bluuebunny/crossref_metadata_2025 huggingface dataset:
Extracts of germinated wheat and barley reduce postprandial hyperglycemia by inhibiting α‐glucosidase in diabetic db/db mice
Authors: Jang Hae-Dong , Zhou Bing , Kwon Young-In
Published: June 2021

Extracts of germinated wheat and barley (EGW and EGB) were investigated as a potent cereal‐derived α–glucosidase inhibitor in diabetic db/db mice. They were prepared by soaking in distilled water with 0.5% tea catechin and 1% chitinoligosaccharide for 24 hr, extracting with autoclaving at 121 °C for 15 min, and freeze drying. The in vitro inhibitory activity of EGW and EGB on intestinal α–glucosidase, sucrase, and maltase was relatively weak compared to acarbose used as a positive control. However, sugar loading test with starch, sucrose, and maltose showed that significantly reduced postprandial increases in blood glucose, indicating that they may act as an intestinal α–glucosidase, sucrase, and maltase inhibitor. These results suggests that EGW and EGB could be used in the development of nutraceutical foods to control the blood glucose levels of diabetic patients by inhibiting intestinal α–glucosidase with reduced side effects.

Grant Funding Source : ASN
Comparison of Chemopreventive Properties of Zanthoxylum armatum and Hibiscus sabdariffa on colorectal adenocarcinoma cell (COLO 201) through apoptosis process
Authors: Khatiwada Janak R , Williams Leonard L , Davis Shurrita
Published: June 2021

Comparison of Chemopreventive Properties of Zanthoxylum armatum and Hibiscus sabdariffa on colorectal adenocarcinoma cell (COLO 201) through apoptosis process

Various concentrations of Zanthoxylum armatum and Hibiscus sabdariffa are used in this experiment to determine the optimum dose for cell cytotoxicity effects on COLO 201 cells. Antioxidant index of both phytochemicals were determine by using antioxidant assay kit. Annexin‐v, caspases 3/7 and in situ cell proliferation methods were used to determine the apoptosis induction through the flow cytometry. Results of this experiment indicated that lower concentration (5 mM/g) of both phytochemicals in combination induced significant amount of cell deaths in two stages of cell cycle (apoptosis 53.79 and early apoptosis 18.11%) compared to other treatment groups and control. Findings of this experiment support that the traditional use of these two herbs in the treatment of various chronic diseases is justified. These findings also suggest that the synergistic effect of these phytochemicals may reduce the incidence of colon tumors and can develop as a chemopreventive agent.
Dietary ellagic acid regulates serum cholesterol and retards atherosclerotic lesions in apoE knockout mice
Authors: Park Sin-Hye , Kang Young-Hee
Published: June 2021

Atherosclerosis is a major portion of death rates all over the world. Atherosclerosis comes up with hypercholesterolemia, hypertension, and heart attacks. The aim of this study is to investigate an atherogenic activity of ellagic acid in apoE knockout (KO) mice. ApoE KO mice are highly susceptible to spontaneous hypercholesterolemia and atherosclerosis. In this study apoE KO mice were fed Paigen's atherogenic diets for 10 weeks to induce atherosclerosis. Serum levels of triglycerides (TG), total cholesterols (TC) and LDL were much higher in apoE KO mice than wild‐type mice, indicating that atherosclerosis and hypercholesterolemia were induced in apoE KO mice. However, the levels of TG, TC and LDL‐cholesterol plummeted and HDL‐cholesterol level was enhanced with an oral supplementation of 10 mg/kg ellagic acid daily for 10 weeks. Atherosclerotic lesions in the aortas of apoE KO mice were ameliorated by treating 10 mg/kg ellagic acid, as assessed by H&E staining. Moreover, the cholesterol handling biomarkers of peripheral ABCA1, ABCG1 and SR‐B1 of mouse peritoneal macrophages were boosted by supplementing ellagic acid to apoE KO mice. Hepatic SR‐B1 and apoA‐1 level of apoE KO mice was also enhanced by 10 mg/kg ellagic acid. Therefore, dietary ellagic acid may be an atheroprotective agent against atherosclerosis by regulating peripheral cholesterol metabolism.
Population dynamics of leukocytes in tissues during immune activation of the chicken immune system by E. coli
Authors: Iseri Vanessa J. , Klasing Kirk C.
Published: June 2021

Immune tissues must have appropriate levels of nutrients to support changes in leukocyte numbers during an immune response, but quantitative aspects are poorly characterized. Examined cellular changes in immune tissues of adult Hyline chickens injected IV with dead E. coli. Bone marrow, spleen and thymus were isolated and total leukocytes and subpopulations analyzed in order to follow the cellular dynamics. No effect of E. coli on absolute weight of bone marrow or thymus (P >0.25, P=0.08). Liver weight increased at 24hr and 5d post‐injection (PI) compared to d0 (P=0.003, P=0.004). Spleen weight increased at 24hr and 5d PI compared to d0 (P=0.013, P=0.0002). Total number of leukocytes in bone marrow increased at 5d PI compared to baseline (P=0.006). Bone marrow subpopulations, CD8+, Bu1+, and KUL01+ numbers increased at 5d PI compared to baseline (P=0.001, P=0.02, P=0.03). In thymus, CD8+ numbers increased at 24hr and 5d PI compared to baseline (P=0.03). There was a trend for total leukocyte, CD4+, and CD4+CD8+ numbers to increase at 24hr and 5d PI compared to baseline (P=0.05, P=0.06, P=0.07). In spleen, Bu1+ number increased on 5d PI compare to baseline (P=0.04). When converted to a whole body basis the increases reported, corrected for the decreases in blood levels reported previously indicate that cell numbers, thus nutrient needs, are greater during systemic activation compared to naïve birds (maintenance).

