetic parameters were significantly greater at thermophilic temperatures compared to 40 °C, but few statistically significant differences were observed among these parameters for the thermophilic anaerobic digesters. This model allows for the direct comparison of different operating conditions (e.g., temperature) on anaerobic digestion performance in mitigating the quantity of ARGs in wastewater solids and could be used to design full-scale anaerobic digesters to specifically treat for ARGs as a "pollutant" of concern.

The assumptions underlying quantitative microbial risk assessment (QMRA) are simple and biologically plausible, but QMRA predictions have never been validated for many pathogens. The objective of this study was to validate QMRA predictions against epidemiological measurements from outbreaks of waterborne gastrointestinal disease. I screened 2,000 papers and identified 12 outbreaks with the necessary data: disease rates measured using epidemiological methods and pathogen concentrations measured in the source water. Eight of the 12 outbreaks were caused by Cryptosporidium, three by Giardia, and one by norovirus. Disease rates varied from 5.5 × 10-6 to 1.1 × 10-2 cases/person-day, and reported pathogen concentrations varied from 1.2 × 10-4 to 8.6 × 102 per liter. I used these concentrations with single-hit dose-response models for all three pathogens to conduct QMRA, producing both point and interval predictions of disease rates for each outbreak. Comparison of QMRA predictions to epidemiological measurements showed good agreement; interval predictions contained measured disease rates for 9 of 12 outbreaks, with point predictions off by factors of 1.0-120 (median = 4.8). Furthermore, 11 outbreaks occurred at mean doses of less than 1 pathogen per exposure. Measured disease rates for these outbreaks were clearly consistent with a single-hit model, and not with a "two-hit" threshold model. These results demonstrate the validity of QMRA for predicting disease rates due to Cryptosporidium and Giardia.

Background: Spray irrigation for land-applying livestock manure is increasing in the United States as farms become larger and economies of scale make manure irrigation affordable. Human health risks from exposure to zoonotic pathogens aerosolized during manure irrigation are not well understood. Objectives: We aimed to a) estimate human health risks due to aerosolized zoonotic pathogens downwind of spray-irrigated dairy manure; and b) determine which factors (e.g., distance, weather conditions) have the greatest influence on risk estimates. Methods: We sampled downwind air concentrations of manure-borne fecal indicators and zoonotic pathogens during 21 full-scale dairy manure irrigation events at three farms. We fit these data to hierarchical empirical models and used model outputs in a quantitative microbial risk assessment (QMRA) to estimate risk [probability of acute gastrointestinal illness (AGI)] for individuals exposed to spray-irrigated dairy manure containing Campylobacter jejuni, enterohemorrhagic Escherichia coli (EHEC), or Salmonella spp. Results: Median risk estimates from Monte Carlo simulations ranged from 10−5 to 10−2 and decreased with distance from the source. Risk estimates for Salmonella or EHEC-related AGI were most sensitive to the assumed level of pathogen prevalence in dairy manure, while risk estimates for C. jejuni were not sensitive to any single variable. Airborne microbe concentrations were negatively associated with distance and positively associated with wind speed, both of which were retained in models as a significant predictor more often than relative humidity, solar irradiation, or temperature. Conclusions: Our model-based estimates suggest that reducing pathogen prevalence and concentration in source manure would reduce the risk of AGI from exposure to manure irrigation, and that increasing the distance from irrigated manure (i.e., setbacks) and limiting irrigation to times of low wind speed may also reduce risk. https://doi.org/10.1289/EHP283

