bluuebunny/crossref_metadata_2025_split huggingface dataset:
Inflammation Biomarkers in Older Adult Rats Consuming Moderate Levels of Ethanol
Authors: Morales Lani , Quigley Jacqueline , Pascua Jeremy , Pinkowski Natalie , Almaiman Lama , Brasser Susan M. , Hong Mee Young 
Published: April 2020

Previous epidemiological studies and experimental data from rodent models have reported a non‐linear relationship between consumption of alcohol and cardiovascular disease risk and colorectal health. These studies indicate that light‐to‐moderate alcohol consumption, in contrast to excessive alcohol use, may provide cardiovascular health benefits as well as a protective effects on risk for colorectal cancer. Although prior studies have examined the effects of moderate ethanol consumption primarily in young adult animals, it is unknown if the results can be extrapolated to older adults or if these effects may be more pronounced in aging animals due to declining indices of health.. This study examined the effects of moderate ethanol consumption on lipid profile and colonic gene expression in older rats. Twenty‐four non deprived middle aged (420 days old) Wistar rats (n=12/group) were allowed to voluntarily consume a 20% v/v ethanol solution on alternate days for 13 weeks or were given access to water as their sole source of fluid (non‐ethanol exposed control). Moderate ethanol consumption significantly increased HDL cholesterol levels (P = 0.016) and ApoA1 (P = 0.048) compared to control. There were no differences in serum triglyceride, total cholesterol, LDL cholesterol, oxidized LDL and glucose between the groups. Blood C‐reactive protein (CRP) was decreased significantly (P = 0.034) in ethanol‐exposed animals. Blood high‐mobility group protein 1 (HMGB1) showed a decreased trend in the ethanol group (P = 0.083). In colon, gene expression of aldehyde dehydrogenase (Aldh) displayed an increased trend in ethanol‐exposed rats (P = 0.073). Colonic superoxide dismutase (Sod) was upregulated with ethanol intervention (P = 0.050). Colonic gene expression of cyclooxygenase 2 (Cox‐2), RelA, b‐catenin, CD68, and Pparr were not different between the groups. These results suggest that moderate consumption of ethanol in older rats may potentially contribute to improved cardiovascular and colorectal cancer risk by improving blood cholesterol, decreasing blood markers of inflammation in and increasing antioxidant enzyme expression in colon.Support or Funding InformationNIH AA023291
Effects of the Astrocyte Endozepine Octadecaneuropeptide (ODN) on Hypoglycemia Patterns of Ventromedial Hypothalamic Nucleus (VMN) Metabolic‐Sensory Neuron Transmitter Signaling and Energy Sensor Activation in Male and Female Rats
Authors: Mahmood ASM Hasan , Uddin Md. Main , Ali Md. Haider , Napit Prabhat Ranjan , Briski Karen P. 
Published: April 2020

The astrocyte gliotransmitter octadecaneuropeptide (ODN) is a cleavage product of diazepam‐binding inhibitor (DBI) and an endogenous central benzodiazepine receptor ligand. ODN regulation of feeding responses to excess or diminished glucose availability in the brain infers ability to modulate neural metabolic‐sensory function [Lanfray et al., 2013]. The ventromedial hypothalamic nucleus (VMN), a principal component of the neural glucostatic network, integrates nutrient and other cues to shape glucose counter‐regulation. Our studies show that VMN gluco‐stimulatory nitric oxide (NO) and gluco‐inhibitory g‐aminobutyric acid (GABA) neurons express the ultra‐sensitive energy sensor 5′‐AMP‐activated protein kinase (AMPK). This research examined the premise that ODN exerts sex‐specific effects on hypoglycemic patterns of VMN nitrergic and/or GABAergic nerve cell sensor activation and neurotransmitter signaling. Testes‐intact male and ovariectomized, estradiol‐replaced female rats were pretreated by intracerebroventricular administration of the ODN octapeptide (OP) fragment or vehicle prior to insulin injection. VMN NO and GABA cells were identified by immunocytochemistry prior to combinatory laser‐catapult dissection/high‐sensitivity Western blotting. Results identified nitrergic cells exhibiting OP‐reversible effects of hypoglycemia on expression profiles of the NO marker protein neuronal nitric oxide synthase (nNOS), AMPK, and phospho (activated)‐AMPK in distinctive regions of the male and female VMN. In each sex, OP effects on GABA neuron AMPK protein content and activation were also observed at specific levels of the VMN. Outcomes provide novel evidence that ODN may critically impact VMN substrate fuel screening. Further studies are needed to characterize neuroanatomical connectivity of OP‐sensitive nitrergic and GABAergic neurons within central metabolic pathways, and to examine whether metabolic cues that regulate VMN astrocyte ODN release, astrocyte reactivity to ODN‐controlling signals, and metabolic‐sensory nerve cell receptivity to ODN are sex‐dimorphic.Support or Funding InformationNIH DK 109382
Genome‐Wide Methylation in Patterns in Idiopathic Pulmonary Arterial Hypertension
Authors: Gunn Brady Luke , Memar Shayan , Markand Shanu , Abeyrathna Prasanna 
Published: April 2020

