bluuebunny/crossref_metadata_2025 huggingface dataset:
An evidence‐based approach to public policy and advocacy efforts (630.5)
Authors: Ziegler Paula , Schofield Marsha , Khan Mujahed , Blankenship Jeanne , Wickstrom Donna , Tuma Pepin , Steiber Alison
Published: June 2021

The objective is to demonstrate the use of an evidence‐based approach to the Academy of Nutrition and Dietetics’ (AND) strategic efforts to shape food and nutrition policies. Evidence Analysis Library (EAL) at AND is used to advocate for policies with third party payers, government officials, regulatory agencies and other decision‐makers. AND identified public priority areas through a national member process. EAL content was used to support messaging for advocacy work around priority areas, e.g. Maternal and Child Nutrition, Healthy Aging and Medical Nutrition Therapy (MNT). Child Nutrition Reauthorization‐Healthy Hunger Free Kids Act and Farm Bill legislation were supported by 19 EAL projects. Older Adults and Nutrition analysis supported legislation, including Affordable Care Act and Older Americans Act. Pediatric Weight Management supported private payer coverage for childhood obesity through Alliance for a Healthier Generation’s Healthier Generation Benefit. MNT provided evidence‐base for National Coverage Determination request to Center for Medicare Services to expand the Medicare Part B MNT benefit. Outcomes of the EAL have been used as the foundation for comments on proposed rules, regulatory initiatives and third party payer coverage policies. Content from the EAL has been translated into talking points, educational materials and communication pieces for stakeholders and policy‐makers.

Grant Funding Source: Supported by the Academy of Nutrition and Dietetics
Effects of extended sleep on female athlete's performance and nutrition (633.2)
Authors: Famodu Oluremi , Bryner Randall , Montgomery‐Downs Hawley , Olfert Melissa
Published: June 2021

The purpose of this study was to investigate the effects of one extra hour of sleep on physical performance and nutrition behaviors on college female track athletes. Eleven females (age 20.6 ± 1.6 yrs) maintained normal sleep habits for a one week. Baseline data was followed by a one week sleep extension period (night time and napping). Anaerobic power and fatigue was measured using the Wingate Test. The ASA 24‐hour food‐recall questionnaire was used to test nutrient intake. Reaction time, perceived stress, and mood were measured using the Psychomotor Vigilance Task (PVT), Cohen's Stress Test, and Profile of Mood States (POMS), respectively. Preliminary data was collected on 11 female subjects. Subjects increased total sleep from baseline (423.5 ± 44.4 minutes, 443.9 ± 55.2 minutes respectively), however this was not significant (z = ‐1.2480, p = 0.2258). No significant differences were seen in maximum power (z = 0.00, p = 1.0000) and fatigue (z = 0.0657, p = 0.9476) following sleep extension. Although not significant, mean PVT reaction time increased (z = ‐0.8213, p = 0.4115) and POMS scores improved (z = 0.7560, p = 0.4496) following more sleep. The athletes showed no significant effects in caloric (z = 0.1313, p = 0.8968) or total fat intake (z = ‐0.2627, p = 0.7928) after sleep extension. Improvements in physical performance and nutritional behaviors were not seen after sleep extension in college female track athletes.

Grant Funding Source: WVU Experimental Station Hatch Funding
Early gut microbiota composition ő a possible target to reduce the risk of developing neuropsychiatric disorders? (637.11)
Authors: Isolauri Erika , Pärtty Anna , Kalliomäki Marko , Endo Akihito , Salminen Seppo
Published: June 2021

Background: Although genetic factors can determine a propensity for neuropsychiatric disorders, environmental exposures, particularly early microbial contact, may shape the risk during a critical period of maturation.

Objective: To address whether early gut microbiota composition and its modification by probiotics could be related to the increasing emergence of attention deficit hyperactivity disorder (ADHD) and Asperger syndrome (AS).

Methods: 75 infants who were randomized to receive Lactobacillus rhamnosus GG (ATCC 53103) or placebo prenatally and during the first six months of life were followed‐up for 13 years. Gut microbiota was assessed at the age of 3 weeks, 3 and 6 months and 13 years using several molecular methods. ADHD and AS were diagnosed by a child neurologist or psychiatrist on the basis of ICD‐10 diagnostic criteria.

