bluuebunny/crossref_metadata_2025_split huggingface dataset:
High‐tryptophan Diet Reduces Weight Gain in Rats via Indole‐3‐propionic Acid, a Gut Microbiota Metabolite
Authors: Konopelski Piotr , Konop Marek , Gawrys Marta , Piotr Podsadni , Ufnal Marcin 
Published: June 2021

Tryptophan is an essential amino acid. Paradoxically, both tryptophan‐low and tryptophan‐rich diets have been found to decrease weight gain in rats, however, the mechanisms are not clear.Energy and water‐electrolyte balance were evaluated in 12‐week‐old, male, Sprague Dawley rats maintained on either control diet (TC) or tryptophan‐high (TH) or tryptophan‐low (TL) or control diet with neomycin, an antibiotic (TCA), for 10 days. Feces and blood levels of tryptophan and its bacterial metabolites were evaluated using chromatography coupled with mass spectrometry.TL rats showed reduction in body weight. TH showed body weight gain, however, it was significantly lower than in TC and TCA rats. TL rats showed significantly lower blood level of tryptophan and its bacterial derivatives including indoxyl sulfate and indole‐3‐propionic acid (I‐3‐P). TH rats had significantly higher blood tryptophan level comparing to TL rats. I‐3‐P in feces and in portal blood of TH rats was significantly higher comparing to TC, TL and TCA rats. In a separate series of experiments rats treated daily i.p. with I‐3‐P showed significantly smaller weight gain than rats treated with the vehicle.Our study suggests that tryptophan‐rich diet has a negative effect on energy balance via I‐3‐P, a gut bacteria metabolite of tryptophan. In contrast, tryptophan‐deficient diet may reduce weight due to the deficiency of the essential amino acid and decreased anabolism.Support or Funding InformationSupported by the Ministry of Science and Higher Education Republic of Poland, Diamond grant DI2016 007346This abstract is from the Experimental Biology 2019 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
Elevated Plasma Endothelin‐1 is Associated with Reduced Weight Loss Post‐Vertical Sleeve Gastrectomy
Authors: Jenkins Haley N , Williams London J , Dungey Adam S , Vick Kenneth D , Grayson Bernadette E , Speed Joshua S 
Published: June 2021

Current research supports that obesity and insulin resistance are positively correlated with plasma endothelin‐1 (ET‐1) levels; however, the mechanisms leading to this increase in ET‐1 are not fully understood. Similarly, while some physiological effects of ET‐1 have been characterized, the full complexity of this hormone has yet to be described in tissues outside of the vascular system. To date, one of the best treatments available for morbid obesity, cardiovascular disease, and metabolic syndrome is bariatric surgery to quickly reduce body fat and the factors associated with obesity‐related disease. We hypothesize that vertical sleeve gastrectomy (VSG) will reduce plasma ET‐1 levels. This was tested by measuring plasma ET‐1 levels from twelve obese patients before VSG, 6 weeks after, and 6 months after surgery. The results indicate that 6 weeks following VSG, plasma ET‐1 levels increased by 24%; however, after 6 months, there was a 27% decrease compared to baseline. Average weight loss in this cohort was 11.3±2.4 percent body weight after 6 weeks and 21.4±5.7 percent body weight after 6 months. Interestingly, we observed an inverse relationship between baseline plasma ET‐1 and percent body weight loss 3 months (R2=0.45, p<0.05) and 6 months (R2=0.49, p=0.01) post bariatric surgery. Our results indicate that VSG reduces plasma ET‐1 levels, a possible mechanism for improved metabolic risk in these patients. These data also suggest that ET‐1 may inhibit weight loss or serve as a predictor of weight loss following bariatric surgery.Support or Funding InformationThis work is supported by NHLBI grant R00 HL127178 to JSS.This abstract is from the Experimental Biology 2019 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
Early Life Stress‐Induced Increases in Adiposity in Mice are Prevented by the Adipose Tissue‐Specific Abrogation of Neuropeptide Y Receptor 2
Authors: Leachman Jacqueline Renee , Dalmasso Carolina , Jones Tucker , Seward Tanya , Taylor Bradley K , Loria Analia S 
Published: June 2021

