bluuebunny/crossref_metadata_2025 huggingface dataset:
Chickens genetically selected for high or low juvenile body weight display differences in glucose tolerance and insulin sensitivity
Authors: Sumners Lindsay , Zhao Xiaoling , Zhang Wei , Honaker Christa , Siegel Paul , Gilbert Elizabeth
Published: June 2021

Chickens selected for high (HWS) or low (LWS) juvenile body weight (BW) for 55 generations differ by 12‐fold in BW at 56 days of age and display differences in appetite and body composition. We hypothesized that HWS display impaired glucose tolerance and are relatively insulin resistant. At 58 days of age, overnight fasted HWS and LWS (N = 12) were orally administered glucose (2 g/kg BW) and whole blood glucose (BG) was measured at 0, 5, 15, 30, 60, 120, 240 and 300 min relative to glucose infusion. At 66 days, the same fasted chickens were injected intra‐peritoneally with 80 μg insulin/kg BW and BG was measured at 0, 5, 15, 30, 60, 120, and 180 min relative to injection. The area under the curve for the glucose tolerance test was greater (P = 0.05) in HWS than LWS, suggestive of impaired glucose tolerance in HWS. For the insulin sensitivity test, there was an effect of genetic line (HWS >; LWS; P < 0.0001) and the interaction of line by time (P = 0.04) on BG, where HWS values were greater than LWS from 0 to 60 min and lower at 120 min. These data suggest that selection for juvenile body weight in chickens leads to differences in glucose homeostasis and insulin sensitivity.
Multiphoton imaging reveals a novel leukocyte recruitment paradigm in the inflamed glomerulus
Authors: Hickey Michael , Devi Sapna , Li Anqi , Westhorpe Clare , Kitching A. Richard
Published: June 2021

In contrast to many capillary beds, the glomerulus supports leukocyte recruitment during inflammation. However, the behaviour of leukocytes after their recruitment to the glomerulus is unknown. In this study, we used multiphoton microscopy to examine leukocyte behaviour in the glomerular microvasculature under steady‐state and inflamed conditions. In uninflamed glomeruli, neutrophils and monocytes were retained within capillaries for several minutes, during which time leukocytes were either static or migrated. Inflammation increased the duration of leukocyte retention, an effect that was eliminated via inhibition of Mac‐1. In addition, both static and migratory neutrophils generated oxidants in inflamed glomeruli. This work establishes a new paradigm for glomerular inflammation, showing that acute inflammation does not induce leukocyte recruitment per se, instead increasing the duration of leukocyte retention in the glomerulus, during which time leukocytes adopt either static or migratory behaviors. These findings describe a novel form of multicellular intravascular patrolling that may contribute to the susceptibility of the glomerulus to inflammation. Supported by the NHMRC (Australia)
Drug delivery system based on PLGA nanoparticles containing Tamoxifen as antitumor agent
Authors: Oliveira Naiara A , Silva Jaqueline R , Azevedo Ricardo B , Gomes Anderson J
Published: June 2021

Tamoxifen (TMX) is an anti‐estrogen drug used in breast cancer treatment, however this drug has diverse side‐effects, including development of liver and endometrial cancers, thromboembolic disorders and resistance in target cells. Hence, an interesting alternative for overcoming these advantages is encapsulating TMX in biodegradable nanoparticles, such as the ones made of poly (lactic‐co‐glycolic acid) (PLGA). NP‐TMX were made by emulsion and solvent evaporation method, showing encapsulation efficiency (%EE = 87%), diameter of 160 nm measured by dynamic light scattering and positive zeta potential of +12.5 mV. A stability test was developed during 7 days in 37 ºC and revealed stability for 96 hours. Cell viability was assayed by MTT test in 4T1 cell line demonstrating an increased action by NP‐TMX TMX (90%) rather than in NIH‐3T3 (76%) being a promise drug delivery system. Supported by FAPDF, CAPES, CNPq, FINATEC and DPP‐UnB.
Impacts of Silver Nanoparticles on Bacterial Multidrug Efflux Pump
Authors: HE GUI , Zhang Wei , Chen Huizhong , Saliba Sam , Thorpe Conner , Rogers Eugene , Hsie Shufeng
Published: June 2021

Although silver nanoparticles (NPs) has shown antibacterial function, however, its impacts on drug resistance are not clear. Here, we report the impacts of silver NPs on multidrug efflux pump (MDRP) of Enterobacter cloacae. Antibacterial tests were performed with CLSI broth dilution method. E. cloacae strains with and without MDRP, EmmdR were utilized. Also, ciprofloxacin, benzalkonium, ethidium bromide, and trimethoprim were utilized for susceptibility tests. The drug efflux alteration of EmmdR after exposure to silver NPs was determined. Also, we tested the antibiotic‐H+ antiport alteration after exposure to silver NPs by using a fluorescence quenching assay. The expression of emmdR were measured by using a real‐time PCR method.

Susceptibility tests showed a silver NPs (< 50nm) enhancing the inhibition of antibiotics such as ciprofloxacin and benzalkonium on the growth of E. cloacae. The efflux of ethidium bromide was decreased after exposure to silver NPs. The ciprofloxacin‐H+ antiport activity was decreased after exposure to the silver NPs also. Also, the expression of emmdR did show a significant increase after exposure to the silver NPs.

