bluuebunny/crossref_metadata_2025 huggingface dataset:
Cerebralcare Granule®attenuates blood‐brain barrier disruption after middle cerebral artery occlusion in rats
Authors: Han Jing-Yan , Huang Ping , Liu Yu-Ying , Hu Bai-He , Chang Xing , Zhao Xin-Rong , Li Quan , Wei Xiao-Hong , Wang Chuan-She , Fan Jing-Yu , Zhou Chang-Man
Published: June 2021

Cerebralcare Granule®(CG) is a compound Chinese Medicine. In this study, we examined the effect of CG on the blood‐brain barrier (BBB) disruption in a focal cerebral ischemia/reperfusion rat model. Male Sprague‐Dawly rats (250 to 300 g) were subjected to 1h middle cerebral artery occlusion (MCAO). CG (0.4 g/kg and 0.8 g/kg) was administrated orally 3h after reperfusion for the first time and then once daily up to 6 days. The results showed that Evans blue extravasation, brain water content, albumin leakage, infarction volumes and neurological deficits were increased in MCAO rats, and were attenuated significantly by CG treatment. Magnetic Resonance Imaging and electron microscopy further confirmed the brain edema reduction in CG‐treated rats. Treatment with CG improved cerebral blood flow. Western blot analysis and confocal microscopy show that the tight junction proteins claudin‐5, Junctional adhesion molecule‐1, occludin and zonula occluden‐1 between endothelial cells were significantly degradated, but the protein expression of caveolin‐1, the principal marker of caveolae in endothelial cells, increased after ischemia, all of which were alleviated by CG. In conclusion, post‐treatment with CG significantly reduced BBB permeability and brain edema, which were correlated with preventing the tight junction proteins degradation and inhibiting caveolin‐1 expression in the endothelial cells.
Arginase 2 activation via subcellular trafficking: A novel mechanism for vascular endothelial dysfunction in atherosclerosis
Authors: Pandey Deepesh , Bhunia Anil , Romer Lewis , Berkowitz Dan
Published: June 2021

Endothelial dysfunction is widely considered a hallmark for initiation and progression of atherosclerosis. Oxidized low density lipoprotein (Ox‐LDL) and its receptor LOX‐1 play major role to elicit endothelial dysfunction in atherosclerosis. However the mechanism by which this occurs is still unclear and is the main objective of our study. In our study using cultured human aortic endothelial cells we found that Ox‐LDL exposure releases a pre‐existing pool of Arg2 into the cytosol from mitochondria. This, in turn, diminishes the concentration of cytosolic L‐arginine and leads to eNOS uncoupling. Ox‐LDL‐evoked changes in cytosolic Arg2 activity are muted by Rho‐kinase inhibition, and do not occur at all in LOX‐1 null endothelial cells. In addition, inhibition or siRNA knockdown of mitochondrial processing peptidase (MPP) reduces the cytosolic abundance and activity of Arg2 following Ox‐LDL stimulation. Furthermore, blocking MPP attenuates Ox‐LDL mediated changes in EC ROS and NO. Ox‐LDL signaling to MPP appears to involve MT1‐MMP and p27kip since siRNA knockdown of these proteins attenuates the cytosolic translocation event. Hence, inhibition of Arg2 translocation to cytosol by targeting MPP and Rho Kinase pathway could be a novel therapeutic target for the treatment of atherosclerosis and other cardiovascular diseases.
MKP1 inhibits myocardial TNF‐alpha expression and improves cardiac function in endotoxemia
Authors: Lu Xiangru , Zhang Ting , Arnold Paul , Liu Yusen , Feng Qingping
Published: June 2021

The objective of this study was to investigate the role of mitogen‐activated protein kinase phosphatase‐1 (MKP1) pathway in myocardial tumor necrosis factor (TNF)‐alpha expression and cardiac function during endotoxemia. Experiments were carried out in a mouse model of endotoxemia and in cultured neonatal cardiomyocytes. Lipopolysaccharide (LPS) increased MKP1 expression in the heart and in cultured neonatal cardiomyocytes. LPS‐induced ERK1/2 and p38 phosphorylation in the myocardium was prolonged in MKP1−/− mice. Myocardial TNF‐alpha mRNA and protein levels were enhanced in MKP1−/− compared to WT mice in endotoxemia, leading to a further decrease in cardiac function. Inhibition of Rac1 and PAK1 by a dominant negative Rac1 adenovirus (Ad‐Rac1N17) and PAK1 siRNA, respectively blocked LPS‐induced MKP1 expression in cardiomyocytes. PAK1 siRNA also decreased p38 and c‐Jun N‐terminal kinase (JNK) activation, and TNF‐alpha expression induced by LPS. Furthermore, deficiency in either Rac1 or JNK1 decreased myocardial MKP1 expression in endotoxemic mice. In conclusion, LPS increases myocardial MKP1 expression via JNK1. MKP1 attenuates ERK1/2 and p38 activation, inhibits myocardial TNF‐alpha expression and improves cardiac function in endotoxemia. Thus, MKP1 represents an important negative feedback mechanism limiting pro‐inflammatory response in the heart during sepsis.
Inhibition of miR‐205 impairs the wound‐healing process in human corneal epithelial cells by stimulating Kir4.1(KCNJ10)
Authors: Lin Daohong , Halivolic Adna , Bellner Lars , yue peng
Published: June 2021

