Mixed-effects ANOVA Research Articles

Research on physical health assessment and intervention strategies for college students based on biomarkers

Effective physical health assessments and intervention strategies are crucial for college students, who often face unique stressors that can impact their physiological health. In this analysis, we explore how biomarkers can be utilized to assess physical health and the efficacy of targeted interventions for this demographic. Hypotheses were developed based on the three intervention groups: (1) a face-to-face intervention group receiving a tailored wellness program, (2) an Internet of Things (IoT) gadgets intervention group using a health management assessment, and (3) a control group with only baseline and follow-up measurements. College students (n = 204) were randomized into these groups. Biomarkers related to stress and inflammation (e.g., cortisol levels, high-sensitivity C-reactive protein (hs-CRP)) were measured at baseline, post-intervention (10 weeks), and follow-up (36 weeks). Psychological and physical health outcomes were also assessed using standardized questionnaires. The Mixed-Effects Models, ANOVA, and Structural Equation Modeling (SEM) were used to analyze the differences between groups, changes in biomarkers over time, and the relationships between psychological, physiological, and lifestyle variables. Results indicated that the face-to-face wellness group demonstrated significantly better physical health outcomes compared to both the IoT gadgets group and control group, with diet modification showing the highest effectiveness, followed by physical activity and stress management interventions.

Effects of hydromorphone alone and combined with medetomidine-vatinoxan or dexmedetomidine on alfaxalone induction dose and mean arterial pressure in dogs anesthetized with sevoflurane

ObjectiveTo evaluate the dose of alfaxalone needed for induction of anesthesia following sedation with either medetomidine-vatinoxan plus hydromorphone (MVH), dexmedetomidine plus hydromorphone (DH), or hydromorphone (H) alone. A secondary objective included evaluating selected cardiopulmonary variables before, during, and after sedation and general anesthesia with sevoflurane in healthy dogs. Study designProspective, randomized, masked, crossover. AnimalsEight healthy Beagle dogs, 3–4 years old. MethodsAll dogs received three intramuscular (IM) treatments: H (0.1 mg kg-1), DH (0.005 mg kg -1 + 0.1 mg kg-1, respectively), or MVH (0.01 mg kg -1 + 0.2 mg kg -1 + 0.1 mg kg -1, respectively) at least 6 days apart. General anesthesia was induced with alfaxalone 20 minutes after treatment administration and maintained for 60 minutes with 2.8% sevoflurane (expired). Sedation scores, selected cardiopulmonary variables, and recovery scores were measured before, during, and after anesthesia at selected timepoints. Mixed effects ANOVA and ANOVA on ranks were used to evaluate differences between treatments, time, and their interaction, and Tukey-Kramer method was used for post-hoc analysis (p < 0.05). Data are presented as mean ± SD or median (range). ResultsDogs given MVH required a lower dose of alfaxalone for induction of anesthesia (0.77 ± 0.4 mg kg-1) compared to DH and H (1.16 ± 0.34 mg kg-1; p = 0.02, 1.13 ± 0.18 mg kg-1; p = 0.02), and had a higher incidence (50%; p = 0.038) and longer duration [median; 10 (0–35) minutes; ] of hypotension during sevoflurane anesthesia compared to H [0%; 0 (0–0) minutes; p = 0.040] but not DH (p = 0.272). Conclusions and clinical relevancePremedication with MVH provided the greatest alfaxalone-sparing effect. However, this treatment was associated with lower arterial pressures and clinically-relevant hypotension. Off-label use of medetomidine-vatinoxan before sevoflurane-based anesthesia should be used with caution due to a high incidence of hypotension.

Effect of close-up metabolizable protein supply on colostrum yield, composition, and immunoglobulin G concentration and associations with prepartum metabolic indicators of Holstein cows

The prepartum diet as well as individual metabolic status of the cow influences colostrum parameters. The objectives of this study were to 1) investigate the effect of increasing prepartum dietary MP supply on colostrum yield, composition, and immunoglobulin G (IgG) concentration, and 2) identify prepartum metabolic indicators associated with these outcomes. Multiparous Holstein cows (n = 96) were blocked by expected calving date and randomly assigned to 1 of 2 prepartum diets formulated to contain a control (CON; 85 g of MP/kg DM; 1,175 g of MP/d) or high (HI; 113 g of MP/kg DM; 1,603 g of MP/d) level of MP starting at 28 d before expected calving. Both prepartum diets were formulated to supply Met and Lys at an equal amount of 1.24 and 3.84 g/Mcal of metabolizable energy (ME), respectively. Metabolic indicators were determined in serum (albumin, glutamate dehydrogenase, cholesterol, aspartate transaminase, total protein, total bilirubin, and IgG) or plasma (Ca, glucose, fatty acids, BHB, and urea nitrogen) twice weekly in a subset of cows (n = 60). Colostrum was harvested at 3.6 ± 2.4 h from calving and yield as well as concentrations of IgG, fat, protein, and Ca were determined. Cows were retrospectively grouped based on the typical volume of colostrum needed for 2 colostrum meals (<6 or ≥ 6 kg), IgG concentration (<100 or ≥ 100 g/L), as well as the median concentrations of fat (<4.4 or ≥ 4.4%), protein (<16.5 or ≥ 16.5%), Ca (<0.21 or ≥ 0.21%), and total colostrum ME (<8.65 or ≥ 8.65 Mcal). Data were analyzed using mixed effects ANOVA, with repeated measures where applicable. Feeding HI tended to increase colostrum yield in cows entering parity 2 (9.4 vs. 7.2 ± 0.9 kg), but treatment did not affect yield from cows entering parity ≥3 (5.1 vs. 6.4 ± 1.0 kg). Supply of MP did not affect concentrations of IgG, fat, protein, or Ca. Cows that produced ≥ 6 kg vs. those producing <6 kg of colostrum had lower plasma concentrations of glucose. Metabolic indicators were not associated with IgG group. Colostrum fat ≥4.4% was associated with cows having lower prepartum concentrations of glucose, total protein, albumin, and aspartate transaminase activity. Colostrum protein ≥ 16.5% was associated with lower circulating serum IgG and elevated cholesterol. Elevated glucose as well as lower cholesterol and BHB concentrations were associated with colostrum Ca ≥ 0.21%. Further, higher albumin and fatty acids as well as lower glucose concentrations were associated with a greater colostrum energy output. In conclusion, increasing prepartum MP supply tended to increase colostrum yield in cows entering parity 2, but did not affect the composition or IgG concentration. The observed associations between metabolic indicators and colostrum parameters suggest that slight adjustment in metabolism during late gestation might be necessary to support colostrogenesis, but the causality of these relationships should be considered.

