bluuebunny/crossref_metadata_2025 huggingface dataset:
The effect of rutin on overactive contractility of bladder smooth muscle in the model of cyclophosphamide‐induced cystitis in rats (690.15)
Authors: Celebi Fikret , Gelen Volkaan , Cınar Dursun , Sengul Emin
Published: June 2021

The effect of rutin on overactive contractility of bladder smooth muscle in the model of cyclophosphamide‐induced cystitis in rats Fikret Çelebi, D.Ali Cınar, Emin Sengül, Volkan Gelen.

Department of Physiology, Faculty of Veterinary Medicine, University of Atatürk, Erzurum, Turkey Cyclophosphamide (CYP) is a widely used chemotherapeutic agent in the treatment of cancer and renal disease.However, CYP often leads to hemorrhagic cystitis. Hemorrhagic cystitis increases the frequency and amplitude of spontaneous contractions in the bladder muscles.

A flavonoid derivative, rutin, has been reported to have antimicrobial, antihepatotoxic, antiviral, and anti‐hypertensives effects. The aim of this study was to investigate the effects of rutin, which is known to have a potent antioxidant and anti‐inflammatory effect, on bladder smooth muscle contractility in a cyclophosphamide (CYP)‐induced cystitis rat model.

24 adult female Sprague Dawley rats (200‐250 g) were used throughout experiments. Rats were divided into four groups (1: control, 2: CYP, 3: routine and 4: routine + CYP ).

Bladder smooth muscle strips isolated from groups, were placed in isolated organ bath. Contraction response curves of smooth muscles of the bladder induced by ACh (10‐4M) and KCl (60 mM), and spontaneous contractions of smooth muscles of the bladder obtained.

The amplitüde and frequency of conrtactions were higher in CYP induced cystitis group than control group (p< 0,05). Group 4 (routine + CYP ) has lower amplitüde and frequency than CYP induced cystitis group (p< 0,05).As a result of this study, the spasmolytic effect of the rutin was found on bladder smooth muscle contractility in a cyclophosphamide (CYP)‐induced cystitis rat model.
Aldosterone increases renal expression of TGFβ in a rat model of acute cyclosporine nephrotoxicity (CsA‐Nx) (690.16)
Authors: Ramírez Victoria , Cruz Cristino , Loredo Lilia , Bobadilla Norma
Published: June 2021

Previously, we demonstrated that spironolactone (mineralocorticoid receptor antagonist) prevented renal dysfunction and reduced structural injury (SI) in rats with chronic CsA Nx, suggesting a main role of aldosterone. This nephropathy however, is only induced in the rat when renin angiotensin aldosterone system (RAAS) is activated by a low salt diet. Here, we evaluated the aldosterone contribution in SI induced by CsA in absence of RAAS activation. Forty male Wistar rats were divided into: vehicle, aldosterone infused (Aldo, 0.7mg/Kg), CsA (5mg/Kg sc) and CsA+Aldo groups. All rats were fed with a standard salt diet and followed for 14 days. Mean arterial pressure, renal blood flow, creatinine clearance, were measured, as well as lipid peroxidation as an oxidative stress marker. mRNA and protein levels of TGFβ, pSMAD3/SMAD3, endothelin and endothelin receptors (ETA and ETB) were assessed. Hypoperfusion and renal dysfunction were observed in the three experimental groups, but only in CsA+Aldo group, oxidative stress, endothelin and TGFβ signaling were significantly increased. These results suggest that structural injury induced by CsA is at least in part, mediated by aldosterone in the absence of RAAS activation by a low sodium diet.

Grant Funding Source: National council of science and technology of Mexico grants numbers 155700 to VRG and 181267 to NAB
Expression of candidate normalization gene mRNAs in rat models of fibrosis and diabetes (690.19)
Authors: Glascock Christopher , Horn Todd , Plato Craig
Published: June 2021

Tissue mRNA expression provides mechanistic insight into disease processes and therapeutic interventions, and is often quantitated by normalization to internal “housekeeping” gene expression. Normalization gene mRNA expression is often assumed to vary minimally across different experimental conditions, although emerging evidence suggests otherwise. The current study sought to evaluate candidate normalization gene mRNA expression in healthy and diseased renal cortical tissue from widely‐studied rat models: Unilateral Ureteral Occlusion (UUO), Ischemia‐Reperfusion (IR), 5/6 Nephrectomy (NPX), and Diabetic Nephropathy (ZSF1). We used xMAPTM technology (Affymetrix, Inc.) to evaluate 27 genes: Rplp0, Ppia, Gapdh, Ppib, Tnf, Il6, Actb, Rpl13a, Hprt1, B2m, Sdha, Tgfb1, Tbp, Rpl19, Rps18, Pgk1, Atp6v1a, Ldha, Rpl32, Rps3, Hmbs, Tfrc, Txn2, Polr2a, Rps23, Rps20, and Gusb. Expression data was then subjected to the BestKeeper (www.gene‐quantification.de/bestkeeper.html) and NormFinder (http://moma.dk/normfinder‐software) methods for repeated pair‐wise correlation analysis to identify genes with the least variable expression. Rps23, Rpl32, Ppia were identified by both analyses as being among the 10 most stable expressions, with each being more stable than the often‐employed Hprt1 and Gapdh in NPX and UUO. Fibrotic (Col1a1, Col3a1, CTGF, a‐SMA, FN1) and inflammatory (MCP1, TGFb) biomarker mRNA expression was then compared using Hprt1 and the three new candidate normalization genes in each of the above model systems. Significant changes in fold‐expression of disease tissue mRNA biomarkers were detected as a function of normalization gene. We conclude that normalization genes need to be optimized based on model system studied, and may greatly influence interpretation of mRNA expression.
Attenuation of age‐dependent chronic kidney disease by soy (691.1)
Authors: Grunz‐Borgmann Elizabeth , Nichols LaNita , Valliyodan Babu , Kerley Monty , Fritsche Kevin , Parrish Alan
Published: June 2021