In conclusion, this study discovered that silver NPs is able to improve the antimicrobial effectiveness of antibiotics in E. cloacae. Also, the silver NPs affect expression and function of EmmdR in E. cloacae.
Cloning and functional characterization of CsγTRPS from Coriandrum sativum
Authors: Galata Mariana , Mahmoud Soheil
Published: June 2021

Plant terpenes are a large and diverse class of naturally‐derived compounds, highly valued in the medicinal, perfume and culinary industries, to name a few. The seeds of Coriandrum sativum (coriander) produce essential oil rich in monoterpenes, volatile C10 terpenes. After Illumina transcriptome sequencing, several terpene synthase (TPS) candidate gene homologs were identified. In this study, one of these candidates, CsγTRPS, was cloned and functionally characterized in an effort to find TPS genes that describe C. sativum seed essential oil. CsγTRPS, which encodes a ca. 60 kDa protein, was expressed in bacteria and the recombinant protein purified by Ni‐NTA agarose affinity chromatography. Enzymatic assays with geranyl diphosphate (GPP), the linear precursor to monoterpenes, revealed that this protein catalyzes the conversion of GPP to an important monoterpene constituent of coriander essential oil, and a few minor monoterpene products. These experiments facilitate future studies concerning essential oil production, and other processes, in C. sativum. They also enable efforts to improve the essential oil of coriander through metabolic engineering or plant breeding. This project was supported by NSERC funding to Dr. Soheil Mahmoud.
High solar salt and refined salt supplementation affects blood pressure differently in Dahl salt‐sensitive rats
Authors: Lee Bog‐Hieu , Yang Ae‐Ri , Kim Mi Young , Seo Mi‐na , Han Chan‐ Kyu
Published: June 2021

Solar salts(SS) are sun‐dried sea salts containing less Na and more minerals like Ca, Mg, and K compared to refined salts(RS) with 99.9% NaCl. The study was conducted to investigate the effects of high levels of solar and refined salts on blood pressure in Dahl salt‐sensitive rats for 15 weeks. Fifty‐eight 13‐week‐old male Dahl rats were assigned to five groups; rat chow (CON), 4% SS(SS4), 4% RS (RS4), 8% SS(SS8), 8%RS (RS8). Blood pressure was measured by the tail cuff method. Body weight and blood pressure were measured weekly. Feed intake was the highest in SS8 and the lowest in CON, but FER and PER was vice versa (p<0.0001). Body weight and gain were the lowest in RS8 followed by SS8 and indifferent between CON and 4% salt groups. In case of systolic blood pressure (SBP), it was higher in order of RS8 (198.7mmHg) >; SS8(174.5mmHg) >; RS4(172.5mmHg)>; SS4(163.7mmHg) >;CON (136.5mmHg) (p<0.001). SBP of RS8 was increased 73 mmHg compared to 9.6 mmHg for CON group at 15 weeks. While diastolic blood pressure (DBP) have shown similar tendency to SBP, but the statistical significance was observed only in CON (92.5mmHg) and RS8(116.1mmHg) groups. In addition, heart and kidney weights of supplemented groups were heavier than those of CON(p<0.001). From the findings, both solar and refined salts raised SBP and DBP, but solar salts did not increase as much as refined salts, showing SS8 was similar BP to RS4 group. It suggests that solar salts, sun‐dried sea salts seem to curb the elevation of blood pressure than refined salts.

Grant Funding Source: This research was supported by grants from the Globalization of Korean Foods R & D program, funded by the Ministry of Food, Agriculture, Forestry and Fisheries, Republic of Korea
DASH‐style diet is effective in patients with treated hypertension and diastolic heart failure independent of change in body weight – a pilot study
Authors: Seymour E Mitchell , Wells Joanna , Han‐Markey Theresa , Soni Tanu , Burant Charles , Hummel Scott L
Published: June 2021

“Diastolic heart failure” (DHF) is the most common type of heart failure over age 65 and is frequently associated with hypertension. The “Dietary Approaches to Stop Hypertension” diet can reduce blood pressure, but its effects in patients with DHF are unknown. This project tested the sodium‐restricted DASH diet in patients with hypertensive DHF using three weeks of controlled‐feeding with effort to maintain body weight. The experimental diet achieved DASH guidelines for nutrient intake without a significant change in total calories. DASH increased six‐minute walk distance and K+ excretion while it reduced Na+ excretion, systolic and diastolic blood pressure, post‐walk test dyspnea, and urinary 8‐ isoprostane. Of over 200 metabolites tested, few were significantly altered; changes in macronutrient metabolism were unlikely associated with clinical findings. Instead, altered micronutrient intake and phytochemicals from fruits/vegetables may be instrumental. Three weeks of the sodium‐restricted DASH diet significantly reduced blood pressure and oxidative stress in subjects with hypertensive DHF. This pilot study revealed endpoints affected early in a DASH diet intervention in this population, independent of changes in body weight. Furthermore, the study provides a model for longer DASH interventions aimed to affect cardiac geometry and function. Supported by NIH UL1TR000433 and K23HL109176.
Collaborative Engaged Research: Rethinking Food Decisionmaking and Nutrition Education
Authors: Gillespie Ardyth Marie Harris , Craig B. Susie
Published: June 2021

Methodology for collaborative engaged research (CER) has evolved through experimentation with integrating research into nutrition education programs for changing food and eating practices and improving community food systems. The collaborative engaged research (CER) approach is characterized by an asset‐based approach which focuses on strengths and opportunities rather than problems. CER projects take a family and community development perspective for creating sustainable food systems which nourish and nurture children across generations. The CER process builds on aspirations and identifies opportunities. CER is lead by interdisciplinary teams which include family and community food system stakeholders, practitioners, and academic researchers. Complex and multifaceted, CER facilitates dynamic communication patterns and develops relationships among community food system stakeholders (bonding social capital) and with external resources (bridging social capital) which builds individual, family, and community capacity for thoughtful personal and collective food decision‐making.
Growth Hormone Receptor In Human Subcutaneous Adipose Tissue: Gluteal Versus Abdominal Depots
Authors: Karastergiou Kalypso , Bredella Miriam A , Smith Steven R , Miller Karen K , Fried Susan K
Published: June 2021

