Crossover Design Research Articles

483 DMI, gastrointestinal tract fermentation and permeability, and white blood cell responses in beef heifers provided post-ruminal yeast cell wall extract following feed restriction

Abstract The objectives of this study were to 1) evaluate gastrointestinal fermentation characteristics and permeability, and white blood cell concentrations as beef heifers recovered from a feed restriction (FR) challenge, and 2) determine whether post-ruminal provision of a yeast cell wall extract (YCW) modulated responses. Ruminally cannulated beef heifers (n = 8) were used in a 2 × 2 crossover design. Treatments (TRT) included an abomasal infusion of a placebo (PLC; maltodextrin) or YCW derived from Saccharomyces cerevisiae (2 g/dose) infused at 0600 and 1800 h. Heifers were subject to 5-d of FR (offering 30% of ad libitum DMI) followed by five 5-d recovery phases (REC1, REC2, REC3, REC4, and REC5). Daily DMI, and rumen, fecal, and blood samples were collected on d 5 of FR, REC1, REC3, and REC5 with an additional collection on d 2 of REC1. Total gastrointestinal and post-ruminal permeability were measured using a ruminal dose of Cr-EDTA and an abomasal dose of Co-EDTA, respectively, during FR, REC1, REC3, and REC5 with concurrent urine collection. Data were analyzed to determine the effects of YCW provision, day, and the 2-way interaction. There were no TRT or TRT×day effects for any variables except DMI (interaction, P = 0.04), where DMI did not differ during the FR challenge; however, heifers provided YCW had a greater increase by REC1 compared with PLC heifers without any means differing. DMI progressively increased from FR, to REC1, REC2, and REC5. Ruminal acetate concentration increased (P < 0.01) from FR (45.4 mmol/L) to REC1 d5 (60.1 mmol/L) and increased further to REC3 (67.3 mmol/L) with no change thereafter (68.3 mmol/L in REC5). Ruminal propionate concentration increased (P < 0.01) from FR (14.5 mmol/L) to REC1 d2 (27.7 mmol/L) and was not different thereafter. Similarly, butyrate concentration increased (P < 0.01) from FR (7.9 mmol/L) to REC1 (d 5; 15.0 mmol/L) and was not different thereafter. Fecal acetate, propionate, and butyrate concentrations were not affected by day (P ≥ 0.12). Total gastrointestinal and post-ruminal permeability were greatest during FR (P < 0.01; 2.6% and 2.2% of the infused marker was excreted in urine, respectively) and decreased to 1.6% and 1.1%, respectively, in REC3 followed by an increase in REC5 for total (2.0% of infused) but not post-ruminal (1.0% of infused) permeability. The concentration of neutrophils (P < 0.01) was greatest on REC1 d2 (4.84 ×106 μmol/mL) and eosinophil concentration (P < 0.01) was greatest during FR (0.53 ×106 μmol/mL), least on d 2 of REC1 and increased on d 5 of REC1 but was not different thereafter. WBC (P < 0.01) gradually decreased (P = 0.02) from FR and REC1 d2 to REC3 and REC5. Post-ruminal provision of YCW may accelerate increases in DMI early in the recovery from FR, yet treatment did not modulate ruminal or fecal SCFA, blood concentrations, or gastrointestinal permeability.

A Deployable Viscoelastic Coagulation Monitor Enables Point-of-Care Assessment of Coagulopathy in Swine With Polytrauma.

