An assessment of a biosensor system for the quantification of microcystins in freshwater cyanobacterial blooms.

Microcystin-producing cyanobacterial blooms are a global issue threatening drinking water supplies and recreation on lakes and beaches. Direct measurement of microcystins is the only way to ensure waters have concentrations below guideline concentrations; however, analyzing water for microcystins takes several hours to days to obtain data. We tested LightDeck Diagnostics' bead beater cell lysis and two versions of the quantification system designed to give microcystin concentrations within 20 min and compared it to the standard freeze-thaw cycle lysis method and ELISA quantification. The bead beater lyser was only 30 % effective at extracting microcystins compared to freeze-thaw. When considering freeze-thaw samples analyzed in 2021, there was good agreement between ELISA and LightDeck version 2 (n = 152; R2 = 0.868), but the LightDeck slightly underestimated microcystins (slope of 0.862). However, we found poor relationships between LightDeck version 2 and ELISA in 2022 (n = 49, slopes 0.60 to 1.6; R2 < 0.6) and LightDeck version 1 (slope = 1.77 but also a high number of less than quantifiable concentrations). After the quantification issues are resolved, combining the LightDeck system with an already-proven rapid lysis method (such as microwaving) will allow beach managers and water treatment operators to make quicker, well-informed decisions.

Development, validation and reliability of a questionnaire to evaluate coaches' and players' perceptions, learning, and resources regarding the ruck in rugby union.

Background: The lack of a reliable research tool for assessing the attitudes, behaviours, and learning resources of rugby coaches and players regarding the ruck event is a significant gap in rugby research. Objectives: This study aimed to adapt an existing questionnaire focused on the tackle event and to validate and establish the reliability of the instrument. The questionnaire explores the attitudes, behaviours, and learning resources of rugby coaches and players, with a focus on the ruck event and its impact on coach and player development, as well as coaching practices. Methods: A seven-step design process was followed to validate the questionnaire's content, construct, clarity, and relevance. A panel of 12 experts evaluated the questionnaire, followed by a test-retest procedure involving 15 coaches and 16 players, highlighting the effectiveness of this questionnaire, and emphasising its potential to generate data that can impact the field of rugby coaching and player development. Results: The questionnaire was deemed appropriate and clear by the expert panel, with an average completion time of 22 minutes. Moderate to good agreement was observed among players (ICC Agreement = 0.71) and coaches (ICC Agreement = 0.88), with high response consistency (ICC Consistency = 0.71 for players and 0.87 for coaches). Significant agreement was also found in Kendall's W scores (players = 0.85, coaches = 0.93, p<0.01). Conclusion: This study presents a developed questionnaire noted for its clarity, reliability, and consistency. It serves as a valuable tool for future rugby research, with the potential to impact coach and player development significantly.

Optimizing Concussion Care Seeking: Using Machine Learning to Predict Delayed Concussion Reporting.

BACKGROUND: Early medical attention after concussion may minimize symptom duration and burden; however, many concussions are undiagnosed or have a delay in diagnosis after injury. Many concussion symptoms (eg, headache, dizziness) are not visible, meaning that early identification is often contingent on individuals reporting their injury to medical staff. A fundamental understanding of the types and levels of factors that explain when concussions are reported can help identify promising directions for intervention. PURPOSE: To identify individual and institutional factors that predict immediate (vs delayed) injury reporting. STUDY DESIGN: Case-control study; Level of evidence, 3. METHODS: This study was a secondary analysis of data from the Concussion Assessment, Research and Education (CARE) Consortium study. The sample included 3213 collegiate athletes and military service academy cadets who were diagnosed with a concussion during the study period. Participants were from 27 civilian institutions and 3 military institutions in the United States. Machine learning techniques were used to build models predicting who would report an injury immediately after a concussive event (measured by an athletic trainer denoting the injury as being reported "immediately" or "at a delay"), including both individual athlete/cadet and institutional characteristics. RESULTS: In the sample as a whole, combining individual factors enabled prediction of reporting immediacy, with mean accuracies between 55.8% and 62.6%, depending on classifier type and sample subset; adding institutional factors improved reporting prediction accuracies by 1 to 6 percentage points. At the individual level, injury-related altered mental status and loss of consciousness were most predictive of immediate reporting, which may be the result of observable signs leading to the injury report being externally mediated. At the institutional level, important attributes included athletic department annual revenue and ratio of athletes to athletic trainers. CONCLUSION: Further study is needed on the pathways through which institutional decisions about resource allocation, including decisions about sports medicine staffing, may contribute to reporting immediacy. More broadly, the relatively low accuracy of the machine learning models tested suggests the importance of continued expansion in how reporting is understood and facilitated.