Central fat accumulation is associated with increased cardiometabolic risk and reduced growth hormone (GH) secretion. GH administration in obese premenopausal women reduced abdominal fatness and systemic inflammation, but had no effect on gluteal fat (Bredella MA et al, Eur J Endocrinol, 2012). To gain insight into the mechanisms underlying the depot‐specific effects of GH, we examined the expression of the GH receptor (GHR), as well as genes that GH may target in the lipolytic pathway, in samples of abdominal and gluteal subcutaneous adipose tissue from premenopausal women (n=16, BMI 27.3±4.0 kg/m2) and age‐matched men (n=28, BMI 26.8±6.1 kg/m2). GHR expression (by qPCR) was significantly lower in the gluteal compared to the abdominal depot (p=0.01). In both sites, GHR mRNA levels correlated significantly with ATGL (abdominal: r=0.62, p=0.0001, gluteal: r=0.61, p=0.0004) and perilipin expression (abdominal: r=0.63, p<0.0001, gluteal: r=0.69, p<0.0001). In the abdominal, but not in the gluteal depot, GHR also correlated with mRNA levels of HSL (r=0.68, p=0.0001), CIDEA (r=0.51, p=0.003) and CIDEC (r=0.47, p=0.006). These associations were statistically independent of BMI, age and sex. We conclude that depot differences in GHR expression may underlie the in vivo effect of GH administration to reduce specifically abdominal adipose tissue mass.
Dietary phenylalanine requirement of adult Labrador Retrievers using the direct oxidation approach
Authors: Shoveller Anna Kate , Fortener Lisa , Davenport Gary
Published: June 2021

Historically, the determination of amino acid (AA) requirements in dogs have been done in growing dogs and have been invasive in nature, requiring long periods of confinement and exposure to diets deficient in AA, which result in a reduction in the health and well being of dogs during the study period. Therefore, we were interested in examining whether carbon oxidation methods, with short periods of exposure to diets deficient in a single AA, could be successfully applied to determine AA requirements in dogs. Four adult Labrador Retrievers consumed 7 levels of phenylalanine (Phe), and excess tyrosine (Tyr) in random order and in combination with kibble that was formulated to be deficient in Phe and adequate in dietary energy (3730 kcal/kg as fed) and fed at ~13.5 g kg−1 BW d−1. Dogs were fed test diets for two days prior to conducting the tracer studies. The Phe requirement was determined by applying a 2‐phase linear regression crossover model to tracer oxidation (F13CO2). The mean Phe requirement was 3.27 g/kg diet (upper 95% CI= 4.96 g/kg diet) on a dry matter basis. The current NRC estimate of the adequate intake for Phe is 4.50 g/kg diet and based on growth and nitrogen balance in growing dogs. This magnitude of difference is similar to that seen when estimates of AA requirements in humans using nitrogen balance vs. carbon oxidation methods are compared.
Lipoic acid‐based strategy to treat hypertriglyceridemia
Authors: Yi Xiaohua , Pashaj Anjeza , Brauer Vanessa M. , Moreau Regis
Published: June 2021

Epidemiologic and intervention studies show that lowering blood triglycerides (TG) decreases the risk for coronary heart disease, liver disease, and type‐2 diabetes. Yet over 65 million Americans have abnormally high blood TG (hypertriglyceridemia). This paradox points to a major limitation in the effectiveness of current TG‐lowering strategies. Using the Zucker Diabetic Fatty (ZDF, fa/fa) rat as a model of hypertriglyceridemia, we found that (R)‐ alpha‐lipoic acid (LA), a naturally occurring food compound, prevents the rapid rise in plasma VLDL‐TG that predictably occurs between week 7 and week 8 of age in pair‐fed control rats. The objectives of the study are two‐fold i) determine to what extent dietary LA corrects hypertriglyceridemia once blood TG have become elevated, and ii) elucidate how LA acts to lower blood TG. Feeding LA to hypertriglyceridemic ZDF rats restored near normal blood TG when compared to pair‐fed controls. Preliminary evidence supports a role of fibroblast growth factor 21 in mediating the metabolic effect of LA. Elucidating the novel mechanism by which LA lowers TG will stimulate interest in its therapeutic use and justify further research on potential additive or synergistic effects of combined therapy with LA.

Grant Funding Source: UNL ARD Hatch Act, Tobacco Settlement Funds, and Layman Seed Award
The role of Aurora‐A in curcumin‐enhanced chemosensitivity of human breast non‐metastatic cancer MCF‐7 and highly metastatic MDA‐MB‐231 cells
Authors: Su Chun‐Li , Yen Cheng‐Hsin , Liu Hsiao‐Sheng
Published: June 2021

Curcumin, an active compound in turmeric and curry, has been revealed to induce tumor apoptosis and inhibit tumor proliferation and metastasis. Overexpression of Aurora‐A, a mitosis‐related serine‐threonine kinase, increases drug resistance and promotes lung metastasis of breast cancer cells. Our previous results indicate that curcumin suppresses Aurora‐A kinase activity, induces S and G2/M mitotic arrest, and decreases proliferation of MCF‐7 and MDA‐MB‐231 cells. In the present study, combination therapy of curcumin with FDA‐approved anti‐cancer drugs (ixabepilone, cisplatin, vinorelbine, or everolimus) overrode chemoresistance in MDA‐MB‐231 cells. Addition of curcumin also enhanced cytotoxicity of ixabepilone in MCF‐7 cells. Induction of apoptosis and G2/M arrest, the consequences of Aurora‐A inhibition, were involved. However, addition of curcumin exhibited an antagonism effect in cisplatin, vinorelbine, or everolimus‐treated MCF‐7 cells due to not being able to suppress Aurora‐A kinase activity to a very low level. Therefore, the intake of diets rich in curcumin or curcumin‐containing supplements should be taken into consideration.