Absence of pre-hospital coagulation tests challenges prompt management of hemostasis after trauma. The Viscoelastic Coagulation Monitor (VCM, Entegrion, Durham, NC) is a hand-held coagulation test for point-of-care. We evaluated VCM in a translational swine polytrauma model, hypothesizing that VCM correlates with a laboratory reference method, the TEG 5000 (Haemonetics, Boston, MA), and can identify coagulopathic phenotypes relevant to trauma. Our secondary hypothesis was that pre-warming of VCM disposable test cartridges using a heating plate versus pre-warming of cartridges by carrying the cartridge in the user's pocket does not significantly alter results. This study was conducted in tandem with a parent study involving anesthetized, mechanically ventilated swine (n = 20; 54 ± 5 kg) that encountered traumatic brain injury, pulmonary contusion and hemorrhage, or combination/polytrauma injury. Blood was collected at baseline, post-injury, post-shock, post-transfusion, and 6-, 24-, and 48 h post-injury to perform VCM at point-of-care. Within-group effect of time was assessed. Spearman correlation examined linear relations between VCM and standard laboratory-based coagulation tests; as well as lactate, ionized calcium, and body temperature. Logistic regression examined predictiveness of VCM to identify coagulopathic phenotypes, with receiver operator characteristic curves generated to assess diagnostic capability. At a subset of timepoints, necessity of pre-warming the VCM test cartridge using a heating plate versus pre-warming the cartridge by placement in the user's pocket was assessed by conducting simultaneous tests on two separate instruments, with results analyzed by paired t-test with crossover design. VCM revealed time-dependent changes in clotting time, clot formation time (CFT), alpha, maximum clot firmness (MCF), and lysis index (LI30). All VCM metrics correlated with the respective TEG 5000 metrics, with strongest correlation for VCM MCF with TEG MA (rhos = 0.77, P < .0001) and VCM LI30 with TEG LY30 (rhos = -0.76, P < .0001). VCM demonstrated good (area under the curve >0.70) to excellent (area under the curve >0.90) diagnostic accuracy in detection of low platelet count (MCF), low hematocrit (clotting time, clot formation time, alpha, and MCF), low fibrinogen (MCF), and high fibrinogen (alpha, MCF). There was no statistically or clinically relevant effect of cartridge warming method on results. In a trauma model, VCM detected significant changes in coagulation at point-of-care in a simplified portable form factor. VCM could enable informed hemostasis management in pre-hospital settings where coagulations tests are unavailable, pending further validation in clinical trials.

Large-scale brain connectivity changes following the administration of lysergic acid diethylamide, d-amphetamine, and 3,4-methylenedioxyamphetamine.

Psychedelics have recently attracted significant attention for their potential to mitigate symptoms associated with various psychiatric disorders. However, the precise neurobiological mechanisms responsible for these effects remain incompletely understood. A valuable approach to gaining insights into the specific mechanisms of action involves comparing psychedelics with substances that have partially overlapping neurophysiological effects, i.e., modulating the same neurotransmitter systems. Imaging data were obtained from the clinical trial NCT03019822, which explored the acute effects of lysergic acid diethylamide (LSD), d-amphetamine, and 3,4-methylenedioxymethamphetamine (MDMA) in 28 healthy volunteers. The clinical trial employed a double-blind, placebo-controlled, crossover design. Herein, various resting-state connectivity measures were examined, including within-network connectivity (integrity), between-network connectivity (segregation), seed-based connectivity of resting-state networks, and global connectivity. Differences between placebo and the active conditions were assessed using repeated-measures ANOVA, followed by post-hoc pairwise t-tests. Changes in voxel-wise seed-based connectivity were correlated with serotonin 2 A receptor density maps. Compared to placebo, all substances reduced integrity in several networks, indicating both common and unique effects. While LSD uniquely reduced integrity in the default-mode network (DMN), the amphetamines, in contrast to our expectations, reduced integrity in more networks than LSD. However, LSD exhibited more pronounced segregation effects, characterized solely by decreases, in contrast to the amphetamines, which also induced increases. Across all substances, seed-based connectivity mostly increased between networks, with LSD demonstrating more pronounced effects than both amphetamines. Finally, while all substances decreased global connectivity in visual areas, compared to placebo, LSD specifically increased global connectivity in the basal ganglia and thalamus. These findings advance our understanding of the distinctive neurobiological effects of psychedelics, prompting further exploration of their therapeutic potential.

Hydrolyzed collagen supplementation prior to resistance exercise augments collagen synthesis in a dose-response manner in resistance-trained, middle-aged men.

Resistance exercise (RE) increases collagen synthesis in young and older men, while hydrolyzed collagen (HC) ingestion improves this response to RE in a dose-response manner in young men. However, the collagen synthesis response to RE with and without HC in middle-aged men is unknown. Eight resistance-trained men (age: 49±8 years; height: 1.78±0.02m; mass: 90±4kg) took part in this double-blind, crossover design study, and undertook 4×10 repetitions of lower-limb RE at maximum load, after consuming 0g, 15g, or 30g vitamin C-enriched HC. We analyzed venous blood samples for N-terminal propeptide of type 1 pro-collagen (PINP), β-isomerized C-terminal telopeptide of type 1 collagen (β-CTx) and 18 collagen amino acids throughout all three interventions. The serum PINP concentration×time area-under-the-curve (AUC) was higher following 30g (169±28 µg/mL×h) than 15g (134±23 µg/mL×h, P<0.05) HC ingestion, and both 15g and 30g were higher than 0g HC (96±23 µg/mL×h, P<0.05). RE with 0g HC showed no change in serum PINP concentration. The AUCs for glycine, proline, hydroxyproline, alanine, arginine, lysine, serine, leucine, valine and isoleucine were greater with 30g than 15g and 0g HC ingestion (P<0.05), and greater with 15g than 0g HC ingestion (P<0.05). Plasma β-CTx concentration decreased after RE independently of HC dose. Our study suggests connective tissue anabolic resistance to RE in middle-aged men but ingesting 15g HC rescues the collagen synthesis response, and 30g augments that response further. This dose-response is associated with the increased bioavailability of collagen amino acids in the blood, which stimulate collagen synthesis.