Chemical genetics has arisen as a powerful approach for identifying novel anti-cancer agents. However, a major bottleneck of this approach is identifying the targets of lead compounds that arise from screens. Here, we coupled the synthesis and screening of fragment-based cysteine-reactive covalent ligands with activity-based protein profiling (ABPP) chemoproteomic approaches to identify compounds that impair colorectal cancer pathogenicity and map the druggable hotspots targeted by these hits. Through this coupled approach, we discovered a cysteine-reactive acrylamide DKM 3-30 that significantly impaired colorectal cancer cell pathogenicity through targeting C1101 on reticulon 4 (RTN4). While little is known about the role of RTN4 in colorectal cancer, this protein has been established as a critical mediator of endoplasmic reticulum tubular network formation. We show here that covalent modification of C1101 on RTN4 by DKM 3-30 or genetic knockdown of RTN4 impairs endoplasmic reticulum and nuclear envelope morphology as well as colorectal cancer pathogenicity. We thus put forth RTN4 as a potential novel colorectal cancer therapeutic target and reveal a unique druggable hotspot within RTN4 that can be targeted by covalent ligands to impair colorectal cancer pathogenicity. Our results underscore the utility of coupling the screening of fragment-based covalent ligands with isoTOP-ABPP platforms for mining the proteome for novel druggable nodes that can be targeted for cancer therapy.

Summary Many natural products that show therapeutic activities are often difficult to synthesize or isolate and have unknown targets, hindering their development as drugs. Identifying druggable hotspots targeted by covalently acting anti-cancer natural products can enable pharmacological interrogation of these sites with more synthetically tractable compounds. Here, we used chemoproteomic platforms to discover that the anti-cancer natural product withaferin A targets C377 on the regulatory subunit PPP2R1A of the tumor-suppressor protein phosphatase 2A (PP2A) complex leading to activation of PP2A activity, inactivation of AKT, and impaired breast cancer cell proliferation. We developed a more synthetically tractable cysteine-reactive covalent ligand, JNS 1-40, that selectively targets C377 of PPP2R1A to impair breast cancer signaling, proliferation, and in vivo tumor growth. Our study highlights the utility of using chemoproteomics to map druggable hotspots targeted by complex natural products and subsequently interrogating these sites with more synthetically tractable covalent ligands for cancer therapy.

Around the world, municipalities have been making substantial investments into broadband access infrastructure to accelerate the build-out of an urban phenomenon that has become known as a smart or connected city. At the 2016 Optical Fiber Communications Conference, a team contest, the Connected OFCity Challenge, was held to discuss the technological innovations and to examine dependencies and intricacies of a connected city project. The participants, four teams of experts coming from a cross-section of the industry, presented and defended their visions of future applications and innovative architecture and technologies to realize the interconnection. This paper provides a synthesis of the four competitive proposals offered for the contest and their ensuing discussions.

Substantial quantities of antibiotic resistance genes (ARGs) are discharged with treated residual municipal wastewater solids and subsequently applied to soil. The objective of this work was to determine the decay rates for ARGs and class 1 integrons following simulated land application of treated wastewater solids. Treated residual solids from two full-scale treatment plants were applied to sets of triplicate soil microcosms in two independent experiments. Experiment 1 investigated loading rates of 20, 40, and 100 g kg(-1) of residual solids to a sandy soil, while experiment 2 investigated a loading rate of 40 g kg(-1) to a silty-loamy soil. Five ARGs (erm(B), sul1, tet(A), tet(W), and tet(X)), the integrase of class 1 integrons (intI1), 16S rRNA genes, 16S rRNA genes of all Bacteroides spp., and 16S rRNA genes of human-specific Bacteroides spp. were quantified using real-time polymerase chain reaction. ARGs and intI1 quantities declined in most microcosms, with statistically significant (P < 0.05) half-lives varying between 13 d (erm(B), experiment 1, 100 g kg(-1)) and 81 d (intI1, experiment 1, 40 g kg(-1)). These kinetic rates were much slower than have been previously reported for unit operations used to treat wastewater solids (e.g., anaerobic digestion). This research suggests that the design and operation of municipal wastewater treatment facilities with the explicit goal of mitigating the release of ARGs should focus on using technologies within the treatment facility, rather than depending on attenuation subsequent to land application.