Sows and piglets adjust their use of an outdoor paddock with season and piglet age during the first weeks of life in an organic farm

Few studies have examined sows and piglets' use of outdoor access during their first weeks of life, when reared with access to a hut and a pasture as in the organic system. We studied whether the age of the piglets and the season of the year influenced the use of an outdoor area by piglets and the sow, and the use of indoor spaces. For this, the localisation of sows and their piglets inside and outside the hut was analysed on 16 sows and their litters during the summer and 16 others during the winter, by scan sampling, on days 2, 7 and 13 after parturition in an organic herd in Denmark. When inside the hut, the number of piglets in the creep area, next to the sow or other piglets was noted. All parameters were analysed by linear mixed-effects ANOVA models. Sows were less often observed outside the hut in the wintertime and during the mornings compared to later times of the day (P<0.001). Furthermore, the sows increased their outdoor use with the age of their litter (P<0.001). The season also influenced the piglet use of outdoor areas (winter < summer; P<0.001). Piglets that were observed outside for the first time were older (8.9±0.9 d) during winter than during summer (3.5±0.5 d). The proportion of piglets observed outside increased with age, with a significant Season x Age interaction (P<0.001). The proportion of observations where the sows and piglets were outdoors together increased with the piglets’ age (P < 0.001). Sows’ outing was correlated with summer (R=0.3) and winter (R=0.2) temperatures, unlike piglets, where outings were correlated only with summer temperatures (R=0.3, P<0.001). The proportion of piglets observed in the creep area was significantly affected by the age with a Season x Age interaction effect (P<0.001). Piglets were rarely observed lying alone inside the hut. The proportion of piglets in contact with the sow and other piglets inside the pen was influenced by a significant Age x Season interaction effect (P<0.001). We showed that piglets given access to an outside paddock gradually increase their use with age and don’t go outside right away. Their first exit and the proportion of piglets outside depended also on the season. In general, a higher proportion of sows and piglets were observed indoor during the wintertime. Thus, sows and piglets adjust their use of outdoor paddock with season and piglet age during the first weeks of life.

CD44 and CD271 expression and chemoresistance in BRAF V600E/K mutation–positive melanoma.

e21542 Background: Melanoma stem cells (MSC) are thought to play a role in cellular proliferation, migration, and chemotherapeutic resistance. We investigated native expression of several putative MSC biomarkers in human BRAF V600E/K mutation positive melanoma cell lines of different phases—radial growth phase (WM35), vertical growth phase (WM278)—and metastatic (LM-Mel-45, LM-Mel-71) cell lines. We examined cell viability of select cell subpopulations during in vitro treatment with BRAF and MEK inhibitor monotherapy and combination therapy. Methods: Using flow cytometry, we assessed surface expression of putative MSC biomarkers: CD44, CD271, CD133, CD186, and CD20. We sorted each cell line by CD44 and CD271 expression with a predetermined hierarchical gating strategy and compared 4 chemotherapy conditions: control, BRAF inhibition with 1.25 µM dabrafenib, MEK inhibition with 0.125 µM trametinib, and combination therapy. Cells (25,000) were collected directly into chemotherapy treated media and incubated for 72 hours. Cell viability was measured by MTT assay. Statistical analysis used mixed-effects ANOVA and Tukey HSD. Results: No significant expression of CD133, CD186, or CD20 was found in any cell line. Over 99% of live cells were CD44+ in all 4 cell lines. Only WM35 and LM-Mel-71 had both CD44+/CD271+ and CD44+/CD271- subpopulations. CD271 expression was restricted to CD44+ cells. Among live cells, the CD44+/CD271+ subpopulation was 31.2% for WM35 and 14.4% for LM-Mel-71. For WM35, cell viability in CD44+/CD271+ cells was greater than CD44+/CD271- cells for all treatments (p &lt; 0.001). For LM-Mel-71 cells, CD271 expression did not affect cell viability within any treatment (p = 0.64). For chemotherapeutic treatments, cell viability in CD44+/CD271- subpopulation was higher in LM-Mel-45 cells than WM278 (p &lt; 0.03) or LM-Mel-71 (p &lt; 0.002), but not WM35 cells (p &gt; 0.05). Trametinib monotherapy and combination therapy had similar cell viability in all 4 cell lines (p &gt; 0.05). Conclusions: To our knowledge, our study is the first to show differences in phenotypic expression of CD44 and CD271 across nonmetastatic and metastatic melanoma cell lines and suggests co-expression of CD44 and CD271 confers increased chemotherapeutic resistance to in vitro BRAF and MEK inhibitor treatment in radial growth phase but not metastatic melanoma. More research is needed to better understand the role of CD44 and CD271 co-expression in melanoma initiation, progression, and chemotherapeutic resistance. CD271 remains a potential immunotherapeutic drug target in the treatment of BRAF V600E/K mutation positive melanoma.