ObjectiveEpigenomic regulation is important in the pathology of many diseases. Evidence suggests epigenetic modification of individual genes in idiopathic pulmonary arterial hypertension (IPAH), and understanding these epigenomic mechanisms involved in vascular remodeling in IPAH may be beneficial. The objective of the current study was to assess the genome‐wide methylation of IPAH lungs. We hypothesized that genome‐wide methylation would have an essential role in vascular remodeling in IPAH.MethodsLung tissues from 8 age‐matched and sex‐matched IPAH lungs and 8 healthy lungs were analyzed in the current study. Genome‐wide methylation of cytosine was assessed using whole genome bisulfite sequencing. Gene functional pathway analysis was performed on identified differentially methylated genes.ResultsBased on bioinformatics analysis of the methylation sequences, differentially methylated genes were identified in IPAH lungs. Further, there were distinct methylation patterns in the pathophysiology of the IPAH lungs.ConclusionIdentification of epigenetically regulated pathways in IPAH may offer novel therapeutic opportunities for IPAH.Support or Funding InformationATSU Start Up Fund
Use of a spectrophotometric assay to evaluate amino acid activation and tRNA charging
Authors: Mobisa Japheth , Samuel Kyra , Okeke Idiuso , Castaneda Jacquelyn , Trinh Thanh , Enewali Chinazom , Bell Anthony Jerome 
Published: April 2020

Conventional assays that investigate amino acid activation and tRNA‐charging rely on using radiolabeled amino acids (14C) and tRNAs (32P) to detect the presence of activated amino acid adenylate(s) and acylated tRNAs. There are also several assays that monitor activation and acylation products by using (32P) radiolabeled ATP and inorganic pyrophosphate. The radioisotope based assays offer unmatched sensitivity. However, the experiments require the use of hazardous reagents and often laborious protocols. More recently a colorimetric based assay has been developed to detect the level of inorganic free phosphate in aqueous solution. The assay is based on the complexation of malachite green molybdate and inorganic phosphate – the complex absorbs at 620 nm. The assay, referred to as the malachite green assay, has been modified to measure amino acid activation and tRNA charging. Our short‐term goal is to use the malachite green assay to measure the ability of aminoacyl‐tRNA syntheses (aaRSs) to activate and charge analog amino acids (AAs). Analog or “unnatural” AAs are non‐proteinogenic residues beyond the set of natural 20 L‐AAs. Bacteria and fungi use large multi‐subunit enzyme complexes, nonribosomal peptide synthetases (NRPs), to generate peptides composed of analog AAs. The resulting analog peptides typically possess enhanced bioactivity. Analog peptide bioactivity is generally linked with enhanced: i)membrane permeability, ii)protease stability and iii)chemical complexity. These advantages make analog peptide ideal for biological therapeutics. Our long‐term goal is to use the cell free expression system – the PURE system (
Protein synthesis
Using
Recombinant
Elements) to synthesize analog peptides. The resulting peptides will be directed against protein‐protein interactions known to exacerbate disease. Our initial studies focus on using the malachite green assay to screen analog amino acids for activation and tRNA charging using purified aaRS and tRNA (total tRNA and single isoacceptors). Analogs identified via screening will be assembled into a library that will be used to generate analog peptides.Support or Funding InformationUniversity of San Diego
Activation of Bradykinin‐Sensitive Pericardial Afferents Increases Splanchnic Nerve Activity
Authors: Martin Doug , Liu Xuefei , Zheng Hong 
Published: April 2020

