bluuebunny/crossref_metadata_2025 huggingface dataset:
EFFECT OF PATHWAY‐INTERCONNECTORS IN SEB‐INDUCED APOPTOSIS RELATED EVENTS IN HUMAN PBMCS
Authors: Klein Rebecca , Daily Sarah , Jett Marti , Mendis Chanaka
Published: June 2021

Staphylococcus enterotoxin B (SEB) is a bacterium that produces toxins which have been known to cause food poisoning, and have also been linked to toxic shock syndrome. The purpose of this study is to further investigate human peripheral blood mononuclear cells (PBMCs) in order to better understand the apoptosis related events induced by SEB. C‐Jun N‐Terminal Kinase (JNK) has been previously identified to be induced by SEB, and has demonstrated that it inter‐connects multiple signaling cascades. As a crucial pathway inter‐connector in SEB induced human PBMCs, we believe that JNK may possess inhibitory effects on SEB‐induced apoptosis. We will further evaluate the alterations induced by specific inhibitor SP600125, Anthrax[1,9‐cd]pyrazol‐6(2H)‐one, by the known gene expression profile of SEB using RT‐PCR, and then use ELISA to examine the effects of the inhibitors on protein expression patterns. We believe that our work will allow us to better understand the complex interactions of multiple signal transduction pathways induced by SEB.
Relationship between Vitamin D Levels and LDL Cholesterol across Tri‐Ethnic Minorities with and without Type 2 Diabetes
Authors: Zarini Gustavo G , Shaban Lamya , Sukhram Shiryn , Huffman Fatma G
Published: June 2021

The study investigated the relationship between 25‐hydroxyvitamin D [25(OH) D] levels and LDL cholesterol across tri‐ethnic minority groups. A cross‐sectional study was conducted in Cuban (CA=184), Haitian (HA=242), and African Americans (AA=224) with and without type 2 diabetes (T2D). Participants were recruited from multiple sources from Miami‐Dade and Broward Counties, Florida. Dietary variables were collected using a food frequency questionnaire. Serum vitamin D levels were measured with enzyme‐immunoassay kit by absorbance. Plasma LDL cholesterol (LDL‐C) was assayed by enzymatic methods. Analyses included descriptive statistics and analysis of covariance (ANCOVA). AA and HA had significantly lower 25 (OH) D levels compared to CA (p<.001). Subjects with T2D had significantly lower 25 (OH) D and LDL‐C levels than those without T2D respectively (p<.001 and p=.007). ANCOVA showed that after controlling for confounding variables lower 25 (OH) D levels were significantly associated with higher LDL‐C levels in AA and HA with T2D [F(2, 631)=3.22, p<.001 and p=.031] respectively. Findings provide a better understanding of the relationship between vitamin D and LDL‐C for minorities. Future studies should focus on the role of vitamin D supplementation and cardiovascular disease outcomes in different racial/ethnic groups. Funding for this research was provided through an NIH/NIDDK sponsored grant.
ANALYSIS OF FACTORS AFFECTING BONE MINERAL DENSITY WITH DIFFERENT AGE AMONG ADULT WOMEN IN SEOUL AREA, KOREA
Authors: Koo Jaeok , Kim MyungSuk , Na Nam Soon
Published: June 2021

This study was carried out to identify bone mineral density risk factors affecting women's BMD, and to analyze the relationship of BMD and other factors between age by physical measurement and questionnaires. The study subjects, 161 women(20~70 years) living in Seoul were divided into 4 age groups. T‐score BMD was significantly decreased according to increasing age and menopause. The rate of BMD risk subjects was significantly increasing with age and menopause. The rate of risk subjects were 8.3%, 19.4%, 22.2% and 50% by age groups, respectively. T‐score of groups was significantly decreased from 0.87(40 years) to −1.43(60 years). (P<0.001). Also systolic blood pressure were significantly increased from 114.9mmHg(20~30years) to 129.8mmHg(60 years). There were significant negative correlationship between T‐score and age(r=−0.395) menopause(r=−0.350), BMI(r=−0.226) systolic blood pressure (r=−0.153), alcohol drinking(r=0.225) (p<0.01). In conclusion, bone density decreased with increasing to age and BMI and systolic blood pressure, menopause and frequency alcohol consuming. To prevent osteoporosis, over 40 years women needed to be educated to maintain normal weight and the improvement of eating and living habits.
Effect of age and body weight on expression and serum concentration of GH, IGF‐1, and IGFBP in pigs
Authors: Trejo Adriana Morales , Avelar Ernesto , Cervantes Miguel , Araiza Alfonso B.
Published: June 2021

There are no available reports regarding the effect of age and body weight (BW) of pigs on the expression of GH, IGF‐1, and IGFBP. A study was conducted to analyze the relationship between age and BW with serum concentration (SC) of GH and IGF‐1, and relative expression (RE) of GH and IGFBP in pigs. Twenty pigs were sacrificed; 4 at each of the following ages: 1, 30, 60, 120, and 180 d. Samples from hypophysis and liver were collected after pigs were sacrificed, total RNA was purified, and cDNA was synthesized to carry out real time qPCR for GH and IGFBP; 18S rRNA was used for normalization. Twenty more pigs were used to collect blood samples at the ages indicated above; serum was collected to analyze GH and IGF‐1 using ELISA. SC of GH and IGF‐1 at each sampling age were: 16.2, 6.9; 23.0, 12.2; 21.0, 12.8; 19.7, 17.6; and 20.4, 14.7 ng/ml, respectively. RE of GH in hypophysis at each age was: 833, 309, 629, 168, and 76, arbitrary units, respectively; in liver, RE of IGFBP was: 21.3, 9.0, 8.5, 3.8, and 6.7, respectively. SC of GH quadratically (P<0.05) and IGF linearly (P<0.01) increased with age and BW. RE of GH (P<0.01) and IGFBP (P<0.01) linearly reduced with age and BW. There were negative correlations between RE and SC of GH (P<0.05), RE of GH and SC of IGF‐1 (P<0.01), and RE of IGFBP and SC of IGF‐1 (P<0.01). These data show a significant and differentiated effect of age and BW on GE and SC of GH, IGF‐1, and IGFBP. (Financial support by CONACyT, México).
Dark chocolate protects against colon carcinogenesis by favorable alteraion of cell kinetics
Authors: Hong Mee Young , Shelechi Mahshid
Published: June 2021

