Unpaired T-test Research Articles

Oxidative stress in the remaining muscle after volumetric muscle loss contributes to impaired mitochondrial bioenergetics and is attenuated by SS-31

Volumetric muscle loss (VML) injury results in the non-recoverable loss of muscle mass and function. Our previous studies indicate mitochondrial dysfunction in the remaining muscle fibers after VML injury, marked by changes in mitochondrial respiration, membrane potential, and enzyme activities. In particular, mitochondrial membrane potential is hyperpolarized during the first 14-days post-injury, a condition that could lead to the production of reactive oxygen species (ROS). When ROS emissions exceed the antioxidant buffering capacity, cellular damage can ensue. The primary objective of this study was to determine ROS production, ROS emission, and antioxidant buffering capacity of remaining muscle fibers after VML injury in the first month after injury. The secondary objective of this study was to determine if a mitochondrial antioxidant (SS-31) could mitigate mitochondrial dysfunction after VML injury. We hypothesized that VML injury will alter the redox status of the remaining muscle. Study 1: Male C57BL/6J mice (n=40) were randomized to Uninjured or VML-injured cohorts. ROS (H2O2) emission/production and endogenous antioxidant buffering capacity were analyzed fluorometrically during live muscle fiber respiration (Oroboros O2k) at 3-, 7-, 14-, 21-, and 30-days post-injury. Study 2: Male C57BL/6J mice (n=32) were randomized into two groups: VML and VML+SS-31. SS-31 is a mitochondrial-targeted peptide with ROS-scavenging properties. VML+SS-31 received SS-31 (8mg/kg/day) for 14 days, and the VML received equal volume saline injections. At 14- and 28-days post-VML, mitochondrial respiration and electron conductance, ROS emission, and antioxidant buffering capacity were assessed in permeabilized gastrocnemius muscle fibers from the muscle remaining after the initial injury. Group differences were detected through ANOVA and unpaired t-test. ROS emission and antioxidant buffering capacity were different between VML-injured and Uninjured mice through 14-days post-injury, specifically at 3-, 7-, and 14-days, mitochondrial ROS emission was significantly higher (+28%, p<0.001), and antioxidant buffering capacity was less (-28%, p=0.002) in VML mice compared to Uninjured mice. Two weeks of SS-31 treatment lead to greater muscle fiber respiration and electron conductance (p≤0.008) at both 14- and 28-days post-VML. Importantly, at 14-days post-injury, mitochondrial ROS emission was 71% lower in VML+SS-31 than VML. At both 14- and 28-days post-injury, antioxidant buffering capacity was 12% greater in VML+SS-31 (p≤0.003) compared to VML. Oxidative stress is a feature of early VML pathophysiology marked by greater ROS emission and less antioxidant buffering capacity. Attenuating oxidative stress during the first two-weeks post-VML injury improves mitochondrial respiration. W81XWH-20-1-0885 and R01AR078903 to SG and JC. 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.

Value of Platelet to Lymphocyte Ratio in Nasopharyngeal Carcinoma Various Stages.

Nasopharyngeal Carcinoma (NPC) is among the most common head and neck cancer, significantly affecting the growth of human tumors. Therefore, this study aimed to investigate platelet-to-lymphocyte ratio (PLR) as a measure of inflammation in patients with various stages of NPC. This retrospective case-control study was conducted on patients diagnosed with NPC between 2016 and 2020 in Dr. Hasan Sadikin Bandung Hospital, Indonesia. Based on NPC classification criteria, patients were classified into early and advanced groups. Subsequently, the PLR ratio was calculated as the number of platelet divided by the absolute number of lymphocytes in the two groups, and the results were statistically analyzed using Unpaired T-test, Pearson, and Chi-square test. A total of 102 patients with an age range of 40 to 49 years were included in this study. A slight correlation of 0.272 (P<0.001) was observed between PLR and clinical stage, while NPC patients with PLR value > 188 had a 2.7 times higher risk of advanced stage (RR=2.702). The PLR was significantly related to the clinical stage of NPC cancer.

Bone marrow mesenchymal stem cell-derived exosomes shuttle microRNAs to endometrial stromal fibroblasts that promote tissue proliferation /regeneration/ and inhibit differentiation

BackgroundHuman bone marrow-derived stem cells (hBMDSCs) are well characterized mediators of tissue repair and regeneration. An increasing body of evidence indicates that these cells exert their therapeutic effects largely through their paracrine actions rather than clonal expansion and differentiation. Here we studied the role of microRNAs (miRNAs) present in extracellular vesicles (EVs) from hBMDSCs in tissue regeneration and cell differentiation targeting endometrial stromal fibroblasts (eSF).MethodsExtracellular vesicles (EVs) are isolated from hBMDSCs, characterized by transmission electron microscopy (TEM) and nanoparticle tracking analysis (NTA) techniques. Extracted total RNA from EVs was subjected to RNA seq analysis. Transfection and decidualization studies were carried out in endometrial stromal fibroblasts (eSF). Gene expression was analyzed by qRTPCR. Unpaired t-test with Welch’s correction was used for data analysis between two groups.ResultsWe identified several microRNAs (miRNAs) that were highly expressed, including miR-21-5p, miR-100-5p, miR-143-3p and let7. MiR-21 is associated with several signaling pathways involved in tissue regeneration, quiescence, cellular senescence, and fibrosis. Both miR-100-5p and miR-143-3p promoted cell proliferation. MiR-100-5p specifically promoted regenerative processes by upregulating TGF-ß3, VEGFA, MMP7, and HGF. MiR-100-5p blocked differentiation or decidualization as evidenced by morphologic changes and downregulation of decidualization mediators including HOXA10, IGFBP1, PRL, PR-B, and PR.Conclusion EVs delivered to tissues by hBMDSCs contain specific miRNAs that prevent terminal differentiation and drive repair and regeneration. Delivery of microRNAs is a novel treatment paradigm with the potential to replace BMDSCs in cell-free regenerative therapies.