Effect of oxytocin use during colostrum harvest and the association of cow characteristics with colostrum yield and immunoglobulin G concentration in Holstein dairy cows

Our objectives were to determine the effect of oxytocin use during colostrum harvest on colostrum yield and IgG concentration in Holstein dairy cows on a commercial dairy in New York and to describe associations of cow characteristics with these outcomes. Animals were enrolled between July and October 2023 using a randomized block design, with day of enrollment as the unit of randomization. A median (range) of 10 (3-19) cows were enrolled per day. Treatments were (1) 40 IU oxytocin (OXY40), (2) 20 IU oxytocin (OXY20), and (3) an untreated control group (CNTR). Oxytocin was administered i.m. ∼45 s before unit attachment in a rotary parlor. Colostrum weight was measured using the colostrum bucket-embedded scale or a platform scale when the yield was less than the smallest bucket scale. The concentration of IgG ([IgG]) in colostrum was determined using radial immunodiffusion and used to calculate total IgG, and DM (%) was determined by oven drying. Individual cow characteristics such as parity, calf sex, weight, dystocia score, stillbirth, and milk production in wk 4 of lactation; additionally, for multiparous cows, dry period length and previous lactation dry-off linear SCS were collected. Data were analyzed separately for primiparous and multiparous cows using backward stepwise elimination. To produce final mixed effects ANOVA models, primiparous cows (n = 201) were randomized to 35.8% (n = 72) OXY40, 32.8% (n = 66) OXY20, and 31.3% (n = 63) CNTR. Multiparous groups (n = 435) were randomized to 34.7% (n = 151) OXY40, 29.7% (n = 129) OXY20, and 35.6% (n = 155) CNTR. The median (range) colostrum yield was 6.0 (0-20.6) kg and [IgG] was 98.5 (0.1-293.6) g/L in the study population. In primiparous cows, OXY40 had a higher colostrum yield (LSM [95% CI]) of 5.4 (4.9-5.9) kg compared with both OXY20 (4.1 [3.5-4.7] kg) and CNTR (3.8 [3.3-4.3] kg). In multiparous cows, OXY40, OXY20, and CNTR did not differ in colostrum yield (5.9 [5.3-6.5], 5.7 [5.1-6.3], and 5.4 [4.8-6.0] kg, respectively), but colostrum yield was greater in parity 2 compared with all other parities, cows giving birth to male calves, cows with the highest milk production at wk 4 of lactation, and cows with a dry period of >65 d. Oxytocin use did not affect [IgG] in either primiparous or multiparous cows, but [IgG] was highest in cows in parity ≥4 and lowest in cows dry >65 d. In summary, oxytocin use at 40 IU i.m. was associated with a higher colostrum yield in primiparous cows only and no change in [IgG]. Oxytocin use likely addressed disturbed milk ejection and therefore increased colostrum yield in heifers milked for the first time in a rotary parlor. This study confirms cow characteristics associated with colostrum production within a single herd.

One minute of stair climbing and descending reduces postprandial insulin and glucose with three-minutes improving insulin resistance following a mixed meal in young adults: A Randomized Controlled Trial

Glycemic markers, including postprandial glucose, insulin, and insulin sensitivity, are strong predictors of morbidity and mortality in individuals with and without diabetes. Stair-climbing and -descending (SCD) at a comfortable pace for as little as 1 to 3 minutes after a sugary beverage (300 kilocalories; 100% carbohydrate) improves glucose and insulin, with insulin sensitivity improving with as little as 10 minutes. Objective: To determine if benefits to glucose, insulin, and insulin sensitivity are seen with SCD following consumption of a meal with mixed macronutrients. Hypothesis: SCD will improve to glucose, insulin, and insulin sensitivity in a dose response manner following a mixed meal. Methods: In a randomized, controlled, crossover trial, young adults (N=31) in apparent good health performed SCD for 0, 1, 3, and 10 minutes after a mixed meal (650 kilocalories; 53% carbohydrates, 33% fat, and 14% protein). Differences in glucose, insulin, and insulin sensitivity (ISI) from baseline to 30 minutes were analyzed using a mixed-effects ANOVA. Results: A significant fixed-effect was found for change in glucose [F(2.551,67.17)=4.724, p=.0071)], insulin [F(2.692,74.49)=11.28, p&lt;.0001)], and ISI [F(2.127,56.00)=5.848, p=.0042)]. Compared to control, changes in glucose (mean (95%CI)) were significantly lower after 1 min (-14 (-7 to -21) mg/dL, p=.0006), 3 min (-18 (-9 to -28) mg/dL, p=.0007), and 10 min (-10.0 (-1 to -20) mg/dL, p=.0387); changes in insulin were significantly lower after 1 min (-1.8 (-0.8 to -2.8) μIU/mL, p=.0011), 3 min (-2.8 (-1.7 to -4.0) μIU/mL, p&lt;.0001), and 10 min (-1.1 (-0.1 to -2.0) μIU/mL, p=.0329); and changes in ISI were significantly higher after 3 min (2.4 (1.2 to 3.6), p=.0003) and 10 min (1.3 (0.3 to 2.4), p=.0143) but not 1 min (1.2 (0.0 to 2.5), p=.0587). Conclusion: Postprandial glucose and insulin were improved with as little as 1 minute, and insulin resistance improved with as little as 3 minutes, of SCD at a self-selected, comfortable pace, after consumption of a mixed meal in apparently healthy young adults. None. This is the full abstract presented at the American Physiology Summit 2024 meeting and is only available in HTML format. There are no additional versions or additional content available for this abstract. Physiology was not involved in the peer review process.