Colonic tumors arise from perturbations of the homeostasis among cell proliferation and apoptosis. Cocoa and chocolate products have among the highest concentrations of polyphenols and an increasing body of epidemiologic evidence supports that polyphenol‐rich diets promote health and attenuate or delay the onset of various diseases, including colon cancer. We determined whether dark chocolate protects against colon cancer by favorable alteration of cell kinetics (cell proliferation and apoptosis). Forty eight male Spague Dawley rats were assigned to receive diets containing control (no chocolate), regular or bloomed dark chocolate (10% of diet) and injected with saline or azoxymethane (2 times at wk 3 and 4). Cell proliferation and apoptosis were determined using Ki‐67 antibody and TUNEL assay, respectively. Carcinogen treatment increased proliferation index (P=0.003) and expanded proliferation zone (P=0.007). Dark chocolate diets, regardless regular or bloomed, lowered proliferation index (P=0.001) especially in lower 1/3 of colonic crypt (P<0.001) compared to control. Apoptosis was not significantly different among diets. These results suggest that cell proliferation was one of the key responses to carcinogen as a function of diet. Dark chocolate reduces risk of colon cancer by inhibiting cell proliferation rather than alteration of apoptosis. Supported by CHNR08‐810 and SDSU NUTR 302L class.
ApolipoproteinA1 promote resolution of lung inflammation and fibrosis in experimental silicosis
Authors: park sung woo , Lee Eun‐hee , lee eun‐joo , Park choon sik
Published: June 2021

Background

Inhalation of silica particles induces silicosis, inflammatory lung disease and eventually generating multiple fibrotic silicotic nodules. Silica is often used as experimental lung fibrosis model.

Objectives

To evaluate Intranasal treatment of Apolipoprotein A1(ApoA1) have prevent and/or resolving effect on the developing experimental lung silicosis.

Methods

BALB/C mice were assigned into four groups. Silica suspension(SiO2; 20mg/50ml in saline) is intratracheally instilled at Day 0. ApoA1 was treated at Day 7,8,9 (APOA1_P group) to investigate its prevention effect of silicosis. ApoA1 was treated at Day 16, 17, 18(APOA1_R group) to evaluate its therapeutic effect. The Mice were sacrificed at day 30. Histology, collagen assay and cytokine measurement including TGF‐b1 were performed. Apoptosis were analyzed using TUNEL assay.

Measurement and Results

Both ApoA1_P and ApoA1_R group have significant reduce the silica‐induced increased in number of inflammatory cells, silicotic nodule fractions and collagen deposition in sham‐ treated mice. TGF‐b1 level in BALF is significantly decreased in both ApoA1 treated group compare to Silica group. TUNEL assay shows that silica induced increased apoptotic cell numbers are significantly reduced in ApoA1 treated lung.

Conclusions

Local treatment with Apo A1 have preventive and pro‐resolving effect on the experimental lung silicosis.
Influence of Nutrient Timing Following Resistance Exercise on RQ and Glucose Two Hours After Supplement Ingestion in Young Men with a Family History of Type 2 Diabetes
Authors: Gutierrez Jean Lorraine , Gribok Andrei , Stevens Michelle , Rumpler William , Tadros Maryann , DiPietro Loretta
Published: June 2021

Objective

To examine the affect of post‐resistance exercise supplement timing on glucose and respiratory quotient (RQ) for two hours after nutrient ingestion.

Methods

Young male participants (n=6) underwent two 48‐h stays in a whole‐room calorimeter. The first 24‐h served as a control day, and on the second day participants completed a 40‐min novel resistance exercise protocol and consumed 600g of 1% chocolate milk either immediately after (IPEN) or 3‐h after completing resistance exercise (3h‐PEN). RQ was measured continuously throughout the 48‐h period and glucose concentrations were determined by a continuous glucose monitoring system.

Results

Mean RQ following the exercise bout, as compared to the same 2‐h time period of the control day, was significantly lower following IPEN (0.77±0.02 vs. 0.81±0.05; p= 0.02), but was not different after 3h‐PEN condition (0.79±0.02 vs. 0.78±0.02 p= 0.15). Mean glucose during the same period following the exercise day, as compared to the control day, was significantly higher following IPEN (107±10 vs. 96±16 mg·dL−1; p= 0.04), but not after 3h‐PEN (103±3.3 vs. 104±14mg·dL−1; p= 0.79).

Conclusions

Preliminary results indicate that lipid metabolism persists in the presence of higher than normal circulating glucose when a high‐carbohydrate drink is consumed immediately after resistance exercise. This study was funded by the GWU Department of Exercise Science.
Intake of CLA‐fortified butter decreases serum levels of leptin in healthy subjects
Authors: Penedo Leticia , Nunes Juliana C , Gama Marco A S , Quirico‐Santos Thereza , Torres Alexandre Guedes
Published: June 2021

Conjugated linoleic acid (CLA) might promote changes on body composition and on the components of the metabolic syndrome. We aimed to investigate the effect of supplementation of 1g/day of CLA, through the daily intake of a fortified butter, on the lean and fat body mass, and on serum levels of the adipokines adiponectin, leptin, and TNF‐α of healthy young adults. Sixteen men and women aged 23 to 36 years were studied, and the experimental design consisted of a baseline collection, a washout period of 8 weeks and the 8 week supplementation period. Supplementation with CLA did not alter body fat mass (P= 0.1884; one way ANOVA) assessed by dual‐energy X‐ray absorptiometry analysis (DXA). However, consumption of the CLA‐fortified butter reduced (P= 0,0004) serum levels (average ± SD) of leptin, from 8.16 ± 2.99 to 4.80 ± 1.93, but did not change significantly serum levels of adiponectin and TNF‐α. Our results indicate that naturally CLA‐enriched butter promoted benefits concerning the management of serum leptin levels, which might help the control of adiposity and to prevent obesity.