Grant Funding Source: USDA Regional Research project 1013
Etiology of anemia in preschool children: A systematic literature review
Authors: Ariza-Nieto Magnolia , Carlson Gwen , Aragon Ingrid , Kordas Katarzyna , Fonseca-Centeno Zulma Yanira , Ocampo-Tellez Paul Rene , Pachon Helena
Published: June 2021

While 28% of Colombian preschool children were anemic in 2010, only 16% had iron‐deficiency anemia. A review was conducted to understand other potential causes of anemia in this age group, and to inform public health policy and programs in Colombia. Literature review methods were predefined and adjusted as needed, per Cochrane guidelines. Systematic searches of Medline, EMBASE, SciELO, and Cochrane Central were performed using a combination of MeSH terms and key words (anemia/etiology, hemoglobin) and limited by publication type (Humans, Clinical Trial, Randomized Controlled Trial, Case Reports, Controlled Clinical Trial, Multicenter Study), age (birth to <5 y), and publication dates (1990 – 2011). For example, the Medline search yielded 540 abstracts, which were independently evaluated by 2 reviewers. Of the 540 abstracts, 163 met inclusion criteria for anemia etiology. Among these publications, causes included sickle‐cell, thalassemia, nutrient intake, malaria, hemorrhage, helicobacter pylori, parasites and parvovirus B19. Conclusions regarding the potential etiologies of anemia in preschool children for whom iron deficiency is not the principal cause will be made upon completion of the literature review. Funding: ICBF, Pennsylvania State University and CIAT.
Nutritional status of young children with inherited blood disorders in western Kenya
Authors: Tsang Becky , Ruth Laird J. , Sullivan Kevin M. , Williams Tom N. , Suchdev Parminder S.
Published: June 2021

There is demonstrated high prevalence of hemoglobinopathies in Africa; however, it is unknown how blood disorders affect nutrition status. To determine the burden of anemia, iron and vitamin A deficiency, stunting, wasting, and underweight amongst young children in western Kenya with one or more blood disorders (haptoglobin 2‐2, glucose‐6‐phosphate dehydrogenase deficiency, sickle cell hemoglobin, and α‐thalassemia), we conducted a cross sectional survey of 882 children aged 6–35 months, randomly selected from 60 villages. Hemoglobin (Hb), ferritin, transferrin receptor (TfR), C‐reactive protein (CRP), alpha‐1‐acid glycoprotein (AGP), retinol binding protein (RBP), anthropometry, and blood disorders using PCR were measured. Of 861 children with Hb results, 71.7% were anemic (Hb<11 g/dL), 27.2% iron deficient (ferritin<12 μg/L, CRP<10 mg/L), and 16.8% vitamin A deficient (RBP<0.7 μmol/L, CRP<10 mg/L). Of the four blood disorders, anemia burden only differed between α‐thalassemics (82.3%), trait (75.6%), or without (66.8%) (p=0.002). Differences in vitamin A and iron deficiency, stunting, wasting, and underweight were not significant. The crude results suggest that in young children in western Kenya, α‐thalassemia is the only blood disorder associated with nutrition status, with children more likely to be anemic.

Grant Funding Source: Centers for Disease Control and Prevention, Rollins School of Public Health
Vitamin D in Patients with Tuberculosis or TB/HIV coinfection in Tanzania
Authors: Mehta Saurabh , Mugusi Ferdinand M , Bosch Ronald J , Aboud Said , Urassa Willy , Villamor Eduardo , Fawzi Wafaie W
Published: June 2021

This study was conducted to examine vitamin D status in adults with tuberculosis (TB) or TB/HIV co‐infection enrolled in a vitamin supplementation trial (excluding D) in Tanzania and its relationship with mortality, HIV disease progression, and relapse during follow‐up. Serum 25(OH)D levels were measured using mass spectrometry. Binomial and Cox regression methods were used to analyze the associations between vitamin D and outcomes. 62% of patients had low vitamin D status (<30 ng/mL) and 16% were deficient (<20 ng/mL). Hemoglobin levels, CD4 T‐cell counts, and money spent on food were significant correlates of low vitamin D status. In longitudinal analyses, there was no significant relationship between vitamin D status and mortality or HIV disease progression. However, patients with low vitamin D levels had 66% higher risk of relapse during follow‐up (95% CI: 1.04–2.64; p=0.03; adjusted for age, Karnofsky score, hemoglobin, CD4 T‐cell counts, HIV status, viral load, and treatment regimen). In HIV‐uninfected individuals, the risk of relapse was 133% higher (p<0.01). Vitamin D may have an important role in patients with TB and it needs to be examined in randomized clinical trials.

Grant Funding Source : NIH NIAID U01AI045441
Effect of Sprinkles on reducing anemia in young children in Cambodia: a cluster‐randomized effectiveness trial
Authors: Jack Susan , Ou Kevanna ,   Good Food for Children Steering Committee, de la Barra Sophia Leon , Hill Philip , Herbison Peter , Gibson Rosalind
Published: June 2021

Whether anemia can be significantly reduced by delivery of micronutrient powders (Sprinkles) through government health services is uncertain. We evaluated the effectiveness of daily Sprinkles mixed with complementary foods in infants from aged 6 to 11 months (mo) with infant and young child feeding education (IYCFE). We followed 1350 randomly selected children at 6 mo intervals to aged 24 mo via biomarkers and anthropometry to establish whether any observed effects on anemia, deficiencies of Fe, vitamin A, Zn, and growth were sustained. Sprinkles plus IYCFE vs. IYCFE alone reduced any anemia at 12 mo by 24% (hemoglobin (Hb)<110g/L, Incidence Rate Ratio (IRR) 0·76, 95% CI 0·64 to 0·89, P=0·001) and moderate anemia (Hb<100g/L) by 57% (IRR 0·43, 95% CI 0·33 to 0·56, P<0.001). At later follow ups there were no significant differences in any anemia between groups. Iron deficiency was reduced by 57% (IRR 0·43, 95% CI 0·32 to 0·58, P<0.001) and iron deficiency anemia (IDA) and non‐IDA were reduced for moderate anemia at 12 mo. We found a significant increase in mean serum zinc (0·44 μmol/L, 95% CI 0·04 to 0·83, P = 0·028) but no difference in zinc and vitamin A deficiency, or growth at any time. Sprinkles reduced anemia, and increased mean serum zinc in Cambodian infants. These effects were in addition to IYCFE alone, but did not persist beyond the intervention. A2Z, USAID; Cambodia Health Sector Support Projects I & II; WHO Cambodia; GAIN.