Blood flow restriction training does not negatively alter the mechanical strength or histomorphology of uninjured equine superficial digital flexor tendons.

Low load exercise training with blood flow restriction (BFR) has become increasingly used by human physical therapists to prescribe controlled exercise following orthopaedic injury; its effects on the equine superficial digital flexor tendon (SDFT), however, are unknown. To investigate outcomes of pressure specific BFR walking exercise on uninjured equine SDFT biomechanics and histomorphology. Controlled in vivo experiment. Four forelimbs of four horses were exposed to 40 BFR-walk sessions (10-min interval walking) on a treadmill over a 56-day study period with their contralateral forelimbs serving as untreated controls. Similarly, four forelimbs of four control horses were exposed to 40 sham cuff walk sessions. On study Day 56, all horses (n = 8) were humanely euthanised and forelimb SDFTs underwent non-destructive biomechanical testing and corresponding histomorphological analysis. Significance in biomechanical parameters between treatment groups was analysed using a mixed-effects ANOVA with Tukey's post-hoc tests. Statistically significant differences in SDFT stiffness for both first (p = 0.02) and last cycles (p = 0.03) were appreciated within the BFR treated group only, with BFR exposed forelimbs being significantly stiffer than the contralateral unexposed forelimbs. When normalised to cross-sectional area, no significant differences were appreciated among treatment groups in elastic modulus for the first (p = 0.5) or last cycles (p = 0.4). No histological differences were appreciated among treatment groups according to Bonar, Movin, or musculotendinous junction evaluation criteria. Short-term comparisons were performed in a small sample population without correlation to performance outcome measures. Optimal occlusion percentages and walk protocols remain unknown. This study demonstrated no negative impact of BFR on mechanical strength of the equine SDFT; however, evidence suggests that BFR results in increased tendon stiffness based on biomechanical testing and subsequent calculations. No consistent detrimental histomorphological changes were seen.

Effects of a ketogenic diet on motor function and motor unit number estimation in aged C57BL/6 mice

ObjectivePathological, age-related loss of muscle function, commonly referred to as sarcopenia, contributes to loss of mobility, impaired independence, as well as increased risk of adverse health events. Sarcopenia has been attributed to changes in both neural and muscular integrity during aging. Current treatment options are primarily limited to exercise and dietary protein fortification, but the therapeutic impact of these approaches are often inadequate. Prior work has suggested that a ketogenic diet (KD) might improve healthspan and lifespan in aging mice. Thus, we sought to investigate the effects of a KD on neuromuscular indices of sarcopenia in aged C57BL/6 mice. DesignA randomized, controlled pre-clinical experiment consisting of longitudinal assessments performed starting at 22-months of age (baseline) as well as 2, 6 and 10 weeks after the start of a KD vs. regular chow intervention. SettingPreclinical laboratory study. Sample sizeThirty-six 22-month-old mice were randomized into 2 dietary groups: KD [n = 22 (13 female and 9 male)], and regular chow [n = 15 (7 female and 8 male)]. MeasurementsMeasures included body mass, hindlimb and all limb grip strength, rotarod for motor performance, plantarflexion muscle contractility, motor unit number estimations (MUNE), and repetitive nerve stimulation (RNS) as an index of neuromuscular junction transmission efficacy recorded from the gastrocnemius muscle. At end point, muscle wet weight and blood samples were collected to assess blood beta-hydroxybutyrate levels. Statistical analysisPrimary analyses were two-way mixed effects ANOVA (diet and time × diet) to determine the effect of a KD on indices of motor function (grip, rotarod) and indices of motor unit (MUNE) and muscle (contractility) function. ResultsBeta-hydroxybutyrate (BHB) was significantly higher at 10 weeks in mice on a KD vs control group (0.83 ± 0.44 mmol/l versus 0.42 ± 0.21 mmol/l, η2 = 0.265, unpaired t-test, p = 0.0060). Mice on the KD intervention demonstrated significantly increased hindlimb grip strength (diet, p = 0.0001; time × diet, p = 0.0030), all limb grip strength (diet, p = 0.0005; time × diet, p = 0.0523), and rotarod latency to fall (diet, p = 0.0126; time × diet, p = 0.0021). Mice treated with the KD intervention also demonstrated increased MUNE (diet, p = 0.0465; time × diet, p = 0.0064), but no difference in muscle contractility (diet, p = 0.5248; time × diet, p = 0.5836) or RNS (diet, p = 0.3562; time × diet, p = 0.9871). ConclusionKD intervention improved neuromuscular and motor function in aged mice. This pre-clinical work suggests that further research is needed to assess the efficacy and physiological effects of a KD on indices of sarcopenia.

One minute of stair climbing and descending reduces postprandial insulin and glucose with 3-min improving insulin resistance following a mixed meal in young adults: A randomized controlled crossover trial

