bluuebunny/crossref_metadata_2025 huggingface dataset:
Effects of Telmisartan and Treadmill Running on Bone Mineral Density in Spontaneously Hypertensive Rats
Authors: Bissoli Nazare , Medeiros Ana Raquel , Nonaka Keico , Pereira Rosa Maria , Ruffoni Leandro , Takayama Liliam , Birocale Marcos Antonio
Published: June 2021

Introduction

In the present study, we compared telmisartan, an angiotensin AT1 receptor blocker (ARB), and treadmill running exercise training on bone mineral density (BMD) and biomechanical properties of spontaneously hypertensive rats (SHR). This study was designed to assess changes in BMD and bone biomechanical properties in male SHR treated with telmisartan (T) or undergoing physical activity.

Methods

We used treadmill running (18 m/min/60min/d) and T (5 mg/kg/d) by a 8 weeks period in 3‐month‐old male SHR that were divided randomly into the following 3 groups: Sedentary (S), T and Exercise (E). At the end of protocol, BMD of femur and fifth lumbar vertebra was detected using DXA, and biomechanical properties of these bones were measured by the three‐point bending test. Blood pressure of all groups was measured using a tail‐cuffmanometer.

Results

T and treadmill running, compared to sedentary regimen, reduced blood pressure, but T did not improve bone characteristics, and instead reduced both BMD and bone biomechanical properties. Treadmill running in the study design kept the bone characteristics evaluated and, nevertheless, when comparing to telmisartan use, was effective in maintaining the health of the bone.

Conclusions

Our data showed that T negatively affects bone mass and quality evaluated in spontaneously hypertensive rats and is necessary more studies to better evaluate the effect of this drug on the bone proprieties.
Effects of Maternal Separation on Behavior and Cardiovascular Function in the Wistar‐Kyoto Rat
Authors: Rana Samir , McCoy Chelsea , Pugh Phyllis , Wyss J. Michael , Clinton Sarah , Kerman Ilan
Published: June 2021

Early‐life experiences can have long‐term effects on cardiovascular function and are likely mediated by a host of factors, such as inborn stress sensitivity and epigenetic alterations. To begin addressing this issue we exposed Wistar‐Kyoto (WKY) rats to 180 min. maternal separation (MS) from postnatal day (P)1 to P14, while control rats experienced neonatal handling (NH; 15 min separation). Animals were weaned at P21, developed to adulthood undisturbed, assessed depressive‐ and anxiety‐ like behaviors followed by continuous blood pressure monitoring using radiotelemetry. Upon completion of cardiovascular recordings, whole genome methylation was quantified within select brain regions. Results of our behavioral analyses showed that MS‐exposed rats exhibited decreased depression‐ and anxiety‐ like behaviors along with augmented social behaviors in adulthood. Cardiovascular recordings revealed that MS resulted in the decrease of resting heart rate throughout life, which was accompanied by an increase in baroreflex sensitivity. These protective baseline effects were accompanied by improved cardiovascular recovery to social stress in the MS‐exposed rats. These behavioral and cardiovascular changes were also correlated with an increase in whole genome methylation within the hippocampus, but not other brain regions. Our data demonstrate protective behavioral and cardiovascular effects of MS in WKY rats, which may be related to their inborn stress reactivity. Epigenetic alterations within the hippocampus may contribute to the emergence of these effects.

Support: R00MH081927 (IAK), R01MH105447 (SMC), 13PRE16940050 (SR), P30 NS47466 (UAB core)
Temporally‐controlled Cell Labelling Reveals Transdifferentiation of Smooth Muscle Cells during Atherogenesis
Authors: Feil Susanne , Fehrenbacher Birgit , Schaller Martin , Feil Robert
Published: June 2021

Atherosclerosis and its secondary disorders, heart attack and stroke, are the major causes of death in the western world, but their pathogenesis is poorly understood. One question is how vascular smooth muscle cells (VSMCs) contribute to atherogenesis. In this study, temporally‐controlled Cre/lox‐mediated cell fate mapping was used to investigate the role of VSMCs in atherosclerotic plaque formation in apolipoprotein E‐deficient mice. Prior to development of atherosclerosis, VSMCs were genetically labelled and their fate was followed during disease progression. Our in vivo analyses showed that distinct plaque regions are composed of clonally growing VSMC‐derived cells, which drastically changed their phenotype. These phenotypic changes were further evaluated by expression profiling of cultured VSMCs as an in vitro model of atherogenesis. Both the synthetic/dedifferentiated VSMCs in cell culture and the VSMC‐derived plaque cells in vivo lost typical smooth muscle markers (e.g., SM alpha‐actin) and instead expressed macrophage markers (e.g., Mac‐2 and CD68) and proteins associated with cellular growth and stress. Interestingly, we found that cGMP signalling via cGMP‐dependent protein kinase I might be involved in the development of the VSMC‐derived plaque cells. These findings provide strong in vivo evidence for smooth muscle‐to‐macrophage transdifferentiation and support important roles of cGMP and VSMC plasticity in atherogenesis. Targeting this type of VSMC phenotypic conversion might be a novel strategy for the treatment of atherosclerosis as well as other diseases with a smooth muscle component.
The commercial supply Sprague‐Dawley rat in Taiwan (2012~2013) with spontaneously mild hypertension may affect the degree of inflammation
Authors: Lue Sheng‐I , Lin Shiuan‐Yea , Fu Yaw‐Syan , Lu Tzong‐Shi
Published: June 2021