Sympathetic drive plays a key role in control of cardiovascular function. The heart performs an important sensory function via a variety of cardiac receptors that modulate sympathetic drive. Bradykinin‐sensitive sympathetic afferent nerves from the heart can trigger marked increases in efferent sympathetic nerve activity to some cardiovascular targets such as the kidney. We reported previously that pericardial injection of bradykinin increased venous tone and cardiac output. The splanchnic vascular compartment plays an important role in control of venous return and cardiac output. This work tested the hypothesis that activation of bradykinin‐sensitive pericardial afferents increases postganglionic splanchnic sympathetic nerve activity.Sprague Dawley rats were anesthetized with a cocktail of alpha chloralose (80 mg/kg) and urethane (800 mg/kg). Catheters were placed in the femoral artery and vein to record arterial pressure (MAP) and heart rate (HR). A PE10 catheter was placed in the pericardial space for the injection of bradykinin (BK), an agent known to stimulate cardiac sympathetic afferents. The chest was closed and the rat allowed to breathe spontaneously. The splanchnic sympathetic nerve was isolated via a dorsal approach and placed on recording electrodes. After stabilization, the blood pressure, heart rate, and integrated splanchnic nerve activity (SPLNA) responses to pericardial injection of BK or vehicle were recorded. Responses were also recorded for intravenous injection of dimethylphenylpiperazinium (DMPP 150 ug/kg), a ganglionic stimulant, and a ganglion blocker (hexamethonium 20 mg/kg).Pericardial injection of saline vehicle had little effect (MAP: 0.1±0.4 mm Hg; HR: 1.2±1.4 bpm; SPLNA 1±2%). In contrast, intravenous injection of a ganglionic stimulant, DMPP, increased MAP (23±1 mm Hg) and SPLNA (95±46%). Pericardial injection of BK (10 ug/kg) increased MAP (21±3 mm Hg), HR (28±7 bpm) and SPLNA (93±37%). Ganglionic blockade with hexamethonium reduced SPLNA (−75±6%) and abolished the responses to pericardial injection of BK (MAP: −1±3 mm Hg; HR: −2±1 bpm; SPLNA: 7±30 %).We interpret these data to indicate that activation of bradykinin‐sensitive pericardial afferents increases postganglionic splanchnic sympathetic nerve activity. This finding is consistent with the view that activation of the cardiac sympathetic afferent reflex elicits a venoconstrictor response to increase venous return from the splanchnic vascular compartment.Support or Funding InformationSupported by NIH R01 HL136741‐03 (DM), R01 DK114663‐1 (HZ), and the Basic Biomedical Sciences program of the Sanford School of Medicine, University of South Dakota.
Wheat Germ Acid Phosphatase Activity in High Percentages of Organic Solvents
Authors: Moorman Veronica R. , Kindl Evan D. , Summers Fiona N. 
Published: April 2020

While enzymes canonically operate in aqueous environments, several have been shown to also function in non‐aqueous solvents and/or solvent mixes. Phosphatases, for example, have been previously shown to hydrolyze phosphates in mixtures containing modest amounts of certain organic solvents. Here, detecting the fluorescent dephosphorylation product naphthol AS‐MX with fluorescence spectroscopy (ex: 388 nm, em: 512 nm) has shown commercial wheat germ acid phosphatase to be active in high amounts (up to ~70% by volume) of 1,4‐dioxane, 1,2‐ dimethoxyethane, 2‐methoxyethanol, dimethyl sulfoxide, and acetonitrile when the aqueous component was comprised of Tris buffer (pH 7). These solvent mixtures were also physically characterized to better understand differences in both relative naphthol AS‐MX fluorescence and wheat germ acid phosphatase activity across the different solvent mixtures investigated. These results suggest that phosphatase activity can be detected with less water that previously published.Support or Funding InformationAcknowledgment is made to the Donors of the American Chemical Society Petroleum Research Fund and to the National Science Foundation’s Research Experience for Undergraduates program for partial support of this research.
Relationship Between Team Dynamics and Performance in Solo‐Then‐Collaborative (Hybrid) High‐Stakes Testing in Pre‐Clerkship Medical Students
Authors: Yarbrough Tracy Lynnette , Ely Susan 
Published: April 2020