Alteration of the bile acid pool attenuates neuroinflammation in the EAE mouse model of Multiple Sclerosis

Bile acids are critical signaling molecules that mediate gut crosstalk with other organs, including the brain. The primary bile acids, such as chenodeoxycholic acid (CDCA), are synthesized in the liver and modified by gut bacteria to generate secondary bile acids like the 12α-hydroxylated deoxycholic acid (DCA). Bile acids are effciently absorbed in the distal ileum, and recent studies have implicated them in neuroinflammation and the pathophysiology of several neurological diseases such as Multiple Sclerosis (MS). However, the specific effects of different types of bile acids on MS have not been thoroughly investigated. We hypothesize that bile acids differentially affect neuroinflammation and that treatment with cholestyramine (bile acid sequestrant) modulates astrocyte activation in a mouse model of MS. Methods: Primary astrocyte cultures were prepared from 6-10-week-old C57BL/6 mice and incubated with cytokines in the presence and absence of bile acids. The expression of inflammatory and astrocyte activation markers was assessed by real-time qPCR. The effects of blocking bile acid absorption on MS were examined using the established Experimental Autoimmune Encephalomyelitis (EAE) mouse model of MS in 8-week-old female mice (C57BL/6). Mice were fed either a standard chow diet or a standard chow diet supplemented with 2% cholestyramine, a bile acid sequestrant, to block bile acid absorption. Statistical significance between groups was determined by an unpaired T-test. Results: Astrocytes treated with cytokines (TNF-α, 10 ng/mL, IL1-β, 10 ng/mL, 24 h) significantly increased the mRNA expression of TNF-α and IL1-β and markers of astrocyte activation, lipocalin 2 (LCN2), and Serpina3n. Co-treatment with cytokines and CDCA but not DCA (50 μM, 24 h) significantly decreased TNF-α (~56%, p&lt;0.01), IL1-β (~40%, p&lt;0.01), LCN2 (~26%, p&lt;.001), and Serpina3n (~44%, p&lt;.001), vs cytokines alone. The taurine-conjugate, TCDCA, also resulted in significant decreases in the expression of TNF-α (~72%, p&lt;0.001), IL1-β (~53%, p&lt;.001), LCN2 (~30%, p&lt;0.05), and Serpina3n (~16%, p&lt;0.01), suggesting an anti-inflammatory effect specific to CDCA and its taurine conjugate on astrocytes. Cholestyramine treatment in EAE mice was associated with significant reductions in the mRNA expression of TNF-α (~40%, p&lt;0.05) and Serpina3n (~30%, p= 0.217) in the frontal cortex of the brain. Together, these data imply that altering the composition of the bile acid pool may be a viable therapeutic strategy for MS patients by attenuating neuroinflammation. University of Illinois at Chicago, JBVMC, Chicago, IL. 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.

CDC-WONDER Database Analysis of Maternal Morbidity in Florida [ID 2683410

INTRODUCTION: The CDC's WONDER Natality database was updated in 2016 to include comprehensive pregnancy data, encompassing maternal morbidity, a growing concern in the United States. Using the WONDER Natality database, we can better understand risk factors and trends associated with maternal morbidity. The aim of this research is to investigate the variations in maternal morbidity in Florida, focusing on maternal demographics and pregnancy characteristics. METHODS: This cross-sectional descriptive study used results from the CDC’s WONDER Natality 2016–2021 expanded-results database. Five categories of maternal morbidity were analyzed. Maternal morbidity and pregnancy characteristics were analyzed for statistical significance using χ2 and unpaired t-tests. RESULTS: 92,680 (1.04%) births in Census Region 3 (the South) were found to be associated with maternal morbidity. This prompted an in-depth examination of this region, with a particular focus on Florida. Notably, Asian mothers experienced the highest rate of maternal morbidity (1.49%), whereas Black mothers faced the lowest statistical rate (0.67%). Asian mothers experiencing maternal morbidity tended to be older, have lower pre-pregnancy body mass indexes, more prenatal visits, and shorter intervals since their last live births. CONCLUSION: This study underscores statistically significant maternal morbidity disparities in Florida, with Asian mothers being particularly vulnerable. It identifies several pregnancy characteristics linked to increased morbidity. Further exploration, including a county-level analysis in Florida and examination of other states in Census Region 3, could reveal or consolidate emerging trends. Moreover, the absence of a universally accepted definition for severe maternal morbidity calls for the establishment of a clear standard to guide future research and obstetric practices.