During 2012‐2013, the commercial supply of SD rats in Taiwan with a very high variation in blood pressure condition, and a larger percentage of those animals with mild hypertension. For the intravenous (i.v.) application of capsaicin (Cap), substance P (SP), or histamine may induce endothelial gaps formation in the venules of airway mucosa, resulting in an extensive plasma leakage in normotensive SD rats. The aim of this study is focus on the mild hypertensive effects on neurogenic and immunogenic inflammation. The femoral artery of the commercial supply SDrat was cannulated for monitoring arterial blood pressure, and femoral vein was cannulated for injecting Cap, SP, histamine, or vehicle. The plasma leakage of trachea was evaluated by Evans blue extravasation. Five min after application of Evans blue and blood pressure recording, rats were perfused with saline and phosphate‐buffered paraformaldehyde. Evans blue content in the trachea was extracted with formamide and determined with an ELISA reader at 620nm. There is a significant negative linear regression on the degree of plasma leakage of trachea with blood pressure in neurogenic inflammation, but a positive regression in immunogenic inflammation. It is suggested a desensitization on neurogenic inflammation in airway of spontaneously hypertensive SD rat, but rat with hypertension may promote immunogenic inflammation that increased the histamine‐induced plasma‐leakage in airway.
Ski Modulates Myofibroblast Motility via Downregulation of MMP2 and Paxillin
Authors: Kavosh Morvarid , Filomeno Krista , Rattan Sunil , Dixon Ian
Published: June 2021

Cardiac fibrosis is component of a number of cardiovascular diseases, including myocardial infarction. Excessive formation of extracellular matrix (ECM) occurs in activated cardiac (myo)fibroblasts that reside in the infarct scar or those moving from adjacent viable tissue. We propose that Ski, an endogenous repressor of TGF‐β1, regulates ECM remodeling and cellular motility by influencing MMP function, and specifically MMP‐2. Moreover, Ski alters cardiac fibroblast motility by altering the expression of paxillin (a focal adhesion associated protein) and its kinases including FAK (Tyr 397) and PYK2 (Y402). Primary adult rat fibroblasts (P1) were subjected to either exogenous Ski overexpression by adenoviral Ski (Ad‐Ski) or Ad‐Lac‐Z control with the multiplicity of infection (MOI) of 50 and 150. Ski overexpressing cells exhibited significantly lower MMP‐2 secretion and significantly decreased MMP‐2 activity as detected using gelatin zymography. Using Transwell plates, Ski overexpression was associated with a significant decrease in migration of cells in the presence of a chemoattractant in the lower well. Moreover, reduced motility of P1 cells vs. control was observed by Ski overexpression group via scratch assay. Furthermore, downregulation of paxillin was detected in Ski overexpressing cells lysates vs. Lac‐Z and control samples. Both FAK (Tyr 397) and PYK2 (Y402) were decreased in Ski overexpressing cells vs control. We suggest that Ski may exert multiple effects eg, MMP2 and paxillin, which affects motility and adhesion, and thus represents a putative mechanism for modulation of myofibroblast function and cardiac fibrosis.
Effects of Voluntary Exercise Preconditioning on Ang II‐Induced Cardiovascular Pathophysiology in the Mouse
Authors: Wilson Brittany , Forrester Steven , Kawata Keisuke , Kim Boa , Park Joon‐Young
Published: June 2021

Purpose

To determine the effect of five weeks of voluntary wheel exercise on cardiovascular pathophenotypes caused by angiotensin II.

Methods

Twenty‐one C57BL/6J mice were assigned into 4 groups; Sedentary Saline (SS), Sedentary AngII (SAII), Voluntary Wheel Saline (VWS), Voluntary Wheel AngII (VWAII). VW mice had access ad libitum to an exercise wheel for five weeks. AngII was infused at a rate of 700 ug/kg/day for the following two weeks. Echocardiography was performed using Vevo 2100 in parasternal short axis view on the mice after two weeks AngII or Saline infusion.

Results

Heart weight (HW) was significantly higher in VWS and SAII (161.0 & 164.8 mg) compared to SS (139.8 mg; P<.05). HW to body weight (mg/g) was significantly higher in SAII compared to SS (6.1 vs 5.2 mg/g; P<.05). There was a strong pattern that VWAII showed higher HW:BW compared to VWS (6.4 vs 5.7 mg/g; P=.052). Posterior wall thickness in diastole (PWd) was significantly greater in VWAII compared to VWS (1.2 vs 0.9mm; P<.05). Ejection Fraction (EF) was significantly higher in VWS compared to SS (59.3 vs 54.6%, P<.05). VWS showed higher EF compared to VWAII (59.3 vs. 55.4%; P=.053). Fractional Shortening (FS) was significantly higher in VWS compared to SS (30.9 vs 27.8%; P<.05). VWS had significantly higher FS compared to VWAII (30.9 vs 28.2%; P<.05).

Conclusion

AngII, regardless of exercise preconditioning increased HW:BW ratio. Five weeks of VW exercise preconditioning rescued the deleterious effect of AngII on EF and FS compared to SAII mice. Future studies are warranted to identify molecular mechanism of protective effect of exercise on AngII induced hypertrophy.

AHA Scientist Development Grant 12SDG12070327 255 (to J.‐Y. Park)
Knockout of Toll‐like Receptors 2 and 4 Prevents Renal Ischemia‐Reperfusion‐Induced Cardiac Hypertrophy
Authors: Trentin‐Sonoda Mayra , Cirino‐Silva Rogério , Vieira Abrahão Mariana , Monnerat‐Cahli Gustavo , Medei Emiliano , Sorelli Carneiro‐Ramos Marcela
Published: June 2021

Background

It is known that renal ischemia reperfusion (I/R) generates a systemic inflammatory state, promoting molecular and morphological changes in the heart tissue, but the mechanisms remain unknown. TLRs are largely studied in the heart and are known to participate of inflammatory process, moreover it has been demonstrated their role in cardiac hypertrophy (CH) development.

Aim

The aim of this study was to investigate whether cardiac TLRs pathways are involved in renal I/R‐induced CH.