Grant Funding Source : A2Z, USAID; Cambodia Health Sector Support Projects I & II; WHO Cambodia; GAIN
Effects of acute and chronic almond consumption on glycemia in adults at risk for diabetes
Authors: Tan Sze Yen , Mattes Richard
Published: June 2021

This study examined the effects of almond consumption on acute glycemic control in adults at risk for diabetes and assessed whether the effects are sustained during 4 weeks of daily almond consumption. There were 5 study arms which entailed consuming 43g of almonds with breakfast or lunch, and as a morning or afternoon snack, or no almonds. At baseline, participants attended a 490‐min acute‐feeding session, were provided with standard meals and almonds, and blood glucose and insulin were measured at designated intervals. They were then instructed to eat almonds daily for the following 4 weeks before repeating the acute‐feeding session. During the initial acute‐feeding session, blood glucose changed significantly (time & interaction effects, P<0.001), but insulin changes did not differ between groups. Incremental AUC (iAUC) showed lower 60‐min postprandial glycemia with almonds eaten as snacks (P<0.05). Similar trends were observed when they were consumed with meals. Repeated consumption of almonds over 4‐weeks did not alter the 490‐min glycemia trends and 60‐min postprandial iAUC, but the statistical significance diminished. Body weight increased after 4 weeks but not differentially between groups. Body fat remained unchanged. Our study suggests that ingesting almonds as a snack may assist in acute glycemic control, and this effect and body weight are minimally affected by prolonged consumption.

This study was funded by the Almond Board of California.
Unique microRNA expression patterns associated with macrophage infiltration following skeletal muscle injury
Authors: Melton David Wayne , Gelfond Jonathan , Chen Yongxin , McManus Linda , Shireman Paula
Published: June 2021

Skeletal muscle regeneration following injury requires macrophages. MicroRNAs (miRNAs) are vital to macrophage biology and may be key regulators of muscle regeneration. We compared temporal expression of miRNA in cardiotoxin‐injured muscle derived from wild type (WT, C57Bl/6J) and CCR2−/− mice to identify macrophage‐specific miRNA patterns. MiRNA expression was determined on muscle RNA by qRT‐PCR in TaqMan arrays. Compared to uninjured muscle, log‐fold changes in miRNA expression at 1, 3, or 7 days following injury (n=4 mice/group/timepoint) were determined. Flow cytometry confirmed impaired macrophage recruitment in CCR2−/− mice. The temporal expression patterns of 60 miRNAs were significantly different between strains, including miRNAs known to be highly expressed in macrophages (e.g., miR‐146a, miR‐106b, and miR‐221). Interestingly, 15 miRNAs with no known relationship to macrophages were matched with the temporal expression patterns of macrophage‐related miRNAs. These 15 miRNAs may be unique, macrophage‐related miRNA that regulate the muscle regenerative response. This study furthers our knowledge of miRNA regulation of the regenerative response in injured skeletal muscle and identifies potential miRNA targets for advancing tissue regeneration following traumatic injury. Funding: Veterans Administration and NIH (HL110743 and HL074236).
Status of muscle signaling pathways in the early pathogenesis of dystroglycanopathy‐type muscular dystrophy
Authors: Beedle Aaron M , Melick Garrett , Ball Charlotte , Fortunato Marisa
Published: June 2021

A subset of muscular dystrophies are caused by abnormal glycosylation of α‐dystroglycan. Without its proper sugar structure, α‐dystroglycan binding to extracellular matrix proteins (e.g. laminin, perlecan) is compromised, disrupting the structural integrity of muscle and other tissues. Mutations in the FCMD gene, encoding fukutin, cause severe and milder dystroglycanopathies, Fukuyama congenital muscular dystrophy (FCMD) and limb‐girdle muscular dystrophy type 2M (LGMD 2M), respectively. Using fukutin conditional knockout mice, we have evidence that severe congenital muscular dystrophy phenotypes involve abnormal skeletal muscle development and/or differentiation. We will present a molecular, biochemical and histological approach, screening select muscle signaling pathways in young knockout mice, to identify abnormalities in skeletal muscle development/differentiation. These findings bring new insight to potential mechanisms underlying early disease pathogenesis and severe phenotypic outcomes in dystroglycanopathy‐type congenital muscular dystrophies.

This work has been supported by the Muscular Dystrophy Association (Development Grant) and the University of Georgia, College of Pharmacy.
Antagonism of 5‐HT2A receptors attenuates the effects of amphetamine and 3,4‐methylenedioxymethamphetamine on wakefulness and dopamine overflow
Authors: Murnane Kevin Sean , Howell Leonard Lee
Published: June 2021

The neuropharmacology of the sleep and arousal systems involves a complex interplay between multiple neurotransmitter systems. Previous work has downplayed the role of dopamine in sleep and wakefulness, at least in part, because dopamine neurons do not appear to change their firing rates across arousal states. In the present study, we examined the wake promoting effects of amphetamine and 3,4‐methylenedioxymethamphetamine (MDMA) in rhesus monkeys using noninvasive telemetric monitoring. To evaluate the importance of dopamine, we used in vivo microdialysis targeting the caudate nucleus and pretreatment experiments with the selective 5‐HT2A receptor antagonist M100907. We targeted 5‐HT2A receptors because previous studies found that antagonizing this receptor attenuates some of the behavioral and neurochemical effects of amphetamine and MDMA. We found that amphetamine and MDMA significantly and dose‐dependently increased wakefulness. The effects of amphetamine were more pronounced than those of MDMA, but this may have been the result of different durations of action. Antagonism of 5‐HT2A receptors attenuated the effects of amphetamine and MDMA on both wakefulness and dopamine overflow. These findings suggest that increased dopamine neurotransmission may be necessary for the wake promoting effects of amphetamines. These studies were supported by USPHS grants (DA 00517 and RR 00165).
The Effect of Chronic Morphine Treatment on the Abundance of Cell Signaling Proteins in the Guinea Pig Longitudinal Muscle‐Myenteric Plexus
Authors: Thompson Ben G , Li Peng , Thayne Kathleen , Masterson Jackie , Taylor David A
Published: June 2021