The aim of this study is to explore the role of miR‐205, the second most abundant microRNA expressed in human cornea epithelial cells (HCE) in modulating healing process of the cell following the scratch (wound). The scratch of HCE cells increased significantly the expression of miR‐205. Moreover, inhibition of miR‐205 with antagomer (pZip‐miR‐205) significantly impairs the healing process of HCE cells. However, inhibition of endogenous K channels with Ba2+ partially restored the rate of healing process in HCE cells transfected with pZip‐miR‐205, suggesting that the inhibition of HCE healing process by miR‐205 antagomer was partially the result of stimulation of K channel activity. The patch‐clamp experiments further demonstrated that expression of miR‐205 antagomer increased the endogenous K currents in HCE cells in comparison to the cells transfected with vector alone. Analyzing the seed sequence of miR‐205 reveals that 3′UTR of KCNJ10 contains potential complementary matching sequence of miR‐205. Moreover, Western blot analysis demonstrated expression of pZip‐miR‐205 increased the protein expression of KCNJ10 but not KCNJ16 in HCE. Also, Western blot showed that expression of KCNJ10 was decreased 24 hrs in the post‐wound HCE cells. Finally, the role of KCNJ 10 in modulating the healing process of HCEC was further demonstrated by experiments in which down‐regulation of the K channel with kcnj10‐shRNA mimic the effect of Ba2+ and partially restored the healing process of HCE cells transfected with miR‐205 antagomer. We conclude that miR‐205 plays an important role in facilitating the wound‐healing process in HCE, an effect was partially achieved by inhibiting the inwardly‐rectifying K channel.
Prenatal dexamethasone produces altered cardiovascular and autonomic responses to psychological stress
Authors: Polson Jaimie W , Bisterbosch Heimerich M , Panahi S Ehsan , Dampney Roger AL
Published: June 2021

Raised glucocorticoid levels during gestation are known to increase the risk of high blood pressure (BP) in the offspring in later life, possibly due to altered autonomic regulation. We compared the cardiovascular responses and spontaneous baroreflex gain (sBRG) changes to 15 min of repeated air puffs in rat offspring following raised gestational glucocorticoid produced by dexamethasone (DEX, 200 μg/kg) administration in the pregnant dam, and controls. At 10 months of age, the offspring were implanted with telemetry probes and BP was recorded in the awake, freely moving rat. BP, heart rate (HR), and sBRG were determined before, during and after rats were subjected to air puff stress. At rest, we found no difference in BP between groups, while HR (control, 315±8 vs DEX, 336±5 bpm, p<0.05) was higher and sBRG (2.03±0.18 vs 1.14±0.05 ms/mmHg, p<0.05) was lower in the DEX group. During air puff stress, BP and HR increased similarly in both groups. sBRG was reduced more in DEX than controls (by 20±5 vs 50±7%, p<0.05). After stress, BP and HR recovered more slowly in DEX rats. At 30 min post stress, a second “delayed” stress response was observed in which BP and HR increased again. The duration of this response was greater in DEX than controls (20±4.9 vs 27±1.2 min, p<0.05). Our results indicate that cardiovascular and sBRG responses to stress are exaggerated following prenatal exposure to DEX.
Altered expression of FosB/ΔFosB in central autonomic regions in hypertensive obese Zucker rats
Authors: Fu G. Jeffery , Schreihofer Ann M
Published: June 2021

Adult obese Zucker rats (OZR) have elevated sympathetic nerve activity and blood pressure vs. lean Zucker rats (LZR). We determined whether autonomic regulatory regions in the brain are chronically activated in OZR vs. LZR by immunohistochemical examination of FosB/ΔFosB expression. Male rats (14 wks) were perfused with paraformaldehyde and brains were sectioned coronally. One in six sections were processed for FosB expression (Santa Cruz goat anti‐FosB) via a nickel diaminobenzidine (DAB) reaction. A subset was processed to reveal catecholaminergic neurons (Chemicon mouse anti‐tyrosine hydroxylase) via a DAB reaction. In nucleus tractus solitarius (−13.1 to −13.5 mm bregma) FosB expression was higher (85–134%) in OZR (7) vs. LZR (6) at all levels. In rostral ventrolateral medulla (−12.0 to −12.6 mm bregma) FosB expression was enhanced at the most caudal level (−12.6 mm) in OZR with comparable expression in other levels in OZR (5) vs. LZR (5). In NTS and RVLM there was no FosB expression in catecholaminergic neurons in OZR or LZR. In the hypothalamic paraventricular nucleus (PVN) OZR (5) had less FosB expression (94–200%) vs. LZR (5). Similarly, in the anterior hypothalamic nucleus OZR(6) had less (18–44%) expression than LZR(6). These data suggest that the chronic activations of multiple autonomic regions are altered in OZR vs. LZR, and future studies are needed to identify the phenotypes of the activated neurons.
In vivo MEMRI reveals persistent activation of the brain autonomic areas by an acute systemic angiotensin II injection
Authors: Zubcevic Jasenka , Perez Pablo D , Jun Joo Yun , Carvajal Jessica Marulanda , Raizada Mohan K , Febo Marcelo
Published: June 2021

Introduction

Angiotensin II (ANGII) acts centrally to activate the sympathetic nervous system in hypertension. Here, we show that even a single systemic ANGII injection causes long‐lasting neuronal activation in the autonomic brain areas.

Methods

In vivo mapping of basal and ANGII‐evoked neuronal activity was carried out in the Wistar‐Kyoto (WKY) rat using manganese‐enhanced MRI (MEMRI) at 4.7Tesla. Rats were treated with manganese chloride (MnCl2 30 mM solution, i.p.;16–20 hrs prior to the MRI), and T1‐weighted images were obtained 16–20 hrs after a single ANGII injection (0.32μg/kg i.p.). Coronal slice scans (positioned caudally from end of the cerebellum/medulla rostrally towards the hypothalamus) were manually segmented using itkSNAP, and data analyzed for differences in normalized signal intensity.

Results

Acute ANGII injection caused an immediate pressor response (ÄSBP=~20mmHg) which normalized 2 hours following the injection. Despite this, ANGII evoked persistent neuronal activation in the central amygdala, hypothalamus (PVN, preoptic and lateral hypothalamic areas), and brainstem (NTS and RVLM).