Methods/Results

C57BL/6J, TLR2‐/‐ and TLR4‐/‐ mice were subjected to unilateral 60 min renal ischemia followed by reperfusion for 5, 8, 12 and 15 days. A serum peak of inflammatory cytokines was observed after 5 days of reperfusion. Heart weight/tibia length (HW/TL) ratio was increased after 12 days of reperfusion, as well as CH markers BNP and alpha‐actin. Cardiac electrical activity showed longer QT, QTc and action potential duration after 15 days. Cardiac TLRs, NF‐kB, MyD88, HSP60 and 70 mRNA levels were increased in the 15‐days group. After 15 days of reperfusion, the absence of TLRs prevented CH development, as reflected by similar values of HW/TL compared to transgenic non‐ischemic mice, as well as lower expression of HSP and NF‐kB, suggesting that renal IR‐induced increase of these molecules was prevented.

Conclusions

These results indicate that (i) TLRs play a key role on renal IR‐induced CH; (ii) prevention of hypertrophic response was independent of renal function recovery; (iii) TLR2 and TLR4 selectively regulate the systemic inflammatory profile.
Resveratrol Attenuates Vasocontractile Responses to Phenylephrine and Potassium Chloride via a Nitric Oxide Synthase– and Endothelium–Independent Mechanism
Authors: Holder Peter , Denniss Steven , Rush James
Published: June 2021

This study examined the effect of acute pre‐incubation without or with resveratrol (RSV) at increasing in‐bath concentrations (0, 20, 50, or 100 µM) on the dose‐dependent vasocontractile response stimulated by either the α1‐adrenergic receptor agonist phenylephrine (PE; ‐9.0 to ‐5.0 LogM) or potassium chloride (KCl; 0 to 60 mM) in isometrically‐mounted common carotid artery rings isolated from 20‐30 week old Spontaneously Hypertensive rats (SHR) and normotensive Wistar Kyoto rats (WKY). In both SHR and WKY, the contractile dose‐response curve to PE was increasingly blunted by preincubation with 20, 50, and 100 µM RSV, as evidenced by a RSV concentration‐dependent rightward shift of EC50 and decrease in maximum amplitude and area under the curve (p<0.05). The contractile dose‐response curve to KCl was similarly affected by preincubation with increasing concentrations of RSV (p<0.05). Neither nitric oxide synthase (NOS) inhibition nor endothelium removal was found to affect the ability of 50 µM RSV to attenuate either PE‐ or KCl‐stimulated contraction in SHR or WKY. These data indicate that RSV can attenuate vasocontractile activity stimulated by either receptor‐mediated (PE) activation or voltage‐mediated (KCl) activation via a vascular wall cell‐signaling mechanism that is independent of NOS or the endothelium. This suggests that RSV may act to attenuate contraction in the vascular wall by directly affecting central pathways of contractile activation, such as vascular smooth muscle calcium handling and/or sensitivity.

Funded by NSERC Canada
Congenital Malformations of the Young Children in a Metropolis
Authors: Gatauova Madina , Abduldayeva Aigul
Published: June 2021

During this period various malformations has been only 253 children, of which, in 2012 ‐ 124 versus 2011 ‐ 129. In the analysis of medical history patients were identified as follows. LBW births according to WHO criteria with low and extremely low birth weight and gestation less than 34 weeks: 200 were born at term: 53. In 76% of cases dominated the boys. In the maternity ward, these children received treatment for: cerebral ischemia ‐ 84.5%, congenital pneumonia ‐ 55% of the disease specific to the perinatal period ‐ 23.6%. A high percentage of cerebral ischemia due to the fact that neurological disorders combined with almost all diseases especially in LBW and includes a number of symptoms characteristic of all clinical entities neonatal period. This is due as well to the immaturity of the nervous system of infants, the breakdown of adaptive mechanisms, as well as the background on which the pregnancy develops. In the analysis of extragenital pathology revealed anemia in parturientwomen ‐ 43.6%, endocrine diseases ‐12% and 11.7% of kidney disease. Diseases of the cardiovascular system ‐ 3%, and congenital heart disease accounted for 0.1%; respiratory diseases ‐ 4.6% and 0.1% chronic brucellosis. Unobservable and noninvestigated mothers was 0.5%. Major position constitute congenital heart 30%. In particular, the most common cardiac malformations of ventricular septal defect and patent ducts arteriosus. This is followed by congenital malformations of the nervous system and bone and joint system representing 20% of all vices. In the analysis of somatic diseases: malnutrition was diagnosed in 15.8%; cerebral palsy, hyperkinetic form 11%. Bronchopulmonary dysplasia in 19%.Thus, assistance in line with WHO recommendations to help reduce birth defects children.
Allosteric Modulation of CB1 by Pregnenolone
Authors: Spaeth Justin , Camacho Alex , Rios Brigitte , McKinney Tariq , Miller Aaron
Published: June 2021

The endocannabinoid system plays a role in diverse conditions such as anxiety, addiction, eating and memory disorders. Endocannabinoids are produced by postsynaptic neurons and activate receptors on presynaptic neurons in order to decrease neurotransmitter release. In the brain, the most important receptor activated by endocannabinoids is cannabinoid receptor type 1 (CB1). Tetrahydrocannabinol (THC), the active ingredient in marijuana, also activates this receptor. THC and similar drugs have therapeutic potential in the treatment of pain, Alzheimer's disease, anxiety, arthritis, and cancer. A downside to the medicinal use of THC is that it also induces psychotropic effects. Recently, it was discovered that pregnenolone binds to CB1, where it acts as an allosteric modulator that decreases the effects of THC. An allosteric modulator is a molecule that modifies receptor function by binding somewhere other than the active site. Discovery of allosteric modulators is significant because it means that there may be additional ways to target CB1 that have a reduced rate of psychotropic effects. The Messmer SMART (Students Modeling a Research Topic) Team has created a model of CB1 bound to pregnenolone using 3D printing technology. Our model highlights the amino acids E133 and R409, which form hydrogen bonds with pregnenolone and are required for its binding to the allosteric site of CB1. Studying the interaction between CB1 and pregnenolone will allow for a greater understanding of the interaction between synaptic function and pregnenolone levels as well as the design of additional allosteric modulators for testing as therapeutics. This program is supported by a grant from NIH‐CTSA.
Repeatability of Superficial Femoral Artery Flow Mediated Dilation as induced by Calf Exercise
Authors: King Trevor , Schmitter Sarah , Pyke Kyra
Published: June 2021