Chronic exposure to morphine produces heterologous tolerance in the longitudinal muscle/myenteric plexus (LM/MP) of the guinea pig. A decrease in the α3 subunit of the sodium pump was suggested as one mechanism since it develops over a time course similar to the reduced function. However, a large number of cell signaling proteins have also been proposed as contributors to the adaptation process. We examined the abundance of several different proteins using quantitative western blot analysis in homogenates of LM/MP after chronic morphine exposure. We found no change in the abundance of the mu opioid receptor protein either 4 or 7 days after pellet implantation when the loss of function occurs and becomes maximal. We also found no change in the abundance of the α1 subunit isoform of the sodium pump, PKCε, beta actin or GAPDH at any time period from 1–14 days after exposure. In contrast, we observed an increase in the abundance of calcineurin, GRK2, and Giα2 and a decrease in the abundance of PKCγ and the alpha3 subunit isoform at 7 days after implantation. The lack of correlation of changes in protein abundance with functional changes suggests that the heterologous tolerance that develops after chronic morphine treatment may involve multiple cellular mechanisms including the modulation of cell excitability and normal physiology which result from changes in the abundance of proteins in several signaling pathways.
Knockdown of Natriuretic Peptide Receptor‐ A Enhances Receptor‐C Expression and Signaling in Vascular Smooth Muscle Cells (VSMC)
Authors: Li Yuan , Madiraju Padma , Anand-Srivastava Madhu B.
Published: June 2021

Natriuretic peptide receptor‐A (NPR‐A) knockout mice were reported to exhibit an increased blood pressure which may also be attributed to the upregulation of NPR‐C and associated signaling, however, the interaction between the two receptors was not investigated. In the present study, we investigated the effect of knockdown of NPR‐A using NPR‐A antisense (AS) on the expression of NPR‐C and adenylyl cyclase (AC) signaling in VSMC. Treatment of VSMC with NPR‐A AS decreased the expression of NPR‐A and enhanced the expression of NPR‐C but not of AT1 and M2 receptor. In addition, siRNA‐NPR‐A also upregulated NPR‐C. The re‐expression of NPR‐A in AS‐treated cells reversed the enhanced expression of NPR‐C to control levels. In addition, the receptor‐mediated inhibitions of AC activity and Giα expression were enhanced in AS‐treated cells, whereas NPR‐A‐mediated cGMP formation was significantly reduced. Pertussis toxin treatment attenuated AS‐induced enhanced inhibitions of AC to control levels. Furthermore, the enhanced levels of NPR‐C and Giα proteins were reversed to control levels by 8Br‐cGMP and PD98059. In addition, 8Br‐cGMP also attenuated AS‐induced enhanced ERK1/2 phosphorylation to control levels. These results demonstrate that knockdown of NPR‐A upregulates the expression of NPR‐C, Giα proteins and NPR‐C‐linked AC signaling and suggest a cross‐talk between NPR‐A and NPR‐C(Supported by CIHR).
Moderate exercise improves the metabolic response to a fat/sugar overload but exacerbates cardiac damage
Authors: Pohlman Roberta L , Alghamri Mahmoud S , Bechara Ana Carolina C , Hartzler Lynn , Mirkin L David , Farah Vera , Morris Mariana
Published: June 2021

A western diet, high in fat and sugar, causes metabolic and cardiovascular dysfunction. The objective was to determine the effect of moderate swimming exercise (1 hr/d, 3 d/wk, n = 9–10/group) on the metabolic and cardiac responses to a high fat/sugar diet in mice. Groups are: Control (CON); Sedentary (SED, fat/fructose) and Exercise (EX, fat/fructose). Chow (60% fat) and fructose water (10%) were given ad libitum for 8 wk. Parameters measured were body fat, heart function (echocardiography, Echo), glucose, glucose tolerance (GTT) and liver glycogen. Mice gained body fat (up to 45%) on the fat/fructose diet with no difference between groups. Metabolic function was improved in the EX group as determined by glucose (SED>EX p <0.01) and GTT (SED>EX, p< 0.05). Insulin was lower in EX than SED, suggesting improvement in function. The diet increased liver glycogen, an effect which was improved in the EX group. Echo measurements showed impairment in cardiac function in the EX group. Relative wall thickness was higher in EX vs SED (0.35±0.01 vs 0.29 ±0.01, p<0.01). Fractional shortening was reduced in EX group (EX 0.31±0.01 vs SED 0.36±0.01, p < 0.01). This resulted in a reduced ejection fraction of ~37% vs 49% (EX vs SED). Data demonstrate that a moderate exercise paradigm combined with a high fat/sugar diet improves metabolic function but is detrimental to heart function in morbidly obese mice. Supported by NIH R01 HL093567
Manipulating IP3R‐mediated calcium release in permeabilized endothelial cell tubes of resistance arteries
Authors: Socha Matthew J. , Domeier Timothy L. , Segal Steven S.
Published: June 2021