Enhancing Sleep Quantity and Quality with a Novel GABAergic Dietary Supplement

Purpose: The primary objective of this investigation was to evaluate the effects of BH_SLP_01 (Bonafide Health Sleep Product) on sleep quality and brain wave patterns using a caffeine-induced insomnia model, and on sleep duration and latency using a pentobarbital-induced sleep model. Methods: Five male BALB/c mice were allocated per treatment arm (age: 8 weeks, body mass: 180 ± 20 g). Treatment arms included control, caffeine or phenobarbital (active) control, and BH_SLP_01. Caffeine treatment model. To induce sleep disturbance, 7.5 mg/kg caffeine was injected intraperitoneally at the 15th minute (min) of recordings, and second injections (saline or combinations) were performed at the 30th minute. Brain electrical activity was monitored and recorded for a total of two hours using electrocorticography (ECoG) recording. Serum melatonin, serotonin, and dopamine were measured by ELISA. Brain levels of MDA were measured by HPLC, and protein concentrations were determined using Western Blot analysis. Phenobarbital treatment model Groups were administered saline or BH_SLP_01, and then, 45 min later, pentobarbital (42 mg/kg) was injected. After injection, sleep latency and duration were measured. Study treatments were compared using an ANOVA and Student's unpaired t-test. Significance levels were set at p&lt;0.05. Results: Analysis of ECoG recording data revealed notable findings. In the caffeine-pretreated group, a significant increase in amplitude was observed during the 30-45 min interval, compared to control. Caffeine administration also significantly increased frequency at 75-90 and 90-105 min compared to control (p&lt;0.05). Conversely, the group treated with BH_SLP_01 displayed no significant changes in amplitude and frequency, except for a significant change in frequency at 90-105 min compared to control (p&lt;0.05). Both sleep duration and sleep latency were significantly improved for BH_SLP_01 compared to control and phenobarbital control (p&lt;0.05). In addition, serotonin, dopamine, melatonin, GABAA R2, GABAB R1, GABAB R2, 5-HT1A, GluA1, GluN1, GluN2A, Bax, Bcl-2, and Caspase-3 levels were significantly improved by BH_SLP_01 compared to control and caffeine control (p&lt;0.05). Conclusion: The results of these studies provide evidence for the effcacy of BH_SLP_01 to improve impaired sleep, leading to notable enhancements in markers of deep sleep. This conclusion is supported by the significant upregulation of GABA, glutamine receptors, and endogenous melatonin, all of which collectively contribute to the augmentation of both sleep quantity and quality. These findings underscore the potential of BH_SLP_01 as a promising therapeutic intervention for optimizing sleep parameters and promoting overall sleep wellness. This research was supported by Bonafide Health, LLC. 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.

A Retrospective Study Comparing the Surgical Results of Platysma Myocutaneous Flap, Buccal Pad of Fat, and Nasolabial Flap for Reconstruction After Fibrotomy in Cases of Oral Submucous Fibrosis.

To evaluate the effectiveness of nasolabial flap (NLF), a buccal pad of fat flap (BFP), and platysma myocutaneous flap (PMF) for reconstruction following fibrotomy for individuals with oral submucous fibrosis (OSMF). A retrospective study was conducted among patients diagnosed with grade III and IV OSMFin the Departmentof Oral and Maxillofacial Surgery at Sharad Pawar Dental Collegebetween January 2016 and August 2018. The essential patient information was obtained from the Medical Record Department (MRD) at Acharya Vinoba Bhave Rural Hospital (AVBRH), Datta Meghe Institute of Medical Sciences (DMIMS) Sawangi (Meghe) Wardha. The patients were categorized into three groups: the NLF, the BFP, and the PMF groups. Each group had 16 patients, and factors such as interincisal width, diminished burning sensation in the mouth, inter-commissure distance, and flap necrosis were compared pre- and post-operatively. Student's unpaired t-test and chi-square test were employed for statistical analysis. Mean interincisal mouth-opening increased from pre-operative 4.79 to 41.42 mm post-operatively in the NLF group, BFP group from 6 to 39.42 mm and in the PMF group from 9.26 to 39.34 mm with p value=0.0001. NLF group showed complete and partial resolution of the burning sensation of the mouth at 93.75% and 6.25%, BFP at 62.25% and 32.75% while in PMF it was 68.5% and 31.25% respectively. One year postoperatively 3.28 mm increase in inter-commissure width was observed in the NLF group with a marginal increase in the PMF group and a negligible increase in the BFP group. 18.75% partial flap necrosis was seen in BFP, 18.75% in the PMF group, and 6.25% in the NFL group. All the flaps are efficacious in treating OSMF, however, NLF stands ahead with its higher reliability owing to its excellent blood supply.

Cerebrovascular Carbon Dioxide Reactivity Is Intact in Chronic Kidney Disease

Introduction: Chronic Kidney Disease (CKD) is associated with a substantially elevated risk of stroke. One factor that might contribute to this heightened risk is an impairment in cerebrovascular carbon dioxide reactivity (CVR), the mechanism through which cerebral vessels dilate and constrict in response to fluctuations in arterial carbon dioxide tension (PaCO2). CVR is known to be impaired in other chronic disease states associated with stroke risk; thus, we aimed to determine if CVR is also impaired in patients with moderate to severe CKD (stages III-IV). We hypothesized that CKD patients would exhibit impaired CVR compared to a control group (CON) matched for age, hypertension, and diabetes status. Methods: 9 CKD participants (age=56 ± 17y, eGFR=39 ± 4 mL/min/1.73m2), and 9 CON participants (age=61 ± 6y) underwent a single experimental protocol in which PaCO2 was manipulated via 2-min of hypercapnia (5% CO2) followed by 2-min hypocapnia (voluntary hyperventilation) while heart rate (via ECG), blood pressure (via finger photoplethysmography), middle cerebral artery blood velocity (MCAv; via transcranial Doppler ultrasonography), and end-tidal carbon dioxide (etCO2) were measured continuously. CVR was quantified as the slope of the linear relationship between breath-by-breath etCO2 and MCAv while accounting for blood pressure as a covariate. Hyper-, hypo- and, integrated (hyper + hypo) CVR was compared between groups via a 2-tailed, unpaired T-test. Results: We observed no differences in CVR between groups. Hypercapnic CVR: CKD = 1.4 ± 1.0 cm/s/mmHg, CON = 1.5 ± 0.8 cm/s/mmHg, Hypocapnic CVR: CKD = 1.5 ± 0.8 cm/s/mmHg, CON = 1.5 ± 0.8 cm/s/mmHg, Integrated CVR: CKD = 1.7 ± 1.2 cm/s/mmHg, CON = 1.7 ± 0.6 cm/s/mmHg, P ≥ 0.90. Conclusions: Contrary to our hypothesis, CVR remains intact in CKD stages III-IV. These findings suggest that other mechanisms contribute to the elevated stroke risk observed in this population. Funding: This work was supported by the American Heart Association (Career Development Award #929040, PI: Sprick), and NIH grants R01HL135183 and R33AT010457. 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.