BackgroundGlycemic markers, including postprandial glucose, insulin, and insulin resistance, are strong predictors of morbidity and mortality in individuals with and without diabetes. Stair-climbing and -descending (SCD) at a comfortable pace for 3 minutes after a sugary beverage (300 kilocalories; 100% carbohydrate) lowers insulin, with insulin sensitivity improving in 10 minutes. If similar benefits are seen following consumption of a mixed meal is unknown. We hypothesize SCD will improve these markers in a dose-response manner following a mixed meal. MethodsIn a randomized, controlled, crossover trial, young adults (N = 31) performed SCD for 0 (seated control), 1, 3, and 10 minutes after a mixed meal (650 kilocalories; 53% carbohydrates, 33% fat, and 14% protein). Differences in glucose, insulin, and insulin sensitivity (ISI) from baseline to 30 min were analyzed using a mixed-effects ANOVA. ResultsA significant fixed-effect was found for change in glucose [F(2.551,67.17) = 4.724,p = 0.007)], insulin [F(2.692,74.49) = 11.28,p < 0.001)], and ISI [F(2.127,56.00) = 5.848,p = 0.004)]. Compared to the seated control (0 minutes), changes in glucose were lower after 1 minute (−14.0 (−7.2)mg/dL,p < 0.001), 3 minutes (−18.4 (−7.0)mg/dL,p = 0.0007), and 10 minutes (−10.0 (−8.1)mg/dL,p = 0.039); changes in insulin were lower after 1 minute (−1.8 (−0.9)μIU/mL,p = 0.0011), 3 minutes (−2.8 (−0.9)μIU/mL,p < 0.001), and 10 minutes (−1.1 (−0.9)μIU/mL,p = 0.033); and changes in ISI were significantly higher after 3 minutes (2.4 (1.5),p < 0.001) and 10 minutes (1.3 (1.6),p = 0.014) but not 1 minute (1.2 (1.5),p = 0.059). ConclusionPostprandial glucose and insulin improved with 1 minute, and insulin resistance improved with 3 minutes, of SCD at a self-selected, comfortable pace, after consumption of a mixed meal in apparently healthy young adults. ProtocolTrial registration: ClinicalTrials.gov Identifier: NCT04232475.

161 Effects of Sericea Lespedeza Supplementation on Steers Grazing Wild-Type Endophyte-Infected Tall Fescue

Abstract Fescue toxicosis poses a significant challenge to the United States beef industry, resulting in considerable economic losses. This syndrome arises from the consumption of tall fescue (Schedonorus arundinaceous) infected with the wild-type endophyte (WE; Epichloe coenophiala), which produces toxic alkaloids. The most effective solution for mitigating fescue toxicosis is renovating WE tall fescue pastures to novel endophyte-infected (NE) tall fescue. However, pasture renovation is costly and may not be feasible under certain conditions, with the economic profitability taking up to 3 yr. While the potential of condensed tannins (CTs) in specific leguminous forages to reduce the absorption of toxic alkaloids is recognized, their use as supplements remains underexplored. This study aims were to investigate the potential of CT-containing feed supplements, specifically sericea lespedeza (Lespedeza cuneata) pellets, in reducing the post-ingestive effects of fescue toxicosis in steers. Over two summers,12 steers were assigned to either Schedonorus arundinaceous (LES) or LES mixed with polyethylene glycol (LES + PEG) as a positive control for 12 wk. Body weight, hair retention score, rectal and extremities temperatures, cortisol concentrations, and lumen area of caudal arteries were collected at 4-wk intervals. Data were analyzed using a mixed-effect ANOVA test in SAS Studio 3.5. Steers on the LES diet exhibited significantly less (P &amp;lt; 0.05) hair retention scores compared with those on the LES + PEG diet. In 2021, steers on the LES diet showed greater (P = 0.0101) ADG, while in 2022, there was no difference (P = 0.9916) in ADG between the treatments. Steers on the LES diet displayed greater (P = 0.0332) tail skin temperatures and decreased (P &amp;lt; 0.0001) rectal temperatures compared with those on the LES + PEG diet. Hair cortisol concentrations were greater (P &amp;lt; 0.05) in steers on the LES + PEG diet than in those on the LES diet. Additionally, steers supplemented with the LES + PEG diet showed a 21.2% smaller caudal artery lumen area compared with those on the LES diet (P &amp;lt; 0.01). These findings underscore the potential of CTs in reducing the post-ingestive effects of fescue toxicosis. However, further research is needed to explore the complex mechanisms underlying these effects, establish the long-term efficacy of such interventions, and extend the investigation to include a broader range of physiological parameters affecting livestock health and performance.

Evaluation of the analgesic efficacy of grapiprant compared with robenacoxib in cats undergoing elective ovariohysterectomy in a prospective, randomized, masked, non-inferiority clinical trial.

The main objective of this study was to compare the postoperative analgesic effects of grapiprant with those of robenacoxib in cats undergoing ovariohysterectomy (OVH). In total, 37 female cats (age range 4 months-10 years, weighing ⩾2.5 kg) were enrolled in a prospective, randomized, masked, non-inferiority (NI) clinical trial. Cats received oral robenacoxib (1 mg/kg) or grapiprant (2 mg/kg) 2 h before OVH. Analgesia was assessed via the Feline Grimace Scale (FGS), the Glasgow Composite Measure Pain Scale-Feline (CMPS-F), von Frey monofilaments (vFFs) and pressure algometry (ALG) 2 h before treatment administration, at extubation, and 2, 4, 6, 8, 18 and 24 hours after extubation. Hydromorphone (<8 h postoperatively) or buprenorphine (>18 h postoperatively) were administered to cats with scores of ⩾5/20 on CMPS-F and/or ⩾4/10 on FGS. NI margins for CMPS-F and vFFs were set at 3 and -0.2, respectively. A mixed-effect ANOVA was used for FGS scores (P <0.05). Data are reported as mean ± SEM. The data from 33 cats were analyzed. The upper limit of the 95% confidence interval (CI) (0.35) was less than the NI margin of 3 for CMPS-F, and the lower limit of the 95% CI (0.055) was greater than the NI margin of -0.2 for vFFs, indicating NI of grapiprant. The FGS scores were greater than baseline at extubation for both treatments (1.65 ± 0.63; P = 0.001); however, there was no difference between treatments. There was no difference between treatments, nor treatment by time interaction, for vFFs (P <0.001). The CMPS-F scores for both treatments were higher at extubation but returned to baseline after 4 h (P <0.001). For ALG, there was no difference in treatment or treatment by time interaction. The robenacoxib group had lower pressure readings at extubation and 6 h compared with baseline. These results indicate that grapiprant was non-inferior to robenacoxib for mitigating postsurgical pain in cats after OVH performed via ventral celiotomy. The impact of grapiprant for analgesia in OVH via the flank is unknown.