Clinically important interactions of macrolides and tetracyclines with dietary interventions-a systematic review with meta-analyses.

Effective management of drug-food interactions is crucial for enhancing antibiotics' efficacy/safety. Adhering to PRISMA guidelines, we conducted a systematic review to assess the impact of dietary interventions on the bioavailability of 15 macrolides and 10 tetracyclines. We included studies examining the influence of food, beverages, antacids, and mineral supplements on the pharmacokinetic parameters of orally administered macrolides and tetracyclines. We searched Medline (via PubMed), Embase and Cochrane Library databases up to December 2022. Risk of bias was assessed using Cochrane and NIH tools. Quantitative analyses were conducted if two or more comparable food-effect studies were available; otherwise, a qualitative summary was provided. We included 120 studies from 97 reports. Meta-analyses were conducted for 8 macrolides and 4 tetracyclines, with qualitative synthesis for 10 and 9, respectively. About 64% of the studies were open-label, crossover designs. Our assessment found that 37% of the studies had a high risk of bias, while only 6% had low risk. Food significantly affected 10 of 13 macrolides (77%) and 6 of 7 tetracyclines (86%). High positive effects on bioavailability were seen with extended-release azithromycin and clarithromycin, and erythromycin estolate. High negative impacts were observed with erythromycin propionate and stearate, azithromycin capsules, demeclocycline and omadacycline. Antacids and mineral supplements significantly decreased tetracyclines absorption. Milk and grapefruit juice showed variable impacts on absorption. Interactions depend on antibiotics' physicochemical characteristics, intervention type, drug formulation and potential patient factors. The quality of evidence was rated low due to outdated studies, methodological diversity and unequal data availability.

Inhalation of Hydrogen-rich Gas before Acute Exercise Alleviates Exercise Fatigue: A Randomized Crossover Study.

Hydrogen, as an antioxidant, may have the potential to mitigate fatigue and improve selected oxidative stress markers induced by strenuous exercise. This study focused on a previously unexplored approach involving pre-exercise inhalation of hydrogen-rich gas (HRG). Twenty-four healthy adult men first completed pre-laboratories to determine maximum cycling power (Wmax) and maximum cycling time (Tmax). Then they were subjected to ride Tmax at 80% Wmax and 60-70 rpm on cycle ergometers after inhaled HRG or placebo gas (air) for 60-minute in a double-blind, counterbalanced, randomized, and crossover design. The cycling frequency in the fatigue modeling process and the rating of perceived exertion (RPE) at the beginning and end of the ride were recorded. Before gas inhalation and after fatigue modeling, visual analog scale (VAS) for fatigue and counter-movement jump (CMJ) were tested, and blood samples were obtained. The results showed that compared to a placebo, HRG inhalation induced significant improvement in VAS, RPE, the cycling frequency during the last 30 seconds in the fatigue modeling process, the ability to inhibit hydroxyl radicals, and serum lactate after exercise (p<0.028), but not in CMJ height and glutathione peroxidase activity. The cycling frequency during the last 30 seconds of all other segments in the fatigue modeling process was within the range of 60-70 rpm. In conclusion, HRG inhalation prior to acute exercise can alleviate exercise-induced fatigue, maintain functional performance, and improve hydroxyl radical and lactate levels.

The interplay of brain neurotransmission and mental fatigue: A research protocol.