Age is an important predictor of chronic kidney disease (CKD), with a projected incidence of 11% in the adult population of the United States. Common pathways leading to CKD involve renal fibrosis and glomerular disease. Previous studies from our lab show that CKD is detectable in aging male Fisher 344 rats at 16 months of age. Therefore, we fed our rats a 10% low‐fat protein diet containing soy protein or a control diet of casein beginning at 16 months to test whether a dietary intervention of soy, begun after initiation and detection of CKD, would slow progression. The rats were fed the diet for 4 months before sacrifice. The animals on the soy diet had improved kidney function as compared to the casein diet animals, including a decrease in proteinuria, decreased urinary Kim‐1 and improved glomerular filtration rate (GFR). We found that the soy diet significantly reduced several renal fibrosis markers, including total hydroxyproline content and expression of several collagen genes, including collagen, type 1, alpha 1, collagen alpha‐2(I) chain, collagen alpha‐1(III) chain and collagen alpha‐1(IV) chain. Lastly, the soy diet animals also showed a decrease in chemokine (C‐C motif) ligands 5, 17, 19 and 21 (CCL5, CCL17, CCL19 and CCL21), which are linked to the pathogenesis of fibrosis. Taken together, this data shows that even after CKD is detectable a high soy protein diet can significantly attenuate age‐dependent renal dysfunction.
α (E)‐catenin regulates Bmp7 in tubular epithelial cells (691.3)
Authors: Nichols LaNita , Ślusarz Anna , Grunz‐Borgmann Elizabeth , Parrish Alan
Published: June 2021

Acute kidney injury (AKI) most often occurs in a background of chronic kidney disease, both of which increase with age. Several studies have shown increased incidence and severity of AKI in aged patients. Previously, our laboratory has shown that aging is associated with a loss of α‐catenin expression in rat proximal tubular epithelium. To understand the consequences of reduced α‐catenin expression, stable α (E)‐catenin knockdown (C2) and non‐targeted control (NT3) cell lines were generated in the NRK‐52E rat proximal tubular epithelial cell line. C2 cells are marked by a significant decrease in wound repair due to alterations in cell migration. Interestingly, expression of a bone morphogenic protein, Bmp7 is significantly reduced in C2 cells. The loss of Bmp7 expression is also associated with alterations in downstream signaling. In NT3 cells, a strong nuclear staining of phospho‐Smad1/5/8 is seen, while a diffuse cytoplasmic signal is seen in C2 cells. However, shortly after addition of exogenous Bmp7, C2 cells display a similar nuclear localization as control. We then examined the role of Bmp7 in repair. Addition of exogenous Bmp7 to C2 cells rescued wound healing to levels comparable to control NT3 cells. Conversely, a Bmp7 antagonist, LDN193189, inhibits wound healing in NT3. Taken together, these results demonstrate that α‐catenin plays a role in the regulation of Bmp7.

Grant Funding Source: supported by NIH RO1AAG034154
BEA‐induced acute papillary necrosis does not attenuate the enhanced RIHP response to saline volume expansion in rats (692.5)
Authors: Sánchez‐Briones M.Eugenia , López‐Rodríguez J. Francisco , Calvo Turrubiartes Miriam , LLamazares‐Azuara Lilia , Rodríguez‐Martínez Manuel
Published: June 2021

The role of a functional renal papilla in renal interstitial hydrostatic pressure (RIHP) response to saline volume expansion (SVE, 5% bw, ~ 30 min, 0.9%) was assessed in male rats pretreated with either 0.9 % saline (CTR, 200 µl, i.a.) or 2‐bromoethylamine (BEA, 200 mg/kg, i.a.). After 7 days, RIHP (subcapsular, mmHg), renal cortical and medullary blood flows (RCBF/RMBF, L‐D, APU), urinary water (V, ml/ min/ 100 g bw), sodium (UNaV, µEq/ min/100g bw) and nitrogen oxides excretions (UNOxV, Griess, nmol/min/100g), Ht (%) and [Pr]p (g%) were measured in anesthetized animals with hormonal clamp, kidney denervation and renal perfusion pressure control at 100 mmHg (CTR: n = 9, bw = 345 ± 7 g, Uosm = 1940 ± 187 mOSM vs. BEA: n = 12, bw = 303 ± 9α g, Uosm = 371 ± 43α mOSM) during hydropenia (H), SVE, and maintained SVE (mSVE). The results are mean ± SEM. α p<0.05 between groups in the same period, * p < 0.05 vs. H in the same group.

CTR

BEA

H
SVE
mSVE
H
SVE
mSVE

Ht
51.3±0.4
45.0±*0.9
44.2±*0.8
52.6±0.7
44.0±*1.0
44.0±*1.2

[Pr]p
5.1±0.2
3.2±*0.1
2.8±*0.2
5.2±0.2
3.3±*0.2
3.2±*0.2

RIHP
4.2±0.3
27.2±*4.3
26.0±*4.8
4.1±0.6
26.0±*1.5
26.0±*2.2

UNOxV
0.46±0.1
1.8±*0.4
2.7±*0.6
0.44±0.1
1.8±*0.5
2.6±*0.7

RMBF
431±89
464±101
473±95
178±Î±54
249±Î±54
283±Î±65

UNaV
0.019±0.005
1.07±*0.16
2.53±*0.42
0.20±Î±0.06
0.86±*0.12
1.56±Î±*0.16

Conclusions: In rats, a nonfunctional renal papilla does not attenuate enhanced RIHP, UNOxV and diuretic responses to SVE, though decreases ~35% the late natriuretic response to SVE. Apparently, RMBF is a necessary but not sufficient factor for RIHP response to SVE.

Grant Funding Source: Supported by IPL's financial resources
Oxidative stress and vascular reactivity changes in thoracic and abdominal aorta expose at the hyperbaric oxygenation (693.1)
Authors: Acevedo‐Villavicencio Nallely , Kormanovsky Alexander , Villanueva‐Lopez Cleva , Ramirez‐Sanchez Israel , Lara‐Padilla Eleazar , Guevara‐Balcazar Gustavo , Castillo‐Hernandez Maria
Published: June 2021

During the hyperbaric oxygenation therapy (HBO) has been described some changes like hypoglycemia, low blood pressure, etc. The mean idea of this work is to analyze if the HBO induce some changes in the vascular reactivity and oxidative stress in healthy rats. Methods: Male rats were used (3 months old) and divided in 2 groups: with and without HBO.Rats were exposed to pure oxygen at 2 absolute atmospheres during 60 min for 3, 5, 10 and 20 sessions, after that, we obtain thoracic and abdominal aorta and were used some of the aortic rings for the isolated organ system where we made concentration response curves; and other aortic rings were used to analyze the oxidative stress. Results: changes were observed in the contractile effect to phenyleprine in both segments after HBO, without any changes in the relaxant response. On the other hand thoracic oxidative response were higher after 3 sessions of HBO, but in an interesting way we can observe that antioxidant enzyme decreased after this time; those results suggest that the antioxidant system increase the response trying to metabolize O2.