In the brachial artery, a sustained (via handgrip exercise) shear stress stimulus elicits flow mediated dilation (FMD) which is stable and may relate to vascular health. The legs experience different patterns of shear stress than the arms, and are prone to different manifestations of cardiovascular disease, however a protocol to assess leg FMD as stimulated by exercise has yet to be established. The study purpose was to determine the repeatability of superficial femoral artery (SFA) FMD created using a sustained shear stress stimulus via calf exercise (LEX). 19 subjects (9 females) (age: 24 ± 2) performed 4 trials of dynamic LEX for 6 minutes to reach an individually determined shear rate (SR=blood velocity/artery diameter) target in a specially designed machine. SFA blood velocity and diameter were assessed via echo and Doppler ultrasound. FMD was quantified as the % change from baseline to peak 1 min mean diameter during LEX. Values are mean ± SD. There were no differences between trials in mean SR produced during the last 5 min of LEX (p = 0.17) or during the minute with the highest SR during LEX (p = 0.73). Trial 1 (T1) LEX FMD was significantly lower than T3, but not T2 or T4 (T1: 3.6 ± 2.7 %, T2: 3.9 ± 3.0 %, T3: 4.9 ± 3.4 %, T4: 4.5 ± 2.6 % FMD, p = 0.023). There was a trend for declining baseline diameter from T1 to T4 (impact of Trial p = 0.049), but no trials were significantly different (pairwise posthoc testing all p > 0.10), and there was no difference in peak 1 min diameter between trials (p=0.74). The interpretation of %FMD is complicated by changing baseline diameter, however these results suggest that when shear stress is controlled, the artery may dilate to a consistent peak diameter. Further research is required to determine the reasons for the instability in baseline SFA diameter and the utility of SFA LEX FMD. NSERC CFI
Evidence of a Prolonged Post‐Mental Stress Decline in Flow‐Mediated Dilation in Young Healthy Subjects
Authors: Plotnick Meghan , Pyke Kyra
Published: June 2021

A temporary depression in reactive hyperemia stimulated flow‐mediated dilation (RH‐FMD) has been observed in young healthy individuals after an acute bout of mental stress. While it has been shown that a post‐stress reduction in RH‐FMD persists for 90 minutes in middle‐aged participants, it is unclear whether the impairment is similarly prolonged in a young healthy population. The objective of this pilot study was therefore to examine brachial artery RH‐FMD at 90 minutes post‐acute mental stress in young healthy volunteers. Doppler and echo ultrasound were used to obtain brachial artery diameter and velocity respectively. The shear stress stimulus was characterized as the shear rate (SR = velocity/diameter) area under the curve (SR‐AUC) during the first 30 seconds following forearm cuff deflation. Data are mean ± SD. Four men and two women aged 22.2 ± 3.7 years participated. RH‐FMD was assessed at baseline and 90 minutes following a stress task based on the Trier Social Stress Test (TSST). The average subjective stress rating for the TSST was 6.7 ± 1.8 (on a subjective scale of 1‐10). SR‐AUC was not significantly different at baseline (2304.9 ± 1055.2) compared to 90 minutes post‐mental stress (3022.9 ± 1224.7; P = 0.171). RH‐FMD was significantly lower in participants at 90 minutes post‐stress (6.7 ± 5.5%) compared to baseline (9.3 ± 3.7%; P = 0.025). These preliminary findings suggest that acute mental stress can have a prolonged (90 minute) detrimental effect on endothelial function in young healthy adults. Funded by NSERC and CFI.
Excess Alcohol Consumption and Cardiac Fibrosis
Authors: McCalister Aaron , Mouton Alan , Kay McKenzie , Gardner Jason
Published: June 2021

Background

Alcoholic Cardiomyopathy (ACM) is characterized by chronic alcohol consumption resulting in ventricular dilatation and cardiac fibrosis (CF). This CF is due to an increase in extracellular matrix components, including Collagens I and III.

Statement of Purpose

The purpose of this study is to determine the effect of chronic alcohol consumption on the expression of cardiac proteins Collagen I and III.

Methodology

Adult male Sprague‐Dawley rats were exposed to ethanol vapor to achieve approximately 200 mg/dl blood alcohol levels. Rats were exposed to ethanol for 10 weeks, receiving 14 hours on and 10 hour off daily. Left ventricle and livers were harvested and snap frozen for Western Blot analysis. Tissue samples were homogenized in RIPA buffer. Protein concentration was determined by Bradford Assay. Protein samples were separated by SDS‐PAGE and transferred onto PVDF membranes, which were probed with primary antibodies for CYP2E1, alcohol dehydrogenase (ADH), or collagens I and III. Membranes were then exposed to a luminescence detection reagent for protein detection.

Results

Livers were analyzed for the expression of CYP2E1 and ADH. Cardiac tissues were analyzed for the expression of collagen types I and III. Liver expression of protein CYP2E1 was increased due to alcohol exposure, whereas ADH was decreased. Cardiac expression of collagens was increased by alcohol.