Relative motion splints versus metacarpophalangeal joint blocking splints in the management of trigger finger: Study protocol for a randomized comparative trial.

Evidence supports the use of hand-based metacarpophalangeal joint (MCPJ) blocking splints as an intervention for trigger finger (TF). In practice, finger-based relative motion (RM) splints are also implemented without evidence. This randomized comparative trial (RCT) aims to evaluate implementation of MCPJ blocking and RM splints for effectiveness, function, occupational performance and wearability after 6 weeks of TF management. Priori analysis determined 36 individuals were needed for random assignment to the RM or MCPJ blocking splint groups. Individuals must be aged ≥21 years, and diagnosed with TF involving ≥1 finger. For blinding purposes, the primary author screens for eligibility, fabricates the splints and educates. Therapist A administers the primary outcome measures Week-1 and Week-6-stage of stenosing tenosynovitis and secondary outcome measures- number of triggering events in 10 active fists, visual analog scales (VAS) for pain, splint comfort and satisfaction, Disabilities of the Arm, Shoulder and Hand, and Canadian Occupational Performance Measure. Therapist B in Week-3 instructs participants in deep tissue massage and administers splint wearability VASs. The RM pencil test is used to determine the affected finger(s) MCPJ splint position i.e., more extension or flexion based on participant response. The MCPJ blocking splint holds the MCPJ in a neutral position. Analysis involves a mixed-effects ANOVA to compare Week-1 and Week-6 primary and secondary outcomes. Recruitment and data collection are ongoing. Biomechanically RM splints control tendon excursion and reduce passive tendon tension while allowing unencumbered finger motion and hand function. Hence clinicians use RM splints as an intervention for TF, despite the lack of implementation evidence. This RCT implements a function-focused as well as patient-centered approach with partial blinding of assessors and participants. We anticipate that this study will provide evidence for the implementation of RM splints to manage adults with TF. Clinical trial registration This trial is registered with ClinicalTrials.gov (NCT05763017).

Validation of Noninvasive Derivation of the Central Aortic Pressure Waveform from Fingertip Photoplethysmography Using a Novel Selective Transfer Function Method.

Central aortic pressure waveform analyses can provide clinically relevant information beyond conventional brachial blood pressure (BP) assessment. This waveform can be reproduced noninvasively through application of a generalized transfer function (GTF) on a peripheral waveform, as conventionally performed by applanation tonometry. Photoplethysmography (PPG) is an alternate approach; however, differences in measurement site and modality demand the use of a transfer function (TF) specific for those differences. This study aimed to compare central aortic waveform features generated from radial tonometry (reference) using a proprietary GTF with a central aortic waveform and its features generated from a simultaneous fingertip PPG measurement using a selective method where one of three different TFs is chosen based on the input signal harmonic profile. Brachial BP was measured in triplicate under resting conditions and was used for subsequent calibration. Multiple simultaneous radial tonometry (SphygmoCor CVMS) and fingertip PPG measurements were then performed in individual participants (n = 21, 10 females, age: 39 ± 16 years). Measurements were converted into central aortic waveforms with their respective TFs. Twenty central aortic pressure waveform parameters were compared through correlation analysis, Bland-Altman plots, and a repeated measure mixed-effects ANOVA model. Central aortic waveform shape was compared using the root-mean-squared error (RMSE). Correlation (r) of PPG-derived parameters with radially tonometry-derived central aortic parameters was high ranging from 0.79 to 0.99. Mean differences of pressure-related parameters were within 1.3 mm Hg, and differences of time-related parameters ranged from -2.2 to 3.4%. While some parameters were statistically different, these differences are not physiologically meaningful. Central aortic waveform shape had an average RMSE of 1.8 ± 0.9%. Fingertip PPG-derived central aortic waveform parameters using a novel selective TF were comparable to central aortic waveform features derived from radial tonometry using a previously validated GTF.

Gene expression changes in conjunctival cells associated with contact lens wear and discomfort

PurposeThis study aimed to analyze the differences in the expression of pain-related genes in conjunctival epithelial cells among symptomatic contact lens (CL) wearers (SCLWs), asymptomatic CL wearers (ACLWs), and non-CL wearers (non-CLWs). MethodsFor this study, 60 participants (20 non-CLWs, 40 CLWs) were enrolled. The CLW group comprised 20 ACLWs and 20 SCLWs according to the Contact Lens Dry Eye Questionnaire short form©. Conjunctival cells were collected using impression cytology, and RNA was isolated and used to determine the expression levels of 85 human genes involved in neuropathic and inflammatory pain. The effects of CL wear and discomfort were evaluated using mixed-effects ANOVA with partially nested fixed-effects model. Gene set enrichment analysis was performed to assign biological meaning to sets of differentially expressed genes. ResultsSix genes (CD200, EDN1, GRIN1, PTGS1, P2RX7, and TNF) were significantly upregulated in CLWs compared to non-CLWs. Eleven genes (ADORA1, BDKRB1, CACNA1B, DBH, GRIN1, GRM1, HTR1A, PDYN, PTGS1, P2RX3, and TNF) were downregulated in SCLWs compared to ACLWs. These genes were mainly related to pain, synaptic transmission and signaling, ion transport, calcium transport and concentration, and cell-cell signaling. ConclusionsCL wear modified the expression of pain- and inflammation-related genes in conjunctival epithelial cells. These changes may be in part, along with other mechanisms, responsible for CL discomfort in SCLWs.