The study for the roles of intratracheally administered histamine in the neutrophil-mediated acute lung injury in rats: / 결핵

BACKGROUND: Neutrophils are considered to play critical roles in the development of acute respiratory distress syndrome. Histamine, which is distributed abundantly in lung tissue, increases the rolling of neutrophills via increase of P-selectin expression on the surface of endothelial cells and is known to have some interrelationships with IL-1, IL-8 and TNF-alpha. We studied to investigate the effect of the histamine on the acute lung injury of the rats induced by intratracheal insufflation of TNF-alpha which has less potency to cause lung injury compared to IL-1 in rats. METHODS: We intratracheally instilled saline or TNF(R&D, 500ng), IL-1(R&D, 50ng)or histamine of varius dose(1.1, 11 and 55 microg/kg) with and without TNF separately in Sprague-Dawley rats weighing 270-370 grams. We also intratracheally treated IL-l(50ng) along with histamine(55 microg/kg). In cases, there were synergistic effects induced by histamine on the parameters of TNF-induced acute lung injury, antihistainmes(Sigma, mepyramine as a H1 receptor blockade and ranitidine as a H2 receptor blockade, 10 mg/kg in each)were co-administered intravenously to the rats treated TNF along with histamine(1.1 microg/kg) intratracheally. Then after 5 h we measured lung lavage neutrophil numbers, lavage cytokine-induced neutrophil chemoatt- ractants(CINC), lung myeloperoxidase activity(MPO) and lung leak. We also intratracheally insufflated TNF with/without histamine(11 microg/kg), then after 24 h measured lung leak in rats. Statistical analyses were done by Kruskal-Wallis nonparametric ANOVA test with Dunn's multiple comparison test or by Mann-Whitney U test. RESULTS: We found that rats given TNF, histamine alone(11 and 55 microg/kg), and TNF with histamine(1.1, 11, and 55 microg/kg) intratracheally had increased (P<0.05) lung MPO activity compared with saline-treated control rats. TNF with histamine 11 microg/kg had increased MPO activity (P=0.0251) compared with TNF-treated rats. TNF and TNF with histamine(l.l, 11,, and 55 microg/kg) intratracheally had all increased (P<0.05) lung leak, lavage neutophil numbers and lavage CINC activities compared with saline. TNF with histamine 1.1 microg/kg had increased (P=0.0367) lavage neutrophil numbers compared with TNF treated rats. But there were no additive effect of histamine with TNF compared with TNF alone in acute lung leak on 5 h and 24 h in rats. Treatment of rats with the H1 and H2 antagonists resulted in inhibitions of lavage neutrophil accumulations and lavage CINC activity elevations elicited by co-treated histamine in TNF-induced acute lung injury intratracheally in rats. We also found that rats given IL-1 along with histamine intratracheally did not have increase in lung leak compared with IL-1 treated rats. CONCLUSION: Histamine administered intratracheally did not have synergistic effects on TNF-induced acute lung leak inspite of additive effects on increase in MPO activity and lavage neutrophil numbers in rats. These observations suggest that instilling histamine intratracheally would not play synergistic roles in neutrophil-mediated acute lung injury in rats.

Nrf2 Activator PB125 as a Potential TherapeuticAgent Against COVID-19

Nrf2 is a transcription factor that regulates cellular redox balance and the expression of a wide array of genes involved in immunity and inflammation, including antiviral actions. Nrf2 activity declines with age, making the elderly more susceptible to oxidative stress-mediated diseases, which include type 2 diabetes, chronic inflammation, and viral infections. Published evidence suggests that Nrf2 activity may regulate important mechanisms affecting viral susceptibility and replication. We examined gene expression levels by GeneChip microarray and by RNA-seq assays. We found that the potent Nrf2 activating composition PB125(R) downregulates ACE2 and TMPRSS2 mRNA expression in human liver-derived HepG2 cells. ACE2 is a surface receptor and TMPRSS2 activates the spike protein for SARS-Cov-2 entry into host cells. Furthermore, in endotoxin-stimulated primary human pulmonary artery endothelial cells we report the marked downregulation by PB125 of 36 genes encoding cytokines. These include IL1-beta, IL6, TNF-, the cell adhesion molecules ICAM1, VCAM1, and E-selectin, and a group of IFN-{gamma}-induced genes. Many of these cytokines have been specifically identified in the "cytokine storm" observed in fatal cases of COVID-19, suggesting that Nrf2 activation may significantly decrease the intensity of the storm.