ObjectivesTo characterize student perceptions of team dynamics and the relationship between team dynamics and performance within a novel hybrid exam platform.MethodsMedical students enrolled in pre‐clerkship year 1 at CNUCOM during academic year 2017–2018 were recruited for participation in this study. Data was collected at two timepoints— at the mid‐course exam in the final course of year 1 and at the mid‐course exam in the initial course of year 2. All participants completed a 100‐item selected‐response exam individually and then as part of a pre‐formed student team of 8–9 members. Following the team exam, students completed a 15‐item survey reflecting on their team interactions. In addition, faculty proctors of the team exam during course 1 completed a 15‐item survey reflecting on their observations of team dynamics. Finally, student performance (both individual and team exam scores) were collected for all participants. Team dynamics scores were calculated based on responses to survey questions regarding team interactions during the exam as identified by students and proctors (for course 1). This human subjects protocol was reviewed and approved by the CNUCOM Institutional Review Board.Results95 and 94 students completed the first and second hybrid exam surveys, respectively. Average individual scores (+/− SD) on the first and second exams were 69 (+/11) and 80 (+/− 10), and average team scores on the first and second exams were 92 (+/−3) and 99 (+/−1). At the first exam, student‐derived team dynamics score was highly correlated with team exam score, while team member average individual exam score did not strongly correlate with team exam score. {It is noted that teams for the first exam were formed 8 months prior to the exam and had been used extensively in other team‐based learning sessions.} Conversely, teams utilized in the second exam had been formed less than 1 month prior to the exam and had been functioning as a team for a limited number of sessions prior to the exam—on the second exam, team dynamics score was not well‐correlated with team exam score, and instead team member average individual exam score correlated more strongly with team score. Proctor’s assessment of team dynamics correlated moderately well with students’ assessment of team dynamics in course 1 (r = 0.24).ConclusionsOur data suggest that team dynamics play a role in team performance in hybrid testing utilized for high‐stakes exams among undergraduate medical students. This relationship may depend more on student comfort and familiarity within a team than on the individual knowledge and specific contributions of team members. More investigation on long‐term knowledge retention as related to dynamics within teams is desirable to further characterize outcomes utilizing this novel testing format.
The Effect of Long Distance Running on the Pattern of Production of Salivary Cortisol on Rest Days
Authors: Hill Gabrielle Marie , Baumbach Jordan , Barry Alla 
Published: April 2020

INTRODUCTIONCortisol is a glucocorticoid synthesized from cholesterol by the zona fasciculate of the adrenal cortex and is controlled by way of a negative feedback pathway within the hypothalamic‐pituitary‐adrenal (HPA) system. HPA axis receives strong circadian input and plays an essential role in the regulation of stress responses. Cortisol spikes in the morning upon waking up, and then decreases throughout the day, but exercise modifies this pattern of production. Physical exercise activates HPA axis to maintain proper blood glucose levels during these times of energy expenditure. According to previous studies endurance‐trained athletes possess a significantly higher level of circulating cortisol than power trained athletes immediately after exercise. Cortisol is known to have debilitating effects on factors related to bone mineral density (BMD). Consequently, the chronic increase in cortisol in long‐distance runners can lead to adverse effects such as decreased BMD and can contribute to stress fractures. The purpose of this study is to evaluate cortisol levels during days of rest to determine the possible change in the pattern of cortisol production stimulated by repetitive prolonged previous exposure to the physical activity rather than actual stimulus.METHODSMissouri Southern State University (MSSU) Institutional Review Board approval was gained before data collection. A participant group was gathered including 12 males from the men’s cross‐country team at MSSU in the fall of 2019. Participants completed a demographic questionnaire stating the number of hours per week actively participating in sport specified workouts, average hours of sleep per night, and a list of supplements being consumed. Participants gave saliva samples within pre‐determined time intervals before, during, and after their workouts as well as during the same times during days of rest. Samples were collected on two training days and one rest day a week on three separate weeks for a total of 27 samples per athlete. In addition to saliva samples, a stress assessment was filled out by every athlete each day of collection. All samples were placed in a freezer at 0 degrees Celsius until collections were completed and analyzed. Samples were evaluated via ELISA by a competitive immunoenzymatic colorimetric method for the quantitative determination of cortisol concentration. ELISA absorptions were analyzed with linear regression to calculate cortisol concentrations.CONCLUSIONThe result of this study will assist in a better understanding of the hypothalamic‐pituitary‐adrenal axis function. Furthermore, while allowing the endurance‐trained athlete to efficiently and properly recover, it will aid in reducing the risk of injury and overtraining syndrome.
Suppression of Organismal Death Pathway in the Heart Tissues of Diabetic Mice by NP‐ 6A4, A Peptide Agonist of the Angiotensin II Receptor AT2R
Authors: Pulakat Lakshmi , Belenchia Anthony , Mooney Brian , DeMarco Vincent G. 
Published: April 2020