Angiotensin II-Induced Heart Failure Disrupts Cardiac Sympathetic Innervation

About 26 million people worldwide live with heart failure (HF), and 25% have hypertension (HTN) as the primary cause. These patients have a high risk for cardiac arrhythmias and sudden cardiac death. Autonomic dysfunction and sympathetic hyperactivity accompany these diseases, and sympathetic activation is crucial for triggering fatal arrhythmias. However, changes in cardiac sympathetic innervation in HF are not well understood. We hypothesized that cardiac sympathetic innervation is disrupted in a mouse model of Angiotensin II (AngII)-induced HF. To test this hypothesis, C57BL6 male (N=6, 10) and female (N=6, 10) mice 12-20 weeks old were implanted with minipumps releasing either AngII (712ng/min/Kg, N=6-10) or saline (N=6-10) for 4 weeks. HTN and cardiac function were measured with tail cuff quantification and echocardiography, respectively. Fibrosis was assessed using Trichrome staining. Cardiac sympathetic innervation was quantified using immunohistochemistry for Tyrosine Hydroxylase (TH). Cardiac norepinephrine (NE) was measured by High-Performance Liquid Chromatography with electrochemical detection. Values were compared with a Student’s unpaired t-test unless noted, data were represented as mean ± SD, and significance was defined as p&lt;0.05. AngII-treated mice had significantly increased mean arterial pressure (Saline 91.0±15.6 vs. AngII 152.9±22.3 mmHg, p&lt;0.05, N=6-7) 7 days after minipump implantation. Four weeks of AngII treatment led to HF, as indicated by reduced ejection fraction (Saline 54.8±3.8 vs. AngII 43.1±8.2 %, p&lt;0.05, N=10) and fractional shortening (Saline 14.3±2.6 vs. AngII 10.6±3.3 %, p&lt;0.05, N=10). Cardiac hypertrophy was also observed in AngII-treated mice and was reflected by increased heart weight to body weight ratio (Saline 5.2±0.2 vs. AngII 7.7±1.0, p&lt;0.05, N=6). Fibrosis was detected in the intraventricular septum (IVS) and left ventricle (LV) of AngII-treated mice. TH+ fibers in the LV and IVS were decreased significantly (LV: Saline 0.6±0.2 vs. AngII 0.4±0.1 %; IVS: Saline 0.3±0.1 vs. AngII 0.09±0.04 %, p&lt;0.05, N=6), while no differences were detected in the right ventricle (Saline 1.1±0.2 vs. AngII 0.8±0.3 %, P&gt;0.05, N=6). In addition, the subepicardium-subendocardium (EPI-ENDO) gradient was altered (EPI: Saline 0.9±0.2 vs. AngII 0.8±0.3 %, P&gt;0.05; ENDO: Saline 0.3±0.05 vs. AngII 0.1±0.1 %, p&lt;0.05; EPI:ENDO ratio: Saline 2.7±0.7 vs. AngII 15.7±16.8, p&lt;0.05, Mann-Whitney test, N=6). A significant decrease in NE content accompanied the loss of TH+ fibers in the LV (LV: Saline 3.9±0.4 vs. AngII 1.6±0.2 pmol/mg, p&lt;0.05; RV: Saline 4.4±1.0 vs. AngII 3.8±0.8 pmol/mg, P&gt;0.05, N=4-6). Our data demonstrate that AngII-induced HF disrupts cardiac sympathetic nerve density, distribution, and NE content, confirming our initial hypothesis. This evidence contrasts with sympathetic hyperactivity seen in cardiac diseases. Future studies will assess the overall cardiac innervation and, determine whether the loss of TH+ fibers is due to a reduced sympathetic innervation after cardiac remodeling or the cholinergic transdifferentiation of TH+ fibers. A better understanding of the cardiac sympathetic nervous system in HF is imperative to deciphering the mechanistic changes to cardiac innervation in disease, and may help to develop and implement novel and precise neuromodulatory therapies for patients with autonomic imbalance. Supported by NIH R01 HL146833 to Dr. Habecker. 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.