Financial support: CNPq, FAPERJ, CAPES (Brazil)
Serum lipid levels influence cognitive performance in older vervet monkeys (Chlorocebus aethiops sabaeus) on a long‐term stable diet
Authors: Crast Jessica , Renzi Lisa , Jorgensen Matthew , Grand Alison , Finnie Kelsey , Sousa Jennifer , Kaplan Jay , Jentsch J. David
Published: June 2021

Epidemiological data indicate high comorbidity between diseases linked to serum lipid levels, such as atherosclerosis, and cognitive decline. However, it is difficult to reduce the effects of poor general lifestyle to specific mechanisms that can produce these deficits. Here we used an animal model to determine the relation between serum lipids and cognitive decline in a population whose diet was stable across the lifespan. Serum lipids and cognitive performance were assessed in 61 African Green Monkeys, ranging in age from 3–24 years and maintained on a standard diet (high fiber, low fat, high protein) over the lifespan; female subjects were not pregnant at sampling. Learning and response inhibition were measured using a visual discrimination/reversal task. For those animals in the highest quintile for age (17–24 years), serum low density lipids were negatively associated and serum high density lipids were positively correlated with cognitive performance on all indices tested (p <0.05). These relationships were not evident for younger subjects. Thus, the relation between serum lipids and cognitive performance is not independent of age and is present in individuals who maintain a healthy diet across the lifespan. Research supported by P40 RR019963, RL1 MH 083270, UL1‐DE019580.
Microsatellite Instability in Rhesus Colon Carcinoma
Authors: McArthur Mark Jay , Barnhart Kirstin , Baze Wallace , Bernacky Bruce , Hanley Patrick , Benavides Fernando , Perez Carlos
Published: June 2021

Twenty‐five colon carcinoma cases from a specific pathogen free (SPF) breeding colony of Indian origin rhesus macaques (Macaca mulatta) were examined and evaluated for pathogenesis. Although, no specific mode of inheritance was identified there appeared to be a genetic bias, as several parent‐offspring or sibling‐sibling pairs were identified. In a subset of cases, DNA from the tumors and normal tissues from the same animal and tested for microsatellite instability (MSI) utilizing a panel of rhesus dinucleotides (MML3S7, MML4S1, MML11S1, MML16S3, MML17S1, MML17S2 and MML18S3) and a human mononucleotide (BAT‐25). All tumors tested exhibited some degree of microsatellite instability. Immunohistochemistical staining for mutL homolog 1 (MLH1), mutS homolog 2 (MSH2) and mutS homolog 6 (MSH6) was performed with loss of MLH1 staining being observed in several tumors. Rhesus colon carcinoma in the Bastrop colony with its early onset disease, microsatellite instability and loss of MLH1 may represent model of Lynch syndrome.
Sodium montmorillonite and quaternary ammonium organoclays: effect on fungal and bacterial cell growth and survival
Authors: Hutson Amber Sky , Beall Gary W. , Lewis L. Kevin
Published: June 2021

Clays are composed of alumina, silica, water and sometimes contain metal cations such as Mg2+ or Fe3+. Montmorillonite clays have an unusual crystalline structure; they form as platelets so that an Al3+ octahedron layer is in the middle with two tetrahedral silicate layers on the outside. The plates are turbostratic, meaning that the individual plates have no crystallographic relationship between them. Metal cations can be exchanged easily because of their attraction to the negative charge due to isomorphous substitutions of Mg2+ for Al3+ in the octahedral layer. The clay minerals are only 1 nm in width and up to 1000 nm long. Smectite clays are frequently combined with polymers to create new polymer‐clay nanocomposites with improved characteristics such as increased temperature resistance or strength. Polymer nanocomposites often require that they be surface modified with quaternary ammonium ions to render them compatible with the polymer. These organoclays are becoming more widely studied and utilized as surface bactericides. Two problems are currently associated with this; the nanocomposites may not be persistent or fully effective. In the current study, the toxicity and persistence of Na+ montmorillonite, quaternary ammonium ions, and a clay:quaternary ammonium organoclay in eukaryotic and prokaryotic cells have been investigated. Initial studies using the yeast S. cerevisiae have shown that montmorillonite is not toxic while the quaternary ammonium compound Arquad C‐33W was strongly growth inhibitory.
The effect of estrogen and a high salt diet on arteriolar reactivity to increased tissue oxygen in the spinotrapezius muscle of the female SS rat
Authors: Kunert Mary Pat
Published: June 2021