Mental fatigue (MF) significantly affects both cognitive and physical performance. However, the precise mechanisms, particularly concerning neurotransmission, require further investigation. An implication of the role of dopamine (DA) and noradrenaline (NA) is stated, but empirical evidence for this theory still needs to be provided. To address this gap, we aim to investigate the role of brain neurotransmission in elucidating if, and how prolonged cognitive activity induces MF and its subsequent impact on cognitive performance. This study (registration number: G095422N) will adopt a randomized cross-over design with sixteen healthy participants aged 18-35 years. The sessions include a familiarization, two experimental (DA: 20mg Methylphenidate; NA: 8mg Reboxetine) conditions, and one placebo (lactose tablet: 10mg) condition. A 60-minute individualized Stroop task will be used to investigate whether, and how the onset of MF changes under the influence of reuptake inhibitors. Attention and response inhibition will be assessed before and after the MF-inducing task using a Go/NoGo task. The integration of physiological (electroencephalography, heart rate), behavioral (attention, response inhibition), and subjective indicators (scales and questionnaires) will be used to detect the underlying mechanisms holistically. Data analysis will involve linear mixed models with significance at p<0.05. The integration of diverse techniques and analyses offers a comprehensive perspective on the onset and impact of MF, introducing a novel approach. Future research plans involve extending this protocol to explore the connection between brain neurotransmission and physical fatigue. This protocol will further advance our understanding of the complex interplay between the brain and fatigue.

The Effects of L-Citrulline and Malic Acid on Substrate Utilisation and Lactate Elimination

Endurance athletes often aim to improve their aerobic metabolism. The aim of this pilot study was to examine if malic acid and L-citrulline supplementation can improve aerobic metabolism and lactate elimination. Nine young (23.9 ± 1.9 years) recreational male athletes participated in this study. Following a standardised breakfast and a body composition analysis (InBody720), 6000 mg of citrulline and 3000 mg of malic acid or a placebo of 300 mL of water were consumed on three separate days in a cross-over design using a double-blind method. Sixty minutes after the supplementation, participants completed a ramp bicycle spiroergometer protocol (35 W/3 min) until reaching a respiratory exchange ratio (RER) of 1.1, followed by a 9 min active recovery. Cadence, heart rate (HR), rate of perceived exertion (RPE), respiratory parameters and lactate levels were registered. The RPExHR value was calculated to accurately characterise exhaustion. During the exercise protocol, citrulline supplementation induced significantly lower RER values at 70-105-140 W compared to malic acid and the placebo, respectively. There was no difference in lactate levels neither during rest nor at RER 1.1. RPExHR rate values were significantly lower after malic acid supplementation compared to placebo at 175 and 210 W. Power at RER 1.1 was higher after malic acid (+4 W) and citrulline (+5 W) supplementation. Although the supplementation failed to decrease lactate levels, lower RER and RPE values may indicate a performance-enhancing benefit.

Efficacy of Pomegranate Peel Extract Mouthwash on Gingivitis and C-reactive protein

Background and objective: Gingivitis is characterized as the world’s most predominant inflammatory periodontal disease. Healthy mouth and teeth are obtained by brushing and uses mouthwash. And because of the side effects of chlorhexidine, pomegranate peel extract mouthwash used alternative which are thought to have anti-inflammatory effect. The aim of this study is to prepare a herbal mouthwash and to evaluate the efficacy of pomegranate peel extract mouthwash on plaque index, gingival index, and C-reactive protein on moderate gingivitis patients. Method: this study is a clinical crossover design study, which consists of 25 volunteers with moderate gingivitis, who visited Duhok Dental Training Center. This study started from December 2022 to March 2023. Plaque and gingival index with a sample of saliva for C-reactive protein measuring were taken as a baseline before each mouth wash and after that, the pomegranate peel extract mouthwash was used for two weeks, then three weeks wash out between them, then chlorhexidine mouth wash was used for two weeks (crossover method). All saliva samples were analyzed using (CRP ELISA Test). Results: Twenty-five patients who used pomegranate peel extract mouthwash showed a significant reduction after two weeks used (CRP=162.28, GI=0.54, PI=0.82) and p value (&lt;0.0001) in mean C-reactive protein, gingival index, and plaque index from baseline which was (CRP=669.78, GI=1.88, PI=2.11). Conclusion: Pomegranate peel extract mouthwash had been shown effective against moderate gingivitis in adult patients with no systemic diseases through lowering in mean salivary C-reactive protein, gingival index, and plaque index.