Conclusion

Chronic alcohol consumption may lead to cardiac fibrosis.
Measurement and Analysis of the Dynamics of Erythrocyte Oxygen‐Dependent ATP Release
Authors: Sove Richard , Goldman Daniel , Fraser Graham , Ellis Christopher
Published: June 2021

Erythrocytes play a key role in the distribution of oxygen (O2) supply in the microvasculature through the oxygen saturation (SO2)‐dependent release of adenosine triphosphate (ATP). Previous studies address the magnitude of ATP release, however, little attention is given to the dynamics, which are crucial to understanding this regulatory system. Our goal is to characterize the dynamics of SO2‐dependent ATP release as it applies to O2 mediated blood flow regulation. Previously, we developed a computational model of a device capable of measuring the dynamics of ATP release (Sove PLOS 2013). In the present study we have modified the model to match our current prototype that utilizes a new fabrication approach of embedding a polymethyl methacrylate (PMMA) O2 impermeable barrier with a window for O2 exchange, in a polydimethylsiloxane (PDMS) O2 permeable layer. The model predicts how the PDMS exchange surface between the erythrocyte and gas channels affects the rate of O2 saturation decrease; thus making it possible to optimize the design of the device and to define appropriate hematocrit and flow conditions for in vitro studies. The model predicts that the PDMS layer thickness causes a decrease in the rate of SO2 decrease, and that low hematocrit results in larger SO2 decrease in a quasi‐linear relationship (slope ‐0.12). Despite the predicted SO2 decrease, ATP release is less due to the decreased number of erythrocytes. Our model shows that decreasing flow rate decreases the SO2, which results in a decrease in spatially derived resolution of ATP release time. The model of the microfluidic device aids in the design of future prototypes, facilitates the analysis of experimental results and provides a means to extract information from in vitro studies that cannot be measured directly.
Characterization of Dendritogenesis in Cortical Pyramidal Layer V Neurons in Niemann‐Pick Type C Disease Using Yellow Fluorescent Protein
Authors: Morales Silva Roberto , Alvarado‐Torres John , Boudewyn Lauren , Walkley Steven
Published: June 2021

Niemann‐Pick type C (NPC) disease is an autosomal recessive lysosomal storage disorder affecting mostly children, causing progressive neurological deterioration and death. NPC disease is caused by a mutation in either the NPC1 or NPC2 gene, leading to a loss of functional NPC1 or NPC2 protein. These proteins play a role in lipid egress from late endosomes and lysosomes, a deficiency in either results in intracellular accumulation of unesterified cholesterol and gangliosides. Storage is prominent in neurons, influencing cellular pathology, such as growth of ectopic dendrites on pyramidal neurons. This aberrant growth of dendrites following normal dendritogenesis has been documented in several lysosomal diseases characterized by ganglioside storage. Until recently, the visualization of this process depended on the Golgi method. New techniques, such as endogenous expression of fluorescent proteins, provide an in‐depth analysis of neuronal structure. We hypothesized that the accumulation of gangliosides in NPC disease will cause an initial enhancement of dendritogenesis along with ectopic dendrite growth followed by a subsequent degeneration of dendritic trees later in disease. Subsequently, we generated a transgenic Npc1 murine mouse model whose layer V cortical pyramidal neurons express yellow fluorescent protein (YFP) to investigate the dendritic abnormalities of NPC disease relative to lysosomal storage. Confocal imaging and Neurolucida Software are being used to image YFP+ pyramidal neurons, which will be analyzed for changes in dendritic size, complexity and pathological state.
Pathophysiological and Neurobehavioral Identification of Propionic Acid‐mediated Autism‐mimetic Rat Model
Authors: Lee Seunghoon , Hong Yonggeun
Published: June 2021

ASD is a heritable neurodevelopmental disorder with poorly understood and complex etiology. The purpose of this study is to construct autism mimetic animal models, and to establish how to create autism‐mimetic animal models and the standard indicators, through a comparison between the autism clinical aspects, pathophysiological and neurobehavioral alteration. 22 male SD rats were employed and randomly divided the control and propionic acid (500mg/kg) treated rats groups. The rats were subjected behavioral tests, gene expression, and histological analysis for pathophysiological and neurobehavioral alteration. PPA‐treated rats were significantly reduced exploring activity, non‐aggressive behaviors, whereas enhanced aggressive behavior (**p<0.01). Proinflammatory cytokine, TNF‐a was continuously upregulated PPA‐treated rats hippocampus following time‐dependent (**p<0.01). Expression of GFAP was significantly augmented at day 14 after PPA treatment (**p<0.01). In histological evaluation, PPA‐treated rats were significantly reduced diameter of granular cells and thickness of granular cell layer rather than control rats (**p<0.01). We suggest that PPA administration could induce a critical role of proinflammatory cytokine elevation and abnormal neural cells organization, which reveal autism‐mimic neurobehaviors, including increment of aggressive behavior, reduction of exploring activity, and exclusive and passive behavior.

Funding: 2012R1A1A2005089, 2013R1A2A2A01067169, KGM4611512
Polymorphisms in rs 13181 of ERCC2 gene in the personnel of Stepnogorsk gorno chemiches kombinat, Kazakhstan
Authors: Balmukhanov T.S , Kazymbet P.K. , Toltaev B.S.
Published: June 2021

The possibility of mutations in human somatic cells induced by action of radiation in low doses is a subject of investigations and discussion. The protein encoded by ERCC2 (XPD) gene is involved in transcription‐coupled nucleotide excision repair and is associated with some types of cancer. The aim of this study was to determine the influence of chronic action of radiation in low doses to the level of the single nucleotide polymorphisms in DNA of the personnel of uranium enrichment plant, Akmola, Kazakhstan. The sample set consisted of 234 (52–Kazakhs, 182–Russians) men workers of and the control group includes 289 (129–Kazakhs, 160–Russians). Genotyping was performed by restriction fragments length polymorphism analysis of PCR products of the rs13181 site (T/G) of (T/G) of ERCC2 gene using PstI. In the case‐control studythe allele frequencies and genotypes distribution were compared in the groups of personnel and controlsusing STATISTICA v. 5.0 software.