Estrogen may protect against salt induced microvascular dysfunction. Female SS (Dahl Salt Sensitive, SS/JrHsdMcwi) were divided into 2 subgroups: ovariectomized with (OVX +E) or without (OVX‐E) estrogen replacement. Three days prior to the acute experiment the diet of half of the rats was changed from low salt (LS, 0.4% NaCl) to high salt (HS, 8.0% NaCl). On the day of the acute experiment a transverse arteriole in the spinotrapezius muscle was selected, allowed to equilibrate with 0% O2 in the superfusion solution, and then changes in arteriolar diameter to 21% O2 in the superfusion solution before and after the topical application of HET‐0016 (10μM), a selective inhibitor of 20‐HETE production, were measured. Neither estrogen nor HS affected arteriolar constriction to 21% O2. HET‐0016 significantly attenuated arteriolar constriction to 21% O2 in the SS rat with or without estrogen and on a LS diet. However, the inhibition was greater in the SS rat without estrogen. HET‐0016 did not significantly inhibit the oxygen induced arteriolar constriction in the SS rat with or without estrogen and on a HS diet. These results suggest that a HS diet and estrogen reduce the contribution of 20‐HETE to oxygen induced arteriolar constriction in the spinotrapezius muscle. Funded by an internal UW‐M Research Growth Initiative grant.
Cardiotonic Pills®Post‐treatment Ameliorates Ischemia‐Reperfusion Induced Myocardial Microcirculatory Disturbances and Myocardial Lesions
Authors: Han Jing‐Yan , Wei Xiao‐Hong , Liu Yu‐Ying , Hu Bai‐He , Chang Xin , Zhao Na , Fan Jing‐Yu
Published: June 2021

Cardiotonic Pills® (CP) is a compound preparation of Chinese medicine widely used for treatment of cardiovascular diseases in China. We investigated the ameliorating effects of CP post‐treatment on ischemia‐reperfusion (I/R) induced myocardial microcirculatory disturbances and myocardial lesions. Anesthetized SD rats were subjected to 30 min of left anterior descending coronary artery occlusion and followed by reperfusion for 6 days. In some animals, CP (0.1 g/kg, 0.4 g/kg or 0.8 g/kg) was administrated once a day starting from 3 hrs of reperfusion until day 6. CP post‐treatment significantly attenuated I/R‐induced myocardial microcirculatory disturbance, including the decrease in red blood cell velocity and coronary blood flow, and the increase in albumin leakage. In addition, noticeable changes were observed at day 6 after reperfusion in myocardial infarct size, myocardial fibrosis size, myocardial ultrastructure, the number of myeloperoxidase‐ positive cells, and the expression of transforming growth factor β1 in myocardial tissue, which were significantly ameliorated by post‐treatment with CP. These results suggested that post‐treatment with CP ameliorates the myocardial microcirculatory disturbances, myocardial infarct, and myocardial fibrosis induced by I/R.
Muscarinic‐ and Nicotinic‐Acetylcholine Receptor dependent survival signaling in Human Coronary artery Endothelial Cells
Authors: Smedlund Kathryn Beatrice , Vazquez Guillermo
Published: June 2021

Atherosclerosis is a chronic inflammatory disease of the arterial wall and represents a major cause of death in western societies. The balance between apoptosis and survival signaling pathways is crucial to maintaining endothelial integrity during vascular injury. Not surprisingly, alteration of these processes is associated to atherogenesis. It has recently been shown that non‐neuronal nicotinic acetylcholine receptors (nAchR) are involved in the cholinergic anti‐inflammatory reflex signaling pathway, with potential anti‐atherogenic effects, at least at the macrophage level. Because both mAchR and nAchR are expressed in endothelium, we wanted to explore whether they might also exert a protective role in human coronary artery endothelial cells (HCAECs). Using a biochemical approach we found that AKT activation and the subsequent phosphorylation of its downstream target BAD are controlled by both mAchR and nAchR. Furthermore, stimulation of nAchRs caused phosphorylation of ERK1/2 and STAT3. Finally, activation of both mAchR and nAchR in HCAECs significantly reduced TNFα‐induced apoptosis (TUNEL assay). Altogether, our results suggest a pro‐survival, protective role of muscarinic and nicotinic acetylcholine receptors in HCAECs. These findings are discussed in the context of their potential impact in atherogenesis. Supported by UT‐COM and the American Heart Association (SDG0635250N to GV).
MicroRNA has‐let‐7g inhibits proliferation, adipogenic differentiation and inflammation in C3H10T½ cells
Authors: Yan Xu , Zhu Mei‐Jun , Du Min
Published: June 2021

MicroRNAs regulates gene regulation. We previously observed that let‐7g expression was reduced in the skeletal muscle with enhanced adipogenesis and inflammation, and hypothesized that let‐7g, a conserved microRNA, plays an important role in adipogenesis and associated inflammation. C3H10T½ mesenchymal stem cells stably over‐expressing let‐7g or a scrambled microRNA were selected following lentiviral transfection. The expression of let‐7g was 8 times higher in let‐7g infected cells compared with control cells. Let‐7g cells proliferated at 54.3 ± 10.9% of the rate of control cells. Let‐7g inhibited the mRNA expression of peroxisome proliferator‐activated receptor (PPAR)γ and CCAAT/enhancer binding protein (C/EBP)α, both are markers of adipogenesis, as well as the protein contents of PPARγ (57.1 ± 12.7% lower) and C/EBPα (56.1 ± 15.2% lower). Less adipocytes were formed in let‐7g cells. Let‐7g over‐expression inhibited the expression of inflammatory cytokines, tumor necrosis factor (TNF)α and interleukin (IL)6, demonstrating the anti‐inflammatory effect of let‐7g. In summary, let‐7g inhibits C3H10T½ cell proliferation and suppresses adipogenic differentiation. Let‐7g over‐expression also down‐regulates the expression of inflammatory cytokines. MicroRNA let‐7g may be an important target for the treatments of obesity and other metabolic diseases.
C2C12 cell culture model for investigating Ca2+ ‐dependence of myocyte differentiation
Authors: Jacobs Brittany , Chen Dapeng , Chin Eva R
Published: June 2021