P2X7 receptor knockout does not alter renal function or prevent angiotensin II-induced kidney injury in F344 rats

P2X7 receptors (P2X7) mediate immune and endothelial cell responses to extracellular ATP. Acute pharmacological blockade increases renal blood flow (RBF) and glomerular filtration rate (GFR), suggesting that P2X7 receptor activation promotes tonic vasoconstriction. P2X7 expression is increased in kidney disease and blockade/knockout is renoprotective. Here, we generated a novel P2X7 global knockout (KO) rat on the F344 background, hypothesising enhanced RBF and protection from chronic angiotensin II (ANG II)-induced injury. CRISPR/Cas9 technology introduced an early stop codon into exon 2 of P2rx7. In male and female KO and WT rats, renal P2X7 expression was measured by RTqPCR (n=4-5/group) and western blotting (n=3/group). Bone marrow-derived macrophages (BMDM) were used to assess P2X7 function by measuring interleukin (IL)-1β release (ELISA) after stimulation with lipopolysaccharide (LPS) (1μg/mL, 4h) and ATP (3mM, 1h) (n=5-6/group). In vivo kidney function was assessed in anesthetized rats (thiopental, 50mg/kg, i.p.) at baseline and following serial arterial ligation to induce acute pressure natriuresis (n=9-10/group). Arterial blood pressure (BP) was measured directly, and urine was collected via tube cystotomy. RBF was measured using a Transonic Doppler flow probe placed around the main renal artery, and cortical and medullary flux by laser Doppler needle probes. GFR was determined by FITC-inulin clearance. Urinary sodium was measured by sodium-selective electrode analysis. In a separate experiment, BP was recorded over five consecutive days in conscious male KO and WT rats via implanted radiotelemetry devices (n=7-8/group). Finally, male KO and WT rats (n=9-10/group) were infused with ANG II (250ng/kg/min, s.c.) for 6 weeks. At end, one renal artery was isolated and vascular reactivity to phenylephrine, acetylcholine and sodium nitroprusside determined by wire myography. Bladder urine was collected to measure albumin/creatinine ratio (n=4-7/group), and kidneys were harvested for histological analyses (n=6/group). Data were analysed with unpaired t-test (two groups compared) or two-way ANOVA (two variables), with p&lt;0.05 statistically significant. P2rx7 mutation was confirmed by genome sequencing (Sanger). P2rx7 mRNA abundance was partially decreased in kidney from male (-54%) and female (-25%) KO vs WT. P2X7 protein was detected in kidneys from WT but not from KO rats. In BMDM from WT rats, LPS+ATP stimulation induced IL-1β release; this response was markedly suppressed in KO BMDM (males: 2577±140 vs 413±175pg/mL, p&lt;0.0001; females: 1889±394 vs 265±121pg/mL, p&lt;0.0001). In vivo BP, renal haemodynamics and tubular sodium handling were not different between genotypes for either sex (P&gt;0.05). Radiotelemetry confirmed no difference in BP between KO and WT rats. Following chronic ANG II infusion, KO and WT rats had no renal artery dysfunction but showed similar increases in albuminuria (P=0.3587), renal perivascular fibrosis (Picrosirius Red, P=0.5393), cortical interstitium CD68-positive area (p=0.6546) and tubular casts (Periodic acid-Schiff, P=0.8803), compared with naïve controls. Contrary to our hypothesis, global genetic deletion of P2X7 did not affect renal hemodynamics and we found no significant role for P2X7 in the modulation of tubular sodium reabsorption. Our data do not support a major role of P2X7 in causing renal vascular and tubular injury. This work was supported by Diabetes UK grant 17/0005685, The British Heart Foundation(FS/15/60/31510; RE/18/5/34216), the Medical Research Council (MR/S01053X), and the University of Edinburgh College of Medicine &amp; Veterinary Medicine PhD Studentship. 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.

Treatment of Hyperuricemia with Thlaspi Bursa Pastoris 6CH: A Randomized, Open-Label, Equivalence Trial

Abstract Background: Hyperuricemia (HU) remains a priority research area in every medical science, including complementary and alternative medicine interventions and homeopathy; however, the research evidence is limited. In homeopathy, Thlaspi bursa pastoris (TBP) is considered to be a potential treatment option in HU. Methods: An open-label, randomized, equivalence trial was conducted comparing TBP with individualized homeopathic medicines (IHMs). Serum uric acid (SUA) was the primary outcome; Gout Assessment Questionnaire version 2.0 (GAQ-2) and Measure Yourself Medical Outcome Profile version 2.0 (MYMOP-2) were the secondary outcomes; all measured at baseline and after 6 and 12 weeks. Intention-to-treat analysis was carried out to detect noninferiority and group differences using unpaired t-tests and two-way repeated-measure analysis of variance (ANOVA). Results: There were no differences between the groups at baseline. Reduction in SUA after 6 and 12 weeks was similar between groups (P = 0.784). Group differences in the secondary outcomes were also nonsignificant (all P &gt; 0.05). TBP revealed noninferiority against IHMs, both after 6 weeks and 12 weeks in SUA and MYMOP-2 profile scores, but not in GAQ2 scores. Sulfur and Calcarea carbonica were the most frequently prescribed medicines in the IHMs group. There were no harms or serious adverse events reported from either of the groups. Conclusion: TBP acted noninferior to IHM in HU and both appeared to be equally effective. Rigorous and larger trials are warranted. Trial Registration: This trial was registered in the Clinical Trials Registry – India (CTRI/2018/10/016174; UTN: U1111-1221-8000).

Treadmill walking economy is not affected by body fat and body mass index in adults.