Calcium signaling is integral to endothelial cell (EC) function and is governed by stimuli originating both within the vessel lumen and from surrounding cells. Our goal was to gain insight into the intrinsic properties of inositol trisphosphate receptor (IP3R)‐mediated Ca2+ release of resistance artery endothelium under conditions in which the endoplasmic reticulum functions as an isolated signaling organelle. Intact EC tubes were isolated from superior epigastric arteries of male C57BL/6 mice, permeabilized with saponin, and imaged using laser‐scanning confocal microscopy. Exposure to internal solution containing 100 nM free Ca2+ (with 40 μM Fluo‐4) evoked Ca2+ release events at a frequency of 3.9 ± 0.5 events/EC/min; these typically spread along entire ECs at ~15 μm/s (vs. ~17 μm/s for intact EC tubes). Release events were suppressed (P<0.05) by removing free Ca2+ with 500 μM EGTA (0 events), depleting ER Ca2+ stores with 10 μM thapsigargin (0 events), or by inhibiting IP3R with 100 μM 2‐APB (0.4 ± 0.2 events/EC/min; P<0.05). Remarkably, increasing IP3 levels in the internal solution tended to decrease Ca2+ release events (5 nM: 2.1 ± 0.5; 50 nM: 1.5 ± 0.4; 10 μM: 0.5 ± 0.5 events/EC/min). We conclude that IP3R‐mediated Ca2+ signaling can be manipulated in permeabilized endothelium of resistance arteries to evaluate key determinants of EC function in health and disease. (NIH R37HL041026, R01HL086483, F32HL107050)
β1 integrin and associated focal adhesion proteins in cardiac fibroblast function
Authors: Au Carol G. , Manso Ana Maria , Asfaw Elizabeth K. , Liao Peter L. , Ross Robert S.
Published: June 2021

β1 integrin is a member of the integrin receptor family of adhesion molecules that links cells to the extracellular matrix (ECM). Although cardiac fibroblasts are the major cell type responsible for ECM production in the heart, the role of β1 integrin in cardiac fibroblast function has not been defined. Stimulation of primary adult mouse cardiac fibroblasts with transforming growth factor (TGF)‐β1 (10 ng/mL) increased protein expression of β1 integrin, talin and vinculin, and this was associated with enhanced myofibroblast conversion as delineated by α‐smooth muscle actin levels. Our group has generated cardiac myocyte‐specific knockout mouse models of β1 integrin, talin or vinculin which develop dilated cardiomyopathies characterized by progressive cardiac fibrosis. To delete these genes in cardiac fibroblasts, we utilized an ex vivo Cre‐LoxP strategy whereby cultured fibroblasts were infected with an adeno‐Cre recombinase virus. Fibroblast depletion of β1 integrin destabilized focal adhesion complexes and reduced adhesion to fibronectin, but did not alter TGF‐β1‐ mediated myofibroblast conversion. A similar reduction in adhesion to fibronectin was observed in cardiac fibroblasts depleted of talin. In summary, β1 integrin and associated focal adhesion proteins can modulate cardiac fibroblast function and may therefore influence the development of cardiac fibrosis.

Supported by grants from NIH (HL088390, HL46345) and AHA (10POST3520045)
Alterations in Left Ventricular Preload Dynamically Regulate Cardiac Fibroblast Signaling
Authors: Galantowicz Maarten L , Guggilam Anuradha , Cismowski Mary J , Zhang Xiaojin , West Thomas A , Lucchesi Pamela A
Published: June 2021

Left ventricular (LV) hemodynamic volume overload (VO) inhibits cardiac fibrosis in vivo. However, the effects of hemodynamic load changes on connective tissue growth factor (CTGF) and TGF‐β1induced extracellular matrix (ECM) proteins are poorly understood. This study investigated the impact of load‐driven changes on cardiac fibroblast (CF) phenotype and signaling.

Methods

Sprague‐Dawley rats underwent sham/aortocaval fistula (ACF) surgery to induce VO heart failure for 4 weeks (compensatory stage), followed by a load reversal procedure in a subset of rats. Primary cultured LV CFs were obtained at 5 day post VO reversal, and ECM signaling molecules were studied by protein and gene expression profiling.

Results

Collagen 1 secretion was decreased in both ACF and reversal CFs, while CTGF mRNA expression decreased only in ACF CFs, consistent with an anti‐fibrotic phenotype. Interestingly, ACF and reversal CFs demonstrated increased TGF‐β1 mRNA expression and similar Smad activation in response to exogenous TGF‐ β1. However, β1‐integrin expression and downstream AKT signaling was suppressed in ACF CFs compared to sham, but normalized following VO reversal.

Conclusion

These data suggest that hemodynamic load modulates CF signaling and secretion of ECM proteins. Molecular profiling is necessary to understand how LV hemodynamic changes impact CF‐dependent ECM regulation.
Epigenetics of Transgenerational Transmission of In Utero Nicotine‐Induced Asthma
Authors: Rehan Virender , Liu Jei , Sakurai Reiko , Torday John S
Published: June 2021

Whether the in utero nicotine‐induced asthma risk is transmitted transgenerationally, and whether PPARγ agonists have any effect on this transmission risk are not known. Time‐mated Sprague Dawley rat dams received either placebo or nicotine from day 6 of gestation to postnatal day (PND) 21. Following delivery, at PND21, pulmonary function tests, trachea tension studies, and expression levels of contractility proteins in the lungs and tracheas of generation 1 (F1), F2, and F3 rats were determined. As the putative basis for the transgenerational (TG) effect of nicotine on asthma, epigenetic changes in the lungs and gonads of the F1 offspring and the preventive effect of PPARγ agonist rosiglitazone (RGZ) on these epigenetic changes and TG asthma risk were determined. In comparison to the control group, with perinatal nicotine exposure only to F0 rats, 1) F1 through F3 pups demonstrated the asthma phenotype in a gender‐specific manner, with more pronounced effects in males, along with global DNA methylation increase in the testes, decrease in the ovaries, without any change in the lung. All of these changes were blocked by RGZ administration during the perinatal period.