Conclusion

Even a single systemic ANGII injection results in a lasting effect on the brain autonomic areas. Thus, repeated pro‐hypertensive stimuli in these brain areas may lead to pre‐sympathetic neuronal plasticity, resulting in heightened sympathetic drive and hypertension.
Similar peak VO2 at altitude in native lowland (Han Chinese) and Tibetan highland inhabitants despite different hemoglobin concentration [Hb]
Authors: Simonson Tatum S. , Wei Guan , Wagner Harrieth , Wuren Tanna , Yan Ma , Qin Ga , Wagner Peter D. , Ge RiLi
Published: June 2021

Tibetan highlanders exhibit a unique suite of physiological traits, including markedly decreased [Hb] compared to Andean highlanders and high‐altitude sojourners at the same altitude. We performed exercise studies at 4200m in both native lowland Han Chinese males resident at >4000m for at least the past year (n = 7, [Hb]=19+/−0.77 g/dl) and native highland Tibetan males (n = 21, [Hb]=17.7+/−2.07 g/dl, p<0.02 between groups) to determine whether they differed in exercise capacity and/or one or more step(s) of the oxygen transport cascade. During maximal exercise we measured ventilation, VO2 and VCO2, heart rate, cardiac output, arterial PO2, PCO2, pH, HbP50, and arterial O2 saturation and calculated systemic O2 delivery and lung and muscle diffusional conductances (DL, DM). While peak VO2 and arterial blood gas variables were similar between groups, peak cardiac output and index trended lower in Han (both p < 0.11), which combined with higher [Hb], resulted in similar O2 delivery. DL normalized to lung volume trended higher (p < 0.07) in Tibetans, but DM was not different. These results show that in spite of lower [Hb], and thus O2 carrying capacity, Tibetan males can reach the same peak VO2 as Han, and appear to offset lower [Hb] by higher cardiac output, and possibly, lung diffusing capacity. How these differences relate to recently described changes in hypoxia‐sensitive genes in Tibetans remains to be determined.
Loss of murine TRPV1 function alters the thermoregulatory and ventilatory response to acute hypoxia
Authors: Yuen Nathaniel , Vincent Sandra G , Fisher John T
Published: June 2021

Hypoxic hypometabolism is the process in which small mammals and neonates reduce metabolic rate and body temperature in response to hypoxia. Since the transient receptor potential vanilloid 1 (TRPV1) ion channel has been implicated in the regulation of body temperature, we hypothesized that loss of TRPV1 function would affect the thermoregulatory and ventilatory response to hypoxia. Mutant TRPV1−/− (KO) and wild‐type TRPV1+/+ (WT) mice were exposed to hypoxia (10% oxygen) for 4 hours. Body temperature (Tb), heart rate (HR) and activity (ACT) were recorded using telemetry in conjunction with pattern of breathing measurements using plethysmography. During hypoxia, KO mice displayed a significantly lower mean Tb compared to WT mice (31.20 ± 0.13 °C vs. 31.97 ± 0.27 °C; Mann‐Whitney p=0.040). There were also differences in ventilation (1128 ± 36.4 ml/kg/min vs. 1282 ± 59.2 ml/kg/min; 2‐way ANOVA, p=<0.001), and respiratory frequency (161 ± 2 bpm vs. 179 ± 7 bpm; 2‐way ANOVA, p=<0.001) during hypoxia, as well as inspiratory (0.139 ± 0.002 s vs. 0.125 ± 0.003 s; 2‐way ANOVA, p=<0.001) and expiratory time (0.271 ± 0.008 s vs. 0.242 ± 0.015 s; 2‐way ANOVA, p=<0.001) between KO and WT mice, respectively. These data indicate that TRPV1 is involved in the ventilatory and thermoregulatory response to hypoxia. (Supported by CIHR)
Small crosslink may stabilize P22 tail protein at N‐terminus
Authors: Palmer Chris , Johnson Sam , Dean Dexter , Villafane Robert
Published: June 2021

The goal of this study is to determine if a 13 amino acid sequence (aa) is responsible for maintaining the structural stability of the N‐terminus domain of the P22 tailspike protein (TSP). The phage P22 TSP functions as a homotrimer and contains two major structural domains. The first domain consists of 108aa and functions as a homotrimer, taking on the form of a “dome”. Each of the three identical polypeptide chains contributes 23aa at the center of the dome which has been termed the “stem”. This short stem starts with aa23 at the dome top and travels through the dome interior to aa1 the dome bottom. We have determined through a molecular visualization program that the first 13aa of the stem are located at the adjacent chain. They form a hydrophobic interaction and may crosslink adjacent chains providing the rational for structural stability at the NTD. We have changed two out of the four stem hydrophobic residues to aspartic acid by QuikChange Mutagenesis and have determined by a petri plate assay (the Berget Plate Assay) that V9D change causes a change that destroys NTD function. Biochemical studies are continuing to determine the mechanism for this reaction and to study the remaining three amino acids.
Metabolic responses of the aerobic anoxygenic phototrophic bacterium Roseobacter denitrificans during photoheterotrophic and heterotrophic growth
Authors: Tang Joseph Kuo‐Hsiang
Published: June 2021

Roseobacter denitrifican is one of the first two reported species in Roseobacter clade members, which comprise up to 25% of total marine bacterial community and play essential roles on global carbon cycle. Here, we reported the studies of acetate metabolism of Rsb. denitrificans grown under light, dark and light‐dark cycles by investigating transcriptome dynamics, cell dry weight, pigment production and other physiological changes. Acetate‐grown Rsb. denitrificans produce more pigments and less biomass during heterotrophic growth than during photoheterotrophic growth (Fig. 1). Moreover, while Rsb. denitrificans has been reported to be an obligated photoheterotroph, our studies suggest that the bacterium employs the 3‐hydroxypropionate (3HOP) bi‐cycle for assimilating carbon when growing on acetate and bicarbonate. The 3HOP bicycle uses HCO3− as the inorganic carbon source that does not compete with photorespiration. Further, biofilm was found for acetate/bicarbonate‐grown cultures, but not acetate‐grown cultures, with more being produced during phototrophic growth than during heterotrophic growth.
Adiponectin receptor (ADIPOR1) signaling ameliorates obesity‐dependent cell cycle entry in MCF7 cells
Authors: Connor Michael , Theriau Christopher
Published: June 2021