Antimicrobial effects of blue light therapy against cutibacterium acnes: optimal dosing and impact of serial treatments

BackgroundBlue light therapy (BLT) is an Food & Drug Administration cleared modality used in dermatology as an effective treatment of acne. The primary purpose of this study is to determine if there are dose-dependent antimicrobial effects of BLT against Cutibacterium acnes (C. acnes). MethodsA known strain of C. acnes was grown on chocolate agar in a controlled laboratory environment under anaerobic conditions for 1 week. After 1 week, 2-3 colonies of C. acnes were isolated and transferred to broth medium to incubate for 2 or 7 days. Broth vials (treatment arm) then underwent one of 6 different blue light dosing treatment regimens and a duplicate broth vial served as a control left open to the same environment. The BLT regimens were a single treatment of 25 J/cm2; 50 J/cm2, 75 J/cm2, 100 J/cm2, two serial treatments of 50 J/cm2 separated by 24 hours, or two serial treatments of 75 J/cm2 separated by 24 hours. The Omnilux Blue device (415 nm wavelength) was used for all BLT treatments and delivered, on average, 1.68 ± 0.004 J/min. Following treatment, the control and treatment broth samples were plated on chocolate agar and allowed to grow for 7 days. After 7 days, plates were counted and colony forming units (CFUs) were calculated. Six trials were completed for each BLT dosing regimen based on an a priori power analysis of 6 individual two-sided t-tests. Comparisons in the primary outcome were made via mixed effects ANOVA with replicate as a random effect. ResultsAll BLT treatment regimens resulted in significantly fewer CFUs than their aggregate control plate CFUs (p<0.05 for all). Further, in 2-way comparison of CFUs between BLT treatment groups, a single treatment of 75 J/cm2 did lead to significantly less growth than 25 J/cm2 (p=0.017) and 50 J/cm2 (p=0.017).There were no improved antimicrobial effects with serial treatments when comparing two doses of 50 J/cm2 with a single dose of 100J/cm2, nor were 2 doses of 75 J/cm2 more efficacious than 100 J/cm2. Using the Omnilux Blue device, it took 44.8 minutes to deliver a 75 J/cm2 dose. ConclusionsBlue light therapy is an effective antimicrobial agent against this single virulent strain of C. acnes. Treatment dosing of 75 J/cm2 was identified to be the most effective dose per unit time. Serial treatments did not lead to superior antimicrobial effects over a single, high dose treatment.

The acute effects of running on pelvic floor morphology and function in runners with and without running-induced stress urinary incontinence

Introduction and hypothesisThe aim of this study was to examine the impact of a single running session on pelvic floor morphology and function in female runners, and to compare those with and without running-induced stress urinary incontinence (RI-SUI).MethodsThis cross-sectional, observational study involved two groups: female runners who regularly experienced RI-SUI (n = 19) and runners who did not (n = 20). Pelvic floor muscle (PFM) properties were assessed using intravaginal dynamometry during maximal voluntary contractions (MVC) and during passive tissue elongation. The morphology of the pelvic floor was assessed at rest, during MVC and during maximal Valsalva maneuver (MVM) using 2D and 3D transperineal ultrasound imaging before and after a running protocol. Mixed-effects ANOVA models were used to compare all outcomes between groups and within-groups, including the interaction between group and time. Effect sizes were calculated.ResultsNo changes in PFM function assessed using intravaginal dynamometry were observed in either group after the run. Significant and large within-group differences were observed on ultrasound imaging. Specifically, the area and antero-posterior diameter of the levator hiatus were larger after the run, the bladder neck height was lower after the run, and the levator plate length was longer after the run (p ≤ 0.05). At the peak MVM and MVC, the bladder neck height was lower after the run than before the run (p ≤ 0.05). No between-group differences were observed for any outcomes.ConclusionsRunning appears to cause transient strain of the passive tissues of the female pelvic floor in runners both with and without RI-SUI, whereas no concurrent changes are observed in PFM contractile function.

Abstract 13461: Early Vascular Aging is Associated With Early Cardiac Aging, Upregulation of Left Ventricle Pro-Fibrotic and Inflammatory Genes and Memory Decline in Aged Dahl Salt-Sensitive Rats

Introduction: Blood pressure (BP) begins to increase early in life in Dahl salt-sensitive rats (DSS) that consume a normal salt (NS) diet. Epidemiologic studies have shown that early vascular aging (EVA) is associated with central arterial stiffening (CAS) and hypertension, which develop earlier in life than expected in general population and contribute to cognitive impairment later in life. Hypotheses: (i) DSS rats will develop EVA earlier in life on NS diet, and with advancing age will exhibit early CV aging; (ii) In DSS, CV remodeling and CAS are implicated in memory decline. We tested these hypotheses using a longitudinal repeated measure design in DSS and their parental breed Sprague-Dawley rats (SD) at 3- &amp; 12-mo of age. Methods: Male SD and DSS were kept on NS diet (0.5% NaCl; n=8/group) for entire experiment. Repeated measures of systolic BP (SBP), pulse wave velocity (PWV; an index of CAS), echocardiography, and non-repeated measures of Morris water maze (MWM) to test spatial memory, aortic collagen, elastin (histochemistry), and left ventricle (LV) mRNA expression (qPCR) were assessed at 3- &amp; 12-mo. Data analyses: linear mixed-effect 2-way ANOVA, t-test &amp; linear regression (LR) modeling. A 2-sided p&lt;0.05 was considered significant. Results: SBP, PWV, aortic weight, wall thickness and collagen/elastin ratio were higher in DSS-3 vs. SD-3, and in DSS-12 vs. DSS-3. RWT and LV expression of pro-fibrotic, inflammatory and senescence genes were higher in DSS-12 vs. DSS-3 and vs. SD-12 (Table). DSS-12 spent more time to find a hidden platform in MWM vs. SD-12; their impaired spatial hippocampal memory was positively associated with PWV and LV mass by LR analysis. Conclusions: EVA, indexed as higher BP, PWV and aortic remodeling was associated with early CV aging, manifested by activation of LV senescence, inflammatory and pro-fibrotic genes and by higher rates of age-associated changes in BP, RWT and LV mass in DSS vs. SD. In DSS, early CV aging was associated with memory decline.