BackgroundDiabetes accelerates cardiac dysfunction and aging. Angiotensin II type 2 receptor AT2R is a member of the anti‐inflammatory arm of the renin‐angiotensin‐aldosterone (RAAS) system. NP‐6A4 is an AT2R peptide agonist with the FDA orphan drug designation for pediatric cardiomyopathy. To test the hypothesis that NP‐6A4‐AT2R signaling is cardioprotective in chronic diabetes, we investigated the changes in cardiac proteome of 63‐week old diabetic mice after a 55‐week treatment with NP‐6A4.Methods and ResultsAll studies were conducted in accordance with the FASEB Statement of Principles for the use of Animals in Research and Education. Diabetes was induced in 8‐week old male c57Bl6 mice via intraperitoneal delivery of streptozotocin (STZ: 50mg/kg/day: 5 days; n=11). Fasting insulin levels were decreased in the STZ‐treated mice (~0.2±0.03 ng/mL versus ~0.94±0.08ng/mL in control mice; p<0.05), but fasting glucose was increased (313±29mg/dL p<0.05). STZ‐treated mice also showed a reduction in body weight by ~10–14% compared to control mice (p<0.05). NP‐6A4 treatment (5mg/kg/day; n=5) of STZ‐treated mice did not change these metabolic parameters. Whole proteome analysis of the cardiac tissues of these 63‐week old mice identified a total of 2760 protein groups in 15 samples with the following criteria: >=2 unique peptides per protein, peptide spectrum FDR <1%. For quantification, a threshold minimum of 4 spectral counts (average of each group of replicates) was used. This resulted in a quantified proteome of 1,569 proteins. NP‐6A4 treated mice hearts exhibited 109 significant differences (p<0.01) compared to vehicle treated diabetic mice hearts with 100 proteins showing increased and 9 proteins showing decreased abundance. Analysis of these data using Ingenuity Pathway Analysis (IPA) showed that 32 of deferentially expressed proteins in diabetic mouse heart induced a pathway designated ‘suppression of organismal death’ (Z score −4.706). Comparing NP‐6A4‐treated mice hearts to healthy mice hearts resulted in 311 significant differences (p<0.01), with 253 proteins increased and 58 proteins decreased. Moreover, 82 of these differentially expressed proteins in the NP‐6A4‐treated mice hearts, compared to healthy control mice hearts, also induced the pathway ‘suppression of organismal death’ (Z score‐3.415).ConclusionA 55‐week treatment with NP‐6A4 induced a cardioprotective pathway in diabetic mouse heart that has the potential to suppress organismal death when compared to age‐matched healthy mice or vehicle treated diabetic mice. This observation suggests that continuous treatment with NP‐6A4 in chronic diseases such as diabetes has the potential to confer cardiac protection and prevent organ failure.Support or Funding InformationThis work was supported by NIH NHLBI 1R01HL118376‐01 (LP), and NIH NHLBI 1R01HL138988‐01A1 (LP) and a small grant from Novopyxis Inc. (LP).