To determine whether body fat and body mass index (BMI) affect the energy cost of walking (Cw; J/kg/m), ventilation, and gas exchange data from 205 adults (115 females; percent body fat range = 3.0%-52.8%; BMI range = 17.5-43.2 kg/m2) were obtained at rest and during treadmill walking at 1.34 m/s to calculate gross and net Cw. Linear regression was used to assess relationships between body composition indices, Cw, and standing metabolic rate (SMR). Unpaired t-tests were used to assess differences between sex, and one-way ANOVA was used to assess differences by BMI categories: normal weight, <25.0 kg/m2; overweight, 25.0-29.9 km/m2; and obese, ≥30 kg/m2. Net Cw was not related to body fat percent, fat mass, or BMI (all R2 ≤ 0.011). Furthermore, mean net Cw was similar by sex (male: 2.19 ± 0.30 J/kg/m; female: 2.24 ± 0.37 J/kg/m, p = 0.35) and across BMI categories (normal weight: 2.23 ± 0.36 J/kg/m; overweight: 2.18 ± 0.33 J/kg/m; obese: 2.26 ± 0.31, p = 0.54). Gross Cw and SMR were inversely associated with percent body fat, fat mass, and BMI (all R2 between 0.033 and 0.270; all p ≤ 0.008). In conclusion, Net Cw is not influenced by body fat percentage, total body fat, and BMI and does not differ by sex.

Chronic Estrogen Treatment Promotes Human Microvascular Endothelial Dysfunction Through Sex-Specific Regulation of Ceramide Formation

Risk for cardiovascular disease (CVD) is elevated in individuals assigned female at birth and women of trans experience (assigned male at birth) undergoing long-term estrogen therapy (e.g. contraception, gender-affrmation). We have previously shown that chronic estrogen treatment induces microvascular endothelial dysfunction in human arterioles collected from those assigned male and female at birth. The mechanism via which estrogen causes microvascular damage remains unknown. Estrogen can increase expression of the ceramide-forming enzyme neutral sphingomyelinase (NSmase), and elevated ceramides promote cellular oxidative stress. Together, we hypothesize that estrogen induces microvascular endothelial dysfunction via NSmase-mediated ceramide formation. Human arterioles from otherwise healthy adults were treated with 17β-estradiol (E2; 100nM) +/- NSmase inhibitor (GW4869; 4μM, 24hrs). Vascular response to increasing flow rates was assessed using live videomicroscopy. Sex-specific human umbilical vein endothelial cells were treated with E2+/-GW4869 (48hrs), and H2O2 levels were measured using the fluorescent indicator peroxy yellow 1 (PY1). Endothelial ceramide levels were measured using liquid chromatography, tandem mass spectrometry. Exposure to estrogen impaired dilation to flow in arterioles from those assigned male and female at birth, an effect that was prevented by inhibition of NSmase (%Mean Dilation±SE; E2 vs E2+GW4869; male: 11.20±4.99% vs 48.43±7.15%, n=3 both, p=0.037; female: 1.78±1.99% vs 40.35±8.46%, n=5 both, p=0.006; two-way ANOVA). Estrogen treatment increased endothelial H2O2 production in cells from both sexes which was diminished by GW4869 treatment (%Change from control±SE; E2 vs E2+GW4869; male: 20.90±5.86% vs 2.85±1.99%, n=11 both, p=0.009*; female: 63.33±13.65%, n=10 vs 25.25±7.64%, n=9, p=0.031*, unpaired t-test*). Endothelial long-chain ceramide levels were higher in cells from biological males compared to females (pmol/nmol phosphate; male 5.90±2.19, n=4 vs female 2.18±0.86, n=5; p=0.009*). While estrogen treatment increased cellular ceramides in both sexes, ceramide accumulation was greater in cells from biological males compared to females (%change from baseline: 65.97±53.64% vs 6.08±7.86%, p=0.041*). Together, these results show that estrogen-induced microvascular endothelial dysfunction can be prevented through inhibition of NSmase-mediated ceramide formation. There is also notable sexual dimorphism in endothelial ceramide levels both at baseline and in response to estrogen. This study offers mechanistic insight into how chronic estrogen therapy may contribute to microvascular dysfunction and elevate the risk of CVD in patients. Furthermore, it presents the inhibition of NSmase as a plausible therapeutic approach to safeguard microvascular integrity during estrogen therapy. NHLBI R01HL160752 (J.K.F.), NHLBI K08HL141562 (J.K.F.), and AHA predoctoral fellowship grant 909315 (G.S.). 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.

Angiotensin-(1-7) Mas Receptors are Highly Colocalized to GABAergic Proopiomelanocortin Neurons in the Arcuate Nucleus of the Hypothalamus

Previous studies from our laboratory have shown that circulating levels of angiotensin (Ang)-(1-7), a protective hormone of the renin-angiotensin system, are greatly reduced in obese hypertensive mice. Additionally, restoration of this hormone reduces cardiovascular sympathetic tone and lowers blood pressure in this model suggesting a central mechanism of action. In support of this, our recent data show that the sympatho-inhibitory and blood pressure lowering effects of Ang-(1-7) involve activation of mas receptors (masR) within the arcuate nucleus of the hypothalamus (ARC). More specifically, we have found that masR are highly colocalized to proopiomelanocortin (POMC) neurons in the ARC. Due to the heterogeneous nature of POMC neuron subtypes, we wanted to explore the neurotransmitter phenotype of these POMCmasR neurons. We hypothesized that due to the blood pressure lowering effects of Ang-(1-7), POMCmasR are likely colocalized to inhibitory GABAergic neurons. To test this, we performed RNAscope in-situ hybridization in male C57Bl/6J mice after twelve weeks of either a 60% high fat diet (HFD) or matched control diet (n=12-18/group). We used RNAscope to colocalize expression of POMC, masR, and the vesicular GABA transporter, Vgat, mRNA transcripts within the ARC. Colocalization of POMCmasR and Vgat was quantified and percent-colocalization compared between control and HFD groups. We found that POMCmasR are highly colocalized with the GABAergic marker, Vgat, to a similar extent in control and HFD groups (84±4 vs. 90±2% of neurons, respectively; p=0.294 unpaired t-test). The population of POMC neurons that did not express masR also did not colocalize with Vgat. As an initial step to determine if masR modulates GABAergic signaling in the ARC, we assessed gene expression of glutamate decarboxylase 2 (Gad2; a GABA synthetic enzyme) in the ARC following chronic six-week treatment with systemic Ang-(1-7) versus saline infusion in male mice fed a control diet versus HFD (n=9-13/group). We found that Ang-(1-7) increases ARC Gad2 mRNA in HFD mice (1.03±0.09 control diet vs. 0.93±0.07 HFD vs. 1.23±0.07 HFD plus Ang-(1-7); p=0.023 one-way ANOVA). Supporting specificity of these actions, Ang-(1-7) did not alter POMC mRNA in the ARC. These overall data suggest that masR primarily localize to GABA-expressing POMC neurons in the ARC, and Ang-(1-7) may directly modulate GABA synthesis pathways in this brain region. Further studies are needed to elucidate the functional importance of these ARC POMC GABAergic neurocircuits to the depressor effects of Ang-(1-7) in obesity. Funding: NIH R01 HL156986. 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.