(Supported in part by the National Science Council, Taiwan, No. NSC 98–2313‐B‐003–002‐MY3; the Ministry of Economic Affairs, Taiwan, No. 100‐EC‐17‐A‐17‐S1–152; and the National Taiwan Normal University, Taiwan, No. NTNU100‐D‐06).
Intestinal adaptation to diet‐induced obesity involves an increase in intestinal epithelial stem cells but impaired stem cell function
Authors: Mah Amanda , Van Landeghem Laurianne , Gavin Hannah , Lund P. Kay
Published: June 2021

Rates of renewal of the intestinal epithelium adapt to altered nutrient intake, but the impact of diet‐induced obesity (DIO) is not well defined.. We hypothesized that DIO alters intestinal epithelial renewal and mass, and preferentially affects intestinal epithelial stem cells (IESC). Sox9‐EGFP mice permit visualization and isolation of IESC and other cells based on levels of EGFP‐reporter (IESC: Sox9‐EGFP Low; progenitors: Sox9‐EGFP Sublow; enteroendocrine cells: Sox9‐EGFP High; other differentiated cells: Sox9‐EGFP Negative). Sox9‐EGFP mice were fed high fat diet for 20 weeks to induce DIO, or low fat standard chow. Obese mice were 40% heavier than control and had 15.5% and 10.6% increase in villus height and crypt density, respectively. Histology, labeling of S‐phase cells and flow cytometry revealed that DIO increases Sox9‐EGFP Low IESC and selectively increases proliferating IESC but not progenitors. Intrinsic IESC function assessed in vitro by number of organoids formed revealed significantly fewer IESC‐derived organoids from obese mice. We conclude that in vivo, DIO increases proliferation and number of IESC resulting in increased functional epithelial mass. However DIO impairs intrinsic IESC survival or growth indicating detrimental effects of DIO on IESC that may be compensated by extrinsic in vivo factors that promote IESC hyperproliferation. Grants: NIH DK040247–19, NIH 2‐T32‐DK07686

Grant Funding Source: NIH
New, pathogenesis‐based pharmacologic approaches to gastrointestinal ulcers: Gene therapy with egr‐1 & cell therapy with bone marrow‐derived stem cells
Authors: Szabo Sandor , Deng Xiaoming , Chen Longchuan , Khomenko Tetyana , Sandor Zsuzsanna
Published: June 2021

Our recent studies showed that the immediate‐early gene early growth response (egr‐1) that regulates the expression of angiogenic growth factors (e.g., VEGF, bFGF, PDGF) & neovascularization play a mechanistic role in experimental duodenal ulceration (DU). We thus hypothesized that gene therapy with adenovirus (AV) encoding egr‐1 & local administration of bone marrow‐derived stem cells (BMSC) may accelerate the healing of both upper & lower gastrointestinal ulcers, i.e., DU & ulcerative colitis (UC). Groups of rats were given cysteamine‐HCl (C, 85 mg/100g, x1, gavage) to induce DU or 6% iodoacetamide (IA, 0.1ml/rat, x1, enema) to cause UC. A single dose of either AV‐egr‐1 (2 × 109 pfu/rat) or BMSC (1 × 106 cells/rat) was given intraduodenally at laparotomy in DU rats on the 3rd day after C or intracolonically in UC rats on the 2nd day after IA. The rats were euthanized on the 7th or 14th day after C or IA administration. We found that DU & UC lesion areas were significantly reduced in rats treated with AV‐egr‐1 or BMSC compared to controls in both 7 & 14 days (p < 0.05). Increased levels (2‐ to 4‐folds) of growth factors VEGF, PDGF and bFGF were also seen at days 7 and 14 in the rats treated with AV‐egr‐1 or BMSC compared to the controls.

Conclusions

local egr‐1 gene or stem cell therapies seem to be a new option to achieve a rapid healing of DU & UC, possibly by promoting angiogenesis/vasculogenesis & restoring mucosal integrity.

(This work was supported by the Veterans Health Administration Merit Review Grants to S. Szabo and Zs. Sandor).
Ethanol is a Significant Cofactor in HAART Induced Hepatotoxicity
Authors: Donde Hridgandh , Ghare Smita , Zhang Jingwen , Kirpich Irina , Joshi‐Barve Swati , McClain Craig , Barve Shirish
Published: June 2021

Highly Active Antiretroviral Therapy (HAART) has significantly increased life expectancy of HIV patients; however, there are major side effects which include lipodystrophy and hepatotoxicity. Alcohol abuse is highly prevalent in HIV patients and can be a significant negative cofactor in HAART‐hepatotoxicity. Hence, the mechanisms underlying HAART and alcohol induced hepatotoxicity were investigated. Specifically, effects of HAART drugs (azidothymidine, and Indinavir sulphate) in combination with alcohol were examined both in vitro and in vivo. Individually, alcohol and AZT caused cytotoxicity in H4IIEC3 (a rat hepatoma) cells which was significantly increased in combination. These data indicate that alcohol can induce and further enhance HAART‐cytotoxicity.

Assessment of Alcohol and HAART drug interaction and hepatotoxicity using an animal model of chronic alcohol feeding showed increased visceral adiposity compared to pair‐fed animals, suggesting disturbances in lipid metabolism. Lipodystrophy was seen as hepatic macro and microvesicular steatosis, elevated triglycerides and free fatty acids. These animals also exhibited increased inflammation and hepatic neutrophil infiltration. Overall, our data demonstrate that alcohol exacerbates HAART hepatotoxicity, and act as a significant cofactor in the development of hepatic steatosis and liver injury.
Oxytocin affects nitric oxide and cytokine production by sepsis‐sensitized macrophages
Authors: Oliveira‐Pelegrin Gabriela Ravanelli , Saia Rafael Simone , Cárnio Evelin Capellari , Rocha Maria José Alves
Published: June 2021

Oxytocin (OXT) secretion during cecal ligation puncture (CLP)‐ induced sepsis has not yet been examined. Although immune properties have been attributed to OXT, its effect on CLP‐sensitized macrophages has never been investigated. We analyzed the OXT secretion during CLP and its effect in CLP‐sensitized macrophage cultures. Male Wistar rats were decapitated 4, 6 or 24h after CLP surgery or sham operation and blood, brain and neurohypophyses collected for OXT measurements. In another set of animals we studied the effect of OXT on nitrite, TNF‐α, IL‐1β and IL‐10 production of peritoneal macrophages harvested at 6 and 24h after CLP. In the early phase of sepsis (4–6h), OXT levels increased in plasma and decreased in hypothalamus and neurohypophysis. In the late phase (24h), plasma and neurohypophyseal levels remained basal. In the PVN, the OXT content remained low, but in SON increased. Macrophages of the early phase of sepsis pre‐treated with OXT and LPS‐stimulated showed decreased nitrite, TNF‐α and IL‐1β levels, but no alteration in IL‐10 production. In the late phase they showed reduction only on IL‐1 β. OXT secretion during sepsis may represent a neuroendocrine response contributing to the overall host response to infection by decreasing the proinflammatory response and oxidative stress.

