Multidimensional mass profiles increase confidence in bacterial identification when using low-resolution mass spectrometers.

Field-forward analytical technologies, such as portable mass spectrometry (MS), enable essential capabilities for real-time monitoring and point-of-care diagnostic applications. Significant and recent investments improving the features of miniaturized mass spectrometers enable various new applications outside of small molecule detection. Most notably, the addition of tandem mass spectrometry scans (MS/MS) allows the instrument to isolate and fragment ions and increase the analytical specificity by measuring unique chemical signatures for ions of interest. Notwithstanding these technological advancements, low-cost, portable systems still struggle to confidently identify clinically significant organisms of interest, such as bacteria, viruses, and proteinaceous toxins, due to the limitations in resolving power. To overcome these limitations, we developed a novel multidimensional mass fingerprinting technique that uses tandem mass spectrometry to increase the chemical specificity for low-resolution mass spectral profiles. We demonstrated the method's capabilities for differentiating four different bacteria, including attentuated strains of Yersinia pestis. This approach allowed for the accurate (>92%) identification of each organism at the strain level using de-resolved matrix-assisted laser desorption ionization-time of flight (MALDI-TOF) data to mimic the performance characteristics of miniaturized mass spectrometers. This work demonstrates that low-resolution mass spectrometers, equipped with tandem MS acquisition modes, can accurately identify clinically relevant bacteria. These findings support the future application of these technologies for field-forward and point-of-care applications where high-performance mass spectrometers would be cost-prohibitive or otherwise impractical.

Using Machine Learning to Determine a Suitable Patient Population for Anakinra for the Treatment of COVID-19 Under the Emergency Use Authorization.

A randomized, double-blind, placebo-controlled study (SAVEMORE trial) provided data to support an Emergency Use Authorization (EUA) of anakinra in hospitalized adults with positive results of direct severe acute respiratory syndrome-coronavirus 2 viral testing with pneumonia requiring supplemental oxygen (low- or high-flow oxygen) who are at risk of progressing to severe respiratory failure and likely to have an elevated plasma soluble urokinase plasminogen activator receptor (suPAR). Currently, the suPAR assay is not commercially available in the United States. An alternative method was needed to identify patients that best reflect the population in the clinical trial selected based on suPAR level ≥ 6 ng/mL at baseline. A machine learning approach based on data from the SAVEMORE trial was used to develop a scoring rule to identify patients who are likely to have a suPAR level ≥ 6 ng/mL at baseline. External validation of the scoring rule was conducted with data from a different trial (SAVE). This clinical scoring rule with high positive predictive value, high specificity, reasonable sensitivity, and biological relevance is expected to identify patients who are likely to have an elevated suPAR level ≥ 6 ng/mL at baseline. As such, it is included in the EUA to identify patients that fall within the authorized population for whom the known and potential benefits outweigh the known and potential risks of anakinra.

Gentrification of the (Leisure) Mind: Organizational Justifications and Community Concerns of a Proposed LGBTQ2S Sport and Recreation Center.

In 2015, Toronto's largest LGBTQ2S organization, the city, and a private donor collaborated and proposed what they believed constituted the world's first LGBTQ2S sport and recreation facility in Toronto's Moss Park neighborhood. While conceived as a site that would expand community services in an underserved community, this project was met with criticism from segments of the Moss Park community. Using qualitative interviews with both representatives from the LGBTQ2S organization and local residents, the purpose of this was to analyze justifications behind the project and concerns from the community. In so doing, the authors reflect upon the relationship between sport, recreation, and leisure-based place-making and questions of queer/de-queering politics by exploring the transformative or assimilationist aspects of queer recreation practices, the homonormativity of leisure spaces, and spiritual displacement associated with queer gentrification.

"You Have to Validate It": Experiences of Female Sexual Minority Student-Athletes.

Lesbians in sport may reside in a culture of silence due to the fear of being negatively labeled. Often, ideologies regarding lesbian athletes validate social inequalities through institutional practices. The purpose of this study was to examine the experiences of NCAA female sexual minority student-athletes. Employing purposeful sampling, nine current and former female student-athletes participated in semi-structured interviews. Through a combination of inductive and narrative analyses during the data collection and analysis processes, the findings revealed five higher order themes: (1) climate, (2) validation and norms of behavior, (3), misunderstandings and misconceptions, (4) negotiating identities and risk, and (5) gender ideology and assumptions. Although results of the present study are not generalizable, this study can inform inclusive practices to improve the experiences of sexual minority student-athletes. Further, this study will create awareness regarding the obstacles female sexual minority student-athletes endure on college campuses and within the setting of intercollegiate sport.