Grant Funding Source: COFAA / IPN
Bestrophin‐3 expression in mouse glomeruli (694.1)
Authors: Golubinskaya Veronika , Elvin Johannes , Ebefors Kerstin , Gustafsson Helena , Mallard Carina , Nyström Jenny , Nilsson Holger
Published: June 2021

The bestrophin isoform, bestrophin‐3 (Best3), acts as a calcium‐activated chloride channel in cardiomyocytes and in vascular smooth muscle. It may have cell‐protective functions in other cell types. Best3 may be regulated by cyclic GMP and exists in different splice variants.

We have examined expression and localization of Best3 in mouse kidney on the mRNA level using RT‐PCR and on the protein level by immunohistochemistry and Western blotting.

Staining for Best3 was obtained in mouse glomeruli, and was also detected in cultured mouse podocytes. Western blot confirmed the presence of Best3 protein in kidney cortex. Best3 did not co‐localize with markers for endothelial cells or podocyte foot processes. However, it co‐localized with the intermediate filament nestin, suggesting localization of Best3 in podocyte primary processes and cell bodies. In kidney cortex, in glomerulus‐enriched fractions and in cultured podocytes the two shortest splice variants, but no full‐length mRNA, were detected.

In conclusion, two variants of Best3, lacking some of the predicted transmembrane segments, are expressed in podocytes in mouse kidney, and are likely co‐localized with the intermediate filaments.
Potential role of TRPV4 channels in angiotensin II‐induced endothelial dysfunction (696.2)
Authors: Nishijima Yoshinori , Zheng Xiaodong , Lund Hayley , Mattson David , Zhang David
Published: June 2021

Endothelial TRPV4 is a Ca2+ entry channel contributing to endothelium‐mediated dilation in conduit and resistance arteries. In previous studies, we found that chronic infusion of angiotensin II (AII) reduces endothelium‐mediated dilation in mesenteric arteries (MAs) from wild‐type (WT) but not TRPV4‐KO mice. Here we investigated whether AII impairs endothelium‐dependent dilation by reducing TRPV4 expression or function. TRPV4 protein expression was examined in MAs from AII‐treated WT mice and human coronary artery endothelial cells (HCAECs). AII (600 ng/kg/min for 14 days) did not significantly affect total protein expression of TRPV4 in MAs. In HCAECs, AII (500 nM for 24h) did not alter either total or cell surface TRPV4 expression. We next examined whether AII‐induced endothelial dysfunction is associated with reduced TRPV4 function. HCAECs were incubated with AII for 24h, and TRPV4 agonist‐induced Ca2+ response was examined by a fura‐2 assay. 4α‐PDD‐induced [Ca2+]i increase was markedly reduced after treatment of AII (F340/F380, 1.0±0.1 vs. 1.9±0.2 of control). In conclusion, AII does not significantly alter total protein expression of TRPV4 in MAs or cell surface localization of TRPV4 in HCAECs, but attenuates TRPV4‐mediated Ca2+ response in HCAECs. These results indicate that reduced TRPV4‐mediated Ca2+ signaling may be a potential mechanism underlying AII‐induced endothelial dysfunction.
Effects of allopregnanolone, a positive allosteric modulation of GABAA receptors upon stress and hypertension in Schlager genetic hypertensive mice (697.3)
Authors: Head Geoffrey , Stevenson Emily , Jackson Kristy , Abegaz Beza , Moretti John‐Luis , Gueguen Cindy , Davern Pamela
Published: June 2021

Background: Hypertensive Schlager mice (BPH/2J) have neurogenic hypertension associated with differences in GABAA receptors compared to their normotensive counterparts (BPN/3J). Allopregnanolone is an endogenous neurosteroid reduced by chronic stress and when administered, decreases anxiety by positive allosteric modulation of GABAA receptors.

Aim: To determine if allopregnanolone reduces the pressor effects of stress and basal MAP in BPH/2J mice.

Methods: Male BPN/3J (n=7) and BPH/2J (n=5) mice received vehicle or allopregnanolone (5µg/hour) via subcutaneous minipumps for 2 weeks. Implanted telemetric probes enabled recording of mean arterial pressure (MAP), heart rate (HR) and activity as well as the response to aversive and non‐aversive stress tests and ganglionic blockade with pentolinium before and 7 and 14 days of treatment. Mice were perfused following stress and brains were removed for immunohistochemistry.

Results: Allopregnanolone reduced systolic arterial pressure (‐8.8mmHg, P=0.01) and attenuated the depressor response to pentolinium in BPH/2J mice but had no effect upon in BPN/3J. Allopregnanolone produced marked reductions in the pressor response to both cage switch and feeding stress (‐20%, P<0.01) in BPH/2J whilst increasing the pressor response to aversive stress in BPN/3J (P<0.001). Stress induced Fos counts within the medial amygdala and paraventricular nucleus were higher in untreated BPH/2J compared to BPN/3J. Allopregnanolone reduced Fos expression and treatment abolished the difference between strains.

Conclusions: The selective antihypertensive and stress inhibitory effects of allopregnanolone in BPH/2J hypertensive mice suggests that allosteric modulation of GABAA receptors at the level of the hypothalamus and amygdala may be a major cause of hypertension in this model and may offer a possible new area for the development of therapy.
Genetic control of heart weight in mouse and man (697.4)
Authors: Zhao Wenyuan , Lu Lu , Williams Robert , Chen Yuanjian , Meng Weixin , Sun Yao
Published: June 2021

Heart weight (HW) serves not only measure of cardiac structure and function, and also as an intermediate phenotype for cardiovascular disease outcome. In humans HW is a heritable trait and has been linked to several genetic loci. Genome‐wide association study (GWAS) have identified single‐nucleotide polymorphisms in and near genes that may contribute to variability in HW. The BXD family of recombinant inbred strains comprise a large panel of isogenic but diverse strain derived by crossing C57BL/6J and DBA/2J. This genetic reference panel is a remarkable resource because genotype data (high density SNP markers) is available in GeneNetwork database; and the genomes of both parents have been sequenced. Our goal is to define genetic loci and candidate genes that control HW. There is significant variation in HW among the BXD strains_from 81 to 182 mg. A locus in chromosome 3 is responsive for a significant fraction (70%) of HW variation. The expression of candidate genes in this locus was studied using whole transcriptome analysis and qPCR for the parent strains. Morphometric measurement of the parent strains was used to evaluate the causes of HW variation. Candidate genes were compared with genes identified in GWAS. Several candidate genes in our study related with candidate genes identified in human GWAS. The best candidate genes based on this synthesis of human and mouse data include KCND3, KCNA3, SLC16A4, DDX20, and OVGP1.