(Grant Support: HL75405, HD51857, HD058948, HL55268, and 17RT‐0170).
Increased claudin‐5 increases lung epithelial permeability and is associated with disruption of tight junction assembly
Authors: Overgaard Christian , Mitchell Leslie A. , Ward Christina , Guidot David M. , Koval Michael
Published: June 2021

Chronic alcohol abuse increases the risk of developing the Acute Respiratory Distress Syndrome (ARDS) 3–4‐fold. Based on experimental models, alcohol disrupts alveolar epithelial permeability and is associated with an increase in claudin‐5 (cldn‐5). Here we used viral expression systems to alter cldn‐5 expression by cultured primary rat alveolar epithelial cells (AECs). AECs transduced to express increasing levels of cldn‐5 showed decreased transepithelial resistance (TER), which plateaued at 80% of control levels. Conversely, decreasing cldn‐5 using shRNA increased TER of normal and “alcoholic” AECs by 25–35%, thus improving barrier function. By immunofluorescence, cells expressing increased cldn‐5 showed disrupted localization of cldn‐18 and ZO‐1, where cell junctions had an increase in strand breaks and inward projections (“spikes”) suggesting decreased stability. Analysis of differential Triton X‐100 solubility was used to biochemically confirm these observations. This suggests a model for tight junction assembly, where bidirectional interplay between claudin and scaffold protein composition determine tight junction morphology and function.
Role of sodium pump β1 subunit in adult mouse lung alveolar fluid homeostasis
Authors: Flodby Per , Borok Zea , Gao Danping , Kim Yong Ho , Kim Kwang-Jin , Crandall Edward D
Published: June 2021

Regulation of salt and water balance in the lung involves active alveolar epithelial ion (Na+) transport. The basolateral Na+, K+‐ ATPase (sodium pump) plays a crucial role by creating ionic and osmotic gradients across alveolar epithelium, leading to absorption of salt and water from alveolar spaces. The sodium pump contains three subunits (α,β,γ). The β1 subunit is the most highly expressed β isoform in adult lung, being present in both alveolar epithelial type I (AT1) and type II (AT2) cells. To elucidate the importance of the β1 subunit in adult mouse lung and its relative importance in AT1 vs AT2 cell function, we generated mice with knockout of the β1 subunit gene (Atp1b1) specifically in AT1 cells (Atp1b1Aqp5‐cre mice) or in both AT1 and AT2 cells (Atp1b1Sftpc‐cre mice). Lung transport function was assessed by alveolar fluid clearance (AFC) measurements in anesthetized mice. AFC in Atp1b1Aqp5‐cre mice was reduced 45%, while AFC in Atp1b1Sftpc‐cre mice was reduced almost 80%, compared to floxed controls (Atp1b1F/F). Terbutaline, a β‐adrenergic agonist, increased AFC in Atp1b1Sftpc‐cre mice (from 6.6 to 30.8% per hour) compared to controls (from 29.8 to 36.4% per hour). These data (1) indicate significant contributions to AFC from both AT1 and AT2 cells and (2) suggest the presence of β‐agonist‐inducible mechanisms that compensate for lack of the β1 subunit.

Funding: NIH; Hastings and Whittier Foundations
Muscle size assessment with panoramic ultrasound: measurement reliability among varying body masses
Authors: Salley Eric Joseph , Hayes Erik Steven
Published: June 2021

Panoramic ultrasound (US) has recently been suggested as a reliable means of assessing muscle cross‐sectional area (CSA) with high intra‐day reliability between images. However, neither inter‐day reliability nor intra‐day reliability across populations with significantly different body masses has been examined. Therefore, the purpose of this study was to examine both intra‐ and inter‐day reliability measures of the CSA of the vastus lateralis (VL) in participants with varying body masses. 15 participants (age 20.7 ± 1.1 yrs) were divided into three groups according to mass: group A (59.6 ± 7.4 kg), group B (77.5 ± 8.1 kg), and group C (101.8 ± 8.9 kg). Three images taken at the mid‐thigh were obtained on three different days. The CSA was measured for each image using manual planimetry and the coefficient of variation (CV) was calculated. The intra‐day CV for groups A, B, and C was 1.70 ± 0.60, 1.71 ± 0.70, and 1.67 ± 0.95, respectively. The inter‐day CV was similar for groups A and B (4.07 ± 1.96 and 4.07 ± 2.48 respectively), but slightly higher (5.63 ± 2.36) for group C. These results are similar to previous research, suggesting that intra‐day reliability of panoramic US images measuring CSA is high. However, lower inter‐day reliability of the current method suggests caution should be taken when using panoramic US in longitudinal studies where small changes in muscle size may occur. Funding: TU CR&I
No thermal effect of GSM Electromagnetic Waves Exposure on edl rat skeletal muscles
Authors: Shouaib Sara , Ramadan Wiam , Ibreik Aida , Khachfe Hassan , Jbai Fatima , Joumaa Wissam
Published: June 2021

One of the important challenges developed recently and could affect health is the population exposure to electromagnetic waves (EMW), particularly the ‘Global System of Mobile Communication signals’, GSM signals (1). GSM are emitted from many sources as like cell phones and base stations antenna. We investigate the effects of EMW radiation emitted from GSM mobile phones antenna (frequency at 900 MHz, Eeff = 25 V/m) on edl muscle myofibrillar proteins. Animals body weight was not affected by 6 weeks of continuous 24 hours exposure, but edl absolute muscle mass was increased by 10.5% (control: 0.1744±0.00252, exposed: 0.1927±0.00291, n=12, p<0.05). Since myofibrillar proteins content is directly proportional to muscle mass (2), protein dosage using the Bradford technique was performed and protein concentration was calculated in mg/g of muscle. An increase by 59% in edl myofibrillar protein content after the period of exposure was observed (control: 20.11±1.59, exposed: 31.97±2.81, n=12, p<0.05). The analysis of MHC isoforms by SDS‐PAGE as already described by Talmadge et al. (3) shows an increasing by 431.54% in MHC I, 35.08% in MHC IIa a decreasing by 16.34% in MHC IIb without a significant modification in the percentage of expression of MHC IIx/d (n=12 and p<0.05). Thus, 6 weeks of continuous whole body exposure to EMW can induce a no thermal effect in myofibrillar proteins of fast‐ twitch mammalian skeletal muscles.
Effect of temperature on isoproterenol‐induced increases in left ventricular developed pressure
Authors: Klabunde Richard , LePorte Andrew D. , Wilson Thad E.
Published: June 2021