Adipose tissue produces and secretes hormones, termed adipokines, that have endocrine/paracrine effects on a wide variety of tissues. Two of these adipokines (adiponectin and leptin) have cell cycle effects on breast cancer cells and may underlie the molecular link between obesity and breast cancer. Adiponectin (ADIPO) and leptin (LEP) stoichiometrically antagonize each other in MCF7 cells. Stable 2.6‐fold over expression of ADIPOR1 inhibited LEP‐dependent antagonism of ADIPO in MCF7 cells. This effect of ADIPOR1 was also evident in MCF7 cells exposed to media conditioned by adipocytes from high fat diet fed animals (HFD‐CM). HFD‐CM inhibited AMPK, activated AKT, reduced p27 and ADIPOR1 protein levels in MCF7 cells compared to MCF7 cells exposed to CM from chow diet‐fed animals (CD‐CM). Stable overexpression prevented these HFD‐CM effects suggesting that maintenance of ADIPOR1 signaling may prevent obesity‐dependent breast cancer progression.
Vitamin D supplement use by adult Canadians: sociodemographic correlates
Authors: Barr Susan I. , Green Tim J.
Published: June 2021

The 2011 Recommended Dietary Allowances (RDAs) for vitamin D intake in the absence of sunlight exposure are 15 μg/d (1–70 y) and 20 μg/d (70+ y). In Canada, these intakes are not feasible to obtain from food alone, and in winter months cutaneous vitamin D synthesis is very limited, suggesting the need for supplementation. To assess the prevalence and correlates of vitamin D supplement use subsequent to release of the 2011 RDAs, an on‐line survey was completed in September 2011 by a nationally‐representative sample of 2554 English‐ or French‐speaking adults aged ≥19 y. The survey assessed use of vitamin D supplements, health perceptions, and sociodemographic variables. Characteristics associated with supplement use in univariate analysis were used to determine multivariable‐adjusted odds ratios for supplement use. Overall, 47.9% reported use of a supplement with vitamin D; users’ median (interquartile range) supplemental intake was 10 (5, 25) μg/d. Significant adjusted odds ratios (95% CI >; 1.0) were detected for age group (19–30, 31–50, 51–70, 71+ y), female sex, speaking English (vs. French), having an osteoporosis diagnosis, perceived lactose intolerance, and university education. Adjusted odds ratio for marital status, ethnicity and income level were not significant. Certain population subgroups may benefit from targeted information about vitamin D supplementation.

Supported by the UBC Vitamin Research Fund and the Canadian Agri‐Science Cluster Initiative.
Development of a Nutrition Quotient (NQ) Equation Modeling for Children and the Evaluation of its Construct Validity in Korea
Authors: Kim Hye‐Young P. , Kwon Sehyug , Lee Jung‐Sug , Choi Young‐Sun , Chung Hae Rang , Kwak Tong‐Kyung , Park Juyeon , Lee Hye‐Jin , Kang Myung‐Hee
Published: June 2021

The objective of this study was to develop and evaluate the construct validity of a nutrition quotient (NQ) for children. In our previous study, the food behavior checklist (FBC) for children's NQ, consisting of 19 items, was grouped into a 5‐factor structure according to the exploratory factor analysis. In this study, the construct validity of the NQ was assessed using a confirmatory factor analysis. Elementary school students (n=1,393) from six large cities completed the NQ test. Indicator tests suggested an adequate model fit (goodness of fit index = 0.9613; adjusted GFI = 0.95; standardized root mean square residual = 0.0464; chi‐square test statistics of <0.001 p‐value, 82.1), and item loadings were significant for all subscales (p <0.05). The standardized path coefficients were used as the weights of the items. The NQ and the factor scores of the student were calculated by the obtained weights of the questionnaire items. Logistic regression was applied to find the significant factors in order to affect a specific nutrient status. The receiver operation characteristic curve analyses were performed in order to find diagnostic cut‐off points of the five factors. The FBC for children's NQ would be a handy and suitable instrument for evaluating dietary behaviors of Korean children. (This research was supported by a grant from Amway Korea Ltd. in 2011.)
Dietary behavior and Nutrition Knowledge of Korean Elementary Students
Authors: Chang Kyung Ja , Jang Jin‐sook
Published: June 2021

The purpose of this study was to investigate the dietary behavior and nutrition knowledge of senior elementary school students to provide some information on balanced dietary life and nutrition education. The subjects were 348 students (184 boys, 164 girls) who were in their fourth, fifth and sixth grades in elementary school in Incheon metropolitan city, Korea. This cross‐sectional survey was conducted using a questionnaire and data were analyzed with SPSS 12.0 program. As for dietary habit scores, there was no significant difference between boys and girls. But the girls significantly more replied that they had an unbalanced diet compared to the boys. As for eating speed, the boys ate significantly faster than the girls. Regarding the source of nutrition information, 54.9% of the subject acquired the information mainly from parents. Also 57.8% of the subject put confidence in the acquired information and put it into practice, and 50.6% of the subject felt the need for nutrition education. As for the correlation to dietary habit total scores and nutrition knowledge scores, dietary habit scores had a significant positive correlation to nutrition knowledge scores (r=0.178, p<0.001). Therefore, elementary students should receive systematic nutrition education to correct their dietary behavior, and a variety of nutrition education programs should be developed to help them acquire accurate nutrition knowledge in collaboration with their families.
In Vitro Digestive Enzyme Inhibitory Effects of Eight Common Spices and Herbs
Authors: Moss‐Pierce Tijuana , Tu Zheng , Jiang Alan
Published: June 2021