25 High-resolution MRI Reveals Selective Patterns of Hippocampal Subfield Atrophy in Focal Epilepsy

Objective:Hippocampal pathology is a consistent feature in persons with temporal lobe epilepsy (TLE) and a strong biomarker of memory impairment. Histopathological studies have identified selective patterns of cell loss across hippocampal subfields in TLE, the most common being cellular loss in the cornu ammonis 1 (CA1) and dentage gyrus (DG). Structural neuroimaging provides a non-invasive method to understand hippocampal pathology, but traditionally only at a whole-hippocampal level. However, recent methodological advances have enabled the non-invasive quantification of subfield pathology in patients, enabling potential integration into clinical workflow. In this study, we characterize patterns of hippocampal subfield atrophy in patients with TLE and examine the associations between subfield atrophy and clinical characteristics.Participants and Methods:High-resolution T2 and T1-weighted MRI were collected from 31 participants (14 left TLE; 6 right TLE; 11 healthy controls [HC], aged 18-61 years). Reconstructions of hippocampal subfields and estimates of their volumes were derived using the Automated Segmentation of Hippocampal Subfields (ASHS) pipeline. Total hippocampal volume was calculated by combining estimates of the subfields CA1-3, DG, and subiculum. To control for variations in head size, all volume estimates were divided by estimates of total brain volume. To assess disease effects on hippocampal atrophy, hippocampi were recoded as either ipsilateral or contralateral to the side of seizure focus. Two sample t-tests at a whole-hippocampus level were used to test for ipsilateral and contralateral volume loss in patients relative to HC. To assess whether we replicated the selective histopathological patterns of subfield atrophy, we carried out mixed-effects ANOVA, coding for an interaction between diagnostic group and hippocampal subfield. Finally, to assess effects of disease load, non-parametric correlations were performed between subfield volume and age of first seizure and duration of illness.Results:Patients had significantly smaller total ipsilateral hippocampal volume compared with HC (d=1.23, p&lt;.005). Contralateral hippocampus did not significantly differ between TLE and HC. Examining individual subfields for the ipsilateral hemisphere revealed significant main-effects for group (F(1, 29)=8.2, p&lt;0.01), subfields (F(4, 115)=550.5, p&lt;0.005), and their interaction (F(4, 115)=8.1, p&lt;0.001). Post-hoc tests revealed that TLE had significantly smaller volume in the ipsilateral CA1 (d=-2.0, p&lt;0.001) and DG (d = -1.4, p&lt;0.005). Longer duration of illness was associated with smaller volume of ipsilateral CA2 (p=-0.492, p&lt;0.05) and larger volume of contralateral whole-hippocampus (p=0.689, p&lt;0.001), CA1 (p=0.614, p &lt; 0.005), and DG (p=0.450, p&lt;0.05).Conclusions:Histopathological characterization after surgery has revealed important associations between hippocampal subfield cell loss and memory impairments in patients with TLE. Here we demonstrate that non-invasive neuroimaging can detect a pattern of subfield atrophy in TLE (i.e., CA1/DG) that matches the most common form of histopathologically-observed hippocampal sclerosis in TLE (HS Type 1) and has been linked directly to both verbal and visuospatial memory impairment. Finally, we found evidence that longer disease duration is associated with larger contralateral hippocampal volume, driven by increases in CA1 and DG. This may reflect subfield-specific functional reorganization to the unaffected brain tissue, a compensatory effect which may have important implications for patient function and successful treatment outcomes.

O033 Is Sleeping and Waking Important for the Cortisol Awakening Response?

Abstract Introduction The rapid rise in cortisol after waking is known as the cortisol awakening response (CAR). However, as part of its normal circadian rhythm cortisol also rises in the morning independent of awakening. If CAR is a direct response to awakening, it should be eliminated in conditions of sleep deprivation (SD). Given that an atypical CAR is associated with several psychiatric and medical disorders, understanding the factors that influence CAR is of clinical relevance. This study explored cortisol during the normal waking period prior, during, and post-SD. Methods N=21 (11F 22.80±4.40y) completed 62h of SD in the laboratory. Salivary cortisol was collected across 5-time-points (07:00h, 07:15h, 07:30h, and 07:45h) on 2-pre-at-home-study days, 4-in-lab-study days, and 2-post-at-home-study days. Mixed-effects ANOVAs tested for fixed effects of day, time, and their interactions on cortisol, and cortisol area under the curve (AUCi, AUCg). Results SD produced significant effects of time*day (p&amp;lt;0.001). Cortisol levels at +30 and +45min post-awakening were lower on SD days. Cortisol levels on awakening (0mins) did not differ between any study days. There were significant effects of day (p&amp;lt;0.001). AUCi and AUCg were greater on in-laboratory-baseline and recovery days compared to other study days. Discussion This study found CAR is not present during periods of SD when there is no ‘waking’ from sleep. It showed for the first time that CAR recovers to baseline levels following recovery sleep. While the role of the CAR remains unclear this study identified factors that future research should consider to advance understanding of its purpose.