The goal of this project was to determine the effects of elevated temperature on preload‐dependent and independent regulation of left ventricular developed pressure (LVDP) in Langendorff‐perfused, electrically paced (420 bpm), Sprague‐Dawley rat hearts. LVDP responses to steady‐state isoproterenol (ISO) infusions (10−8 M) were determined at 37, 38, 39, and 40ºC. Preload‐dependent LVDP was determined at 37 and 40ºC. ISO‐induced LVDP and preload‐dependent LVDP time controls were conducted in a separate group at 37ºC. The percent increase in LVDP during ISO infusion significantly decreased to 42±6(SE) mmHg at 40ºC, compared to 55±9, 55±6, and 53±7 mmHg at 37, 38, and 39ºC, respectively. No significant differences were observed in corresponding time controls (50±6, 47±3, 56±4, and 56±5 mmHg). Preload‐dependent LVDP decreased across the experimental protocol, but there were no temperature effects. These data indicate that intrinsic heart contractility is not altered by moderate heating but is compromised at very high temperatures (40ºC). Furthermore, local temperature does not alter the inherent preload‐dependent LVDP.

Funded by a NIOSH Education and Research Center Grant #T42/OH008432‐05.
Cardiac mitochondrial proteomic expression in inbred rat strains divergent in survival time after hemorrhage (STaH)
Authors: Klemcke Harold G. , DeKroon Robert M. , Mocanu Mihaela , Robinette Jennifer B. , Alzate Oscar
Published: June 2021

We have previously identified inbred rat strains different in STaH (DA = FHH > BN/Mcwi). In efforts to identify cellular mechanisms and ultimately genes important to STaH, we measured cardiac mitochondrial proteins (MP) in rats from these three inbred strains (BN/Mcwi, DA, and FHH). Rats were euthanized (n = 12/strain), hearts were removed, and mitochondria were isolated. MP expression was determined using 2D DIGE‐based Quantitative Intact Proteomics. Among all strains, 1476 MP spots were detected. MP differed among strains [DA vs BN/Mcwi, n= 485 MP differed; DA vs FHH, n= 314; BN/Mcwi vs FHH, n= 430; p ≤ 0.04]. Eighty‐one MP spots were isolated and identified (IMP) using matrix‐assisted laser desorption ionization time of flight mass spectrometry. Of these, 38 distinct proteins were present and 43 IMP represented multiple posttranslational modifications of 24 of these distinct proteins. For DA rats (longest STaH), 36 IMP were increased (P ≤ .036) whereas 30 IMP were decreased (p ≤ 0.03) compared with BN/Mcwi (shortest STaH). IMP were associated with nucleic acid, lipid, and carbohydrate metabolism and with energy production. Such data indicate multiple differences exist in cardiac mitochondrial proteins among these inbred rat strains, and suggest the possibility that such protein differences might be among those cellular mechanisms influencing differences in STaH. (Funding by USAMRMC)
Comparison of clock gene expression across skeletal muscles of different origins and functions
Authors: Weekman Erica Marie , Zhang Xiping , Esser Karyn A
Published: June 2021

Circadian rhythms are approximate 24 hour cycles and are associated with many physiological processes in the body. The molecular clock mechanism that underlies circadian rhythms has been found to exist in all cells in both central and peripheral tissues. Our lab has demonstrated that clock genes oscillate in gastrocnemius muscle but nothing is known about clock gene expression in muscles that have different developmental origins and different functions. In this study we compared clock gene expression in gastrocnemius, tongue and diaphragm muscles that were collected from mice every 4 hr for 28 hr. RNA was isolated from each tissue and real time polymerase chain reaction (PCR) was performed to determine mRNA levels of select clock and clock‐controlled genes. The results indicate that core clock genes, Bmal1 and Per2, oscillate with a similar phase across all three muscles. However we found that other clock genes, including Per1 and Cry2 oscillated in gastrocnemius and diaphragm but not tongue. Previously characterized clock controlled genes, MyoD and PGC1 á were found not to oscillate in either diaphragm or tongue. These findings confirm that some core clock genes are oscillating in diverse skeletal muscle but identified that not all components of the clock are conserved.

Support for this work is from NIH R01AR055246 to KAE.
Prior eccentric exercise modifies diving‐induced inflammatory gene expression in rats
Authors: Eftedal Ingrid , Jørgensen Arve , Flatberg Arnar
Published: June 2021

Decompression sickness (DCS) after diving is a major risk of injury to divers, aviators and space travelers. DCS is associated with gas bubbles formed during decompression, but the processes linking bubbles to DCS are unknown. Exercise before diving reduces DCS risk. The effect of exercise on DCS could be caused by reduced bubble formation and/or to the induction of genetic pathways that affect biological responses to bubbles.

Study objective

To examine the effects of eccentric exercise on vascular gene expression and gas bubble production after diving.

Methods

Three groups of female Sprague Dawley rats were used; two diving groups with or without exercise and one control group, n=9 in each group. The exercise group did one bout of 100 min downhill treadmill running before diving, and saturation diving was simulated in an air‐filled pressure chamber at 709 kPa for 50 min. Diving‐induced stress was measured by ultrasonic detection of bubbles in the pulmonary artery and by full genome mRNA analysis of the aorta. All procedures were approved by the Animal Research Ethics Authority.