Excess calorie intake can lead to obesity as well as insulin resistance; the latter is associated with increased risk of coronary heart disease and type II diabetes. Inhibition of pancreatic lipase, α‐glucosidase and α‐amylase is one of the therapeutic approaches for delaying fat and carbohydrate digestion. We examined the potential inhibitory effects of common spices and herbs, i.e., cinnamon, clove, coriander, fenugreek, ginger, oregano, rosemary and sage, on digestive enzyme activity in vitro. Each spice was extracted in water: ethanol solution. Lipase, α‐glucosidase and α‐amylase inhibitory activities were determined by IC50 values. For lipase, cinnamon has the highest inhibitory activity with IC50 of 47.8 μg/ml, followed by clove >; sage >; rosemary >;ginger >; oregano >; coriander >; fenugreek. Cinnamon also has the highest α‐glucosidase inhibitory effect among tested samples, existing IC50 of 1.4 μg/ml. IC50 values ranged from 186 to 2,810 μg/ml for α‐amylase, in which clove (IC50=186) exhibited the strongest inhibitory effect. In conclusion, cinnamon and clove spice extracts were shown to be the most effective in inhibiting digestive enzyme activity. This research is supported by McCormick and Company.
Sorghum extracts reduced hepatic cholesterol biosynthesis in mice fed high cholesterol diet
Authors: Kim Se hee , Park Yongsoon
Published: June 2021

The purpose of the present study was to investigate that the sorghum extract reduces hepatic cholesterol biosynthesis. Male C57BL/6 mice were fed either a modified AIN‐93G diet or a modified AIN‐93G diet with 2% cholesterol for 8 weeks. During the last 4 weeks of diets, normal control group was orally administered saline (NC) and 2% cholesterol diet groups were received either saline (HC) or 600mg sorghum extract per kg body weight (S). There were no significant differences in daily intake, body weight, and weights of various organs. However, liver weight was significantly higher in mice fed HC than NC and S groups. Levels of serum total‐ and LDL‐cholesterol were significantly lower in NC and S groups than HC group, while levels of HDL‐cholesterol were significantly higher in NC and S groups than HC group. Serum levels of triglyceride were not significantly different, but hepatic triglyceride and total‐cholesterol levels were significantly lower in S and NC groups than HC group. The expression of AMPK, phosphor‐AMPK, HMGCR and SREBP2, were significantly lower in S group than HC group. However, there were no significant differences in the expression of FAS and CYP7A1 between HC and S groups. In conclusion, our results suggested that sorghum extract significantly reduced blood levels of cholesterol by down‐regulation of the hepatic cholesterol biosynthesis.
Protein turnover in pregnant pigs when feeding limiting amounts of amino acids
Authors: Moehn Soenke , Rafii Mahroukh , Pencharz Paul B , Ball Ronald O
Published: June 2021

Experiments to determine the amino acid (AA) requirements of pigs for threonine, lysine, tryptophan and isoleucine in early and late pregnancy were evaluated to determine which factors caused changes in protein turnover during pregnancy in pigs when the test AA intake was below requirements. In all experiments, each of 6 to 7 sows received 6 diets with graded levels of test AA in both early and late gestation. Protein turnover was determined based on the 13C enrichment in expired CO2 and plasma free phenylalanine (Phe) after oral L[1–13C]Phe dosing using a stochastic model. The observations (max n=188) were evaluated using mixed models (SAS). The final models were correlated to the data with r2 = 0.82 (protein synthesis, S) to r2 = 0.92 (Phe oxidation, Ox, g/d). Increasing AA intake decreased Ox (P=0.001) and increased Phe retention (RPhe, P=0.001) but did not affect S, breakdown (B) or Phe flux. Increasing BW and gestational age decreased Phe flux and Ox, S and B (P<0.02) and increased RPhe (P=0.001). In parity 3, values for Phe kinetics were greatest (P < 0.05), except for RPhe, which decreased linearly (P = 0.001) with sow age. Increasing litter size increased B (P=0.001) but did not affect other Phe kinetic parameters. Phe kinetics in pregnant pigs given limiting AA intakes were more affected by physical attributes, i.e. sow age (parity), BW, weight gain and gestational age than dietary attributes.
G‐protein coupled receptor profiling: an omics approach to study receptors and cell signaling
Authors: Wilderman Andrea Scarpa , Snead Aaron , Murray Fiona , Aroonsakool Nakon , Insel Paul A.
Published: June 2021

G‐protein‐coupled receptors (GPCRs) are the largest family of signaling receptors; ~350 non‐chemosensory GPCRs are expressed in mice, rats and humans. GPCRs and their signaling pathways are targets for 30–50% of marketed drugs. However, it is not known which and how many GPCRs are expressed by particular cell types. We thus undertook GPCR expression profiling on murine cell lines and primary rat and human cells and to determine if expression profiles differ between normal and diseased cells. We used the Taqman Gene Signature Array (Life Technologies) to define the GPCR profile of S49 murine lymphoma cells; we found that this Array has greater sensitivity for detection of GPCRs than did a genome‐wide array (R2=0.349 for GPCRs with high expression, R2=0.0107 for GPCRs with low expression). Using this GPCR array in cells from various tissues (e.g., lymphoid cells, fibroblasts, pulmonary artery smooth muscle cells and renal epithelial cells), we found that orphan GPCRs (ones without physiologic agonists) are among the highest expressed GPCRs. Importantly, for multiple cell types, we identified many GPCRs that were not previously known to be expressed. The increased sensitivity and high‐throughput format of the GPCR array defines new, potential physiologic regulators as well as targets and theraputic leads, including potential roles for orphan GPCRs. Supported by the Lymphoma and Leukemia Society, NIH, and Roche.
Parents’ perceptions of the importance and availability of nutrition and physical activity offerings in out‐of‐school time programs
Authors: Economos Christina D. , Angstrom Heather , Anzman‐Frasca Stephanie , Dolan Peter R. , Newman Molly B. , Sharma Shanti , Nelson Miriam E.
Published: June 2021