Intracellular calcium (Ca2+) is required for myocyte differentiation into myotubes and has been shown to differentially regulate slow and oxidative skeletal muscle genes. The purpose of this study was to develop a cell culture model to examine the calcineurin and Ca2+ ‐calmodulin –dependent kinase II (CamKII) dependence of slow, fast, oxidative and glycolytic gene expression. C2C12 myogenic cells were differentiated in low serum media for 3d prior to treatment with 0.1 μM A23187 in the presence or absence of vehicle (Veh), calcineurin inhibitor cyclosporin A (CsA), CamKII inhibitor KN93 or its negative control KN92. After 5d drug treatment, cells were lysed, mRNA isolated and gene expression determined by quantitative RT‐PCR for Troponin I slow (TnIs; slow gene), myoglobin (Mb; oxidative gene), Troponin I fast (TnIf; fast gene) and muscle creatine kinase (MCK; glycolytic marker). 18S was used as control and expression levels calculated using the ΔΔCt method. TnIs, Mb, TnIf and MCK were markedly upregulated in differentiated myotubes, increasing 25866‐, 62870‐, 41627‐ and 59789‐ fold, respectively. CsA decreased the level of TnIs, Mb and TnIf expression, indicating calcineurin‐dependence of differentiation‐induced gene expression. Changes in gene expression with A23187 differed based on Veh for the inhibitor: no changes with ETOH+A23187 but all genes decreased with DMSO+A23187. DMSO+A23187 reduced TnIs (0.57±0.3), Mb (0.22±0.2), TnIf (0.48±0.4) and MCK (0.32±0.3) expression. This decrease was prevented by KN93 for Mb, TnIf and MCK but only Mb was not altered by KN92, indicating CamKII –dependence only of Mb down‐regulation. These data suggest a complex interaction of pharmacological agents in regulating gene expression in differentiated myotubes and indicate that well‐controlled experiments must be run to investigate the Ca2+ ‐dependence of muscle gene expression.
Liver afferents contribute to water drinking‐induced sympathetic activation in human subjects
Authors: May Marcus , Gueler Faikah , Barg‐Hock Hannelore , Engeli Stefan , Brand Andre , Sweep Fred , Tank Jens , Jordan Jens
Published: June 2021

Water drinking acutely increases sympathetic activity in human subjects. In animals, the response appears to be mediated through transient receptor potential channel TRPV4 activation on osmosensitive hepatic spinal afferents, the so called osmopressor response. We hypothesized that hepatic denervation attenuates water drinking‐induced sympathetic activation. We studied 20 liver transplant recipients (44±2.6 years, 1.2±0.1 years post transplant) as model of hepatic denervation and 20 kidney transplant recipients (43±2.6 years, 0.8±0.1 years post transplant) as drug matched control group. Before and after 500 ml water ingestion, we monitored brachial and finger blood pressure, ECG, and thoracic bioimpedance and obtained venous blood samples for catecholamine analysis. 30–40 minutes after water drinking, plasma norepinephrine had changed 0.0096±0.073 nmol/l in liver and 0.214±0.066 nmol/l in kidney transplant recipients (p<0.05 between groups). While blood pressure and systemic vascular resistance increased in both groups, the responses tended to be attenuated in liver transplant recipients. Our findings support the idea that osmosensitive hepatic afferents are involved in water‐drinking induced sympathetic activation in human subjects.
Spatiotemporal analysis of renal autoregulation
Authors: Scully Christopher G. , Cupples William A. , Gorbach Alexander M. , Braam Branko , Chon Ki H.
Published: June 2021

Laser speckle perfusion imaging was used to study vascular interactions among nephrons in relation to renal autoregulation on physiologically relevant spatial and temporal scales. Surface images of stabilized rat kidneys were collected with a moorFLPI imager (Moor Instruments, UK) at a sampling rate of 1 Hz. High resolution time‐frequency spectra (TFS) were generated for each pixel after application of a 2D spatial filter. The maximum amplitude from each TFS within the two autoregulation frequency ranges (tubuloglomerular feedback: 0.02 – 0.05 Hz and the myogenic mechanism: 0.1 – 0.3 Hz) were found at each time point, and the corresponding instantaneous frequency and phase were extracted. Frequency maps at each time point were constructed to visualize how autoregulation is organized across the kidney surface. Time‐varying, wide‐radius synchronization in the form of frequency‐locked pixels was observed. The reported methods can be used to study the synchronization of nephrons and the autoregulatory components involved. It can also be used to study pathophysiological consequences of loss of synchronization. This research was supported by CIHR MOP‐102694 and the Intramural Research Program of the National Institute of Biomedical Imaging and Bioengineering, NIH.
Acute mineralocorticoid receptor blockade produces rapid renal vasodilation with no impairment of autoregulation in vivo
Authors: Hamza Shereen M , Hildebrandt Drew A , Hall John E
Published: June 2021

Chronic blockade of the mineralocorticoid receptor (MR) significantly attenuates obesity‐induced rise in mean arterial pressure (MAP), sodium reabsorption and glomerular filtration rate (GFR). The mechanisms and immediate time‐course underlying these responses are not known but may involve rapid, non‐genomic actions of aldosterone at the MR. We determined the immediate renal hemodynamic response to MR blockade in anesthetized Sprague‐Dawley rats instrumented for measurement of MAP and renal blood flow (RBF). Animals were infused with the MR antagonist potassium canrenoate (20mg/kg i.v) or saline. Within 1 min of MR blockade, MAP fell (−9.1±3.6 mmHg, n=5, p<0.001) which was absent in saline controls (−0.4±1.6 mmHg, n=4). RBF rose (0.3±0.1 mL/min, n=5, p<0.001) with no change with saline alone (−0.1±0.1 mL/min, n=4). Renal arterial conductance (RBF/MAP) rose only with MR blockade (0.01±0.003 mLmin−1 · mmHg−1, n=5, p<0.001; saline −0.0003±0.001 mLmin−1 · mmHg−1, n=4) reflecting renal vasodilatation. Autoregulatory index was comparable in both groups (canrenoate 0.34±0.2 vs saline 0.21±0.1). We demonstrate rapid renal vasodilation in response to MR blockade in vivo suggesting differences between acute and chronic modulation of renal hemodynamics by aldosterone. NIH HL51972, CIHR
D1 Dopamine Receptor Modulation of Breathing in Young Hypothyroid Female Hamsters
Authors: Schlenker Evelyn , Wilson Marysa
Published: June 2021