Financial support: FAPESP, CNPq/PROCAD
mTOR Pathway Mediates Diazoxide Preconditioning in Cultured Neurons
Authors: Dutta Somhrita , Wappler Edina , Katz Paige S , Carvalho Cristina I , Katakam Prasad V G , Busija David W
Published: June 2021

Diazoxide (DZ), a mitochondrial ATP‐sensitive potassium channel opener, is protective in models of brain ischemia but the signaling pathway is not clear. The mammalian target of Rapamycin (mTOR) pathway is a major regulator of protein synthesis and has been shown to be involved in protection during cerebral ischemia. We tested the hypothesis that DZ protects neurons via mechanisms involving the mTOR pathway. Cultured cortical neurons were treated with DZ, and Rapamycin (R) or Wortmannin (W) and exposed to 3 hours of oxygen‐glucose deprivation (OGD) followed by reoxygenation. Viability, mitochondrial membrane potential (Ψm), of neurons and western blots of cell homogenates were performed. OGD reduced viability (100% to 46.7% ± 0.9, p<0.05) and DZ partially rescued the viability (64.9% ± 4.5, p<0.05). W, R, DZ + R, DZ + W, DZ + R + W, however, did not alter viability after OGD. Ψm decreased after OGD (to 49.9% ± 9.5, p<0.05 vs. Ctrl) compared to untreated controls (Ctrl). DZ reduced Ψm under normoxic conditions (65.3% ± 6.2, p<0.05 vs. Ctrl) and further decreased it after OGD (26.2% ± 2.9, p<0.05 vs. Ctrl). R or W did not affect Ψm at normoxia but decreased it with OGD (50.9% ± 8.4, p<0.05 and 53.9% ± 8.9, p<0.05; respectively; both vs. Ctrl). Immuno‐blot analysis showed decreased phosphorylated S6‐ Kinase (phospho‐S6K) expression (29.9% ± 6.9, p<0.05) following OGD that was rescued by DZ (206.5% ± 52.8, p<0.05 vs. OGD). DZ + R, DZ+W, DZ + R + W decreased phospho‐S6K when compared to Ctrl. Thus, DZ preconditioning is mediated by mTOR pathway by phosphorylation of S6K. Supported by HL‐030260, HL‐065380, HL077731, & HL093554.
Cerebral White Matter CD4+ T cell Infiltration is Associated with Hypertension in a rat model of HELLP Syndrome
Authors: Wallace Kedra , Morris Rachael , Moseley Janae , Evans Stephanie , LaMarca Babbette
Published: June 2021

HELLP syndrome (hemolysis, elevated liver enzymes, low platelets) and preeclampsia (PE) (reduction in uterine perfusion pressure ‐RUPP), are hypertensive disorders that can occur during pregnancy with severe symptoms and consequences. The development of hypertension (HTN) may increase ischemia and inflammation in the maternal brain. We hypothesize that HTN in response to placental ischemia or HELLP increases CD4+ T cells expression in the maternal brain.

On gestational day (GD) 12, sEndoglin (7ug/kg) and sFlt‐1 (4.7ug/kg) infusion began via miniosmotic pump into normal pregnant (NP) rats to induce HELLP. On GD14 RUPP rats had the lower abdominal aorta and ovarian arteries physically constricted. On GD18 carotid catheters were inserted (n=5/group), GD19 mean arterial pressure (MAP), plasma, tissue and maternal brains were collected.

MAP increased from 96.2 ± 3.6 in NP rats to 127.8 ± 3.8 (p=.05) in RUPP rats; to 113.4±3.2mmHg in HELLP rats (p=.05). RUPP and HELLP rats had increased circulating CD4+ T cells compared to NP rats (p=.01, p=.05). CD4+ T cells along the corpus callosum in RUPP (136.1 OD) and HELLP rats (151.9 OD) were increased compared to NP (114.5 OD, p=.05, p=.05).

These data indicate that CD4+ T cells are increased in both the circulation and the brain and may therefore be one potential link in the neuropathophysiology of HTN associated with PE and HELLP syndrome.
Early life events effect on myelin gene expression
Authors: Castilloveitia‐Vega Gloria Luz , Pedre Xiomara , Casaccia Patrizia
Published: June 2021

Myelin is crucial for proper CNS function. It can be affected in diseases like multiple sclerosis and psychiatric disorders. Previous studies reported that children raised in abusive environment had change in white matter tracts; we therefore asked whether traumatic early life events have long lasting changes on myelin gene expression. To address our hypothesis we used an animal model of early weaning and maternal deprivation in mice. We then allowed the mice to develop until 5 or 8 weeks of age, when we isolated RNA from different brain regions and used qPCR to compare myelin gene expression in the prefrontal cortex, involved in social interaction, and in the corpus callosum, involved in interhemispheric communication. We detected a trend for decreased Mag and Mbp gene expression in early weaned mice in the prefrontal cortex, but not in the corpus callosum. To define whether decreased myelin gene expression was associated with impaired differentiation of progenitors, we measured the transcript levels of progenitor markers Pdgfr‐α and Cspg4, myelin gene activator Sox10 and myelin gene inhibitor Hes5. We found no difference for progenitor markers in the 5 week old group, but an increase in the 8 week old group. This may suggest that early life events have long lasting effects on myelin gene expression by impairing the differentiation of progenitor cells and predispose to demyelinating disorders. Funding by R01NS42925.
Renal molecular reponses elicited by electrical stimulation of sympathetic renal nerve in wistar rats
Authors: Pontes Roberto Braz , Garcia Michelle L , Veiga Glaucia RL , Oliveira‐Sales Elizabeth B , Mirian A B , Campos Ruy R , Bergamaschi Cassia T
Published: June 2021