Results

Diving alone activated expression of 18 genes, whereas eccentric exercise reduced expression of 12 of these and upregulated an additional 25 genes. Specifically, elevated cytokine‐cytokine receptor expression and decreased expression of anti‐fibrinolytic Serpine1 was found in the exercise group. There were no differences in bubble formation between the groups.

Conclusions

Whereas diving induces cytokine and oxygen‐responsive gene expression, the protective effect of eccentric exercise before diving may be attributed to altered expression of genes involved in inflammatory responses.

This study was supported by Samarbeidsorganet HMN/NTNU, Kontaktutvalget St. Olavs Hospital/NTNU, and FUGE Midt‐ Norge, Norway.
High doses of pseudoephedrine taken 2 hours prior to “diving” to 132 feet of seawater (5 ATA) while breathing pure O2 increases risk for CNS oxygen toxicity in unanesthetized rats
Authors: Pilla Raffaele , Held Heather E. , Landon Carol S. , Dean Jay
Published: June 2021

Pseudoephedrine (PSE) salts (hydrochloride and sulfate) are commonly used as a decongestant by scuba divers. Anecdotal evidence collected by Divers Alert Network (DAN) suggests that use of PSE prior to making a high dose O2 dive (Nitrox or a rebreather using pure O2) increases the risk of CNS oxygen toxicity (CNS‐OT; i.e., seizures). We hypothesized that high doses of PSE reduces latency to seizure (LS) in unanesthetized rats breathing a lethal dose of HBO2 (5 ATA). Adult male rats were implanted with a radio‐transmitter for recording EEG, body temperature (TB), and activity. After ≥7 day recovery, a rat was given saline solution (control) or a dose of PSE hydrochloride intragastrically via gavage tube 2hr prior to “diving” (mg PSE/Kg): 0 (control), 80, 100, 120, 160 & 320 (9 rats/group). The rat was placed into a sealed chamber inside a hyperbaric chamber, which were pressurized in parallel with pure O2 and air, respectively, to 5 ATA until the onset of seizure. Our results show a significant dose dependent decrease in LS, compared to the control, at doses of 100–320 mg/Kg. We conclude that high doses of PSE should be avoided in deep dives breathing pure O2 as it increases the risk for CNS‐OT (DAN).
Adiponectin expression in skeletal muscle cells in response to hypertrophic and atrophic stimuli
Authors: Goto Katsumasa , Ohno Yoshitaka , Goto Ayumi , Ikuta Akihiro , Sugiura Takao , Ohira Yoshinobu , Yoshioka Toshitada
Published: June 2021

Adiponectin, one of adipocytokines, has insulin‐sensitizing effects. Recently, several reports showed that adiponectin protein expresses in skeletal muscle cells. However, the role of adiponectin in skeletal muscle is still unclear. In the present study, we examined the changes in adiponectin expression in skeletal muscles in responsible to hypertrophic, as well as atrophic, stimuli. The expression of adiponectin in mouse soleus muscles (C57BL/6J) was evaluated. Functional overloading and unloading on soleus was induced by synergist ablation for 4 weeks and hindlimb suspension for 2 weeks, respectively. The expression of adiponectin was also examined in both growth and differentiation phases of C2C12 cells. In C2C12, the expression of adiponectin mRNA was up‐regulated during differentiation. Increment in the protein and mRNA of adiponection was observed following overloading and reloading after the suspension. Results indicated that the trained skeletal muscles might secrete adiponectin. This study was supported, in part, by Grant‐in‐Aid for Scientific Research (B, 20300218, KG; A, 22240071, TY; S, 19100009, YO) from Japan Society for the Promotion of Science.
Methylxanthine reversal of opioid‐induced respiratory depression in the in situ neonatal rat working heart‐brainstem preparation
Authors: Ferguson Erin , Ciechanski Patrick , Roy Arijit , Ballanyi Klaus , Wilson Richard
Published: June 2021

Methylxanthines, such as caffeine and theophylline, are commonly given to premature infants to counter apneas of prematurity, but the mechanism by which these drugs prevent apnea is poorly understood. Methylxanthines act as nonselective adenosine receptor antagonists at lower concentrations and nonselective phosphodiesterase inhibitors at higher concentrations. Previous work in en bloc and slice preparations has demonstrated that >1 mM theophylline can reverse breathing suppressed by the μ‐opioid receptor agonist DAMGO. Using the in situ neonatal rat working heart‐brainstem preparation (a more intact preparation than previously employed) we show that theophylline restores DAMGO‐suppressed respiration in doses <1 mM, starting to effectively restore respiration at 250 μM. Denervation of the carotid bodies did not alter the response, suggesting that the response to theophylline is mediated centrally. These experiments suggest that in vivo, block of adenosine receptors may be involved in countering opioid depression. Funded by CIHR and Hotchkiss Brain Institute (EVF)
Cardiac effects of the disruption and reestablishment of social bonds in the male prairie vole
Authors: Wardwell Joshua , McNeal Neal , Scotti Melissa-Ann L , Preihs Kristin , Grippo Angela J
Published: June 2021

Social isolation and the disruption of social bonds have been shown to adversely affect cardiac function in both humans and prairie voles. To the best of our knowledge, the possible reversal of these effects through the reestablishment of male/female bonds has not been explored using the prairie vole model. In the present study, male prairie voles were implanted with wireless electrocardiographic transmitters. The animals were allowed a recovery period and then paired with a novel female for 5 days. After this bonding period, animals were separated for 5 days then reintroduced to the same female for a period of at least 10 days. Electrocardiographic parameters were monitored throughout all phases of the experiment and compared to baseline variables. Our preliminary results show a slight increase in resting heart rate during the pairing and isolation periods, along with a marked decrease in resting heart rate throughout the course of the repairing period, eventually falling to levels similar to basal resting heart rates before initial pairing. These findings support the possible reversal of the deleterious effects of social isolation through the reestablishment of social bonds and can be utilized to underline the importance of social support for human patients with cardiac dysfunction. Support: MH077581, NIU Graduate College