This study investigated whether air-drying beds reduce antibiotic resistance gene (ARG) concentrations in residual municipal wastewater solids. Three laboratory-scale drying beds were operated for a period of nearly 100 days. Real-time PCR was used to quantify 16S rRNA genes, 16S rRNA genes specific to fecal bacteria (AllBac) and human fecal bacteria (HF183), the integrase gene of class 1 integrons (intI1), and five ARGs representing a cross-section of antibiotic classes and resistance mechanisms (erm(B), sul1, tet(A), tet(W), and tet(X)). Air-drying beds were capable of reducing all gene target concentrations by 1 to 5 orders of magnitude, and the nature of this reduction was consistent with both a net decrease in the number of bacterial cells and a lack of selection within the microbial community. Half-lives varied between 1.5 d (HF183) and 5.4 d (tet(X)) during the first 20 d of treatment. After the first 20 d of treatment, however, half-lives varied between 8.6 d (tet(X)) and 19.3 d (AllBac), and 16S rRNA gene, intI1, and sul1 concentrations did not change (P0.05). These results demonstrate that air-drying beds can reduce ARG and intI1 concentrations in residual municipal wastewater solids within timeframes typical of operating practices.

Precise collimation of the beam halo is required in the International Linear Collider (ILC) to prevent beam losses near the interaction region that could cause unacceptable backgrounds for the physics detector. The necessarily small apertures of the collimators lead to transverse wakefields that may result in beam deflections and increased emittance. A set of collimator wakefield measurements has previously been performed in the ASSET region of the SLAC Linac. We report on the next phase of this programme, which is carried out at the recently commissioned End Station A (ESA) test facility at SLAC. Measurements of resistive and geometric wakefields using tapered collimators are compared with model predictions from MAFIA and GdfidL and with analytic calculations.

In vitro mechanotransduction studies, uncovering the basic science of the response of cells to mechanical forces, are essential for progress in tissue engineering and its clinical application. Many varying investigations have described a multitude of cell responses, however as the precise nature and magnitude of the stresses applied are infrequently reported and rarely validated, the experiments are often not comparable, limiting research progress. This thesis provides physical and biological validation of a widely available fluid stimulation device, a see-saw rocker, as an In vitro model for cyclic fluid shear stress mechanotransduction. This allows linkage between precisely characterised stimuli and cell monolayer response in a convenient six-well plate format. Computational fluid dynamic models of one well were analysed extensively to generate convergent, stable and consistent predictions of the cyclic fluid velocity vectors at a rocking frequency of 0.5 Hz, accounting for the free surface. Validation was provided by comparison with flow velocities measured experimentally using particle image velocimetry. Qualitative flow behaviour was matched and quantitative analysis showed good agreement at representative locations and time points. A maximum shear stress of 0.22Pa was estimated near the well edge, and time-average shear stress ranged between 0.029 and 0.068Pa, within the envelope of previous musculoskeletal In vitro fluid flow investigations. The CFD model was extended to explore changes in culture medium viscosity, rocking frequency and the robustness to position on the rocking platform. Shear stress magnitude was shown to increase almost linearly with an increase in the viscosity of culture medium. Compared with 0.5 Hz, models at 0.083 and 1:167 Hz, the operational limits of the see-saw rocker, indicated a change in shear stress patterns at the cell layer, and a reduction and increase in mean shear stress respectively. At the platform edge at 0.5 Hz, a 1.67-fold increase in time-average shear stress was identified. Extensive biological validations using human tenocytes underlined the versatility of the simple In vitro device. The application of fluid-induced shear stress at 0.5 Hz under varying regimes up to 0.714Pa caused a significant increase in secreted collagen (p < 0.05) compared to static controls. Tenocytes stimulated at a shear stress magnitude of 1.023Pa secreted significantly less collagen compared to static controls. The potential for a local maximum in the relationship between collagen secretion rate and shear stress was identified, indicating a change from anabolic to catabolic behaviour. Collagen biochemical assay results were echoed with antibody stains for proteins, where a co-localisation of connexin-32 with collagen type-I was also identified. A custom algorithm showed that four hours of fluid-induced shear stress of 0:033Pa intermittently applied to tenocytes encouraged alignment and elongation over an eight day period in comparison to static controls. Primary cilia were identified in human tenocyte cultures and bovine flexor tendon tissue; however primary cilium abrogation In vitro using chloral hydrate proved detrimental to cell viability. Collaborative investigations identified that ERK signalling and c-Fos transcription factor expression peaked after the application of 0.012Pa at 0.083 Hz for 20 minutes and anabolic collagen gene expression relative quantities increased after 48 hours of rocking at 0.083 Hz. In conclusion, validated shear stresses within a six-well plate, induced by cyclic flow from a see-saw rocker, provides an exceptional model for the In vitro study of dynamic fluid shear stress mechanobiology. Biological investigations have been linked to precise applied shear stress, creating a foundation for understanding the complex relationship between tenocytes and fluid-induced shear stress In vitro. Using this model, research is repeatable, comparable and accurately attributed to shear stress, accelerating the scientific advancement of musculoskeletal mechanobiology.