Recombinant expression, purification, and structural analysis of two ectodomains of Syndecan-1.

Syndecan-1 (SDC-1) is an integral membrane heparin sulfate proteoglycan that is involved in inflammatory response, cell-signaling, cell proliferation, and numerous other cell-matrix interactions. Like the other members of the syndecan family, very little is known about structural conformations and dynamics of SDC-1. A majority of interactions occur through the extracellular ectodomain, therefore we have dedicated our research efforts to the study this specific portion of SDC-1. The ectodomain is often shed from the cell surface due to various stimuli. The released fragment has already been used as a useful biomarker for prognosis of some diseases and cancers. SDC-1 can be cleaved in different locations depending on the sheddase, generating soluble shed ectodomains that can be carried away in blood sera. In this study, we focus specifically on two main cleavage fragments that can be generated. We show the first successful expression and purification of recombinant SDC-1 ectodomains. Production of SDC-1 in E. coli allows the production of the core protein without risking heterogeneous post-translational modifications such as glycosylation, allowing a certain level of control over protein homogeneity that is not possible in mammalian expression. An expression vector was used to generate two different fusion proteins consisting of a His-tag and a TEV cleavage site for the removal of the fusion partner. SDS-PAGE was used to track the expression as well as the purification. Masses of the isolated proteins were determined using mass spectrometry and the purity and homogeneity were evaluated by solution NMR.

Campus recreational participation and COVID-19: impact on college student health and well-being.

Objective: Campus recreational sport activities impact college student health and well-being in a variety of domains. This multi-institutional study examined the participation of students in campus recreation during the pandemic and explored the relationship between student participation and their demographic markers, COVID-19 experience, and perceptions of risk, health and safety. Methods: Results from a survey of 1,815 American college students indicated the presence of statistical differences in sport participation based on Gender, Race, State, Personal COVID-19 Experience, Campus Safety Efforts, and Risk Perception Score within various areas of analysis. Results: The impact and meaning of these cross-sectional results are discussed with reference to the ongoing pandemic and student health. Recommendations are presented within the context of this vital programming area both during and beyond the current crisis. Conclusion: These results are especially important given the health benefits associated with participation in recreational sport as tools to mitigate against the unprecedented consequences of the pandemic.

Establishing age-stratified red blood cell fatty acid reference ranges using model-based clustering and iterative application of the harris-boyd method.

BACKGROUND: The current assessment of nutritional status and diagnosis of essential fatty acids deficiency (EFAD) utilizes the analysis of long-chain fatty acids (LCFAs) in serum or plasma; however, these concentrations do not represent habitual LCFA intake. LCFAs in red blood cells (RBCs) are less prone to intra-individual variability and exclude the need for fasting, which is unrealistic in pediatric populations. Our study objective was to characterize the RBC LCFA profiles in pediatric and adult reference populations and establish age-specific reference intervals (RIs). METHODS: Twenty-one LCFAs in RBCs were measured in 523 pediatric and adult controls by gas chromatography-mass spectrometry. Model-based clustering was used to identify possible age subgroups. After removing outliers by the Tukey method, initial age subgroups were then compared using the Harris-Boyd method in an iterative manner. RIs (95%), with confidence intervals (90%), in the final age groups were established using parametric or non-parametric statistics. RESULTS: Our data showed heterogeneous changes in the concentrations of most LCFAs and the EFAD biomarkers (mead acid, Triene/Tetraene ratio) during infancy. Model-based clustering identified six initial age subgroups per fatty acid, on average. Our application of the iterative Harris-Boyd method decreased the average number of age groups to three per fatty acid, with 13 total unique age cut-offs. Finally, using these age groups, we established age-specific RIs for 21 fatty acids, six group totals, and the Triene/Tetraene ratio. CONCLUSION: Our study revealed significant age-dependent changes in RBC fatty acid profiles warranting separate pediatric and adults RIs. Model-based clustering and the iterative application of the Harris-Boyd method were successfully used to establish RBC fatty acid RIs for an objective assessment of long-term nutritional status in pediatric and adult populations.