Previously we noted in 8 month old female hamsters hypothyroid (H) for 5 months that administration of the dopamine D1 receptor antagonist SCH 23390 (SCH) in saline increased ventilation (VE) while animals were exposed to air, hypoxia and air following hypoxia (PH) relative to saline. These ventilatory responses to SCH were dissimilar to those observed in euthyroid age‐matched females (E). The increased VE's were due to increased tidal volumes (VT), but not frequency of breathing (F). In the present study effects of SCH on VE were studied in 15 four month old female hamsters rendered H for two months by administration of propylthiouracil in drinking water. Fifteen age and gender matched female E hamsters served as controls. Ventilation was measured in conscious hamsters using plethysmography. Relative to saline, 0.025 mg/kg SCH decreased VE during exposure of the H hamsters to air, PH and hypercapnia by decreasing F (due to increases of inspiratory & expiratory times), without affecting VT. Similar decreases in VE occurred in E hamsters exposed to air and hypercapnia. Hypoxic responsiveness, lower in H saline‐treated hypothyroid hamsters, was increased by SCH to levels comparable to those in E hamsters. No effects of SCH were noted on hypercapnic responsiveness. Thus, age and the length of hypothyroidism affect D1 receptor modulation of ventilation in female hamsters.

Funding: COBRE Pilot Grant
Feasibility of Stent Fracture and Expansion in the Palliation of Branch Pulmonary Artery Stenosis
Authors: Bates Melissa , Hembrook Kara , Smith Jenna , Anagnostopoulos Petros , Lamers Luke
Published: June 2021

Branch pulmonary artery stenosis (BPAS) is a common consequence of the repair of congenital heart disease in pediatric patients. Although a small stent may be placed to relieve the BPAS, this intervention is temporary as these patients will eventually outgrow the stent. The purpose of our study was to investigate the feasibility of a proposed method of catheter‐based palliation of BPAS with 1) placement of a small stainless steel stent followed by 2) repeat catheterizations with sequential dilation and eventual fracturing of the stent and 3) restenting of the BPAS with a larger, expandable stent. We first compared the bench‐top fracture pattern of the 5mm Paramount Mini GPS stent with the in‐vivo fracture pattern in a piglet model of BPAS in which a 4 mm Gortex band was placed around the left pulmonary artery at 3 weeks of age. The 5 mm stent fractured on its horizontal axis at 12 mm and 12 ATM and the vertical axis at 14 mm and 16 ATM. In‐vivo, a 7 mm Paramount stent was placed two weeks post‐surgical creation of the BPAS and expansion was performed at 7 and 11 weeks (73kg). This stent also fractured on it horizontal axis after dilation to 12 mm diameter but was unable to be fractured on the vertical axis despite high pressure balloon dilation to 16 mm and 20 ATM. The stent substantially for‐shortened without vertical fracture, documented with fluoroscopy and necropsy. In subsequent studies to be presented here, we will examine the impact of restenting and subsequent stent expansion on vascular growth and right heart function.
Discriminative Stimulus Effects of Naltrexone in Rats with Limited Access to Sucrose
Authors: Jewett David , Twaroski Kyleigh , Barlow Molly , Badzinski Hannah , Pagel Breanna , Levine Allen
Published: June 2021

In most operant paradigms, large doses of naloxone are typically required to establish naloxone as a discriminative stimulus in opioid‐naïve subjects. Chronic, but limited access to sucrose increases endorphin and dopamine function in rats. We wondered if rats given limited access to sucrose could be trained to recognize doses of naltrexone (NTX) that are typically not discriminable in operant paradigms. Prior to discrimination training, rats were trained to lever press and were then given access to sucrose (25% or 32%) for 12 hours a day for two weeks. Rats were then given daily injections of either saline or NTX (3.2 mg/kg, 15 min PT) 1 hour after sucrose access. After the training session subjects were returned to their home cage for the remainder of the 12 hour sucrose‐access period. Subjects learned to discriminate NTX from saline in a mean of 72 sessions (Md = 63, range 27‐135 sessions). No significant differences in acquisition of the NTX‐saline discrimination or initial NTX dose‐effect functions were noted between sucrose concentrations. These results provide further support for the notion that chronic sucrose consumption increases endogenous endorphin function.
Identification of Protein Complexes in mAKAP Signalosome by Mass Spectrometry
Authors: Suryavanshi Santosh , Singh Sonal , Bark Steven , McConnell Bradley
Published: June 2021

A‐Kinase Anchoring Proteins (AKAPs) are scaffolding proteins that bind protein kinase A (PKA). AKAPs can also bind other kinases, phosphatases, membrane receptors, ion channels and/or substrate proteins to mediate complex signaling pathways. Understanding of such AKAP signalosomes would be crucial to study crosstalk among different intracellular signaling pathways in the heart. Our lab works with two AKAPs, muscle‐specific AKAP, mAKAP (AKAP6, AKAP100) and Gravin (AKAP12, AKAP250). mAKAP is present on the nuclear envelope of cardiomyocytes and regulates multitude of cardiac hypertrophic pathways. Our lab has previously published that polymorphisms in human mAKAP significantly alter its binding with PKA and phosphodiestrase4D3 (PDE4D3) that might contribute to increased risk of heart failure. Therefore our aim is to identify all protein complexes in wild‐type (WT) and polymorphic mAKAP signalosomes. To study this, we performed immunoprecipitation (IP) experiments by expressing flag‐tagged WT and polymorphic mAKAP in HEK cells. By silver staining, we confirmed that our pulled‐down IP samples express other proteins along with mAKAP. Our next step would be to prepare samples for mass spectrometric analyses by trypsin digestion and analyze the data for possible binding partners of mAKAP. We are also looking into identifying binding partners of gravin by the above mentioned method. In conclusion, unambiguous identification of protein complexes in mAKAP (and gravin) using proteomics‐based approach would substantially help us to recognize their role in regulating cardiac physiology.
Buthionine Sulfoximine, an Inhibitor of Glutathione Biosynthesis, Induces Expression of Soluble Epoxide Hydrolase and Cellular Hypertrophy Markers in a Rat Cardiomyoblast Cell Line: Roles of The NF‐κB And MAPK Signalling Pathways
Authors: Abdelhamid Ghada , El‐Kadi Ayman
Published: June 2021