The differences in allele frequencies and genotypes distribution in the groups of personnel and controls were evaluated neither in Kazakh nor in Russian groups. At the next stage comparing of allele frequencies and genotypes distribution was performed in the subgroups of the workers divided according accumulated dose: less than 40 mZv, 40‐100 mZv.

The differences were found in genotype distribution only in the group of 50 Russian workers with accumulated dose 40–100 mZv compared with 100 of controls: χ2=6.796, p=0.033, OR=1.363, 95%CI=0.844‐2.201. The group of persons who accumulated dose more than 100 mZv encountered only 5 persons and was not included in calculations.
The T Protein: Vertebrae Fit to a T
Authors: Tiffany Karen , Tiffany K , Anderson N , Arnholt A , Baumgartner I , Butt A , DeBuhr O , Dyke S , Griffin M , Hu J , Janecek E , Kalmer I , Ketelhohn L , Lawniczak J , Minerva N , Naas A , Roddy M , Satchie A , Squires E , Wandsnider M , Wankowski J , Wilde A , Zietlow E , Pickart Michael
Published: June 2021

Congenital vertebral malformations (CVMs) comprise a group of spinal abnormalities that include alterations in vertebral shape or number. Evidence suggests CVMs have a genetic link, possibly resulting from mutations in multiple genes. One candidate gene is T. T protein, a transcription factor found in a variety of animals including humans, is essential for correct embryonic development and guides the development of bone and cartilage from embryonic mesodermal tissue. T protein accumulates in the nuclei of notochord cells, interacts with DNA at specific genes, and acts as a genetic switch to activate the genes. T protein binds to the major and minor grooves of DNA as a dimer. Mutations in T (turning “off” the T protein switch) are hypothesized to result in defects in spinal development. The Cedarburg SMART (Students Modeling A Research Topic) Team has designed a partial model of T protein using 3D printing technology to investigate its structure‐function relationship, focusing primarily on the residues important for dimerization of T (Pro125, Asp126, and Pro128) and for binding DNA (Arg67). A 3D model could indicate how the location of the mutations may impact the function of T. T could consequently be a potential target for the development of treatment or prevention options. Program supported by a grant from NIH‐CTSA.
acetylcysteine prevents maximal exercise‐induced increase in circulating miR‐126 expression and angiogenic mediators in intermittent claudication patients
Authors: da Silva Junior Natan , Roseguini Bruno , Chehuen Marcel , Fernandes Tiago , Mota Gloria , Forjaz Claudia , Wolosker Nelson , Oliveira Edilamar
Published: June 2021

In intermittent claudication (IC) patients, maximal exercise increases systemic levels of pro‐angiogenic mediators. The mechanisms responsible for this response are unknown, but oxidative stress may play a role. This study investigated the effects of maximal exercise and antioxidant treatment with N‐acetylcysteine (NAC)on the expression of miR‐126 (an endothelial‐specific microRNA that regulates angiogenesis) and its targets in IC patients. Following a double‐blinded randomized crossover design, 10 patients received oral NAC (1800 mg/day for 4 days plus 2700 mg prior to experimental session) and placebo (PL) before undergoing a treadmill exercise test to maximal leg pain. Blood samples were taken at rest, 1 hour after the ingestion of the last dose, and 30 min after the exercise test. Whole blood miR‐126, PI3KR2, VEGF and eNOS expression were determined by real‐time PCR. Treatment with NAC did not alter baseline expression of miR‐126 and its downstream targets. After maximal exercise, the expression miR‐126 (Fold change PL 1.79 vs NAC 0.79), eNOS (Fold change PL 1.73 vs NAC 1.14) and VEGF (Fold change PL 1.45 vs NAC 0.71) increased relative to baseline (p<0.05), and this response was blunted after NAC. Similarly, PI3KR2 decreased significantly (p<0.05) after exercise with placebo while no change was observed with NAC (Fold change PL 0.56 vs NAC 1.08).In conclusion, oral treatment with NAC prevents maximal exercise‐induced increase in circulating miR‐126 expression and other angiogenic mediators in IC patients. Funding: FAPESP 2013/05883‐8
An investigation into practical exercise strategies to offset the negative impact of 12h of sitting on glycaemic control
Authors: Brooks Naomi , Henderson Deborah , Tipton Kevin , Hamilton David Lee
Published: June 2021

Recent evidence suggests that there is a dose‐response association between sitting time and mortality. It has been reported that breaking up sitting time improves glycaemic control. The AIM of this study was to compare 12h sitting (SIT) with two interventions to influence sedentary time – a bout of resistance exercise prior to 12h sitting (RE) and 6min walk/hour of 12h sitting (WALK).

Methods

Healthy, active males took part in 3 trials. SIT: 12h sitting without any physical activity. RE: one bout of Resistance Exercise (4 x 10 reps; 75% 1RM) followed by 12h sitting. WALK: 12h sitting with bouts of walking (6min/hr; 40‐59% VO2max). Caloric intake remained similar between trials. The morning following each trial (12h post), blood samples collected at baseline and 15, 30, 45, 90 and 120 mins following an Oral Glucose Tolerance Test drink. Glucose and insulin measures [MATSUDA, HOMA‐IR, Area Under Curve (AUC)] were analysed.