Out‐of‐school time (OST) programs offer opportunities for obesity prevention as millions of children, many of whom are at increased risk for obesity, participate in these programs. Since standards for nutrition and physical activity (PA) vary by OST organization, identifying and understanding gaps, opportunities, and parents’ perceptions of OST program offerings are important. An online survey was conducted in 2012 among 600 parents of children aged 5–12 in the US, to gather perceptions of the importance and availability of healthy snacks and PA at the following OST organizations: Boy Scouts, Girl Scouts, National Council of La Raza, National 4‐H Council, National Urban League, Pop Warner, US Youth Soccer, and YMCA of the USA. Most parents reported it was important that children have access to water (95%), fruits and vegetables (78%) and PA (97%). A minority (26–29%) viewed sweetened beverages and less healthy snacks as important. Only about half (57%) of parents reported that water was available, and fewer reported fruits and vegetables (16%) and PA (33%) availability at every OST meeting. 62% of parents agreed that OST organizations share responsibility to create healthy settings for children. The discrepancies between parental perceptions of the importance of OST healthy nutrition and PA, and what is currently available must be addressed in developing interventions to reverse the child obesity epidemic. RWJF
GUCY2C hormone deficiency contributes to diet‐induced colorectal tumorigenesis
Authors: Lin Jieru Egeria , Colon‐Gonzalez Francheska , Kim Gilbert Won , Li Peng , Blomain Erik Scott , Marszalowicz Glen , Stoecker Brian Arthur , Waldman Scott Arthur
Published: June 2021

While diet and obesity increase the risk for colorectal cancer (CRC), mechanisms underlying this association remain undefined. Guanylyl cyclase C (GUCY2C) is the intestinal receptor for the paracrine hormone guanylin. GUCY2C silencing, reflecting loss of guanylin expression, universally contributes to colorectal cancer initiation and progression. Indeed, silencing the GUCY2C tumor suppressor accelerates the cell cycle, increases DNA damage, and reprograms cell metabolism, increasing genetic and carcinogen‐induced colorectal cancer. Here, we reveal that excess calorie ingestion underlying obesity recapitulates mechanisms underlying CRC by eliminating guanylin expression, silencing the GUCY2C tumor suppressor, promoting intestinal tumorigenesis. Suppression of guanylin is reversibly modulated by ingested calories, rather than by the endocrine, adipokine or inflammatory milieu of obesity. Moreover, transgenic expression of guanylin, which is resistant to diet‐induced suppression, prevented colorectal cancer induced by excess calories in mice. Recent FDA approval of an oral GUYC2C ligand highlights the translational opportunities for GUCY2C hormone supplementation in preventing CRC in obesity.
Modulation of indoleamine 2,3‐dioxygenase pathway by a combination therapy strategy targeting myeloid derived suppressor cell function in lung cancer
Authors: Schafer Cara , Jin Tong Huan , Sawant Anandi , Roth Justin , Grant Stefan , Ponnazhagan Selvarangan , Deshane Jessy
Published: June 2021

Immunosuppression in the tumor microenvironment (TME) has dampened the efficacy of current front line combination chemotherapies for lung cancer thus providing limited improvement in patient survival. Myeloid Derived Suppressor Cells (MDSCs) are the major contributors of immunosuppression by producing Reactive Oxygen Species (ROS) and by modulating amino acid metabolism in the TME. Indoleamine 2,3‐dioxygenase (IDO) is an intracellular enzyme that is a key regulator of tryptophan metabolism and is involved in immunosuppression.

We have previously shown that a combination strategy using gemcitabine, a front line therapy for lung cancer and inhibitor of MDSC expansion, along with a Superoxide Dismutase (SOD) mimetic, which targets ROS in the TME, enhanced anti‐tumor immunity in mice and prolonged survival compared to controls. We hypothesized that this combination therapy will modulate IDO expression and activity and will diminish tumor burden. By flow cytometry and Western blot analyses, we demonstrate a significant reduction in IDO+ immunosuppressive cell types within tumor and in overall IDO expression in tumor and lung tissues, respectively, of tumor challenged mice treated with combination therapy compared to controls. Our preliminary studies suggest that this novel combination therapy may be effective in modulating the IDO pathway and in promoting long‐term immunity against lung cancer.

NCI CA 13148–39 UAB CCC CDGP
Induction of neuronal nitric oxide synthase (nNOS) in livers of mice treated with toxic doses of acetaminophen
Authors: Agarwal Rakhee , Banerjee Sudip , Hinson Jack A
Published: June 2021

In overdose acetaminophen (APAP) is hepatotoxic. Toxicity occurs by metabolism to a reactive metabolite (NAPQI). Following low doses NAPQI is detoxified by GSH; however, in overdose GSH is depleted and NAPQI covalently binds to proteins. This is followed by oxygen/nitrogen stress (protein nitration) and mitochondrial dysfunction. Nitration is by peroxynitrite formed from nitric oxide and superoxide. Toxicity and nitration in hepatocytes were inhibited by two nNOS inhibitors. In mice the principal nitrated protein was MnSOD. In nNOS knockout mice toxicity was delayed with less nitration compared to wildtype mice. We have now examined livers for the presence of nNOS in APAP toxicity. In livers of saline treated mice nNOS was not observed by Western blot analysis. However, in livers of APAP treated mice nNOS was induced. Low levels of nNOS were detected as early as 2 h following APAP. By 4 h nNOS was significantly induced and by 8 h it was dramatically induced. Its relative induction correlated with relative increases in protein nitration of MnSOD and development of toxicity (serum ALT). As anticipated inducible NOS (iNOS) was also induced following a hepatotoxic dose of APAP but its induction appeared to be later than induction of nNOS. These data indicate that nNOS is induced in livers of APAP treated mice. Supported by NIH grant R01 DK079008.
Olaparib, a PARP inhibitor approved for human testing, prevents allergen‐induced airway inflammation and hyperresponsiveness in a mouse model of asthma and reduces proliferation of human CD3/C28‐stimulated CD4+ T cells
Authors: Ghonim Mohamed A. , Naura Amarjit S , Rodriguez Paulo , Al Khami Amir , Hernandez Claudia , Mansy Moselhy S , Ochoa Augusto , Boulares Hamid
Published: June 2021

