Integrated plasma proteomic and single-cell immune signaling network signatures demarcate mild, moderate, and severe COVID-19

The biological determinants underlying the range of coronavirus 2019 (COVID-19) clinical manifestations are not fully understood. Here, over 1,400 plasma proteins and 2,600 single-cell immune features comprising cell phenotype, endogenous signaling activity, and signaling responses to inflammatory ligands are cross-sectionally assessed in peripheral blood from 97 patients with mild, moderate, and severe COVID-19 and 40 uninfected patients. Using an integrated computational approach to analyze the combined plasma and single-cell proteomic data, we identify and independently validate a multi-variate model classifying COVID-19 severity (multi-class area under the curve [AUC]training = 0.799, p = 4.2e-6; multi-class AUCvalidation = 0.773, p = 7.7e-6). Examination of informative model features reveals biological signatures of COVID-19 severity, including the dysregulation of JAK/STAT, MAPK/mTOR, and nuclear factor κB (NF-κB) immune signaling networks in addition to recapitulating known hallmarks of COVID-19. These results provide a set of early determinants of COVID-19 severity that may point to therapeutic targets for prevention and/or treatment of COVID-19 progression

RELATIONSHIP BETWEEN 24-H VOID FREQUENCY, HYDRATION STATUS, AND INGESTED BEVERAGE COMPOSITION IN HEALTHY MALES

Matthew A. Tucker, Matthew S. Ganio, Jenna M. Burchfield & Nicole E. Moyen Department of Health, Human Performance, and Recreation, University of Arkansas, Fayetteville, Arkansas Void frequency (VF) increases with increased fluid intake; however, the relationship between VF and hydration status with controlled fluid intake is unknown. Further, the influence of different beverage types on VF is unclear. PURPOSE: To investigate the relationship between hydration status and VF in healthy males when given a fixed volume of different beverages. METHODS: In a randomized order, 32 healthy males (age 23.7 ± 4.9 y; mass 75.9 ± 11.7 kg) completed four weeks of testing consuming various beverage treatments (Txt) over 24-h with total daily fluid intake equal to 35 ml/kg body mass (including fluids from beverages and moisture content of a prescribed diet). Txt were: water only (A), water+caloric cola (B), water+non-caloric cola (C), or water+caloric cola+non-caloric cola+orange juice (D). Volumes of fluid were split evenly between beverages within each 24-h treatment. 24-h urine osmolality (24-Uosm), specific gravity (24-USG), volume (24-Uvol) and VF were measured during 24-h of intervention. 24-h urine samples were classified as euhydrated (24-USG ≤1.020 and 24-Uosm ≤700 mOsm/kg) or hypohydrated (24-USG \u3e1.020 and 24-Uosm \u3e700 mOsm/kg). RESULTS: As designed, total fluid intake from all sources was the same across treatments (2659 ± 400 ml; p\u3e0.05). There were no differences between treatments in 24-Uosm or 24-Uvol (grand mean: 607 ± 211 mOsm/kg and 1536 ± 593 ml, respectively; p\u3e0.05). VF over 24-h was not different between Txt A, B, C, and D (5.2 ± 1.7, 5.3 ± 1.8, 5.6 ± 2.0, and 5.1 ± 1.8, respectively; p\u3e0.05). Independent of beverage treatment, VF significantly correlated with 24-USG (r= -.47; posm (r= -.46; pSG and 24-Uosm values, a receiver operating curve analysis produced an area under the curve of 73.7% with a 95% CI [64.9, 82.5] (pCONCLUSION: These results indicate that VF is significantly correlated with hydration status, such that higher values of 24-USG and 24-Uosm are inversely related to VF. Further, urinary measures of hydration status and VF were not different between beverage treatments, suggesting that ingestion of different beverages does not affect voiding behavior in healthy males. Funded by The Coca-Cola Company