Oral Intake of Linseed Oil Inhibits Skin Barrier Dysfunction in Obese Mice.

Obesity is not only a risk factor for lifestyle-related diseases but also causes skin barrier dysfunction, which leads to a reduced quality of life due to dryness, itching, and scratching, and thus requires appropriate treatment. However, there are no studies on this issue. Therefore, this study aimed to examine whether oral intake of linseed oil is effective for skin barrier function in obesity and to confirm how the effect is demonstrated. TSOD mice received either sterile distilled water (Control group) or linseed oil (Omega group), containing a high level of omega-3 fatty acids, including α-linolenic acid, orally for eight weeks. Mice were then irradiated with ultraviolet B (UVB) and three days later, transepidermal water loss (TEWL), which is the primary outcome of skin barrier function, was measured and gross skin appearance was observed. Hematoxylin and eosin (HE) staining and Ki-67 immunostaining were performed on skin samples. mRNA expression levels of the inflammatory markers Tnfα, Cox2, Mcp1, and Hmox1 were measured by real-time reverse transcriptase-polymerase chain reaction (RT-PCR). We also performed fatty acid analysis of skin and erythrocytes by gas chromatography.Statistical analysis was performed using unpaired Student's t-test and Pearson's correlation analysis. Compared with the Control group, the Omega group exhibited lower TEWL values and little skin erythema. Histological analysis revealed thinner epidermis and fewer Ki-67 positive cells. Additionally, in the Omega group, mRNA levels of four inflammation-related genes were lower, α-linolenic acid levels in both skin and erythrocytes were higher, and a lower n-6/n-3 ratio was observed.And α-linolenic acid levels in the skin were negatively correlated with the expression levels of inflammation-related genes. Oral intake of linseed oil was found to inhibit skin barrier dysfunction in obesity. This effect was mediated by α-linolenic acid, a major component of linseed oil with anti-inflammatory properties, which was taken up by erythrocytes and supplied to the skin. Therefore, oral intake of linseed oil is expected to be a useful therapeutic method forskin barrier dysfunction in obesity.

Linking IL-6 and hsCRP among Indian patients with myocardial infarction

The association between serum interleukin-6 (IL-6) and highly sensitive C - reactive protein (hsCRP) as predictors of the risk factors for Myocardial Infarction. The study included a total of 50 patients with Myocardial Infarction, aged between 25 to 74 years. The levels of hsCRP were measured using the immunoturbidimetry method, while Interleukin 6 was estimated using the sandwich ELISA method. Statistical analysis was conducted using SPSS version 21.0, with p values calculated using Quartile ratio, ANOVA unpaired t-test, and Kaplan-Meier Curve Method. A p-value of less than 0.05 was considered statistically significant. All participants underwent a questionnaire, physical examination, medical history assessment, and laboratory tests. The results of the study showed that there was a significant correlation between IL-6 and hsCRP levels in the Quartile groups, as well as with lipid profiles. The Kaplan-Meier method also demonstrated a significant association between IL-6 and hsCRP levels in participants. The comparison of biomarkers further supported these findings. Thus, data shows that elevated levels of hsCRP and IL-6 could serve as valuable diagnostic markers for predicting Acute Myocardial Infarction. Our study strongly suggests that IL-6 could be a powerful marker in evaluating the Myocardial Infarction.

Differences in mental index value in patients with type II diabetes mellitus using panoramic radiography

Objectives: This research is aimed to determine the mandibular cortical thickness by the Mental Index value using panoramic radiography in patients with type 2 DM based on length of suffering ≤ 5 years and &gt; 5 years. Materials and Methods: This research is using unpaired comparative analytical design with a cross-sectional stratified random sampling method and unpaired T-test statistic. Results: 34 samples of patients with type 2 DM showed that the mean mental index was 4.219 ± 1.223. The two groups did not have a difference in the meaning of the MI value. The MI of the group with a shorter history of suffering (≤ 5 years) was 4.227 ± 1.063 and the group with a longer history of suffering (&gt;5 years) was 4.211 ± 1.399. There were 8 people who experienced thinning bone from the whole age group, the most were from the age group of 51-60 years old. Conclusion: Mental Index (MI) value is greater or exceeds the normal value (≥ 3,1 mm) in both groups of patients with type 2 DM based on length of suffering ≤ 5 years and &gt; 5 years. Keywords: Diabetes mellitus type 2, Mental Index, Mandibular Cortical Bone, Panoramic Radiography, Bone Mineral Density.