Evidence suggests that up‐regulation of soluble epoxide hydrolase (sEH) is associated with the development of cardiac hypertrophy. However, the up‐regulation mechanism is still unknown. In this study, we treated H9C2 cells with buthionine sulfoximine (BSO) to explore whether oxidative stress up‐regulates sEH gene expression and to identify the molecular mechanisms behind this up‐regulatory response. We found that BSO up‐regulated sEH at mRNA levels leading to increase in the cellular hypertrophic markers, atrial natriuretic peptide (ANP) and brain natriuretic peptide (BNP). Furthermore, BSO increased the cytosolic phosphorylated IκB‐α and translocation of NF‐κB p50 subunits. This level of translocation was paralleled by an increase in the DNA‐binding activity of NF‐κB P50 subunits. To understand the role of mitogen‐Activated Protein Kinases (MAPKs) pathway in BSO‐mediated induction of sEH mRNA, we examined the role of extracellular signal‐regulated kinase (ERK), c‐JunN‐terminal kinase (JNK) and p38 MAPK. Indeed, treatment with the ERK inhibitor, PD98059, partially blocked the activation of IκB‐α and translocation of NF‐κB p50 subunits induced by BSO. Moreover, pre‐treatment with ERK inhibitor and MEK inhibitors, U0126 and PD0325901, significantly inhibited BSO‐mediated induction of sEH and cellular hypertrophic markers gene expression. These results demonstrated that MAPK/NF‐κB signaling pathways is involved in BSO‐mediated induction of sEH gene expression. Furthermore, our findings provide strong link between sEH‐induced cardiac dysfunction and involvement of NF‐κB in the development of cellular hypertrophy.
The Cardioprotective Effect of Dexamethasone through Activation of RISK Pathway
Authors: Ma Yina , Wang Jinli , Techiryan George , Sun Wanqing , Wang Yanqing , Li Ji
Published: June 2021

Background

Treatment of ischemic heart disease, mainly focuses on returning blood flow back to the ischemic area. However, irreversible cell damage caused by reperfusion is the main risk of these approaches. The reperfusion injury salvage kinase (RISK) pathway can be targeted to decrease cell death and restore cell function during ischemia/reperfusion (I/R). Glucocorticoids are part of the feedback mechanism in the immune system that inhibits inflammation. Their intracellular effect is mediated by binding to the glucocorticoid receptor (GR). Dexamethasone(DEX) is a corticosteroid medicine used for anti‐inflammation which has a higher affinity to the GR than cortisol.

Methods

The effects of DEX was studied in an acute I/R injury model in which the left anterior descending coronary artery was occluded. Ex vivo mouse heart perfusion Langendorff system was used to measure the effects on glucose receptor 4 (GLUT4) translocation.

Results

‐Significant recovery of cardiac function was observed following DEX pretreatment during I/R injury. DEX activated the protective RISK pathways (includes ERK, AKT, and AMPK) significantly in a time and dose dependent manner. Furthermore, AMPK's translocation into nucleus was found after DEX treatment compared with I/R only. To deeply investigate the reason of AMPK translocation, immunoprecipitation was conducted. Results showed GR could interact with AMPK and recruit Sestrin 2‐LKB1 complex during I/R, which helped AMPK's phosphorylation and translocation during I/R. Moreover, one of AMPK's downstream effects ‐ stimulating GLUT4′s translocation to cell surface ‐ was also triggered after DEX stimulation.

Conclusion

DEX's cardioprotective function is achieved by activating the RISK protective signaling pathways during I/R.
The Calcium‐Activated Chloride Channel, Anoctamin‐1, is a Critical Regulator of Proliferation in Amphibian Tail Regeneration and This Effect is Mediated via p44/42 MAPK Phosphorylation
Authors: Franklin Brandon , Voss S , Osborn Jeffrey
Published: June 2021

Urodele amphibians have a unique ability among vertebrates to regenerate entire appendages including muscle, bone, neural and connective tissues. A better understanding of the cellular and molecular mechanisms involved in this process is critical for the advancement of translational therapies in regenerative medicine. We have recently shown that chloride channel inhibition, via 4,4′‐Diisothiocyano‐2,2′‐stilbenedisulfonic acid (DIDS), 4,4 '‐Dinitro‐stilbene‐2,2. '‐disulfonic acid (DNDS), T16a(inh)‐A01 or Cl‐ free environment effectively inhibits or delays tail regeneration in larval Ambystoma mexicanum salamanders. Proliferation was assessed via EdU incorporation assay, which revealed that this effect was manifested through regulation of mesenchymal cell proliferation, with all 3 pharmacological treatments reducing proliferation rate in the mesenchyme (Control: 33.5±3.2%; DIDS: 18.13±2.25%; DNDS: 19.64±3.79%; T16a(inh)‐A01: 19.65±1.85% EdU+ cells). There is no effect on proliferation in either neural or epidermal tissues. Western blot analysis reveals that there is a down regulation of p44/42 MAPK phosphorylation in response to anoctamin‐1 blockade (T16a(inh)‐A01). These results indicate that upon amputation the calcium activated chloride channel, anoctamin‐1, is activated and, through unknown intermediary signaling mechanisms, regulates proliferation via p44/42 MAPK. Further studies will investigate the effect of anoctamin‐1 inactivation on intracellular Ca+ dynamics in an attempt to elucidate the pathway between anoctamin‐1 and p44/42 MAPK.