Background: Measurements of polarization observables for the reactions γ⃗ p→K+Λ and γ⃗ p→K+Σ0 have been performed. This is part of a program of measurements designed to study the spectrum of baryon resonances in particular, and nonperturbative QCD in general.\ud Purpose: The accurate measurement of several polarization observables provides tight constraints for phenomenological fits, which allow the study of strangeness in nucleon and nuclear systems. Beam-recoil observables for the γ⃗ p→K+Σ0 reaction have not been reported before now.\ud Method: The measurements were carried out using linearly polarized photon beams incident on a liquid hydrogen target, and the CLAS detector at the Thomas Jefferson National Accelerator Facility. The energy range of the results is 1.71

Bright-red colors in vertebrates are commonly involved in sexual, social, and interspecific signaling [1-8] and are largely produced by ketocarotenoid pigments. In land birds, ketocarotenoids such as astaxanthin are usually metabolically derived via ketolation of dietary yellow carotenoids [9, 10]. However, the molecular basis of this gene-environment mechanism has remained obscure. Here we use the yellowbeak mutation in the zebra finch (Taeniopygia guttata) to investigate the genetic basis of red coloration. Wild-type ketocarotenoids were absent in the beak and tarsus of yellowbeak birds. The yellowbeak mutation mapped to chromosome 8, close to a cluster of cytochrome P450 loci (CYP2J2-like) that are candidates for carotenoid ketolases. The wild-type zebra finch genome was found to have three intact genes in this cluster: CYP2J19A, CYP2J19B, and CYP2J40. In yellowbeak, there are multiple mutations: loss of a complete CYP2J19 gene, a modified remaining CYP2J19 gene (CYP2J19(yb)), and a non-synonymous SNP in CYP2J40. In wild-type birds, CYP2J19 loci are expressed in ketocarotenoid-containing tissues: CYP2J19A only in the retina and CYP2J19B in the beak and tarsus and to a variable extent in the retina. In contrast, expression of CYP2J19(yb) is barely detectable in the beak of yellowbeak birds. CYP2J40 has broad tissue expression and shows no differences between wild-type and yellowbeak. Our results indicate that CYP2J19 genes are strong candidates for the carotenoid ketolase and imply that ketolation occurs in the integument in zebra finches. Since cytochrome P450 enzymes include key detoxification enzymes, our results raise the intriguing possibility that red coloration may be an honest signal of detoxification ability., Current Biology, 26 (11), ISSN:0960-9822, ISSN:1879-0445

Breast cancers possess fundamentally altered metabolism that fuels their pathogenicity. While many metabolic drivers of breast cancers have been identified, the metabolic pathways that mediate breast cancer malignancy and poor prognosis are less well understood. Here, we used a reactivity-based chemoproteomic platform to profile metabolic enzymes that are enriched in breast cancer cell types linked to poor prognosis, including triple-negative breast cancer (TNBC) cells and breast cancer cells that have undergone an epithelial-mesenchymal transition-like state of heightened malignancy. We identified glutathione S-transferase Pi 1 (GSTP1) as a novel TNBC target that controls cancer pathogenicity by regulating glycolytic and lipid metabolism, energetics, and oncogenic signaling pathways through a protein interaction that activates glyceraldehyde-3-phosphate dehydrogenase activity. We show that genetic or pharmacological inactivation of GSTP1 impairs cell survival and tumorigenesis inTNBC cells. We put forth GSTP1 inhibitors as a noveltherapeutic strategy for combatting TNBCs through impairing key cancer metabolism and signaling pathways.