Tradipitant in the Treatment of Motion Sickness: A Randomized, Double-Blind, Placebo-Controlled Study.

Introduction: Novel therapies are needed for the treatment of motion sickness given the inadequate relief and bothersome and dangerous adverse effects of currently approved therapies. Neurokinin-1 (NK1) receptor antagonists have the potential to be effective in improving the symptoms of motion sickness, given the involvement of Substance P in nauseogenic and emetic pathways and the expression of NK1 receptors in the gastrointestinal system. Here we evaluated the efficacy of tradipitant, a novel NK1 receptor antagonist, in preventing motion sickness in variable sea conditions. Methods: A total of 126 adults participated in the Motion Sifnos study. Groups of participants were assigned to one of seven boat trips lasting ~4 h on the Pacific Ocean. Participants were randomized 1:1 to tradipitant 170 mg or placebo and completed the Motion Sickness Severity Scale (MSSS) every 30 min, in addition to other assessments. Severity of motion sickness was assessed with the incidence of vomiting and the MSSS. Results: Participants on tradipitant had a significantly lower incidence of vomiting as compared to those on placebo across all boat trips (tradipitant = 17.5%, placebo = 39.7%, p = 0.0039). For trips exposed to rough sea conditions, the difference in the incidence of vomiting between the groups was more dramatic (tradipitant = 15.79%, placebo = 72.22%, p = 0.0009). Across these trips, motion sickness symptoms were significantly lower in the tradipitant group compared to the placebo group (tradipitant = 3.19, placebo = 4.57, p = 0.0235). Discussion: Tradipitant has the potential to be an effective therapy for the prevention of vomiting and treatment of nausea in people with motion sickness.

EGFR/c-Met and mTOR signaling are predictors of survival in non-small cell lung cancer.

BACKGROUND: EGFR/c-Met activation/amplification and co-expression, mTOR upregulation/activation, and Akt/Wnt signaling upregulation have been individually associated with more aggressive disease and characterized as potential prognostic markers for lung cancer patients. METHODS: Tumors obtained from 109 participants with stage I-IV non-small cell lung cancer (NSCLC) were studied for EGFR/c-Met co-localization as well as for total and active forms of EGFR, c-Met, mTOR, S6K, beta-catenin, and Axin2. Slides were graded by two independent blinded pathologists using a validated scoring system. Protein expression profile correlations were assessed using Pearson correlation and Spearman's rho. Prognosis was assessed using Kaplan-Meier analysis. RESULTS: Protein expression profile analysis revealed significant correlations between EGFR/p-EGFR (p = 0.0412) and p-mTOR/S6K (p = 0.0044). Co-localization of p-EGFR/p-c-Met was associated with increased p-mTOR (p = 0.0006), S6K (p = 0.0018), and p-S6K (p < 0.0001) expression. In contrast, active beta-catenin was not positively correlated with EGFR/c-Met nor any activated proteins. Axin2, a negative regulator of the Wnt pathway, was correlated with EGFR, p-EGFR, p-mTOR, p-S6K, EGFR/c-Met co-localization, and p-EGFR/p-c-Met co-localization (all p-values <0.03). Kaplan-Meier analysis revealed shorter median survival in participants with high expression of Axin2, total beta-catenin, total/p-S6K, total/p-mTOR, EGFR, and EGFR/c-Met co-localization compared with low expression. After controlling for stage of disease at diagnosis, subjects with late-stage disease demonstrated shorter median survival when exhibiting high co-expression of EGFR/c-Met (8.1 month versus 22.3 month, p = 0.050), mTOR (6.7 month versus 22.3 month, p = 0.002), and p-mTOR (8.1 month versus 25.4 month, p = 0.004) compared with low levels. CONCLUSIONS: These findings suggest that increased EGFR/c-Met signaling is correlated with upregulated mTOR/S6K signaling, which may in turn be associated with shorter median survival in late-stage NSCLC.

geoBoundaries: A global database of political administrative boundaries.