Stress has long been associated with the physiology of disease, however, its mechanism of action in obesity remains unknown. Neuropeptide Y (NPY) plays a critical role in the regulation of energy homeostasis acting in the central nervous system as an orexigenic agent resulting in feeding that particularly favors the ingestion of foods rich in carbohydrates. Peripherally, NPY stimulates lipid storage in white adipose tissue (WAT) via NPY receptor 2 (NPY2R) promoting adipocyte precursor proliferation, differentiation of preadipocytes into mature adipocytes, as well as lipogenesis and adipose tissue deposition. Under chronic stress, NPY is co‐released with catecholamines by sympathetic axons to stimulate increased fat storage. We have previously discovered that mice exposed to a model of early life stress, maternal separation and early weaning (MSEW), display increased adiposity at weaning. Thus, we hypothesized that knocking down NPY2R in adipose tissue using the cre‐lox technology could prevent increases in adiposity in mice exposed to MSEW. MSEW was performed by separating adipose tissue‐specific floxed NPYR2 pups from the dam for 4 to 8 hours during postnatal days (PD) 2 to 16, and weaned early on PD 17. Control mice remained undisturbed and were weaned on PD 21. Body weight and echoMRI were performed in all litters at postnatal day 21 and tissue samples were taken for genotyping. Despite no changes in BW, wild type (WT, Y2 floxed) mice exposed to MSEW showed a ~30 % increase in adiposity compared with control WT mice (7.5±0.6 vs. 5.6±0.3 % body fat, p<0.05). However, this increase in adiposity was abrogated in MSEW NPYR2 knock out mice (5.7±0.1 vs. 5.5±0.6 % body fat, p=NS). In addition, WT mice exposed to MSEW showed reduced ratio of the adipose tissue norepinephrine/NPY concentrations and increased NPY2R mRNA expression compared with control mice. Taken together, these data suggest that exposure to postnatal MSEW may induce a shift in co‐transmitter release by catecholaminergic neurons innervating the adipose tissue leading to an increase in adiposity.Support or Funding InformationR01HF135158 to ASL, R01HL135158‐01A1S1 to JRLThis abstract is from the Experimental Biology 2019 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
Exposure to Microbial Infection In Utero Has Long‐Term Effects on the Cardiovascular Response to Angiotensin II and Forebrain Gene Expression in Rats
Authors: Watkins Jacqueline , Von Chamier Maria , Brown Mary B , Hayward Linda 
Published: June 2021

Prenatal stressors, such as low protein diet, maternal smoking, and chronic psychological stress, predispose offspring to cardiovascular disease later in life. Microbial infection during pregnancy also increases the risk of adverse offspring health outcomes, but it remains to be determined whether this form of prenatal “stress” also triggers long‐term changes in offspring cardiovascular health. Thus, the present study was undertaken to evaluate whether the cardiovascular response to angiotensin II (ANGII) is altered in adult offspring born to mothers exposed to a microbial infection late in pregnancy when compared to non‐exposed controls (CON). Using an established rodent model of prenatal maternal infection (INF), dams were exposed to the rat pathogen Mycoplasma pulmonis or to vehicle via intravenous injection, on day 14 of pregnancy. Following natural birth, pups were cross‐fostered to non‐infected dams. Preliminary data (n=6–8/group) demonstrate that INF modestly elevated body weight by ~25 gm, significantly reduced resting heart rate (HR) by ~30 bpm in both sexes (P<0.001), but did not change resting blood pressure (BP) at 14 wks of age compared to CON. When subjected to a subcutaneous injection of ANGII, the rise in BP was significantly elevated in both female (48±6 vs 18±3 mmHg; P<0.002) and male (51±4 vs 39±5 mmHg; P<0.03) INF offspring compared to CON. The compensatory drop in HR was not significant between INF vs CON in either sex, suggesting INF induces baroreflex dysfunction. Gene expression analysis of the amygdala identified significant changes in corticotrophin system gene expression in females but not males. These results indicate that prenatal stress associated with INF significantly alters both cardiovascular function and gene expression in the forebrain later in life, potentially increasing the likelihood for offspring to develop cardiovascular disease when exposed to a second chronic stressor.Support or Funding InformationFunded by FL‐DOH James and Esther King Biomedical Research Grant (LH & MB).This abstract is from the Experimental Biology 2019 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
Regulation of Natriuretic Peptide Receptor‐A Expression in Gene‐targeted Mice via Modulation of Histone 3 Acetylation and Methylation Levels
Authors: Kumar Prerna , Pandya Krishna , Pandey Kailash N. 
Published: June 2021