Background: Measurements of polarization observables for the reactions $\vec{\gamma} p \rightarrow K^+ \Lambda$ and $\vec{\gamma} p \rightarrow K^+ \Sigma^0$ have been performed. This is part of a programme of measurements designed to study the spectrum of baryon resonances. Purpose: The accurate measurement of several polarization observables provides tight constraints for phenomenological fits. Beam-recoil observables for the $\vec{\gamma} p \rightarrow K^+ \Sigma^0$ reaction have not been reported before now. Method: The measurements were carried out using linearly polarized photon beams and the CLAS detector at the Thomas Jefferson National Accelerator Facility. The energy range of the results is 1.71\,GeV $, Comment: 29 pages, 13 figures

These notes were delivered as a series of NIMROD lectures at the Rutherford Appleton Laboratory by the author in February 1976 (RL-76-022). The purpose of these lectures was primarily two-fold: to discuss the classical theory of free point particles, free strings and free membranes from a unified viewpoint; and to present in the process of doing this the rudiments of an intrinsic geometrical calculus that the author has found of immense value in investigating these systems. It is shown how the equations of motion for such classically extended relativistic systems arise in a very simple manner from a principle of stationary action and furthermore how the boundary conditions for finite systems may be derived in a gauge invariant way. Momenta are naturally introduced and the primary constraints that exist in a Hamiltonian description follow simply. Calculations may proceed in an index-free manner until components are required. It is at this stage that one can, if one desires, impose gauge conditions and remove non-independent degrees of freedom. Such methods can be applied in any spacetime of any dimension and metric, and examples are given throughout., Comment: 48 pages, 8 figures

Background and Purpose: The psychometric properties of the Kansas City Cardiomyopathy Questionnaire (KCCQ) have been examined primarily in community-dwelling patients with heart failure (HF). The objective of this research was to examine the properties of the KCCQ administered to patients hospitalized with HF (N = 233). Methods: Confirmatory factor analysis, Cronbach's alphas, and correlations were performed to examine the scale's dimensions, reliability, and validity. Results: Confirmatory factor analysis indicated a 5-factor solution (63.6% of the variance). The Cronbach's alpha levels were greater than .70, except for the self-efficacy dimension (.60). Convergent validity was not verified between the KCCQ and several illness severity measures. Conclusions: The psychometric properties of the KCCQ may be different based on the population in which the KCCQ is administered, which may have clinical implications.

Waterborne pathogens were measured at three beaches in Lake Michigan, environmental factors for predicting pathogen concentrations were identified, and the risk of swimmer infection and illness was estimated. Waterborne pathogens were detected in 96% of samples collected at three Lake Michigan beaches in summer, 2010. Samples were quantified for 22 pathogens in four microbial categories (human viruses, bovine viruses, protozoa, and pathogenic bacteria). All beaches had detections of human and bovine viruses and pathogenic bacteria indicating influence of multiple contamination sources at these beaches. Occurrence ranged from 40 to 87% for human viruses, 65-87% for pathogenic bacteria, and 13-35% for bovine viruses. Enterovirus, adenovirus A, Salmonella spp., Campylobacter jejuni, bovine polyomavirus, and bovine rotavirus A were present most frequently. Variables selected in multiple regression models used to explore environmental factors that influence pathogens included wave direction, cloud cover, currents, and water temperature. Quantitative Microbial Risk Assessment was done for C. jejuni, Salmonella spp., and enteroviruses to estimate risk of infection and illness. Median infection risks for one-time swimming events were approximately 2 × 10(-5), 8 × 10(-6), and 3 × 10(-7) [corrected] for C. jejuni, Salmonella spp., and enteroviruses, respectively. Results highlight the importance of investigating multiple pathogens within multiple categories to avoid underestimating the prevalence and risk of waterborne pathogens.