We present the geoBoundaries Global Administrative Database (geoBoundaries): an online, open license resource of the geographic boundaries of political administrative divisions (i.e., state, county). Contrasted to other resources geoBoundaries (1) provides detailed information on the legal open license for every boundary in the repository, and (2) focuses on provisioning highly precise boundary data to support accurate, replicable scientific inquiry. Further, all data is released in a structured form, allowing for the integration of geoBoundaries with large-scale computational workflows. Our database has records for every country around the world, with up to 5 levels of administrative hierarchy. The database is accessible at http://www.geoboundaries.org, and a static version is archived on the Harvard Dataverse.

Sustainable resolutions for environmental threat of the acid mine drainage.

Acid Mine Drainage (AMD) caused by abandoned mines is an enormous source of negative impact on the environment and the species that inhabit it. The low levels of pH and high concentration of metals and metalloids (copper, gadolinium, lithium, etc.) in mining pits with standing water lead to changing the balance of surrounding organisms and ecosystems. The scale of the issue and the quantity of AMD sites throughout the globe are factors that make AMD a critical environmental threat. Many AMD treatments have been implemented to reduce the negative impact of AMD, with many solutions being very costly and only suited for particular project situations. Policymakers have strong leverage in correcting AMD problems by developing regulations and laws. This study proposes three more sustainable solutions for reducing and eventually eliminating the impact of AMD with less capital investment while also resolving the landfill problem as well. Also, some governmental strategies are suggested for forming collaborative relationships between industry professionals from different perspectives with the goal to resolve the AMD issue through innovative ideas. Implementation of previous strategies and suggested ones, as well as the further involvement of more communities, can enhance the sustainability of life exposed to AMD.

Potential Telomere-Related Pharmacological Targets.

Telomeres function as protective caps at the terminal portion of chromosomes, containing non-coding nucleotide sequence repeats. As part of their protective function, telomeres preserve genomic integrity and minimize chromosomal exposure, thus limiting DNA damage responses. With continued mitotic divisions in normal cells, telomeres progressively shorten until they reach a threshold at a point where they activate senescence or cell death pathways. However, the presence of the enzyme telomerase can provide functional immortality to the cells that have reached or progressed past senescence. In senescent cells that amass several oncogenic mutations, cancer formation can occur due to genomic instability and the induction of telomerase activity. Telomerase has been found to be expressed in over 85% of human tumors and is labeled as a near-universal marker for cancer. Due to this feature being present in a majority of tumors but absent in most somatic cells, telomerase and telomeres have become promising targets for the development of new and effective anticancer therapeutics. In this review, we evaluate novel anticancer targets in development which aim to alter telomerase or telomere function. Additionally, we analyze the progress that has been made, including preclinical studies and clinical trials, with therapeutics directed at telomere-related targets. Furthermore, we review the potential telomere-related therapeutics that are used in combination therapy with more traditional cancer treatments. Throughout the review, topics related to medicinal chemistry are discussed, including drug bioavailability and delivery, chemical structure-activity relationships of select therapies, and the development of a unique telomere assay to analyze compounds affecting telomere elongation.

Circadian Effects on Performance and Effort in Collegiate Swimmers.

Although individual athletic performance generally tends to peak in the evening, individuals who exhibit a strong diurnal preference perform better closer to their circadian peak. Time-of-day performance effects are influenced by circadian phenotype (diurnal preference and chronotype-sleep-wake patterns), homeostatic energy reserves and, potentially, genotype, yet little is known about how these factors influence physiological effort. Here, we investigate the effects of time of day, diurnal preference, chronotype, and PER3 (a circadian clock gene) genotype on both effort and performance in a population of Division I collegiate swimmers (n = 27). Participants competed in 200m time trials at 7:00 and 19:00 and were sampled pre- and post-trial for salivary α-amylase levels (as a measure of physiological effort), allowing for per-individual measures of performance and physiological effort. Hair samples were collected for genotype analysis (a variable-number tandem-repeat (VNTR) and a single nucleotide polymorphism (SNP) in PER3). Our results indicate significant and parallel time-of-day by circadian phenotype effects on swim performance and effort; evening-type swimmers swam on average 6% slower with 50% greater α-amylase levels in the morning than they did in the evening, and morning types required 5-7 times more effort in the evening trial to achieve the same performance result as the morning trial. In addition, our results suggest that these performance effects may be influenced by gene (circadian clock gene PER3 variants) by environment (time of day) interactions. Participants homozygous for the PER34,4 length variant (rs57875989) or who possess a single G-allele at PER3 SNP rs228697 swam 3-6% slower in the morning. Overall, these results suggest that intra-individual variation in athletic performance and effort with time of day is associated with circadian phenotype and PER3 genotype.