One of the most pivotal goals for the treatment of allergic inflammatory diseases such as asthma is the identification of therapeutic drugs that block the release or effects of Th2 cytokines after allergen exposure. Our laboratory has extensively examined the role of poly(ADP‐ribose)polymerase (PARP) in the pathogenesis of asthma‐related lung inflammation and has demonstrated that PARP inhibition confers protection against the onset of the disease. AZD‐2281 (Olaparib) is a novel PARP inhibitor that is being tested in advance clinical trials against several cancers in humans. In this study we examined the efficacy of olaparib against asthma using an animal model of the disease and human immune cells from healthy donors. Our results show that a single administration of olaparib 30 minutes post‐allergen (ovalbumin) challenge conferred a robust reduction in eosinophilia and completely blocked airway hyperresponsiveness to methacholine. Such protection was preceded by a significant impairment of CD4+ T cells recruitment to the lung and suppression of Th2 cytokines (e.g. IL‐4/IL‐5/IL‐6/eotaxin/IP10/M‐CSF/MIG) as well as ovalbumin‐specific IgE. Conversely and unlike other PARP inhibitors such as TIQ‐A, olaparib did not affect IL‐10 levels in the lung. Using immune cells from healthy donors, olaparib markedly reduced proliferation of CD3/CD28‐stimulated T cells. These findings further support the potential of PARP inhibition as a novel therapeutic strategy and olaparib with a likely application in the treatment of asthma.
Tissue specific regulation of CB1/CB2 gene receptors in salt sensitive hypertension of SHR
Authors: Varagic Jasmina , Gallagher Patricia E.
Published: June 2021

A growing body of evidence suggests that endogenous cannabinoids participate in the regulation of blood pressure in salt‐sensitive hypertension. However, there is limited information on the role of cannabinoids in the development of cardiac and renal abnormalities associated with high dietary salt intake. Therefore, the present study evaluated gene expression of the two major cannabinoid receptors (CB1 and CB2) in the heart and kidneys of the spontaneously hypertensive rat (SHR) fed a high salt diet. Male SHRs were given an 8% salt diet (HS; n=7), while their age‐matched controls (C; n=7) received standard chow for 5 weeks. CB1 and CB2 mRNA was measured using reverse transcriptase, real‐time polymerase chain reaction. After 5 weeks, the HS exacerbated hypertension (C: 208 ± 13; HS: 260 ± 11 mmHg, p<0.05) and increased left ventricular weight (C: 980 ± 12; HS: 1163 ± 38 mg, p<0.05), cardiac fibrosis [C: 3.41 ± 0.48; HS: 13.58 ± 3.29 %; p<0.05] and urinary protein excretion (C: 20 ± 2; HS: 160 ± 38 mg/day; p<0.05). CB1 receptor mRNA was 2.4‐fold (p<0.05) and 1.4‐fold (p<0.05) reduced in the heart and kidneys of salt‐loaded SHR, respectively. In contrast, CB2 mRNA was increased 1.8‐fold in kidney (p<0.05), but not in the heart. These results suggest that CB1/CB2 gene receptors are regulated in a tissue‐specific manner in salt‐loaded SHRs and may play a role in the development of end organ damage due to high dietary salt intake.
Orchiectomy increases food intake and perifornical orexin A neuron's activity during glucoprivation
Authors: Takamata Akira , Miyake Kana , Mabuchi Kaori , Morimoto Keiko
Published: June 2021

We examined the effect of orchiectomy on food intake and lateral hypothalamic neuron fs activity during glucoprivation induced by i.v. 2‐deoxy‐D‐glucose (2DG) administration in male rats. Rats (7W) were bilaterally orchiectomized (OrX) or sham‐operated (Sham). 2DG (400 mg/kg) was infused two weeks after the operation and food intake was measured for 3 hours after the injection. The same experiment was performed for immunohistochemical examination of c‐Fos and orexin A expression in the perifornical area (PFA) and arcuate nucleus (Arc). Glucoprivation induced food intake, and food intake in OrX rats was larger than Sham rats, and food intake was inversely correlated with plasma testosterone concentration. Glucoprivation stimulated c‐Fos expression in the orexin A neurons at the PFA, and c‐Fos expression in the Arc. The number and fraction of c‐Fos expressed orexin A neurons was significantly higher in OrX rats than Sham rats. The number of c‐Fos ir cells in the Arc was also higher in OrX rats than Sham rats. These data indicate that testosterone attenuates food intake induced by reduced glucose availability via attenuated perifornical orexin A neuron's activity and Arc neuron's activity.
Cardiovascular Response to an Inhaled β‐Agonist in Patients with Heart failure, Effect of β‐Blockade
Authors: Martinez Marina , Baker Sarah E. , Taylor Bryan , Johnson Bruce D. , Snyder Eric M.
Published: June 2021