Guanylyl cyclase/natriuretic peptide receptor‐A (GC‐A/NPRA) signaling plays a well‐defined role in the regulation of blood pressure and blood volume. Mice lacking functional Npr1 gene (coding for GC‐A/NPRA) exhibit hypertension, kidney disease, and heart failure; however, the mechanisms regulating Npr1 expression are not well understood. The objective of the present study was to determine the effect of class I histone deacetylase (HDAC) inhibitor, mocetinostat (MGCD0103) on the expression of Npr1 via modulation of epigenetic changes. Male and female (16–20 weeks old) Npr1 gene‐knockout heterozygous (1‐copy; Npr1+/−), wild‐type (2‐copy; Npr1+/+) and gene‐duplicated heterozygous (3‐copy; Npr1++/+) mice were injected intraperitoneally with MGCD0103 (2 mg/kg) at alternate days for 2‐weeks. The Western blot result showed that MGCD0103 significantly increased the renal NPRA protein levels in male and female mice compared with vehicle‐treated controls. Plasma cGMP levels were higher in female mice compared with male mice both before and after the treatment with MGCD0103. Renal HDAC activity was significantly higher (p < 0.05) in male mice than female mice; however, treatment with MGCD0103 significantly attenuated (p < 0.05) HDAC activity by 40% in male and ~50% in female animals. The histone acetyltransferase (HAT) activity was lower in untreated 1‐copy male mice compared with wild‐type mice and MGCD0103 treatment significantly increased HAT activity whereas no significant differences were observed in drug‐or vehicle‐treated female mice. Male and female mice showed differential renal acetylation levels of histone 3 at different lysine positions (H3‐K9, H3‐K18, and H3‐K27) and treatment with MGCD0103 significantly enhanced the acetylation levels of H3‐K9 and H3‐K27 in 1‐copy male mice and H3‐K18 in 1‐copy female mice. There was substantial decrease in methylation of suppressive histone marks (H3‐K9 and H3‐K27) in MGCD0103‐treated 1‐copy male and female mice. Male mice exhibited significantly higher systolic blood pressure (SBP) than female mice and treatment with MGCD0103 decreased SBP in both 1‐copy male (106 ± 0.6 vs. vehicle‐treated, 129 ± 1.9; mmHg) and female (97 ± 2.2 vs. vehicle‐treated, 110 ± 2.1 mmHg, p < 0.001) mice. The present results provide direct evidence that MGCD0103 epigenetically regulates NPRA expression in vivo in 1‐copy male and female haplotype mice, via attenuation of HDAC activity and increase in histone H3 acetylation levels. These findings will have important implications in prevention and treatment of hypertension and renal pathophysiological conditions under reduced NPRA signaling.Support or Funding InformationThis work was supported by NIH grants (HL057531 and HL062147) and a partial support from Tulane Carol Lavin Bernick grant award.This abstract is from the Experimental Biology 2019 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
CRISPR‐Cas9 Gene Editing Yields a Novel Rat Model of Cardiometabolic Disease
Authors: Clark Karen C , Wagner Valerie A , Malek Jacob A , Arroyo Alyssa E , Pearson Nicole A , Grobe Justin L , Kwitek Anne E 
Published: June 2021