VALIDITY OF UCHEK IPHONE APPLICATION ON ASSESSING DEHYDRATION FROM URINE SAMPLES

1Christian B. Ridings, 1J.D. Adams, 1Matthew S. Ganio, 1Brendon P. McDermott, 2Dimitris Bougatsas, 1Stavros A. Kavouras, FACSM. 1Human Performance Laboratory, Department of Health, Human Performance, and Recreation, University of Arkansas, Fayetteville, AR; 2Laboratory of Nutrition & Clinical Dietetics, Harokopio University, Athens, Greece The widespread acceptance of smartphones presents an opportunity for the development of mobile health applications and accessories. These devices can help reduce costs and improve accessibility of healthcare. PURPOSE: To determine the validity of the uChek iPhone application on measuring hydration status compared to urine refractometry and urine dipstick applicators. METHODS: Fifty-three urine samples were analyzed for urine specific gravity (USG) by a refractometer under a well-lit area. Urine strips (Multistix 10 SG Reagent Strips) were used to record USG by the Multistix label compared to an 8-color control stick, and was then analyzed by the uChek application. RESULTS: Sensitivity analysis showed that overall accuracy of USG via strips and uChek were 68 and 67.3%, respectively. Specifically, the sensitivity of a positive urine test for dehydration (USG≥1.020) was 55.6% with the strips and 51.9% with the iPhone app. The specificity for detecting hypohydrated persons was 36.4% and 31.8% for the strips and iPhone app, respectively. CONCLUSION: We concluded that the iPhone application uChek is not accurately detecting dehydration mainly due to limitations of the urine strips

PHYSIOLOGICAL VALIDATION OF HYDRATION KNOWLEDGE, ATTITUDES AND BEHAVIOR SURVEY: PRELIMINARY FINDINGS

Aidan P. Fiol1, Whitley C. Atkins1, Rachel B. Grunnert1, Matthew S. Ganio1, Jennifer C. Veilleux2, Brendon P. McDermott,1 FACSM 1Exercise Science Research Center, University of Arkansas, Fayetteville, AR; 2Department of Psychological Science, University of Arkansas, Fayetteville, AR Adequate water consumption is associated with positive short- and long-term health outcomes and improved homeostasis. A validated survey that allows greater understanding of water intake relating to knowledge, attitudes, and behaviors would better connect hydration and overall health research. PURPOSE: Examine the validity of our hydration survey on hydration knowledge, attitudes, and behaviors among adults using urine analysis, and diet and fluid intake. METHODS: Thirty adults (28 ± 9y 167.9 ± 10.9cm, 75.27 ± 18.02kg) were recruited from the northwest Arkansas area and were given the hydration survey to complete. Following completion, participants collected all urine produced for the next 24-hr. Participants self-recorded dietary food and fluid consumption and urinary void frequency for the 24-hr period following survey administration. Urine was analyzed for specific gravity (USG), osmolality (Uosm), 24-hr volume, and void frequency. Survey data was analyzed and Pearson r correlations were conducted on survey results compared to hydration variables. Results are reported as mean ± standard deviation. RESULTS: Mean 24-hr urine volume was 2429.33 ± 1021.96mL, USG was 1.011 ± 0.004, Uosm was 372.72 ± 146.39mOsm·kg-1, and void frequency was 8 ± 3. Average 24-hr fluid intake was 4187.84 ± 1532.71mL. Total Fluid Behavior Index score was not significantly correlated with hydration variables (p≥.268). Total Hydration Facilitator and Barrier scores were not significantly correlated with hydration variables (p≥.224). Total Hydration Knowledge score was not significantly correlated with hydration variables (p≥.086), though a higher number of correct items showed a positive correlation with 24-hr void frequency (r=.363, p=.049). Among participants reaching an optimal hydration cutpoint of 24-hr Uosm\u3c500mOsm·kg-1 (n=23), there was no significant correlation between hydration variables with Fluid Behavior Index score (p≥.149), Hydration Knowledge score (p≥.126), or Hydration Facilitator and Barrier scores (p≥.112) CONCLUSION: Our data demonstrate an inability of our hydration survey knowledge, habits, or barriers to relate to physiological hydration variables in our largely hydrated participants. Larger participant numbers are required for more robust physiological survey validation