Grant Funding Source: NIH
Nicotinamide mononucleotide supplementation reverses vascular endothelial dysfunction and large elastic artery stiffness in old mice (698.10)
Authors: de Picciotto Natalie , Gano Lindsey , Johnson Lawrence , Eng Jason , Weber Stewart , Imai Shin‐ichiro , Mills Kathryn , Seals Douglas , Sindler Amy
Published: June 2021

Reduced bioavailability of nicotinamide adenine dinucleotide (NAD+), a cofactor for the deacetylase sirtuin1 (SIRT1), may contribute to age‐related vascular dysfunction via oxidative stress and reduced nitric oxide (NO). We hypothesized that boosting NAD+ with the precursor nicotinamide mononucleotide (NMN) might reverse these adverse effects of aging. Old (O: 26 mo, n=14) C57 male mice had impaired ex vivo carotid artery endothelium‐dependent dilation (EDD) to acetylcholine (ACh, 60 ± 5 vs. 82 ± 3%) and NO‐mediated EDD (max dilation to ACh ‐ max dilation to ACh + L‐NAME, 37 ± 4 vs. 69 ± 5%), and increased aortic pulse wave velocity (aPWV, 467 ± 18 vs. 359 ± 8 cm/s) and elastic modulus (EM, 5899 ± 731 vs. 3307 ± 288 kPa, all p<0.05) (indices of arterial stiffness) vs. young (Y; 6 mo, n=12). In O treated with NMN (300 mg/kg/d in drinking water, 8 w) (n=10), EDD (89 ± 3%), NO‐mediated EDD (61 ± 5%), aPWV (364 ± 14 cm/s) and EM (3694 ± 315 kPa) all were improved (p<0.05 vs. O). Aorta from O had reduced expression (WB) of SIRT1 (0.7 ± 0.1 vs. 1.0 ± 0.1 AU, p=0.05 vs. Y) and increased nitrotyrosine (NT) (oxidant modification) (8.7 ± 1.1 vs. 1.0 ± 0.3 AU, p<0.05 vs. Y). In O, NMN restored SIRT1 (1.1 ± 0.1 AU) and reduced NT (4.7 ± 0.7 AU) (both p<0.05 vs. O). Increasing NAD+ bioavailability may be a novel therapy for reversing age‐related vascular dysfunction by reducing oxidative stress and restoring NO signaling.

Grant Funding Source: Supported by NIH AG013038 and AG000279
Blood pressure lowering effects of the CFTR‐F508del mutation are associated with reduced arterial reactivity rather than circulating blood volume (700.1)
Authors: Peotta Veronica , Bhandary Prasad , Ogu Ugochi , Volk Kenneth , Roghair Robert
Published: June 2021

CFTR‐F508del mutation is the most common cause of cystic fibrosis. F508del homozygous patients and female F508del carriers have decreased blood pressure (BP). We tested the hypothesis that heterozygote F508del mice have lower BP and reduced aortic contractility. By radiotelemetry, female F508del carriers had significantly lower systolic BP than wild‐type C57 controls (112±1 versus 119±2 mmHg). Although male carriers did not have lower baseline BP, they had significantly lower diastolic pressures after intraperitoneal co‐injection of isoproterenol + epinephrine (60±3 versus 78±8 mmHg). Aortas isolated from male and female F508del carriers had reduced constriction to noradrenaline (0.9±0.2 versus 2.9±0.7 mN). Inhibition of wild‐type CFTR or the inositol triphosphate receptor replicated the phenotype of F508del aortas. CFTR carrier status did not alter circulating blood volume. We conclude the CFTR‐F508del mutation decreases aortic contractility and arterial pressures. As a cAMP‐activated chloride channel that facilitates calcium mobilization, we speculate wild‐type CFTR co‐activation during beta‐adrenergic receptor stimulation buffers the vasodilatory response to catecholamines, and loss of this compensatory vasoconstrictor tone may contribute to the lower BP seen in heterozygote carriers of a CFTR‐F508del mutation.

Grant Funding Source: Supported by NIH R01 HL102659
Glucagon‐like peptide‐1 receptor agonist, exenatide, enhances Ca2+ buffering by SERCA in coronary smooth muscle cells from lean, healthy Ossabaw swine (700.3)
Authors: Dineen Stacey , McKenney Mikaela , Sturek Michael
Published: June 2021

The sarco‐endoplasmic reticulum Ca2+ ATPase (SERCA) maintains optimal SR/ER Ca2+ to avoid ER stress. Exenatide may provide cardioprotection by enhancing SERCA activity, thereby preventing ER stress. Coronary smooth muscle cells were enzymatically isolated from left anterior descending artery of lean, healthy Ossabaw swine. Cytosolic Ca2+ was digitally imaged using the fluorescent Ca2+ indicator, fura‐2. The SR store was released with caffeine in Ca2+‐free physiologic salt solution (PSS). Following caffeine, cells recovered in Ca2+‐free PSS. Cells treated with exenatide demonstrated significantly enhanced Ca2+ buffering ability as shown by the undershoot of the Ca2+ below baseline upon recovery from caffeine; ΔF0/380=‐0.054±0.005 vs. ‐0.03±0.005 (p<0.005) in untreated cells. The role of SERCA was confirmed with blockage by cyclopiazonic acid, which significantly reversed the effect of exenatide. SERCA function was further elucidated by examination of the time required for buffering of Ca2+ to baseline levels. Cells treated with exenatide demonstrated more rapid Ca2+ buffering to 100% recovery to baseline levels: t100=1.7±0.06 min vs. untreated cells, t100=1.9±0.05 min (p<0.005) but not to 50% recovery. The lack of exenatide effect on t50 shows no action on Ca2+ extrusion mechanisms; instead, the predominant action of exenatide is stimulation of SERCA.