Music listening for improvement of sleep in post-acute rehabilitation of adults with acquired brain injury: A feasibility study

ABSTRACT Introduction Acquired brain injury (ABI) often leads to sleep disturbances impacting recovery. Previous research has shown that listening to music at bedtime can have a beneficial effect on sleep quality in various populations, but the impact of bedtime music on sleep problems related to ABI has not been studied. This feasibility study investigates the acceptability and effectiveness of music listening for sleep improvement in ABI patients. Methods In a within-subjects crossover design with repeated measures for 28 days, ABI patients with sleep disturbance were allocated to a control or music condition, switching conditions after 14 days. Acceptability was assessed with questionnaires on compliance and attrition. The effect was evaluated with the Pittsburgh Sleep Quality Index (PSQI) at baseline, day 14 and 28, daily sleep ratings, and actigraphy. Results The intervention was well-liked, and participants listened to the music most of the intervention nights. A descriptive statistical comparison of outcome data on PSQI and daily sleep ratings regarding how well participants slept indicated that participants (n = 4) experienced better sleep with the music intervention. The objective sleep measures reflected a slightly longer time to fall asleep with the music intervention. Discussion The results suggest good acceptability and a potential beneficial effect of the intervention. Music listening may be a safe and effective tool, and a full RCT is recommended to evaluate the efficacy of music listening for sleep improvement in ABI patients. https://www.clinicaltrials.gov, Identification number: NCT05620069.

Testosterone rapidly increases men’s emotion-based dehumanization of a conservatively dressed woman

Past research has found that sexualized women are often dehumanized (i.e., attributed reduced human qualities). However, the mechanisms contributing to such dehumanization remain poorly understood. In this pre-registered experiment involving a within-subject, placebo-controlled, cross-over design, we tested whether testosterone contributes to men’s (N = 120, age range: 18–38 years) dehumanization of women. After administration of intranasal testosterone or placebo gel, men watched a video of a woman wearing either modest (i.e., conservative) or revealing (i.e., sexualized) clothing (between-subjects factor) and then completed three subtle dehumanization tasks, measuring emotion-based, personality-based, and perceptual dehumanization. We hypothesized that testosterone would increase dehumanization, especially for men who watched the “sexualized-clothing” video. Instead, we found that, while men engaged in emotion-based dehumanization toward the sexualized woman both when they had testosterone and placebo, testosterone increased emotion-based dehumanization toward the conservatively dressed woman. Other forms of dehumanization were not affected by testosterone. We also explored whether personality (e.g., dominance) and biological (e.g., CAG repeat polymorphism) traits that have been found to moderate the effects of testosterone on some social behaviors also moderated the effects examined here, but we did not find any significant moderations. Overall, this experiment revealed a novel physiological mechanism affecting emotion-based dehumanization.

Comparison of cooking emissions mitigation between automated and manually operated air quality interventions in one-bedroom apartments

We implemented a crossover study design exposing 15 participants to two indoor air quality conditions in the Well Living Lab. The first condition, the Standard Control Condition, resembled the ventilation and air supply of a typical home in the USA with a manually operated stove hood. The second condition, Advanced Control, had an automated: (i) stove hood, (ii) two portable air cleaners (PAC), and (iii) bathroom exhaust. The PM2.5 sensors were placed in the kitchen, living room, bedroom, and bathroom. Once the sensor detected a PM2.5 level of 15 μg/m3 or higher, an air quality intervention (stove hood, PAC or bathroom exhaust) in that space was activated and turned off when the corresponding PM2.5 sensor had three consecutive readings below 6 μg/m3. Advanced Control in the overall apartment reduced PM2.5 concentration by 40% compared to the Standard Control. The PM2.5 concentration difference between Advanced and Standard Control was ~ 20% in the kitchen. This can be attributed to using the stove hood manually in 66.5% of cooking PM2.5 emission events for 323.6 h compared to 88 h stove hood used in automated mode alongside 61.9 h and 33.7 h of PAC use in living room and bedroom, respectively.

Risk perception increase due to COVID-19 impacted antenatal care utilization among women in an indigenous community

BackgroundRisk perception varies greatly among individuals, affecting their behavior and decision-making in risky situations. The COVID-19 pandemic affected worldwide, but the role of risk perception related to COVID-19 in ethnic minorities in Mexico is unclear. This study quantifies the impact of COVID-related risk perception (susceptibility and severity) and perceived fear on the utilization of antenatal care services among indigenous women in San Cristobal de las Casas, Chiapas, Mexico.MethodsWe conducted a retrospective crossover study between June and December 2021, interviewing 98 women from San Cristóbal de las Casas, Chiapas. In a crossover design, each subject acts as their own control, so we required the participants to have a previous pregnancy experience. A logistic model was used to calculate the odds ratio for the outcome of having an adequate number of antenatal care visits. The analysis considered the period (during or before the pandemic) as well as perceived severity and susceptibility levels as independent variables.ResultsCOVID-19 reduced antenatal care utilization by 50%. During the pandemic, the adjusted odds ratio for attending health antenatal care services was 0.83 (95% CI: 4.8, 14.5) compared to pre pandemics. Adjusted for fear of contagion, the mother’s perception of severity was associated with an increased likelihood of an insufficient number of antenatal visits. OR = 0.25 (95% CI: 0.10, 0.65).ConclusionThe risk perception for COVID-19 decreased the likelihood of receiving an adequate number of antenatal care visits.