Results

Participants were young, fit individuals with VO2max of 57.4 ± 2.0 ml/kg/min and ~ 6 ± 0.8 hrs/week activity. Matsuda and HOMA‐IR were not significantly different between trials and participants remained in the healthy category for each even after 12h sitting. There were no significant differences in AUC for glucose (SIT: 150 ± 31, RE: 141 ± 28, WALK: 179 ± 29), but there was a trend for a reduction in AUC for insulin in WALK compared to SIT (SIT: 5346 ± 745, RE 4685 ± 809, WALK: 4308 ± 678; p<0.09).

Conclusions

Our results demonstrate that neither antecedent resistance exercise nor 6min/hr of brisk walking is sufficient to improve glucose tolerance in the face of 12 hrs of sitting. Further research is needed to optimise exercise strategies to offset the negative effects of prolonged sitting.
Acute Exercise Improves Insulin Action in Parallel with Increases in Markers of Inflammation in Liver from Mice Fed a High Fat Diet
Authors: Peppler Willem , MacPherson Rebbeca , Wright David
Published: June 2021

Exercise training in high fat fed mice is protective against liver inflammation and insulin resistance, which occurs in parallel with attenuation in weight gain. The objective of this study was to determine if acute exercise induces an insulin sensitizing effect in parallel with changes in the inflammatory profile of liver in obese mice. C57BL/6 male mice were fed a low fat diet (LFD, 10% kcals from lard) or high fat diet (HFD, 60% kcals from lard) for 7 weeks. Mice from the HFD group were then subjected to 2 hours of treadmill exercise at 15 m·min‐1. At 2 hours post‐exercise, or cage control, mice were injected with insulin (1U/kg BW) and livers were harvested. After 7 weeks, mice fed the HFD were significantly heavier than mice fed the LFD, had impaired glucose clearance during a glucose tolerance test, and had attenuated insulin‐induced phosphorylation of AKT. In mice fed a HFD, acute exercise recovered the impaired phosphorylation of AKT, suggesting improved insulin action. This was accompanied by increased phosphorylation of the mitogen‐activated protein kinase JNK1 and p38. Further, both socs3 and mcp‐1 mRNA were up regulated, along with increased phosphorylation of stat3. The results from this study suggest that in an obese mouse, acute exercise improves liver insulin action in parallel with increases in inflammatory signaling. Funded by NSERC. DCW is a Tier 2 CRC. REKM is funded by an Alzheimer Society PDF.
Restoration of Liver Function Following Transplantation of Healthy Hepatocytes into the Fah ‐/‐ IL2rg‐/‐ Rat Model
Authors: Rasmussen Shauna , Mallanna Sunil , Lerch‐Gaggl Alexandra , North Paula , Fahrenkrug Scott , Carlson Daniel , Tan Wenfang , Wilson Elizabeth , Duncan Stephen , Geurts Aron
Published: June 2021

Immune‐compromised mouse models of Hereditary Tyrosinemia Type I, which are deficient in fumarylacetoacetate hydrolase (Fah), have been widely appreciated as robust models for hepatocyte xenotransplantation studies. We hypothesized that a similar rat model with its increased size and ease of surgical procedures might provide unique experimental advantages. With the use of transcriptional activator‐like effector nuclease (TALEN) technology, we generated a Fah ‐/‐;Interleukin 2 receptor gamma (IL2rg) ‐/‐ double knockout rat and examined whether this model could undergo transplantation of healthy hepatocytes from either rat or human sources to restore liver function over time. Hepatocytes isolated from a green fluorescent protein (GFP) transgenic rat or commercially available human hepatocytes were injected intrasplenically into Fah ‐/‐ IL2rg ‐/‐ rats. Animals were cycled off/on 2‐[2‐nitro‐4‐(trifluoromethyl)benzoyl] cyclohexane‐1,3‐dione (NTBC) in order to obtain engraftment of the transplanted cells. Correction of liver function and robust engraftment of GFP (+) hepatocytes (2.12‐76.46%; n=25) were seen in the liver after one or three months of NTBC cycling. Human albumin was detected in serum (1µg/ml‐60µg/ml; n=5) of rats transplanted with human hepatocytes. This data supports that our Fah ‐/‐ IL2rg ‐/‐ rat model demonstrates effective engraftment and clonal expansion of transplanted healthy hepatocytes.
The Impact of Hydroxylase Inhibition on TGF‐β1 Induced Fibrosis Associated with Inflammatory Bowel Disease
Authors: C Manresa Mario , Tambuwala Murtaza , Cavadas Miguel , Cheong Alex , Cummins Eoin , Taylor Cormac
Published: June 2021

Fibrosis is a complication of chronic inflammatory disorders such as inflammatory bowel disease (IBD). Fibrosis is responsible for 75% of surgical interventions in IBD. Pharmacological inhibition of O2 sensing hydroxylases ameliorates inflammation in models of IBD. We hypothesized that hydroxylase inhibition may also impact fibrosis in IBD. To test this, the effect of the hydroxylase inhibitor DMOG was investigated. Mice exposed to 2.5% dextran sodium sulphate in drinking water for 5 days and allowed to recover for 14 days, developed colon fibrosis reflected by an altered pattern of submucosal collagen deposition, increased fibroblast infiltration and higher expression of Smad3. Mice treated with DMOG exhibited lower submucosal collagen accumulation, reduced fibroblast infiltration and less Smad3 positive staining. An in vitro model was then used to investigate the mechanism of the anti‐fibrotic action of DMOG. Fibroblasts were treated with TGF‐β1, key mediator of fibrosis. DMOG was given to the cells prior to TGF‐β1, and α‐smooth muscle actin (α‐SMA) and collagen‐I were analyzed as markers of fibrosis. TGF‐β1 induced phenotypic changes associated to higher expression of collagen‐I and α‐SMA. Both markers were reduced in DMOG treated cells, suggesting an inhibition of TGF‐β1 mediated fibroblast activation. We further investigated the impact of DMOG upon TGF‐β1 signalling. DMOG did not affect Smad2/3 (canonical) TGF dependent phosphorylation. In contrast, it reduced TGF‐β1 dependent ERK (non canonical) phosphorylation. In conclusion, hydroxylase inhibition reduces fibrosis in vitro and in vivo, at least in part, through the inhibition of non canonical TGF‐β signalling.
“Hard Core” Acetyl Coenzyme‐A Carboxylase
Authors: Elniff Chris , Bernardi Dayana , Boonyasiti Nina , Derib‐Adugna Ahadou , Einspahr Alana , Grover Sahiba , Kandru Namratha , Lux Nathan , Marre Sai , Dhillon Jansher , Duong Kyra , Hameedi Herra , Razo Julissa , Fogt Laura , Fenton Aron
Published: June 2021