Grant Funding Source: Supported by NIH HL062552, NIH/NCATS CTSI TL1 TR000162
Uterine artery smooth muscle contractions are mediated by PKC and NO signaling mechanisms in diet‐induced obese pregnant rats (700.8)
Authors: Goulopoulou Styliani , Hannan Johanna , Webb R. Clinton
Published: June 2021

Pre‐existing obesity increases the risk of maternal and fetal complications during gestation and is associated with systemic vascular dysfunction. We hypothesized that diet‐induced obesity would impair uterine artery contractile responses in pregnant rats. Female rats were fed for 3 weeks prior to pregnancy and throughout gestation either normal chow (LN‐Preg, n=8) or normal chow plus lard and liquid sucrose (OB‐Preg, n=8). Contractile responses to phenylephrine (PE, 10‐8 – 3x10‐5 M) were measured in isolated uterine arterial segments in late pregnancy, in the presence and absence of inhibitors of ERK1/2 (PD98059, 10‐5 M), Rho kinase (ROCK, Y27632: 10‐6 M), protein kinase C (PKC, BIM: 10‐6 M) and nitric oxide synthase (NOS, L‐NAME: 10‐4 M). OB‐Preg uterine arteries had reduced sensitivity to PE compared to LN‐Preg arteries. Inhibition of PKC reduced sensitivity to PE in OB‐Preg but not in LN‐Preg uterine arteries (‐logEC50, OB‐Preg, PE: 5.75 ± 0.087 vs. PE+BIM: 5.35 ± 0.128, p<0.05; LN‐Preg, PE: 6.07 ± 0.086, PE+BIM: 5.99 ± 0.119, p>0.05), whereas inhibition of ERK1/2 or ROCK reduced sensitivity to PE in both groups. NOS inhibition increased contractile responses to PE in OB‐Preg arteries to a greater extend as compared to LN‐Preg vessels and abolished the differences between groups (‐logEC50 in the presence of L‐NAME, OB‐Preg, 6.19 ± 0.103 vs. LN‐Preg, 6.19 ± 0.079, p>0.05). Pre‐existing obesity reduces uterine artery contractile responses to α1 adrenergic stimulation in late pregnancy and this response is mediated by PKC and NO signaling mechanisms.

Grant Funding Source: Supported by NIH, SWHR
Improvement of body‐fluid dysfunction by Poria cocos in nephrotic syndrome (701.3)
Authors: Lee So Min , Lee Yun Jung , Yoon Jung Joo , Han Byung Hyuk , Kang Dae Gill , Lee Ho Sub
Published: June 2021

Improvement of body‐fluid dysfunction by Poria cocos in nephrotic syndrome

So Min Lee 1,2, Yun Jung Lee1,2, Jung Joo Yoon1,2, Byung Hyuk Han1,2, Dae Gill Kang1,2, and Ho Sub Lee1,2

1College of Oriental Medicine and Professional Graduate School of Oriental Medicine, Wonkwang University, Iksan, Jeonbuk 570‐749, Republic of Korea; 2 Hanbang Body‐fluid Research Center, Wonkwang University, Shinyong‐dong, Iksan, Jeonbuk, 570‐749, Republic of Korea

Abstract Nephrotic syndrome is associated with altered renal handling of water and sodium along with changes of aquaporins (AQP) and renal epithelial Na channel (ENaC).

The dried sclerotia of Poria cocos Wolf (WPC) has been used for the treatment of chronic edema and nephrosis. This study was conducted to evaluate the effects of WPC on puromycin aminonucleoside (PAN)‐induced renal functional derangement and the change of renal AQP2 and ENaC expression. The nephrotic syndrome animal models were constructed by PAN 75 mg/kg injection and then were treated with losartan (30 mg/kg/day) or WPC (200 mg/kg/day) for 7 days. Consequently, WPC group was significantly improved the proteinuria and ascites. In addition, the plasma level of triglyceride, total cholesterol, and low density lipoprotein (LDL)‐cholesterol were significantly decreased in WPC. Besides, WPC group attenuated the PAN‐induced increase in protein, and mRNA levels of AQP2 and ENaC subunit α/β. WPC was significantly suppressed PAN‐induced organic osmolytes regulator such as serum‐ and glucocorticoid‐inducible protein kinase (Sgk1), and sodium‐myo‐inositol cotransporter (SMIT) mRNA expression.

Taken together, WPC improve nephrotic syndrome including proteinuria and ascites, through inhibition of AQP2 and ENaC subunit. Thus, WPC is involved in the body‐fluid regulation via inhibition of water and sodium channels against renal disorder such as edema or nephrosis.
Selected laboratory parameters and seric PRR correlation as prognostic factors in patients with preeclampsia (701.6)
Authors: Lopez‐Islas Ines , Vasquez‐Vasquez Juan Ernesto , Sanchez‐Bautista Sergio Alejandro , Garcia‐Canales Anyel Angelica , Lopez‐Sanchez Pedro
Published: June 2021

Preeclampsia (PE) shows high maternal‐fetal morbidity. Its pathophysiology is multifactorial. Prorrenin/renin receptor (PRR), a new component of the renin‐angiotensin system, has been implicated in hypertensive disorders. Our goal was to determine if seric PRR correlates with selected laboratory parameters as prognostic factors in PE. After ethics committee approval, we selected term pregnant patients with PE from the intensive care unit (n=8), and normal pregnant patients (n= 15). Pregnancy was interrupted within 24 hrs by cesarean incision. Blood samples were collected and analyzed for cholesterol, triglycerides, albumin, calcium, uric acid (UA), Mg, lactate and C‐reactive protein before and after pregnancy termination. Body mass index (BMI) was calculated at the beginning and end of pregnancy. Our results showed that PE patients had elevated serum cholesterol, triglycerides, C‐reactive protein and hypoalbuminemia before and after delivery. BMI, UA and Mg were significantly higher and all PE patients showed significant thrombocytopenia even 24 h after cesarean. Placental weight was significantly lower. Seric PRR was higher in PE patients and positively correlated with albumin, calcium and Mg. These results suggest a close correlation of PRR with several markers and might be used as prognosis factors during PE

Grant Funding Source: Supported by SIP20130546
Effects of genetic strain on stress‐induced weight and body fat loss in rats: application to air pollution research (703.3)
Authors: Johnstone Andrew , Phillips Pamela , Jarema Kimberly , Dheyongera Geoffrey , Gordon Christopher
Published: June 2021