Effect of Salvadora persica on resin-dentin bond stability

BackgroundThe stability of resin–dentin interfaces is still highly questionable. The aim of this study was to evaluate the effect of Salvadora persica on resin–dentin bond durability.Materials and methodsExtracted human third molars were used to provide mid-coronal dentin, which was treated with 20% Salvadora persica extract for 1 min after acid-etching. Microtensile bond strength and interfacial nanoleakage were evaluated after 24 h and 6 months. A three-point flexure test was used to measure the stiffness of completely demineralized dentin sticks before and after treatment with Salvadora persica extract. The hydroxyproline release test was also used to measure collagen degradation by endogenous dentin proteases. Statistical analysis was performed using two-way ANOVA followed by post hoc Bonferroni test and unpaired t-test. P-values < 0.05 were considered statistically significant.ResultsThe use of Salvadora persica as an additional primer with etch-and-rinse adhesive did not affect the immediate bond strengths and nanoleakage (p > 0.05). After 6 months, the bond strength of the control group decreased (p = 0.007), and nanoleakage increased (p = 0.006), while Salvadora persica group showed no significant difference in bond strength and nanoleakage compared to their 24 h groups (p > 0.05). Salvadora persica increased dentin stiffness and decreased collagen degradation (p < 0.001) compared to their controls.ConclusionSalvadora persica extract pretreatment of acid-etched dentin preserved resin–dentin bonded interface for 6 months.Clinical significanceDurability of resin-dentin bonded interfaces is still highly questionable. Endogenous dentinal matrix metalloproteinases play an important role in degradation of dentinal collagen within such interfaces. Salvadora persica may preserve resin-dentin interfaces for longer periods of time contributing to greater clinical success and longevity of resin composite restorations.

The Effect of Torbangun (Coleus amboinicus Lour) Leaf Extract on Antibiotic Resistant Bacteria Escherichia coli pBR322 and Toxicity Tests on Artemia salina Leach

Inappropriate antibiotic use can lead to antibiotic resistance, causing many problems in the treatment of Escherichia coli infection. One way to avoid resistance is to use the traditional medicine Coleus amboinicus Lour, also known as Torbangun in Indonesia. The purpose of this study was to determine the inhibitory ability of 70% ethanol extract of Torbngun leaf against Escherichia coli pBR322 has plasmid pBR322, which is resistant to antibiotics Ampicillin and Tetracycline based on activity and minimum inhibitory concentration. Extraction was carried out using the maceration method. The presence of inhibition zones indicated antibacterial activity. The appearance of the inhibition zone at the minimum concentration indicated the minimum inhibitory concentration. Following this, the cytotoxic activity was evaluated using the brine shrimp lethality test. Data analysis was performed using an unpaired t-test using GraphPad Prism® software. Antibacterial activity tests showed that a 15% torbangun leaf extract concentration had inhibitory power against Escherichia coli pBR322. The toxicity test of torbangun leaf extract against Artemia salina Leach shrimp larvae showed LD50 results at a concentration of 150 ppm.

Effect of Structured Nursing Intervention on Knowledge and Treatment Adherance Among Patients With End Stage Renal Disease in Kano Metropolis

&lt;p&gt;&lt;em&gt;&lt;strong&gt;Background&lt;/strong&gt;: Chronic Renal Failure is a progressive irreversible deterioration in renal function in which the body’s power to maintain metabolic, fluid, and electrolyte balance fails, resulting in uremia which contributes to the patient's dependence upon hemodialysis for the maintenance of the internal milieu and to avoid uremia. Patients with end-stage renal disease (ESRD) are characterized by low levels of physical activity and a continuous decline in physical function. The main aim of the study was to assess the effect of structured nursing intervention on treatment adherence among patients with end-stage renal disease in Kano Metropolis. &lt;/em&gt;&lt;/p&gt;&lt;p&gt;&lt;strong&gt;&lt;em&gt;Methods&lt;/em&gt;&lt;/strong&gt;&lt;em&gt;: This study utilized a two-group pretest-post-test quasi-experimental design. Patients receiving dialysis care at Muhammadu Abdullahi Wase Teaching Hospital and Abubakar Imam Urology Hospital were purposively selected as experimental and control groups respectively. A total of 422 participants were estimated and selected for the study and equally shared among the two groups, i.e. 211 for the control and 211 for the experimental group. All statistical data were analyzed and described using simple frequencies, Chi-square, T-test and regressions. &lt;/em&gt;&lt;/p&gt;&lt;p&gt;&lt;strong&gt;&lt;em&gt;Results&lt;/em&gt;&lt;/strong&gt;&lt;em&gt;: Regarding treatment adherence, the pre-intervention mean score was 52.9 and 50.00, and the post-intervention score was 73.2 and 54.6 in the intervention and control groups, respectively. The effectiveness of the intervention was tested using unpaired t-test which was found to be significantly effective (p&amp;lt;0.005). &lt;/em&gt;&lt;/p&gt;&lt;p&gt;&lt;strong&gt;&lt;em&gt;Conclusion&lt;/em&gt;&lt;/strong&gt;&lt;em&gt;: Based on the findings of the study, the structured nursing intervention was effective in improving the level of knowledge regarding the disease and treatment adherence (p&amp;lt;0.005) among patients with end-stage renal disease in the Kano metropolis. It is therefore recommended that the nephrology nurses, hospital administrators, and the general public join hands to make the services accessible and affordable to these patients to maintain their normal life activities through good adherence to the treatment regimen.&lt;/em&gt;&lt;/p&gt;