Despite strong evidence that the Metabolic Syndrome (MetS) and its defining features (central obesity, dyslipidemia, hypertension, hyperglycemia) are highly heritable, the genetic etiology is complex and relatively few causative genes are known. For this reason, we employ the Lyon Hypertensive (LH) rat, a well‐characterized polygenic inbred MetS model. Previous genetic and computational approaches identified a novel gene (C17h6orf52) as a putative master regulator of gene expression and oxidative phosphorylation with pleiotropic effects on body weight and blood pressure regulation.In this study, two different frameshift mutations were generated independently by CRISPR‐Cas9 gene editing, producing either a 5 base pair deletion or a 10 base pair insertion in exon 2 of C17h6orf52, and females from the resulting mutant rat strains, designated M1 and M2, respectively, were characterized (Table 1). Herein, mutations in C17h6orf52 are shown to significantly increase blood pressure and serum cholesterol in vivo, despite no significant changes in body composition or body weight in LH‐derived female rats.Either mutation appears to cause modest, but significant or nearly significant, increases in blood pressure on a 0.3% NaCl diet. M1−/− females exhibit significantly increased systolic and diastolic blood pressure, while M2−/− females' systolic and diastolic blood pressure is more moderately elevated. This effect is potentiated when animals are administered a high salt diet. Finally, although neither total nor low‐density lipoprotein (LDL) cholesterol is changed in the M1 female rats with respect to their wildtype control, M2 females develop higher serum cholesterol, as well as significantly higher LDL cholesterol.The differences in phenotype between the M1 and M2 mutations were unexpected. Preliminary data indicates that 5 base pair deletion present in the M1−/− rats results in two distinct splicing variants that may be contributing to the observed phenotype differences. Nevertheless, these data suggest that C17h6orf52 is protective against cardiometabolic risk factors, given that C17h6orf52 mRNA is more highly expressed in MetS‐susceptible LH rats than MetS resistant Lyon Normotensive (LN) rats, and the mutation confers a greater burden of MetS features on an already obese and hypertensive rat. The continued study of this rat model of Metabolic Syndrome has the potential to functionally validate an uncharacterized regulatory gene, and provide novel targets for pharmacological intervention in the treatment of obesity.Support or Funding InformationR01HL089895, R21DK089417, T32GM008629, 14GRNT20410043This abstract is from the Experimental Biology 2019 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
Being a First Time Flipper is Challenging and Worthwhile to Increase Student Understanding of Cell Signaling
Authors: Halpin Patricia A. , Gopalan Chaya 
Published: June 2021

To learn more about creating a Flipped Classroom, I attended a full day workshop on Flipped Teaching and Retrieval Practice (FT‐RP) at Southern Illinois University Edwardsville. This instructional strategy was then integrated into my BSCI 735 cell biology class. Two weeks in advance, the 17 students (juniors and seniors) were notified that the class would be flipped and the student expectations for that class meeting were described. Cell signaling was the subject chosen because students often have difficulty understanding the different pathways of steroid and peptide hormones. In advance of the class meeting, students watched a YouTube video on cell signaling and reviewed a PowerPoint outline. At the beginning of the class period, students took a multiple choice quiz on the subject and then participated in two types of cell signaling dramatizations. For the dramatization of the mechanism of action of steroid hormones, each student wore a placard with an assigned role: steroid hormone, plasma membrane, intracellular receptor, nuclear pore, and DNA. With the guidance of the instructor, students participated in the signaling cascade that began with the steroid hormone diffusing through the plasma membrane. The students switched roles and the activity was repeated. For the dramatization of the mechanism of action of peptide hormones, each student wore a placard with an assigned role: ligand, receptor, G protein, adenylate cyclase, plasma membrane, ATP, protein kinase A, intracellular enzymes and ion channels. The students performed the dramatization that concluded with cAMP binding to protein kinase A (PKA). The exercise was repeated twice with students switching roles each time. In the final dramatization, the next level of complexity of the PKA pathway was added and additional roles were assigned: the regulatory and catalytic subunits of PKA, the nuclear pore, and DNA, while enzymes and ion channels moved to the nucleus. This signaling pathway ended with genes being activated, enzymes turned on and ion channels opened. At the end of the class, students took the same quiz to assess what they learned and all scores increased. The flipped teaching was successful as students were able to differentiate the cell signaling pathways of steroid and peptide hormones.Support or Funding InformationAPS‐Teaching Career Enhancement AwardThis abstract is from the Experimental Biology 2019 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
Preliminary assessment of the use of out‐of‐class video presentations for content review in an allied health pathophysiology course
Authors: Walton Kristen LW 
Published: June 2021