HYDRATION BIOMARKERS: CREATING A NEW HYDRATION ASSESSMENT TECHNIQUE

Jenna M. Burchfield, Matthew S. Ganio, Stavros A. Kavouras, FACSM, J. D. Adams1, Melina A. Gonzalez, & Christian B. Ridings; 1Human Performance Laboratory, Department of Health, Human Performance, and Recreation, University of Arkansas, Fayetteville, Arkansas Despite the necessity of euhydration, an optimal, user-friendly, accurate hydration assessment technique fails to exist for the general population. PURPOSE: The purpose of this study was to investigate a new hydration assessment technique, which utilizes no equipment or technical expertise to administer. We hypothesized that the number of urine voids over a 24-hr time period correlates to hydration status. METHODS: Forty-seven healthy individuals (n=24 females) participated (age 22±4 years, body mass 76±17 kg, height 1.64±0.27 m, body fat 20.4±8.7%). Participants abstained from caffeine and alcohol during the experiment. Participants were educated on how to record diet, fluid intake, and void output measurements. For 24-hours participants voided at a normal urge into a standard medical-grade container. Participants indicated volume, time, and urge of each void. Ad libitum fluid and food intake also were recorded over the 24 hrs. Total 24-hr urine volume, osmolality, specific gravity (USG), and color were measured. Subjects were classified as euhydrated (USG≤1.020) or hypohydrated (USG\u3e1.020) according to standard criteria. RESULTS: Euhydrated (n=41) versus dehydrated (n=6) individuals had more voids (6±2 versus 4±1, respectively, P=0.02), greater 24-hr urine volume (2007±863 versus 1193±434 ml, P\u3c0.001), lower urine color (2±1 versus 4±1, P\u3c0.001), lower USG (1.012±0.004 versus 1.023±0.003, P\u3c0.001), and lower osmolality (431±143 versus 841±137 mOsm/kgH20, P\u3c0.001). As hypothesized, void number moderately correlated to hydration status as identified by USG (r=0.37, P\u3c0.001), and osmolality (r=0.38, P\u3c0.001). CONCLUSION: Void number positively correlated with USG and urine osmolality. This indicates that individuals with higher void numbers are more hydrated than those with lower void numbers over a 24-hr time period. Using void number as a hydration biomarker could be an optimal technique for the general public to self-assess hydration, considering it is as simple as counting void numbers over a 24-hr period. Funding provided by the University of Arkansas College of Education & Health Professions and the Office of Research & Economic Development

EFFECT OF SPACER GARMENT ON PHYSIOLOGICAL STRAIN DURING EXERCISE IN THE HEAT WHEN WEARING PROTECTIVE VESTS

J.D. Adams, Brendon P. McDermott, Christian B. Ridings, Lacey Mainer, Matthew S. Ganio, and Stavros A. Kavouras, FACSM. Human Performance Laboratory, Department of Health, Human Performance, and Recreation, University of Arkansas, Fayetteville, AR.; email: [email protected] Numerous cooling technologies have been developed to reduce heat strain when wearing a variety of protective ensembles during military and law enforcement duties. Some cooling vests demonstrate improved sweat evaporation and decreased core temperature when worn along with protective body armor. Spacer garments represent a potentially lighter, cost-efficient method for improved airflow. PURPOSE: To determine if a spacer garment reduces physiological strain during exercise in the heat when wearing a military protective vest or a law enforcement concealable vest. METHODS: Sixteen men (24.5 ± 3.9 yrs.; 179.5 ± 5.6 cm; 84.6 ± 12.3 kg) performed either 2 or 4 trials of treadmill walking (3.1 mph; 2% grade) over 120 min in a hot, dry environment (37°C, 30% rh, wind speed 3.5 m·s). Participants completed trials with either a military patrol vest or a law enforcement concealable vest, with either a spacer garment (Ps; Cs) or no spacer garment (Pc; Cc) in random order. During trials, participants wore Army Combat Uniform pants and physiological measurements that were measured every 5 min included gastrointestinal temperature (TGI), mean skin (Tsk) temperature, heart rate (HR), and sweating rate (SR). RESULTS: In the patrol trial (Ps vs. Pc), no differences in final TGI (38.2 ± 0.4 vs. 38.3 ± 0.4°C), Tsk (35.0 ± 0.9 vs. 35.0 ± 1.0 °C), HR (142 ± 19 vs. 143 ± 23 bpm), or SR (1.15 ± 1.13 vs. 1.54 ± 0.46 L/hr) existed (P\u3e0.05). In the concealable trial (Cs vs. Cc), no differences in final TGI (38.0 ± 0.4 vs. 38.1 ± 0.3°C), Tsk (35.3 ± 1.1 vs. 35.6 ± 0.9°C), HR (132 ± 20 vs. 135 ± 20 bpm), or SR (1.39 ± 0.52 vs.1.37 ± 0.18 L/hr) existed (P\u3e0.05). CONCLUSION: When participants exercised with either a military protective vest or a law enforcement concealable vest while wearing the spacer garment, there were no thermoregulatory differences when compared to control trials. Thus, the passive spacer garment had no effect on the physiological responses during mild exercise in the heat. Funded by Cortac