Exposure to some air pollutants is suspected of contributing to obesity. Hazelton chambers are commonly used in air pollution studies but we found unexpected reductions in body weight and body fat of rats housed in Hazelton chambers under control conditions. We suspect that stress from housing in Hazelton chambers could compromise data from a study on air pollutant‐induced obesity. Male Long Evan (LE), Sprague Dawley (SD), Brown Norway (BN) and Fisher (F344) rats were housed individually in Hazelton chambers for 6 h, 1 day/wk for 3 wks. Using a Bruker NMR system, % body fat was measured before and after exposure and compared to cage controls. Housing in the chambers led to strain‐dependent decreases in %body weight and body fat. Weight loss was greatest in LE and SD (4.4+0.3% and 4.0+0.2%, respectively, on wk 1). BN and F344 were less affected (3.2+0.2% and 3.8+0.3%, respectively, on wk 1). Body fat was reduced by 0.6% in BN and LE rats at 5 d after exposure in the Hazelton Chambers. Overall, housing in Hazelton chambers appears to be a stressor leading to significant weight loss and lipolysis that is strain‐dependent. BN and LE rats may not be the best model to use to study possible links of air pollutants to obesity. This abstract does not necessarily reflect US EPA policy.
Determinants of muscle protein synthesis in severe pediatric burns (703.5)
Authors: Diaz Fuentes Eva , Potter Craig , Cotter Matthew , Lee Jinhyung , Herndon David , Borsheim Elisabet
Published: June 2021

Introduction: Following a major burn, a large increase in skeletal muscle protein breakdown is seen. The simultaneous increase in muscle protein synthesis cannot completely compensate for the breakdown, resulting in net protein loss. The aim of this study was to identify the determinants of skeletal muscle protein fractional synthetic rate (FSR) in severely burned children. Methods: 87 patients with 蠅40% Total Body Surface Area (TBSA) burns were included. Patients participated in stable isotope infusion studies during their acute hospitalization and at 6, 12 and 24 months post injury in order to determine skeletal muscle FSR. Generalized estimating equations with log link normal distribution were applied to account for clustering of patients and control for patient characteristics. Results: Patients (8±6 yr) had large (59±14 % TBSA), deep (47±21% 3rd degree) burns. Time was a significant determinant, with a decrease in FSR [%/h, median (Q1‐Q3)] at 12 months vs. admission [0.08 (0.05‐0.18)vs. 0.12 (0.07‐.0.24)] (P<0.05). Further, FSR was 48% higher in childrenwith co‐existent inhalation injury vs. those with burn injury alone. Inaddition, FSR was 36% lower in male vs female kids, while children >3 years had 30% lower FSR values than younger children. Finally, FSR was significantly lower in patients with burns 蠅60% TBSA. Conclusion: The current data demonstrate that burn size, gender; the presence of inhalation injury and time all affect the rate of skeletal muscle FSR following burn injury. The impact of these findings on net protein balance remains unknown and warrants further investigation.

Grant Funding Source: Supported by NIH P50‐GM60338, RO1‐GM056687, T32‐GM008256, and Shriners grants 84090, 84080, 71006/08
Evaluation of long‐haul shipping stress for beef calves transported from Hawaii to the contiguous United States (703.7)
Authors: Sy Tracy , Richards Tim , Lee C. , Onaga Jonathan , Coetzee Johann , Stock Matt , Stokes Ashley
Published: June 2021

Objectives of this study were to examine physiological parameters of weaned calves being shipped to determine response to long‐haul shipping; also, to determine the animals’ ability to adapt to the stress of shipment. Two shipments of weaned, Angus and Charolaise, female calves (n=48) were transported to Washington or California. Each shipment, the calves were divided and placed into two separate, bi‐level, four‐compartment, custom cowtainers. Vaginal temperatures (T°) were taken every minute. Blood was collected prior to shipment (P), upon arrival (A), and 6 days post‐arrival (PA). Blood work parameters (including substance‐P (SP), hematocrit % (H), bands (B), and white blood cell (WBC) numbers and differentials) were also used to determine physiological status of the animals. Overall T° values did not change significantly. T° only increased above normal range during insertion of temperature sensor, loading, unloading, trucking, and at Kawaihae, Hawaii. SP values showed a significant increase from P to A (p<0.001) and decrease back to normal from A to PA (p<0.001). SP values had no significant change from P to PA (p=0.45). WBC and B showed no significant differences for any time points. HCT significantly decreased from P to A and from P to PA (p<0.001). HCT values stayed within normal range for all stages. The study showed some physiological changes with shipping, however, animals quickly returned to a homeostatic physiological state.

Grant Funding Source: Supported by National Cattlemen's Beef Association
FNDC5 expression in skeletal muscle in chronic heart failure: relevance of inflammatory cytokines (704.1)
Authors: Adams Volker , Gleitsmann Konstanze , Mangner Norman , Bowen T Scott , Schuler Gerhard , Linke Axel
Published: June 2021

Chronic heart failure (CHF) is associated with muscle atrophy. Irisin, a myokine proteolytically processed by the FNDC5 protein and suggested to be PGC‐1α activated, modulates the browning of adipocytes and is related to muscle mass. Therefore, we investigated whether skeletal muscle FNDC5 expression in CHF was reduced and if this was mediated by inflammatory cytokines and/or angiotensin II (Ang‐II).

Methods: Skeletal muscle FNDC5 mRNA/protein and PGC‐1α mRNA expression were analyzed in: Rats with CHF; Mice injected with TNF‐α (24h); Mice infused with Ang‐II (4 wk); C2C12 myotubes exposed to recombinant cytokines or Ang‐II. Results: CHF reduced (p<0.05) FNDC5 protein (1.3±0.2 vs. 0.5±0.1 arb. units) and PGC‐1α mRNA expression (8.2±1.5 vs. 4.7±0.7 arb. units) compared to control. TNF‐α and Ang‐II reduced FNDC5 protein by 28% and 45%, respectively. Incubation of myotubes for 24h with a TNF‐α, IL‐1ß, and IFN‐ γ cocktail reduced (p<0.05) FNDC5 protein expression by 71%, whereas Ang‐II had no effect. PGC‐1α was linearly correlated to FNDC5 in all conditions.