This study evaluated the effects of acute sympathetic renal nerve electrical stimulation (RNES) over the expression of sodium (NHE3 and ENAC) and water (AQP2) transporters in the kidney in the presence or not of AT1 receptors angiotensin II blockade. Male Wistar rat (250g–300g), were divided in four groups: Sham (renal sympathetic nerve activity registered for 1h), RNES for 1h, [15V, 1Hz and 0,5 ms]), Sham+Losartan (30 mg/kg/day) and Losartan + RNES. Electrical stimulation produced increase in the NHE3 (1,6±0,2AU) gene expression and reduction in β‐ENaC (0,6±0,1AU) and AQP2 (0,6±0,1AU) expression. Animals just treated with Losartan did not presented differences at NHE3 expression but shows a reduction in ENAC (0,4±0,1AU) and AQP2 (0,3±0,1UA) expression compared with SHAM animals.. Treatment with Losartan+ RNES had no effect over the studied transporters. RNES produces alterations in sodium and water transporters in the renal tubule which can be involved in increased sodium reabsorption at the proximal tubule and reduced signalization to distal tubule reabsorption and these changes are angiotensin‐dependent.

Supported by CAPES and Cnpq.
Chronic heart failure in male Lewis rats alters corticotropin releasing hormone receptor‐2 (CRH‐R2) mRNA expression in both cardiac and brain tissue
Authors: Hayward Linda F. , Dickerson Erin E , Yoo Jeung‐Ki , Christou Demetra D , Ferreira Leonardo F
Published: June 2021

In heart failure (HF) circulating levels of urocortin, a positive inotrope and potent vasodilator, are elevated but peripheral actions are limited by downregulation of the endogenous receptor CRHR2. Urocortin is also readily transported into the brain where it can be anxiolytic. The present study was undertaken to test the hypothesis that chronic HF is also associated with a central downregulation of CRH‐R2 receptors and may contribute to HF‐associated mood disorders. Male Lewis rats underwent ligation of the left anterior descending coronary artery to induce HF (n=7) or a SHAM (n=6) operation and were studied 16 weeks post‐surgery. HF was characterized by a significant reduction in fractional shortening (29±2 vs 46±2 %, HF vs SHAM). CRH‐R2 mRNA levels from HF animals were found to be decreased in the right ventricle (68±13 % of SHAM) and hypothalamus (75±13% of SHAM) and slightly increased in the dorsal pons (112±5% SHAM), but unchanged in the amygdala and hippocampus. A positive relationship between cardiac CRH‐R2 receptors and hypothalamic CRH‐R2 receptor expression was also identified. Urocortin and CRH mRNA levels in the hypothalamus were found to be down‐regulated in HF (<75%) compared to SHAM. These observations suggest that autonomic and psychological changes associated with HF may be linked to a change in the balance of CRH‐R2 receptors in the hypothalamus and dorsal pons.
Brain Endoplasmic Reticulum (ER) Stress Mediates Lipopolysaccharide‐Induced Central Inflammation and Sympathetic and Cardiovascular Excitation
Authors: Zhang Zhihua , Yu Yang , Wei Shun‐Guang , Felder Robert B.
Published: June 2021

Inflammation in central cardiovascular regulatory regions has recently emerged as an essential contributor to increased sympathetic nerve activity in hypertension and heart failure. However, the mechanisms are poorly understood and are a topic of intense investigation. Endoplasmic reticulum (ER) stress is an important pathophysiological mechanism in various cardiovascular diseases including heart failure and hypertension. We hypothesized that ER stress contributes to the central inflammation and sympathetic excitation induced by intracerebroventricular (ICV) administration of lipopolysaccharide (LPS). In anesthetized rats, ICV injection of LPS (5 μg) increased (p<0.05, vs baseline) renal sympathetic nerve activity, mean blood pressure and heart rate. ICV infusion of the ER stress inducer thapsigargin (TG, 5 μg/hr) elicited similar responses, but with smaller amplitude. LPS significantly increased paraventricular nucleus (PVN) mRNA for tumor necrosis factor‐α, interleukin‐1β, and cyclooxygenase‐2. ICV infusion of the ER stress inhibitor tauroursodeoxycholic acid (TUDCA, 5 μg/hr) significantly attenuated all of these TG‐ and LPS‐induced responses. These data suggest that therapeutic interventions targeting brain ER stress may modulate the sympatho‐excitatory effects of central inflammation in heart failure and hypertension.
Sources of Intravascular ATP during Exercise in Man: Critical Role for Skeletal Muscle Perfusion
Authors: Kirby Brett S , Crecelius Anne R , Richards Jennifer C , Dinenno Frank A
Published: June 2021

The origin of intravascular ATP resultant from exercise in humans is a matter of debate and is suggested to arise from sympathetic nerves, blood sources, endothelial cells, and skeletal myocytes, among others. We tested the hypothesis that acute activation of the sympathetic nervous system (SNA) increases plasma ATP draining skeletal muscle at rest, and that SNA superimposition during exercise further augments ATP. We show that augmenting SNA via −40mmHg lower body negative pressure does not increase ATP at rest (51±8 vs 58±7 nmol/L) nor in conjunction with forearm handgrip exercise (79±11 vs 71±8 nmol/L). Next, we tested the hypothesis that active perfusion of skeletal muscle is essential to increase intravascular ATP during exercise. We identify that complete obstruction of muscle perfusion abolishes exercise‐mediated increases in ATP (90±19 to 49±12 nmol/L), and further that cessation of blood flow prior to exercise completely inhibits the typical rise in ATP draining active muscle (3 vs 61%; obstructed vs intact perfusion). Our collective observations suggest that blood extracellular ATP is unlikely to originate from sympathetic nerves or the contacting muscle itself. We conclude that increased exercising blood flow per se is essential for a rise in plasma ATP and indicates an important role for red blood cells or endothelial cells in augmenting plasma ATP during exercise.