Crocus Sativus Linnaeus (Saffron) intake does not affect physiological and perceptual responses during a repeated sprint test in healthy active young males

The study aimed to investigate the effects of acute ingestion of saffron (SAF) on physiological (i.e., heart rate and blood lactate) and perceptual (i.e., ratings of perceived exertion [RPE] and feeling scale) measures in response to a repeated-sprint ability test (RSS) in healthy young males (N = 22; mean ± SD: age, 21.7 ± 1.24 yrs.). All participants completed two experimental trials with a one-week washout period using a double-blind, placebo-controlled, crossover design. In each session, the participants were randomly chosen to receive either a capsule of saffron (300 mg) (SAF session) or a capsule of lactose (PLB session) two hours before performing the RSS.No significant differences (p > 0.05) were found for heart rate, RPE, and feeling scale between the SAF or PLB sessions at pre- and post-RSS. There were no significant changes (p > 0.05) in peak time, total time, fatigue index, and blood lactate in either the SAF or PLB sessions. Acute SAF ingestion did not significantly improve RSS performance nor physiological and perceptual measures in active young males. Future trials should address the topic by using shortened/prolonged higher doses of SAF on biological, physical, physiological, and perceptual responses to acute and chronic exercise.

Repetitive Transcranial Magnetic Stimulation Modulates Brain Connectivity in Children with Self-limited Epilepsy with Centrotemporal Spikes.

Interictal epileptiform discharges (IEDs) alter brain connectivity in children with epilepsy; this connectivity change may be a mechanism by which epilepsy induces cognitive deficits. Here, we test whether repetitive transcranial magnetic stimulation (rTMS), a non-invasive neuromodulation technique, modulates connectivity and reduces IEDs in children with epilepsy. Nineteen children with self-limited epilepsy with centrotemporal spikes (SeLECTS) participated in a cross-over study comparing the impact of active vs. sham rTMS on IEDs and brain connectivity. SeLECTS is an epilepsy syndrome affecting the motor cortex, and prior studies show that motor cortices become pathologically hyper-connected to frontal and temporal language cortices. Using a crossover design, we compared the effect of single doses of active versus sham motor cortex rTMS. Connectivity, which was quantified by the weighted phase lag index (wPLI), was measured before and after rTMS using single pulses of TMS combined with EEG (spTMS-EEG). Analyses focused on six regions: bilateral motor cortices and bilateral inferior frontal and superior temporal regions. IEDs were counted in the five minutes before and after rTMS. Active, but not sham, rTMS significantly and globally decreased wPLI connectivity between multiple regions, with the greatest reductions seen in the superior temporal region connections in the stimulated hemisphere. Additionally, there was a trend suggesting that rTMS decreases IED frequency. These findings underscore the potential of low-frequency rTMS to target pathologic hyperconnectivity and reduce IEDs in children with SeLECTS and potentially other pediatric epilepsy syndromes, offering a promising avenue for therapeutic intervention.

Digoxin and exercise effects on skeletal muscle Na+,K+-ATPase isoform gene expression in healthy humans.