Conclusion: Skeletal muscle FNDC5 expression is reduced in CHF, which seems mediated by inflammatory cytokines, Ang‐II, and PGC‐1α.

This suggests a reduced FNDC5 expression may act as a protective mechanism in catabolic states, by preserving energy homeostasis via slowing the browning of adipocytes.
Systemic and lower limb hemodynamics following dynamic exercise in type 2 diabetics: effects of exercise intensity and duration (705.12)
Authors: Pellinger Thomas , Neighbors Catherine , Snitzer Jack
Published: June 2021

For individuals with type 2 diabetes mellitus (T2DM), regular exercise is critical for protecting vascular function and reducing cardiovascular disease risk, but the hemodynamic responses to differing doses of acute exercise in type 2 diabetics (T2D) are unclear. Our aim was to compare postexercise (PE) hemodynamics in T2D in response to four different doses of acute dynamic exercise. Two subjects with well‐controlled T2DM (42‐48 yrs; Hemoglobin A1c: 6.6 ± 0.7%) participated in four study days, during which they exercised on an upright cycle ergometer at four different combinations of exercise duration and intensity: 30 min at 40% VO2 peak (30@40), 30 min at 60% VO2 peak (30@60), 60 min at 40% VO2 peak (60@40), and 60 min at 60% VO2 peak (60@60). Heart rate, arterial pressure, and femoral blood flow (Doppler ultrasound) were measured pre‐exercise and every 15 min through 120 min PE. Femoral vascular conductance (FVC) was calculated as flow/pressure. Compared to pre‐exercise baseline, FVC was higher and mean arterial pressure (MAP) was lower throughout 120 min of recovery in all conditions, except for the 30@40 trial. Expressed as percentage change from pre‐exercise baseline, at 30 min PE, FVC was elevated 7.8 ± 1.3% on the 30@40 day, 32.3 ± 1.9% on the 30@60 day, 40.7 ± 4.6% on the 60@40 day, and 43.9 ± 9.0% on the 60@60 day. Compared to pre‐exercise baseline, at 30 min PE, MAP was reduced 0.53 ± 0.6% on the 30@40 day, 2.1 ± 2.9% on the 30@60 day, 4.2 ± 0.2% on the 60@40 day, and 6.1 ± 0.6% on the 60@60 day. These preliminary results suggest a somewhat dose‐dependent elevation of PE skeletal muscle blood flow and reduction in PE MAP in T2D.

Grant Funding Source: Supported by the Salisbury University Faculty Mini‐Grant
The Ironman Triathlon shifts innate/adaptive immune cell populations and increases systemic inflammation (705.3)
Authors: Munoz Colleen , Apicella Jenna , Radom‐Aizik Shlomit , Maresh Carl , Haddad Faddia , Cooper Dan , Nugas Cherryl , Zaldivar Frank , Casa Douglas , Lee Elaine
Published: June 2021

The Ironman Triathlon shifts innate/adaptive immune cell populations and increases systemic inflammation Colleen X. Munoz1, Jenna M. Apicella1, Shlomit Radom‐Aizik2, Carl M. Maresh1, Fadia Haddad2, Dan M Cooper2, Cherryl Nugas2, Frank P Zaldivar2, Douglas J. Casa1, Elaine C. Lee1 1University of Connecticut, Storrs, CT2University of California Irvine, Irvine, CA Exercise and environmental stressors threaten cellular function and integrity. We recently showed that whole body acute stress determines the extent of measurable cellular stress. In this study, we tested the hypothesis that the profile of circulating immune cells, level of systemic inflammation, and chaperone response would reflect the whole body stress of an Ironman. To test this hypothesis, we recruited 40 race participants (29 males, 11 females) and 17 control subjects (9 males, 8 females). Blood was drawn between 2‐3 days before the triathlon (baseline), and during recovery at 1‐, 24‐, and 48‐h after completion. CBC (with differential), multiplex cytometric bead array, and ELISA were used to determine cell counts, inflammatory cytokines, and chaperone HSP70, respectively. Statistical significance was defined by repeated measure ANOVA with Bonferroni‐corrected post hoc tests (p<0.05). Total white blood cell counts increased post‐exercise and returned to normal by 48h post‐race. Neutrophil and monocyte subsets increased post‐exercise and remained elevated even 2 days post‐race, while lymphocyte, eosinophil, and basophil subsets transiently decreased post‐exercise before increasing above normal 24h and 48h later. Inflammatory cytokines IL‐10, IL‐6, and IL‐8 increased >10 fold post‐race and returned to normal by 24h post‐race. HSP70 similarly increased ~2‐fold post‐ race and returned to normal levels by 24h later. The stress of the Ironman event likely shifted the balance of innate/adaptive immune cells, and induced a systemic inflammatory response and chaperone response.

Grant Funding Source: Timex, Inc.
Strain‐dependent variation in transcriptional activation of metabolic and angiogenic genes after acute exercise in mouse skeletal muscle (705.8)
Authors: Kim Seung , Avila Joshua , Massett Michael
Published: June 2021

Repeated bouts of exercise can lead to metabolic and vascular adaptation in skeletal muscle, and the magnitude of the adaptation is determined, in part, by genetic background. The aim of this study was to test the hypothesis that the transcriptional response to an acute bout of exercise would be different between high and low responders to training. Gene expression for seven metabolic and nine angiogenic genes was assessed in plantaris muscles from FVB/NJ (FVB, high responder) and C57BL/6J (B6, low responder), which were harvested before and 0, 3, 6, 12, and 24 h (n=4/each) after an acute bout of exercise on a treadmill (60 min, 18m/min, 10° incline). Significant strain effects were found for mRNA expression of several metabolic and angiogenic genes. The peak of PGC1α mRNA was found at 3 h (p < 0.01) in both strains, showing greater increase (36 %) in FVB than B6. Nrf2 mRNA was increased at 0 h in FVB and 3 h in B6. Nrf1 mRNA was decreased at 3‐24 h in FVB. Interestingly MyoG mRNA changed in opposite directions (P < 0.05) for B6 and FVB; B6 decreased at 6‐24 h and FVB increased at 3‐6 h. VEGFa and eNOS mRNAs were increased at 0 h only in FVB. Unexpectedly both B6 and FVB showed increased TSP1 and reduced HIF1α mRNA at 0 h. These data indicate that genetic background influences the transcriptional activation of metabolic and angiogenic genes in response to an acute bout of exercise in mouse skeletal muscle.