Supported by NIH HL102720
Nrf2‐regulated antioxidant defenses in rodent models of longevity
Authors: Reuland Danielle J , Drake Joshua C , Biela Laurie M , Ehrlicher Sarah E , Peelor Frederick F , Miller Richard A , Miller Benjamin F , Hamilton Karyn L
Published: June 2021

Lifespan is extended by rapamycin treatment, caloric restriction (CR), crowded litter (CL), and in genetic models such as the Snell Dwarf mouse. Various mechanisms have been proposed by which lifespan is extended in these rodent models, including improved antioxidant defenses. The transcription factor nuclear factor (erythroid‐derived 2)‐like 2 (Nrf2) has been suggested to be the “master regulator” of cellular antioxidant defenses. However, whether these rodent longevity models show enhanced Nrf2 activation and antioxidant enzyme expression is unknown. The purpose of this study was to determine the expression of Nrf2 and ARE‐regulated antioxidant enzymes in four rodent models of longevity. Endogenous antioxidant enzyme and Nrf2 expression were not consistently increased in long‐lived animals, with considerable variation existing between models. While Nrf2 and its targets were greater than controls in Snell liver and heart, antioxidant defenses were significantly lower than controls across all tissues in CL animals. CR resulted in increased antioxidant defenses in some, but not all, tissues. Rapamycin did not affect Nrf2‐regulated enzymes, but expression differed between sexes. Not all models of longevity displayed similar Nrf2‐regulated antioxidant enzymes and these data suggest that sex, age, and tissue specific differences must be considered with regard to antioxidant capacity and longevity.
Effects of voluntary exercise on cognition, neurogenesis, and plaque load in a mouse model of Alzheimers disease.
Authors: Maliszewska‐Cyna Ewelina , McLaurin JoAnne , Aubert Isabelle
Published: June 2021

Alzheimer's disease (AD) is a neurodegenerative disorder leading to cognitive impairment and disruption of adult neurogenesis. It has been shown that exercise influences adult neurogenesis, synaptic plasticity, and cognitive functions in animal models. In healthy adults and AD patients, studies are suggesting that exercise has potential benefits on cognitive functions. However, we only begin to understand the global effects that exercise can have on amyloid pathology, neuronal plasticity, and cognition in AD mouse models.

Here we evaluated the effects of moderate voluntary exercise on cognition, neurogenesis, and plaque burden by giving mice access to running wheels for 1 and 2 months and assessing their functional recovery. We used an AD mouse model of amyloid pathology and their non‐transgenic littermates as control. Our results show an improvement in hippocampal‐dependent spatial and non‐spatial memory in transgenic mice exercising for 2 months compared to non‐running transgenic mice. Transgenic exercising animals also showed significantly greater levels of hippocampal adult neurogenesis (increase in BrdU/NeuN positive cells), and increase in survival of newborn cells (increase in BrdU positive cells). Plaque burden, evaluated as the mean plaque size and plaque number, in the hippocampus was not statistically different in runners compared to non‐runners transgenic mice.

In summary, exercise improved spatial working memory in an AD mouse model of amyloidoisis without reducing plaque burden. This improvement was accompanied by an increase in neurogenesis. We conclude that physical voluntary exercise has the potential to improve cognition even in presence of high levels of amyloid pathology.
Hypoxia dysregulates fibrinolytic pathways in cultured human mesothelial cells (HMCs): Implications for intraabdominal adhesion formation
Authors: Currie Brian , Heydrick Stanley J , Stucchi Arthur F
Published: June 2021

Abdominal surgery induces mesothelial injury and the extravasation of a fibrinous exudate into the peritoneum. The ensuing deposition of a fibrin matrix serves as a precursor to adhesions. Our aim was to determine the mechanism(s) by which injury‐induced hypoxia and/or inflammation contribute to the formation of the fibrin matrix, the precursor of adhesions. Cultured HMCs were treated with either an inflammatory cytokine cocktail of TGFβ+IL‐1β+IFN‐γ (C), the hypoxia mimetic dimethyloxaloglycine (D) or both, and mRNA levels for vascular endothelial growth factor (VEGF; extravasation), protein c receptor (PrCR; inhibits Protein C activation), thrombomodulin (TM; inhibits thrombin), and tissue factor (TF; activates thrombin) were measured at 1, 3, 6, 12 & 18 hrs via RT‐PCR. VEGF mRNA was increased by both D (peak: 6hr, 8±2.3 fold vs. control), and C (peak 12 hr: 7±0.1 fold, both p<.01) at all times, and the combination was additive. Hypoxia reduced PrCR mRNA levels to a min at 18 hr (0.5±0.06 fold, p<.05) and elevated TF mRNA levels to a max at 18 hr (3.5±0.9 fold; p<.05). TM mRNA levels were decreased by D and D+C to a min at 18 hr (0.25±0.06 fold, p<.05). These data show that hypoxia impairs fibrinolytic pathways in HMCs and may promote adhesion formation. Funded by Cooper‐Tyler Endowment, Department of Surgery, Boston University Medical Center.
Role of Nrf2 transcription factor in ozone‐induced emphysema in mice
Authors: Limjunyawong Nathachit , Mitzner Wayne
Published: June 2021

Ozone is a potent air pollutant which is known to cause pulmonary oxidative stress and inflammation. It has been shown that repeated exposure in mice to ozone at high concentration can damage lung parenchymal tissue and lead to emphysema. In the current study, we investigated the protective role of nuclear factor‐erythroid 2‐related factor 2 (Nrf2), a nuclear transcription factor that is induced by oxidative stress and is known to activate production of several antioxidant enzymes. Female wild‐type (WT) C57BL/6 and Nrf2‐deficient (Nrf2−/−) mice were exposed to ozone at a concentration of 2 ppm for 3 hours/day, 2 days/week, for 6 weeks in total. Control mice were exposed to room air. Measurements of lung function were performed one day after the last exposure. Results showed that the total lung capacity (volume at 35 cmH2O) was found to be significantly increased in the WT mice exposed ozone to compared to room air (1.465 ± 0.040 mL and 1.144 ± 0.052 mL, respectively). The effect of O3 was even higher in Nrf2−/− mice (1.784± 0.113 mL vs 1.301 ± 0.082 mL). However, no changes were observed in the diffusion capacity of the lung for carbon monoxide and cell profile in bronchoalveolar lavage fluid in either strain. These results indicate that the role of Nrf2 in ozone exposure is more complex than simple protection.