The flipped classroom technique has continued to gain popularity in undergraduate and postgraduate biology and medical education in recent years. This technique generally involves delivery of lecture or other methods of content coverage assigned before class, while class time is instead spent on discussion or other, more active approaches to learning. This technique was implemented for a specific set of content in an upper division undergraduate pathophysiology course. The prerequisite for this course is a one‐semester anatomy and physiology (A&P) course. Although students have to earn a C or better in this prerequisite A&P in order to enroll in the pathophysiology, many students continue to struggle with some major topics in A&P, such as cardiovascular physiology. This makes it difficult to have meaningful discussion of the related pathophysiology without first reviewing content from the prerequisite course. This project aimed to assess the use of pre‐class video lectures as a replacement for in‐class prerequisite course content review. Short (15–30 minute) video lectures were created and made available through the course management system 1–2 class meetings before the related pathophysiology content was covered in class. Students were required to complete an online multiple choice quiz that included questions from the video lecture content. For this preliminary study, data was analyzed for a 19‐minute video review of the cardiovascular system and the related quiz and exam questions. Out of 77 students enrolled in the class, 44 (57%) accessed the video through the course management system prior to the specific deadline; an additional 9 students (12%) accessed the video before the unit exam that included that topic. By the date of the exam, 56 students (73%) had watched the entire video. Five students accessed the video but watched less than half of it. Over 90% of students correctly answered the two multiple choice questions associated with this content in the untimed online quiz (91% for one question, 93% for the other question). A different multiple choice question over this review content appeared on the unit exam; 66% of students answered this question correctly. The average grade on this unit exam, which included cardiovascular pathophysiology as well as three other chapters, was 70.6%. In the previous semester, 62% of students (41 of 66 completing the exam) answered the same multiple choice question correctly, while the average on the full exam was 73.0% . These data support a conclusion that shifting review content to a pre‐class, video lecture format did not strongly impair student learning. Future studies will expand the number of topics analyzed to further assess the impact of pre‐class video lectures for review content on student learning in this course.This abstract is from the Experimental Biology 2019 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
The T‐type Voltage Gated Calcium Channel Cav3.2 is Important for Enteroendocrine Cell Mechanotransduction
Authors: Knutson Kaitlyn , Alcaino Constanza , Nayak Vaishali , Kacmaz Halil , Treichel Anthony J. , Farrugia Gianrico , Beyder Arthur 
Published: June 2021

BACKGROUNDEnteroendocrine cells (EECs) in the gastrointestinal (GI) epithelium are specialized sensors of luminal chemicals and forces. Upon stimulation, EECs release a variety of neurotransmitters and hormones, but cellular mechanisms of excitation‐secretion coupling are poorly understood. We recently discovered that a population of EECs expresses the mechanosensitive ion channel Piezo2, which is necessary for force‐dependent intracellular Ca2+ increase and serotonin release. However, the molecular mechanism that links Piezo2 and serotonin release is unknown. In other sensory systems, the Cacna1h‐encoded t‐type voltage‐gated calcium channel Cav3.2 is important in mechanotransduction for linking mechanical activation with neurotransmitter release.AIMto determine if Cav3.2 is expressed and functionally relevant in EECs.METHODSQuantitative RT‐PCR for Cacna1h mRNA expression was evaluated in colon epithelium of control and NeuroD1 Cre x RiboTag mice, in which actively transcribed mRNA specifically from EECs is tagged with hemagglutinin (HA). NeuroD1 Cre x tdTomato colon cross‐sections were fixed and immunolabeled with anti‐Cav3.2 antibody. Single primary NeuroD1 cre x GCaMP5 EECs were mechanically stimulated and Cav3.2 role in mechanotransduction was tested by pharmacology (Ca2+ free, nickel, mibefradil, ω‐agatoxin IVA) and Cacna1h siRNA knockdown.RESULTSMurine colon epithelium expressed Cav3.2 mRNA. In NeuroD1 Cre x RiboTag mouse, HA co‐localized with EEC marker Chromogranin A (Chga). NeuroD1+ EEC ribosomal trapped mRNA was enriched for markers of epithelium (Vil1), EEC (Chga), and enriched 7.0‐fold for Cav3.2 (n=3, p<0.05 compared to whole epithelium). Immunohistochemistry in NeuroD1 Cre x tdTomato showed that Cav3.2 co‐localized specifically with tdTomato (NeuroD1+) EECs. In primary NeuroD1+ EECs, Cav3.2 inhibition by all pharmacologic agents and siRNA knockdown inhibited mechanically induced Ca2+ response (96±1% for Ca2+ free, 71±9% nickel, 61±6% mibefradil, 80±4% ω‐agatoxin IVA and 96±1% Cav3.2 siRNA, n=3–18, p<0.05 compared to control untreated cells or cells treated with non‐targeted siRNA).CONCLUSIONSCav3.2 is densely and specifically expressed in mouse colon EECs and it is important for regulating Ca2+ in EEC mechanotransduction. Future studies will determine how Cav3.2 channels contribute to EEC electrical excitability, excitation‐secretion coupling, and ultimately EEC Cav3.2 roles in GI physiology.Support or Funding InformationSupported by NIH DK106456, NIH DK084567, and AGA RSAThis abstract is from the Experimental Biology 2019 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.