Supported by Biomedical Sciences Research Group, LLC.
Concurrent Blockade of Muscarinic, Neurokinin‐1, and Serotonergic Receptors in the Ventral Respiratory Column of Intact Goats Does Not Affect Breathing
Authors: Muere Clarissa , Neumueller Suzanne , Olesiak Samantha , Miller Justin , Hodges Matthew , Pan Lawrence , Forster Hubert
Published: June 2021

Past and current studies in our laboratory have found that dialyzing antagonists of muscarinic, neurokinin‐1, or serotonergic receptors in the ventral respiratory column (VRC) of intact goats either increases or does not change breathing. However, we found that receptor antagonism alters the effluent dialysate concentration of one or more neuromodulators. These changes may have compensated for disruption of excitatory transmission in the VRC, preventing a decrease in breathing. Thus, we sought to test the hypothesis that simultaneous blockade of all three receptors with a cocktail of antagonists would elicit compensatory changes in multiple neurochemicals, but would ultimately decrease breathing. We dialyzed a cocktail of antagonists of muscarinic, neurokinin‐1, and serotonergic receptors into the VRC of adult goats, and collected the effluent dialysate for quantification of neurochemicals. Breathing was continuously monitored throughout the dialysis period. Data show that breathing is not significantly different (P>0.05, n=5) between time control studies and dialysis of the triple cocktail. Treatment with the cocktail tended to increase effluent substance P and decrease effluent glutamine, glycine, and GABA concentrations. Our results suggest that attenuation of multiple excitatory receptors in the VRC of intact animals may be compensated for by a reduction in inhibitory transmission to maintain a normal level of breathing.
Tissue‐Specific Abundance of NAMPT, CLOCK, and BMAL1 Proteins in Murine Tissues
Authors: Lupolt Alex , Brandauer Josef
Published: June 2021

Circadian Locomotor Output Cycles Kaput (CLOCK) and Aryl hydrocarbon receptor nuclear translocator‐like (BMAL1) are nuclear transcription factors that regulate protein expression oscillations in peripheral tissues. CLOCK and BMAL1 form a heterodimer complex that initiates the transcription of circadian‐regulated genes like nicotinamide phosphoribosyl transferase (NAMPT), the rate‐limiting enzyme in NAD biosynthesis. The abundance of these molecular clock proteins likely varies in a tissue‐ specific manner. We investigated protein abundance of CLOCK, BMAL1, and NAMPT in mammalian tissues.

10 male ICR mice were euthanized at ∼11:00AM. Various skeletal muscles (quadriceps (Q), gastrocnemius (G), extensor digitorum longus (EDL), soleus (S)), heart (H), liver (L), brown adipose tissue (BAT), visceral adipose tissue (VAT), and subcutaneous adipose tissue (SAT) were removed and immunoblots for NAMPT, BMAL1, and CLOCK were performed.

Relative CLOCK protein abundance was high in liver tissue, while BMAL1 expression was most highly expressed in mouse subcutaneous adipose tissue, visceral adipose tissue and heart. We detected statistically significant elevations in NAMPT protein concentrations in mitochondria‐dense murine tissues (soleus muscle, heart, liver, brown adipose tissue).

In conclusion, relative CLOCK and BMAL 1 protein abundances are not clearly correlated within tissues, even though these proteins form a heterodimer complex to initiate transcription of circadian‐regulated genes. These data could indicate that these proteins have additional functions within the cell that do not require their interaction. Highly oxidative tissues contained the highest amount of NAMPT, which may suggest that circadian variations are especially pronounced in mitochondria‐dense tissues.
Lipid Peroxidation Product, 4‐HNE (4‐Hydroxynonenal) Modulates hERG Channels
Authors: Choi Seong Woo , Lee Hyang‐Ae , Zhang Yin‐Hua , Kim Sung Joon
Published: June 2021

4‐Hydroxynonenal (4‐HNE) is considered as a harmful signaling molecule. The accumulation of 4‐HNE is implicated under oxidative stress such as ischemia/reperfusion injury. Previously we have reported the modulation of several ion channels including hERG K+ channels by exogenous electrophile molecules such as caffeic acid phenethyl ester (CAPE). However, the effect of endogenous electrophilic substances such as 4‐HNE has not been reported yet. Here we investigate the effects of 4‐HNE on hERG current overexpressed in HEK‐293 cells using whole‐cell patch clamp. Acute application of 4‐HNE (>30 mM) gradually but significantly decreased the tail current of hERG (IhERG,tail) within 3 – 10 min. Also, the half‐activation voltage (actV1/2) was shifted to the left. Both changes were not reversed by washout of 4‐HNE. Although lower concentraion of 4‐HNE (10 mM) had no significant acute effect, sustained incubation (1 ‐ 6 h) also induced time‐dependent suppression of IhERG,tail and leftward‐shift of actV1/2. Half‐inactivation voltage (inactV1/2) was affected by neither acute application nor pre‐incubation of 4‐HNE (1 to 100mM). Analysis of action potential duration (APD) was carried out in papillary muscles from rabbit heart using conventional intracellular microelectrode technique. Consisten with the suppression of IhERG,tail, APD was prolonged by perfusion of 100 µM 4‐HNE. Taken together, the suppression of hERG by 4‐HNE may participate in cytotoxic and proarrhythmic effects of lipid peroxidation under oxidative stress.