Asthma, chronic obstructive pulmonary disease, and other respiratory disorders often occur concurrently with heart failure (HF). HF patients with respiratory disease can improve lung function and, therefore, quality of life with β‐agonists. Beta‐agonist use remains controversial as it has been linked with increased mortality risk; however, recent studies that account for co‐morbidities find no association between β‐agonist use and mortality. Previous studies exploring the cardiovascular effect of β‐agonist use in HF are primarily retrospective and focus on heart rate (HR). We have previously demonstrated altered cardiovascular function in response to albuterol in healthy subjects. In the present study we sought to comprehensively assess the cardiovascular effects of albuterol sulfate using cardiac output (Q), stroke volume (SV), HR, systolic, diastolic, and mean arterial blood pressure (SBP, DBP, MAP, respectively) and systemic vascular resistance (SVR). We hypothesized that cardiovascular parameters would be altered 30‐minutes following albuterol administration, and that HF patients using β‐blockers would have an attenuated response. Twenty‐two healthy and twenty‐two HF subjects were recruited (age=56±13 vs. 62±12yrs; height=175±10 vs. 176±9cm; weight=78±15 vs. 93±20; BMI=26±4 vs. 30±6; and BSA=2.08±0 vs. 1.93±0m2, for healthy and HF, respectively, p<0.05 for weight, BMI, and BSA). We found that HR and Q increased in healthy but not HF subjects at 30min following albuterol. Patients who were not on β‐blockers (n=4, 18%) had an increase in SV and a decrease in SVR with albuterol, while there was no change in patients who were on β‐blockers (n=14, 64%). These data suggest that HF patients have an attenuated cardiovascular response to inhaled albuterol, particularly with concurrent β‐blocker usage.
The protection of urocortin 2 against ischemia/reperfusion injury is due to p38 MAPK down‐regulation in an isolated rat heart model
Authors: Zhou Yue , Gao Xiufang , Wang Daying , Lew Karsheng , Richards Arthur , Wang Peipei
Published: June 2021

It is known that Urocortin 2 (UCN2) protects hearts against ischemia/reperfusion injury (I/R). However, the mechanisms underlying this beneficial effect are not fully understood. In this study we investigated the regulations of UCN2 on Akt and MAPK pathways under different conditions. Hearts from adult Sprague‐ Dawley rats were isolated and subject to Normoxia (NOR), I and I/R (20 min zero flow ischemia followed by 60 min reperfusion) with or without UCN2 treatment. Protective effect was as assessed by rate‐pressure product (RPP) recovery and LDH release. Hearts were freeze clamped at the end of perfusion for Western blot analysis. We found that UCN2 protects hearts as indicated by improved RPP (58±5 vs. Cont 31±4 % Baseline, p<0.05) and decreased LDH release (17±9 vs. Cont 170±87 mM, p<0.05). p38 MAPK is significantly down‐regulated by 7.8 and 1.6 fold during I and I/R respectively p<0.05 vs. Cont), but not in NOR. However, UCN2 has no impact on Akt, ERK1/2‐ and JNK‐MAPK pathways under any conditions. These data suggest the protective effect of UCN2 against I/R is due to down‐regulation of p38 MAPK. Inhibition of p38‐associated pro‐inflammation and pro‐apoptosis might be involved in the protection.
Ablation of MMP9 ameliorates epigenetic modifications and mitigates diabetic cardiomyopathy
Authors: Mishra Paras Kumar , Chavali Vishalakshi , Metreveli Naira , Tyagi Suresh C.
Published: June 2021

Mutation of Insulin2 gene (orthologous to human Insulin) activates latent matrix metalloproteinase‐9 (MMP9) that fosters cardiomyopathy. Epigenetic modifications such as DNA methylation through DNA methyl transferases (DNMTs) and histone de‐acetylation by HDAC play crucial role in cardiac dysfunction. However, the epigenetic signature of MMP9 in diabetic hearts is unclear. We hypothesize that abrogation of MMP9 attenuates DNA methylation and histone de‐acetylation in diabetic hearts and ameliorates contractile dysfunction. To test the hypothesis, we created double knock out (DKO: Ins2+/−/MMP9−/−) mice by targeted deletion of MMP9 in Ins2+/− Akita, and determined the levels of DNMT‐1, HDAC, H3K9 and sarco‐endoplasmic reticulum ATPase‐2a (serca‐2a) in WT (C57BL/6J), Ins2+/− (Akita) and DKO hearts by RT‐QPCR, Western blot and confocal microscopy. We also measured contractility of cardiomyocytes using Ion‐optics device. The results revealed that DNMT‐1, HDAC and H3K9 are robust in Akita but alleviated in DKO hearts. Conversely, serca‐2a is down regulated in Akita but induced in DKO. The rate of contraction and relaxation (±dL/dt) is improved in DKO as compared to Akita. We conclude that targeted deletion of MMP9 ameliorates diabetic cardiomyopathy through epigenetic modifications.
Angiotensin II increases arterial stiffness that is prevented by polyphenol S17834
Authors: Fry Jessica L , Weisbrod Robert M , Seta Francesca , Cohen Richard A
Published: June 2021

Arterial stiffness measured by carotid‐femoral pulse wave velocity (PWV) recently emerged as an important risk factor predicting cardiovascular disease events. We previously showed that male C57Bl/6J mice fed a high‐fat (35%) high‐sucrose (16.5%) diet (HFHS) have increased aortic PWV at 2 months, preceding the onset of systolic hypertension at 6 months on diet, indicating that arterial stiffness precedes hypertension in this disease model. To further probe this temporal relationship, we developed a mouse model of arterial stiffness using subpressor angiotensin II (AngII, 140 ng/kg/min via minipump). After 7, 14 and 28 days, mice infused with AngII had significantly increased PWV with no change in systolic blood pressure determined by tail cuff, and there was no change in aortic medial thickness or heart weight/tibia length ratio. At 28‐days there was a modest increase in medial collagen deposition. Concurrent treatment with S17834, a polyphenol that prevents HFHS‐induced PWV, prevented the increase in PWV in AngII‐treated mice. In conclusion, subpressor AngII produces a significant increase in arterial stiffness prior to a measurable increases in systolic blood pressure or medial hypertrophy in mice, suggesting that stiffness is linked to functional changes in the aorta that can be prevented by polyphenols such as S17834. Source: RO1 HL105287, Servier.