In muscle, digoxin inhibits Na+,K+-ATPase (NKA) whereas acute exercise can increase NKA gene expression, consistent with training-induced increased NKA content. We investigated whether oral digoxin increased NKA isoform mRNA expression (qPCR) in muscle at rest, during and post-exercise in 10 healthy adults, who received digoxin (DIG, 0.25mg per day) or placebo (CON) for 14 days, in a randomised, double-blind and cross-over design. Muscle was biopsied at rest, after cycling 20min (10min each at 33%, then 67% ), then to fatigue at 90% and 3h post-exercise. No differences were found between DIG and CON for NKA α1-3 or β1-3 isoform mRNA. Both α1 (354%, P=0.001) and β3 mRNA (P=0.008) were increased 3h post-exercise, with α2 and β1-2 mRNA unchanged, whilst α3 mRNA declined at fatigue (-43%, P=0.045). In resting muscle, total β mRNA (∑(β1+β2+β3)) increased in DIG (60%, P=0.025) and also when transcripts for each isoform were normalised to CON then either summed (P=0.030) or pooled (n=30, P=0.034). In contrast, total α mRNA (∑(α1+α2+α3), P=0.348), normalised then summed (P=0.332), or pooled transcripts (n=30, P=0.717) did not differ with DIG. At rest, NKA α1-2 and β1-2 protein abundances were unchanged by DIG. Post-exercise, α1 and β1-2 proteins were unchanged, but α2 declined at 3h (19%, P=0.020). In conclusion, digoxin did not modify gene expression of individual NKA isoforms at rest or with exercise, indicating NKA gene expression was maintained consistent with protein abundances. However, elevated resting muscle total β mRNA with digoxin suggests a possible underlying β gene-stimulatory effect. HIGHLIGHTS: What is the central question of this study? Na+,K+-ATPase (NKA) in muscle is important for Na+/K+ homeostasis. We investigated whether the NKA-inhibitor digoxin stimulates increased NKA gene expression in muscle and exacerbates NKA gene responses to exercise in healthy adults. What is the main finding and its importance? Digoxin did not modify exercise effects on muscle NKA α1-3 and β1-3 gene transcripts, which comprised increased post-exercise α1 and β3 mRNA and reduced α3 mRNA during exercise. However, in resting muscle, digoxin increased NKA total β isoform mRNA expression. Despite inhibitory-digoxin or acute exercise stressors, NKA gene regulation in muscle is consistent with the maintenance of NKA protein contents.

Prolonged administration of oral phenylbutazone and firocoxib in horses has no impact on selected cytokine and growth factor concentrations in platelet-rich plasma and autologous protein solution.

To determine the effects of prolonged administration of the oral NSAIDs phenylbutazone and firocoxib on concentrations of cytokines and growth factors in platelet-rich plasma (PRP) and autologous protein solution (APS). 6 adult University owned horses. Horses were randomized to receive phenylbutazone (1 g, orally, q 12 h) or firocoxib (57 mg, orally, q 24 h) for 6 days. Blood was obtained and processed for APS (Pro-Stride) and PRP (Restigen) before the administration of NSAIDs and at 7 days (1 day following cessation of NSAIDs). Horses underwent a two-week washout period, during which blood was obtained at 14 days and 21 days. The protocol was repeated with a crossover design. PRP and APS were analyzed for concentrations of platelets, leukocytes, and several cytokines (IL-1β, IL-10, IL-6, IL-8, and tumor necrosis factor-α) and growth factors (PDGF, FGF-2, and TGF-β1) using immunoassays. Plasma was evaluated for drug concentrations. No significant differences existed in concentrations of growth factors and cytokines before or after prolonged administration of NSAIDs. There were significant differences in concentrations of leukocytes and platelets in PRP compared to APS, with higher concentrations of leukocytes at the day 7 time point (T) in APS (phenylbutazone) and in concentrations of platelets in APS at T0 (firocoxib) and in APS at T7 (phenylbutazone). Veterinarians can recommend the administration of these oral NSAIDs prior to obtaining blood for PRP and APS provided a single-day washout period is instituted.

Abstract 39: Tryptophan contributes to the sex-dependent loss of diurnal sodium excretion in BMAL1 knockout rats.

Kidney function is time-of-day-(TOD) dependent irrespective of light, sleep, activity, and meal timing. Recent discoveries reveal that diurnal (day/night) kidney function is maintained by an intrinsic molecular clock. We recently observed that loss of this transcriptional regulatory system impairs functional rhythms in electrolyte excretion in a novel rat model. When the core clock gene, brain and muscle ARNT-like protein 1 (BMAL1) is knocked out of Sprague-Dawley rats we now find that male, but not female, BMAL1 knockout (KO) rats no longer have diurnal changes in both glomerular filtration rate (GFR; Day:1.3±0.1 vs Night:1.2±0.1mL/min/100gBW;t-test p=0.919) along with sodium excretion (UNaV) (Day:819±58 vs Night:756±113 μmol/12h; t-test p=0.439) compared to littermate wildtype controls (CON; GFR Day:1.1±0.1 vs Night:1.6±0.1mL/min/100g; t-test p=0.044; UNaV Day: 849±138 vs Night:1423±102 μmol/12h; t-test p=0.003). LC/MS analysis of plasma from KO rats found that levels of tryptophan (trp) were significantly (2-way ANOVA p Interaction =0.847, p&lt;span style="font-size:10.8333px"&gt;Genotype&lt;/span&gt;=0.041, p TimeofDay =0.952) lower in KO males during the day (KO:77±6, CON:99±7μM; Sidak post-hoc p=0.049) but not night (KO:78±5, CON:99±12μM; Sidak post-hoc p=0.084) with no differences in plasma trp seen in KO females (Day:110±26,107±14μM vs Night:96±8,96±12μM KO and CON respectively). Trp had recently been shown to both regulate and be regulated by the circadian clock as well as increase GFR similar to other aromatic amino acids. This led us to hypothesize that TOD trp availability may underlie the changes seen in both UNaV and GFR in male KO rats. Therefore, we gave a single dose of trp (100 mg/kg,PO) at either 7pm (lights off, ZT12) or 7am (lights on, ZT0). After a 1 wk washout period, rats were dosed again in a crossover design. Male KO rats receiving trp at ZT0 had no significant change from baseline in UNaV (818±58 vs 784±61 μmol/12h) over the following 12hrs; however, a significant increase in UNaV (756±114 vs 1152±146 μmol/12h) following ZT12 dosing was observed (2-Way ANOVA p Interaction =0.038, p TrpTime =0.087, p TimeofDay =0.210; Sidak Post-Hoc: baseline vs ZT12 trp p=0.003) with no significant change seen in female KO or either male or female CON rats. Taken together these results suggest that BMAL1 is necessary for maintaining kidney physiologic rhythms in sodium excretion potentially through changes in trp availability.