Funding source: NIH HL‐10342
Ouabain and Digoxin Preconditioning Protect Cardiac Na/K‐ATPase against Ischemia‐Reperfusion Injury in Langendorff‐perfused Mouse Hearts
Authors: DUAN QIMING , GULATI GAURAV K , PIERRE SANDRINE V
Published: June 2021

Protection against myocardial Ischemia‐Reperfusion (IR) injury by Ischemic PreConditioning (IPC) involves the protection of Na/K‐ATPase activity. The cardiac glycosides (CG) digoxin and ouabain are specific inhibitors of Na/K‐ATPase‐mediated ion transport, but also trigger preconditioning through activation of Na/K‐ATPase signaling. In Langendorff‐perfused mouse hearts, we investigated whether CG preconditioning also protects Na/K‐ATPase.

Forty min of ischemia followed by 30 min of reperfusion decreased the left ventricular developed pressure (LVDP) by 71.4% (60.8 ± 2.3 vs 17.4 ± 1.9 mmHg, P<0.01), induced a lactate dehydrogenase (LDH) release of 5.8 ±1.2 U, and resulted in an infarct of 52.9 ± 5.4% of the area at risk (as measured by tissue staining). Na/K‐ATPase enzyme activity measured by ouabain‐sensitive ATP hydrolysis was decreased by 59% (3.2 ± 0.89 vs 7.8 ± 0.6 umol/h/mg, P<0.05) without change in Na/K‐ATPase α or β subunit content as shown by Western blotting. In hearts exposed to 10 μM ouabain (OPC) or digoxin (DigPC), or 4 cycles of brief ischemia (IPC) prior to prolonged ischemia, LVDP recovery was nearly doubled (P<0.01 vs IR), infarct size was reduced by half (P<0.01 vs IR ), and LDH released was decreased by 70%(P<0.05 vs IR).

These suggest that, like IPC, OPC and its more clinically relevant counterpart DigPC protect cardiac Na/K‐ATPase activity during IR. (NIH P01HL036573)
Novel potential binding partners of the C‐terminal tail of the sodium bicarbonate cotransporter NBCn1
Authors: von Stemann Jakob Hjorth , Lauritzen Gitte , Schröder Jacob Morville , Pedersen Stine Falsig
Published: June 2021

We recently showed that the sodium bicarbonate cotransporter NBCn1 (SLC4A7) is upregulated in human breast carcinomas, in which it is the main pH regulator after acidification. Further, NBCn1 is upregulated after overexpression of the receptor tyrosine kinase ErbB2 in MCF‐7 breast cancer cells. The aim of this work is to identify novel NBCn1 binding partners and address their relevance in breast cancer. We performed GST‐pulldown with the full NBCn1 C‐terminus (NBCn1‐Ct) and GST controls, using lysates from ΔNErbB2‐overexpressing MCF‐7 cells and corresponding vector cells, followed by mass spectrometry. Notably, mitochondrial‐associated proteins were pulled down preferentially in the ΔErbB2+ cells. Other proteins pulled down by NBCn1‐Ct with high specificity included the calcium‐binding proteins S100A13, S100A14 and calmodulin, 14–3‐3 beta, and basigin and Receptor for Activated protein C Kinase 1 (RACK1), both of which are implicated in regulating subcellular localization of other membrane transporters. NBCn1‐Ct also associated directly with several protein kinases and ‐phosphatases including Casein Kinase II, which is predicted to phosphorylate several sites in NBCn1. In conclusion, we identified several potential novel interaction partners of NBCn1‐Ct. Analysis to further validate these findings and address their significance in breast cancer are ongoing.

Funding: Danish Cancer Society, Danish Research Council
Sex Differences in Oxidative Stress Following Uniphrectomy in Antenatal Glucocorticoid Exposed Sheep
Authors: Bi Jianli , Contag Stephen , Su Yixin , Chen Kai , Figueroa Jorge , Chappell Mark , Rose James C.
Published: June 2021

Antenatal glucocorticoid (AG) administration is associated with nephron loss and sex specific reductions in renal function in adult offspring. It is not known if AG induced responses to a second renal insult also reflect sex differences.

Therefore, we determined if AGs increase susceptibility of proximal tubule cells (PTC) to oxidative stress induced by a second hit, further nephron loss (uninephectomy UN) in a sex dependent fashion.

1–1.5 year old rams and ewes exposed to either Betamethasone (BMX) (n=5) or Vehicle (V) (n=5) at 0.6 gestation underwent UN. Kidney cortex and proximal tubule cells (PTC) were obtained from one kidney at UN (1st kidney) and the other at necropsy two weeks later (2nd kidney). PTC were incubated with angiotensin II (Ang II) and the NADPH oxidase components (p47phox and NOX2) in cortex and 8‐isoprostane (8‐PGF) secretion in media were measured. ANOVA was used for data analysis.

Antenatal BMX exposure or UN increased p47 expression in cortex in rams but not in ewes (F=23.1, p=0.003). Expression was higher in BMX exposed rams (F=6.5, P=0.02). NOX2 expression did not change. Ang II stimulated 8‐PGF secretion from PTC of the 1st and 2nd kidneys from BMX rams, but not ewes. The Ang II response was highest in PTCs from the 2nd kidney 139% (F=20.6, p<0.0001) and was blocked by candesartan, but enhanced by PD123319 (F=10.1, P=0.0004). We conclude that the kidneys in BMX exposed male sheep exhibit a greater susceptibility to a secondary insult than those of females. AT1 receptors contribute to Ang II induced oxidative stress, while AT2 receptors may attenuate the stress in PTC. The heightened response in males may reflect the increase in the NADPH oxidase components p47phox.. Supported by HD‐47584 and HD‐17644.