Primary antiphospholipid syndrome associated Liebman-Sachs endocarditis leading to diffuse alveolar hemorrhage: A case report.

A 26 year old female presented for recurrent blood tinged sputum during the previous year with development of frank hemoptysis three days prior to admission. Diffuse alveolar hemorrhage (DAH) was confirmed with serial lavages. The patient had no history of autoimmune disease, vascular thrombosis or pregnancy morbidity including miscarriages or pre-eclampsia. High dose steroids were initiated along with noninvasive ventilatory support. Transthoracic echocardiogram showed severe mitral regurgitation and a vegetation on the mitral valve; transesophageal echocardiogram determined the lesion highly suggestive of Libman-Sachs endocarditis. Blood cultures were negative. Immunological evaluation established the patient was negative for: anti-nuclear antibody, anti-double-stranded DNA antibody, rheumatoid factor, anti-smith antibody, anti-cyclic citrullinated peptide, anti-neutrophil cytoplasmic antibodies, anti-glomerular basement membrane antibodies. Further evaluation revealed elevated levels of anticardiolipin immunoglobulin G and anti-beta 2 glycoprotein immunoglobulin G which continued to increase for months after hospitalization. She was diagnosed with DAH secondary to acute mitral regurgitation caused by Libman-Sachs endocarditis in the presence of primary antiphospholipid antibody syndrome. DAH is an important disease to understand given its high mortality rate. Few case reports relating the presence of Libman-Sachs endocarditis induced by antiphospholipid antibody syndrome leading to DAH have been published. Unique here is the absence of rheumatologic markers thus supporting a diagnosis of primary antiphospholipid antibody syndrome (APS). This patient had no findings associated with rheumatological disorders potentially making this diagnosis easily overlooked. This case further illustrates the importance of evaluating patients with APS presenting with DAH as there are multiple etiologies that lead to this pathology thus different treatment avenues are to be considered during management.

Social motivation and health in college club swimming.

OBJECTIVE: Participation in recreational sport clubs on campus is a popular student activity nationwide. These sport-based organizations provide a host of benefits within recognized dimensions of health and wellness. Understanding participants' motives for engaging in these types of activities can provide insight in design and delivery and enhance participant health. This study focuses on outcomes related to the social motivations for participation in a recreational sport swim club and their potential relationship to social health. PARTICIPANTS: Current members of recreational swimming clubs were contacted for participation in the study from March-April 2016. METHODS: A Leisure Motivation Scale (LMS) survey was sent electronically to 196 collegiate swim clubs nationwide. Aggregate and multivariate analyses from 1011 responses were conducted to examine the social motivation and motivational differences of participants. RESULTS: Social motivations emerged as the predominate motivational construct, indicating important implications for social health improvement through participation. Demographically, results indicated no statistically significant differences in social motivation factors based on participant gender, and statistically significant differences within participant race, university affiliation and practice frequency. CONCLUSIONS: Impacts of these findings are important for practitioners and participants when evaluating the potential these programs have to influence participant social health.

The ventromedial prefrontal cortex in a model of traumatic stress: fear inhibition or contextual processing?

The ventromedial prefrontal cortex (vmPFC) has consistently appeared altered in post-traumatic stress disorder (PTSD). Although the vmPFC is thought to support the extinction of learned fear responses, several findings support a broader role for this structure in the regulation of fear. To further characterize the relationship between vmPFC dysfunction and responses to traumatic stress, we examined the effects of pretraining vmPFC lesions on trauma reactivity and enhanced fear learning in a rodent model of PTSD. In Experiment 1, lesions did not produce differences in shock reactivity during an acute traumatic episode, nor did they alter the strength of the traumatic memory. However, when lesioned animals were subsequently given a single mild aversive stimulus in a novel context, they showed a blunting of the enhanced fear response to this context seen in traumatized animals. In order to address this counterintuitive finding, Experiment 2 assessed whether lesions also attenuated fear responses to discrete tone cues. Enhanced fear for discrete cues following trauma was preserved in lesioned animals, indicating that the deficit observed in Experiment 1 is limited to contextual stimuli. These findings further support the notion that the vmPFC contributes to the regulation of fear through its influence on context learning and contrasts the prevailing view that the vmPFC directly inhibits fear.