Caffeine Gum Improves Reaction Time but Reduces Composure Versus Placebo During the Extra-Time Period of Simulated Soccer Match-Play in Male Semiprofessional Players.

This study aimed to determine whether caffeine gum influenced perceptual-cognitive and physical performance during the extra-time period of simulated soccer match-play. Semiprofessional male soccer players (n = 12, age: 22 ± 3years, stature: 1.78 ± 0.06m, mass: 75 ± 9kg) performed 120-min soccer-specific exercise on two occasions. In a triple-blind, randomized, crossover design, players chewed caffeinated (200mg; caffeine) or control (0mg; placebo) gum for 5min following 90min of soccer-specific exercise. Perceptual-cognitive skills (i.e.,passing accuracy, reaction time, composure, and adaptability) were assessed using a soccer-specific virtual reality simulator, collected pre- and posttrial. Neuromuscular performance (reactive-strength index, vertical jump height, absolute and relative peak power output, and negative vertical displacement) and sprint performance (15 and 30m) were measured at pretrial, half-time, 90min, and posttrial. Caffeine gum attenuated declines in reaction time (pre: 90.8 ± 0.8AU to post: 90.7 ± 0.8AU) by a further 4.2% than placebo (pre: 92.1 ± 0.8AU to post: 88.2 ± 0.8AU; p < .01). Caffeine gum reduced composure by 4.7% (pre: 69.1 ± 0.8AU to post: 65.9 ± 0.8AU) versus placebo (pre: 68.8 ± 0.8AU to post: 68.3 ± 0.8AU; p < .01). Caffeine gum did not influence any other variables (p > .05). Where caffeine gum is consumed by players prior to extra-time, reaction time increases but composure may be compromised, and neuromuscular and sprint performance remain unchanged. Future work should assess caffeine gum mixes with substances like L-theanine that promote a relaxed state under stressful conditions.

Low-carbohydrate diet in children and young people with type 1 diabetes: A randomized controlled trial with cross-over design

AimsWe investigated whether a short period of tightly controlled low-carbohydrate diet (LCD) leads to higher time in range without increasing the associated risks in children and young people with diabetes (CYPwD). MethodsThirty-five (CYPwD) were recruited into this randomized controlled cross-over study (20 female; 20 CSII; age 14.5 ± 2.9 years; HbA1c 48.9 ± 9.4 mmol/mol). The interventions were five and five weeks of ready-made food box deliveries of isocaloric diets in random order: either LCD (94.5 ± 4.7 g/day) or recommended carbohydrate diet (RCD) (191 ± 19.2 g/day). The outcomes were continuous glucose monitoring parameters, anthropometric, laboratory and quality of life (QoL) data. ResultsTime in range was significantly higher in the LCD than in the RCD period (77.1 % vs. 73.8 %, P=0.008). Times in hyperglycemia and average glycaemia were significantly lower in the LCD. There was no difference between the diets in time in hypoglycemia or glycemic variability. The subjects’ body weight and BMI were significantly lower during the LCD. There was no significant difference in the LDL-cholesterol levels. No significant differences were observed in the self-assessed QoL. ConclusionsShort-term LCD led to an improvement of glycemic parameters without increasing time in hypoglycemia, disturbing the lipid profile or negatively affecting the quality of life of CYPwD.