Diseases involving unregulated cell reproduction, (i.e. cancer) can be inhibited through acetyl coenzyme‐A carboxylase (ACC) manipulation, preventing the lipid production that is imperative for cell survival. ACC catalyzes the first step in the fatty acid biosynthesis. ACC contains two dimers, biotin carboxylase (BC) and carboxyltransferase (CT), and a monomer, biotin carboxyl carrier protein (BCCP). Though BC, CT, and BCCP are separate proteins in prokaryotes, they are linked in a single unit within a eukaryotic cell. BCCP contains two sets of four parallel β sheets and a core of hydrophobic residues, both of which stabilize it. Due to its symmetry BCCP can be described as a capped β sandwich. In fatty‐acid biosynthesis, BCCP oscillates between the BC and CT domains to carboxylate biotin at the N1 site on the BCCD region. The surface of BCCP is interspersed with charged amino acids, arginine and lysine which are positive, and aspartate and glutamine which are negative. BC contains three domains: a central ATP grasp, an N‐terminal and a C‐terminal. The BC dimer works through “half‐site reactivity,” where one subunit binds the substrate with catalysis; another releases the carboxylated biotin product. This Olathe North High School SMART Team (Students Modeling a Research Topic) 3D modeling project is attempting to identify surface charges on BCCP that may contribute to binding of this subunit to BC and CT. Funded by Black & Veatch.
A Comparative Study of the 3D Structure of Myoglobin and its Affect on the Physiology of Terrestrial and Aquatic Mammals
Authors: Granberry Allison , Assad Fatima , Briglall Annie , Eusebio Diana , Frempong Hayford , Gajik Peter , Simon Margarita , Valentine Tashina , Steinberger Joseph
Published: June 2021

Sperm whales dive deeper than two kilometers and swim for up to 90 minutes while searching for prey. Underwater, they depend on previously inhaled oxygen molecules. Myoglobin (Mb) contributes to this process by binding and storing oxygen until the time of need, and then releasing it, providing oxygen within muscle cells for aerobic respiration. Although the molecular structure of Mb was solved more than five decades ago, its unique connection to the physiology of underwater lifestyles has only recently come to light. Surprisingly, aquatic and terrestrial mammals differ in the net surface charge (Z) of their Mb protein, as calculated by summing up all the positively and negatively charged surface residues. For instance, sperm whales' Mb (pdb 1MBN) has a Z of +4, while aardvarks' Mb has a Z of +0.25. It thus seems possible that aquatic mammals' larger oxygen supply results from their Mb's greater net surface charge, as the greater the net surface charge leads to more repulsion between Mb proteins and subsequent increase in the amount of Mb which may be packed into each cell. Terrestrial animals have myoglobin with a more neutral net surface charge and therefore cannot increase oxygen storage in muscle by increasing the number of Mb, since the Mb will stick to each other at high concentrations. The Hostos‐Lincoln Academy SMART (Students Modeling A Research Topic) Team created a physical model using 3D printing technology of an alignment of sperm whale Mb pdb file 1MBN and aardvark Mb, pdb file 1EMY. We further compared multiple Mb structures of aquatic and terrestrial mammals to verify the significance of Mb net surface charge to the physiology of deep diving mammals.
TRPV4 contributes to renal injury induced by angiotensin II in mice
Authors: Wang Youping , Zhu Minjun , Cui Lin , Liu Weihong , Wang Xiaoxiao , Shen Si , Xu Hui
Published: June 2021

To determine whether deletion of TRPV4 attenuates angiotensin II (Ang II)‐induced renal injury, Ang II was infused systemically into wild type (WT) and TRPV4‐null mutant (TRPV4‐/‐) mice for 4 weeks. Systolic blood pressure increased in Ang II‐treated WT or TRPV4‐/‐mice with the similar degree (P>0.05). Ang II treatment increased urinary excretion of albumin and 8‐isoprostane, and decreased creatinine clearance in both WT and TRPV4‐/‐mice with a greater magnitude in the former strain (P<0.05). Periodic acid‐Schiff and Masson's trichrome staining showed that kidneys of Ang II‐treated WT mice exhibited more severe glomerulosclerosis and tubulointerstitial injury compared with Ang II‐treated TRPV4‐/‐mice (P<0.05). Hydroxyproline assay and F4/80‐staining showed that renal collagen levels and monocyte/macrophage infiltration were greater in Ang II‐treated WT mice compared with TRPV4‐/‐mice (P<0.05). Thus, our data show that deletion of TRPV4 alleviates renal injury in the absence of alteration in blood pressure in Ang II‐hypertensive mice, indicating that TRPV4 could serve as a novel target for treating renal injury associated with hypertension. [This work was supported by grants from the National Natural Science Foundation of China (No.81170243) and Henan Provincial Science and Technology Innovative Program for Outstanding Scholarship (No. 124200510007)].