Response rates and durability of chemotherapy among 62 patients with metastatic Merkel cell carcinoma.

Cytotoxic chemotherapy is commonly used to treat advanced Merkel cell carcinoma (MCC). However, its efficacy in distant metastatic MCC patients is unclear, in part because most prior reports aggregated these patients with those receiving adjuvant chemotherapy and combined chemoradiation for whom prognosis and outcomes may differ. In this retrospective study, we analyzed detailed records from 62 patients with distant metastatic MCC treated with cytotoxic chemotherapy. Efficacy outcomes including response rate (RR), durability of response (DOR), progression-free survival (PFS), and overall survival (OS) were evaluated. In this cohort, platinum plus etoposide was the most commonly used first-line regimen (69%). RR to first-line chemotherapy was 55% (34/62) with complete responses (CR) in 13% (8/62) and partial responses (PR) in 42% (26/62) while 6% (4/62) had stable disease and 39% (24/62) had progressive disease. Median PFS was 94 days and median OS was 9.5 months from start of chemotherapy. Among responding patients (n = 34), median PFS was 168 days and median DOR was 85 days. Among 30 of the 62 patients who received second-line chemotherapy, RR was 23% (7/30; 1 CR, 6 PR), median PFS was 61 days, and median DOR was 101 days. In summary, first-line chemotherapy is associated with a high RR in metastatic MCC, but responses are typically not durable, and the median PFS is only 3 months. These results suggest rapid emergence of chemoresistance in MCC tumors, and may serve as a useful comparator for immunotherapies currently being explored for metastatic MCC.

Single-fraction radiation therapy in patients with metastatic Merkel cell carcinoma.

Merkel cell carcinoma (MCC) is an aggressive, polyomavirus-associated cancer with limited therapeutic options for metastatic disease. Cytotoxic chemotherapy is associated with high response rates, but responses are seldom durable and toxicity is considerable. Here, we report our experience with palliative single-fraction radiotherapy (SFRT) in patients with metastatic MCC. We conducted retrospective analyses of safety and efficacy outcomes in patients that received SFRT (8 Gy) to MCC metastases between 2010 and 2013. Twenty-six patients were treated with SFRT to 93 MCC tumors located in diverse sites that included skin, lymph nodes, and visceral organs. Objective responses were observed in 94% of the measurable irradiated tumors (86/92). Complete responses were observed in 45% of tumors (including bulky tumors up to 16 cm). "In field" lesion control was durable with no progression in 77% (69/89) of treated tumors during median follow-up of 277 days among 16 living patients. Clinically significant toxicity was seen in only two patients who had transient side effects. An exploratory analysis suggested a higher rate of in-field progression in patients with an immunosuppressive comorbidity or prior recent chemotherapy versus those without (30% and 9%, respectively; P = 0.03). Use of SFRT in palliating MCC patients was associated with an excellent in field control rate and durable responses at treated sites, and with minimal toxicity. SFRT may represent a convenient and appealing alternative to systemic chemotherapy for palliation, for which most patients with oligometastatic MCC are eligible. SFRT may also synergize with emerging systemic immune stimulants by lowering tumor burden and enhancing presentation of viral/tumor antigens.

Beliefs about using an outdoor pool: Understanding perceptions of place in the context of a recreational environment to improve health.

Following the Reasoned Action Approach, an online survey of 74 outdoor pool users illustrates how a theory-based behavioral analysis can be used to identify people's perceptions of a place in their environment. Regression analysis of close-ended data demonstrated that intention "to use the outdoor pool next summer season" was predicted (R(2)=.636, p<.0001) from the weighted combination of attitude (ß=.663, p<.0001) and perceived control (ß=.197, p<.05). The importance of attitude suggests that use can be encouraged by addressing advantages. The content analysis of open-ended elicitation data revealed that users believed that pool use provides physical, social, and emotional advantages and could be made easier by modifying facility parameters. Implications for promoting and designing public health spaces to attract users to aquatic environments and to improve health are discussed.