Grant Funding Source: Supported by NIH grant HL085918 to MPM
Combined exercise training improves cardiovascular and inflammatory parameters in an experimental model of metabolic syndrome and menopause (706.12)
Authors: Conti Filipe , Brito Janaina , Dias Danielle , Bernardes Nathalia , Sanches Iris , Malfitano Christiane , Irigoyen Maria Claudia , De Angelis Kátia
Published: June 2021

The aim of this study was to verify the effects of combined exercise training (aerobic+resistance) in an experimental model of metabolic syndrome and menopause on cardiovascular autonomic modulation and inflammation. Female SHR rats were divided into (n=8/group): hypertensive (H), hypertensive ovariectomized (HO) and hypertensive ovariectomized submitted to fructose overload sedentary (100g/L in drinking water) (FHO) or trained (FHOT). Combined exercise training was performed in ladder+treadmill on alternate days (5x/week, 8 weeks, 40‐60% maximal exercise test). Arterial pressure (AP) signals were directly recorded. Cardiovascular autonomic modulation was evaluated by spectral analysis. Cardiac levels of TNFα and IL‐10 were measured by Elisa. AP was higher in FHO group (174±4 vs. H: 146±4 HO: 164±5 mmHg) and combined exercise training attenuated this dysfunction (FHOT: 158±4 mmHg). FHOT group showed resting bradycardia. The vascular sympathetic modulation (LF‐IP) was higher in FHO group (10.62±2.33 mmHg2) and exercise training normalized this change (4.51±0.32 vs. H: 5.06±0.91; HO: 5.07±0.52 mmHg2). The TNFα was higher in FHO group than H and HO groups and exercise training reduced this parameter. Negative correlation was observed between adiposity tissue and IL‐10 among sedentary groups (r=‐0.6, p<0.05). In conclusion, fructose overload induced impairment in cardiovascular autonomic control and in inflammatory parameters in hypertensive rats undergoing ovarian hormones deprivation. The combined exercise training reverted and/or attenuated these impairments.

Grant Funding Source: FAPESP 2011/15828‐9
Effects of impact‐loaded sport training on bone bending stiffness and BMD in young females (706.16)
Authors: Liang Michael , Robles Patricia , Rocha Jose , Spalding Thomas
Published: June 2021

BLACKGROUND: Participation in high‐impact loading or odd‐impact loading sports is associated with higher bone mineral density (BMD) and bone mass. OBJECTIVE: The objective of this study is to compare the impact of sport training in long‐distance running, soccer and volleyball practice on BMD and bone bending strength in female athletes, 18‐24 years old.

METHODS. 57 trained female athletes participated in this study. They were 22 soccer players (SOC), 22 long‐distance runners (RUN), and 13 volleyball players (VOL). Their ages were 19.1, 20.5 and 21.5 yr for the SOC, RUN and VOL, respectively. Sport training history was: 13, 11 and 10 yr for the SOC, RUN and VOL, respectively. Mean height, weight and BMI were 163 cm (±4.7), 59.8 kg (±4.1) and BMI 22.6 kg/m2 (±1.9), respectively. We also recruited 23 non‐athlete females, age 22.6 ± 0.5 yr, as control subjects. Bone bending stiffness was measured using a Mechanical Response Tissue Analyzer (MRTA, NASA‐ARC, CA), an in vivo technique expressed as EI in Nm2. EI was measured at the ulna and tibia. BMD was measured using a Duel‐Energy X‐ray Absorptiometry (PIXI‐DXA, in g/cm2) at two skeletal sites: heel and wrist.

RESULTS. There are no differences in height, weight and BMI between the groups, except that VOL were significantly heavier and taller than the other groups. Tibia EI (TEI) of the RUN (122.7±89 Nm2) were significantly lower (p<0.05) than VOL (228±89 Nm2), but not SOC (152±89 Nm2) and CON (85±89 Nm2). RUN has the lower ulnar EI (UEI=30.7±14 Nm2) relative to SOC (44±14 Nm2) and VOL (44±14 Nm2). VOL has significantly higher TEI (228.3±89 Nm2, p<0.05) compared with SOC, RUN and CON. Heel and wrist BMD of the SOC and VOL are 7% to 9.5% higher (p<0.05) than the RUN and CON.

CONCLUSION. Volleyball training exhibited greatest osteogenic adaptation in tibial EI compared with RUN and CON, but not in ulnar EI or heel and wrist BMD. Distance running did not induce high osteogenic adaptation in the tibial EI or heel BMD. The mechanism for these changes in bone EI and BMD is unknown.
microRNA‐206 modulates important regulators of gene expression in response to aerobic exercise training in human skeletal muscle (706.3)
Authors: Alves Cleber , Junior José , Fernandes Tiago , Alves Guilherme , Negrão Carlos , Oliveira Edilamar
Published: June 2021

Aerobic exercise training (AET) has been established as an important phenotype capable of altering the human skeletal muscle. MicroRNAs (miRNAs) are regulators of biological processes by modulating gene expression. The microRNA‐206 is skeletal muscle‐specific. Therefore, we evaluate the effects of expression of miR‐206 in response to AET in human skeletal muscle. Twelve healthy volunteers were biopsied pre and post period aerobic exercise training. Moreover, heart rate (HR), mean blood pressure (MBP), maximal exercise capacity (VO2peak) forearm blood flow (FBF) and forearm vascular conductance (FVC) were evaluated, the miRNA‐206, PAX‐7, MYF5, MYOD, MYOG levels were analyzed. Aerobic exercise training was performed for 16 weeks. All variables were re‐assessed following completion of the training period. After physical training, the individuals showed an increase of 97% in human skeletal muscle in microRNA‐206 expression levels. These alterations were accompanied by increased of 55%,84%,65%, 80% in PAX‐7, MYF5, MYOD, MYOG expression level, respectively. However, when analyzed by western blot comparing pre and post period there is decreased of 21% in MYOD and 33% in PAX‐7 protein levels, but MHC and MYOG there were increase of 44% and 40% in protein levels, respectively. Furthermore, decreased of resting bradycardia 12,5%, increased of VO2peak 18%, increased FBF 68% and increased of FVC 63%. These results suggest that in healthy subjects the miRNA miR‐206 is highly expressed after aerobic exercise training can thus locally modulating some signaling ways in muscle regeneration.