Results: At the age of 13 years ADHD and/or AS was diagnosed in 6/35 (17.1%) children in the placebo and none in the probiotic group (p=0.008). The affected children manifested lower numbers of Bifidobacterium species bacteria in feces, in particular less Bifidobacterium longum, than healthy children during the first six months of life (p=0.03, p=0.045, respectively).

Interpretation: The composition of the gut microbiota and probiotic supplementation early in life may have an impact on the development of neuropsychiatric disorders later in childhood; our preliminary study suggests both an intriguing link between gut microbiota and behavior and a potential treatment target.

Grant Funding Source: Supported by the Academy of Finland
Biomarkers of oxidative stress and antioxidant defense system among Guatemalan preschool children attending daycare centers with a common menu offering (639.2)
Authors: Soto‐Méndez María , Aguilera Concepción , Mesa María , Campaña‐Martín Laura , Martín‐Laguna Victoria , Solomons Noel , Schümann Klaus , Gil Angel
Published: June 2021

Objective: To describe and compare urine oxidative stress biomarkers and erythrocyte antioxidant enzymes activity, in preschoolers from 3 daycare centers in Guatemala Methods: 74 (2‐6 y/o) children (36 F / 38 M) attending 3 government‐subsidized daycare centers identified by location, coded as Center (C) and as to unique location: A (semi‐urban n=19); B (marginal‐urban n=23); and C (rural n=32) enrolled to measure oxidative damage to DNA (8‐hydroxydeoxyguanosine {8‐OHdG}) and lipid (15‐Isoprostane F2t {F2‐Iso}) and the erythrocyte activity of Catalase (CAT), Superoxide Dismutase (SOD), and Glutathione Reductase (GSHR) and Peroxidase (GPx) Results: 8‐OHdG and F2‐Iso median values were different between sexes (p=0.03 and p=0.04, respectively), even when calculating the total amount excreted per day and adjusting to weight. GPx also differed by sex (p=0.01). According to location 8‐OHdG and F2‐Iso were different between CA and CB (p<0.01 and p<0.01, respectively). CAT and GSHR were different in CB when compared to CA and CC (p<0.01 in both cases). SOD activity was different among all 3 centers (p<0.01), whereas GPx was different between CB and CC (p=0.02) Conclusion: Differences in oxidative biomarkers between sexes, especially those biomarkers that are involved in lipid oxidation (F2‐Iso and GPx), are seen.

Funded by: Fundación Iberoamericana de Nutrición (FINUT), Spain and The Hildegard Grunow Foundation (HGF), Germany

Grant Funding Source: Supported by Fundación Iberoamericana de Nutrición (FINUT) and The Hildegard Grunow Foundation (HGF)
Nutrient excess promotes accumulation of bone marrow‐derived progenitor cells in adipose tissue (641.12)
Authors: Holden Candice , Wysoczynski Marcin , Sansbury Brian , Hellmann Jason , Zafar Nagma , Gibb Andrew , Bhatnagar Aruni , Hill Bradford
Published: June 2021

Obesity has been implicated as a major risk factor for the development of several chronic diseases, including Type 2 diabetes. Because adipose tissue (AT) must remodel remarkably to accommodate lipid storage under conditions of nutrient excess, we hypothesized that AT stem cells are essential for adipose organ expansion. Therefore, we first sought to determine the source of AT stem cells and to measure changes in AT stem cell populations that occur with obesity. To delineate the contribution of bone marrow (BM)‐derived cells, mice were irradiated and the BM was repopulated with GFP+ donor cells. The mice were then placed on low‐fat (LF) or high‐fat (HF) diets for 12 weeks and changes in inflammatory cell populations and AT stem cells were measured. F4/80+ macrophages increased 2.5‐fold in the visceral AT with 12 weeks of HF diet. Similarly, BM‐derived Sca‐1+/CD29+/CD24+ cells—putative adipocyte stem cells shown to have the capacity to form adipocytes in vivo—increased 2.8‐fold in the AT of mice fed a HF diet. Sca‐1+/Flk+ endothelial progenitor cells also increased 2.6‐fold in the visceral AT. These data suggest that adipocyte and endothelial stem cell populations derive from bone marrow. Their recruitment to adipose tissue could be important for AT remodeling under conditions of caloric excess.
Effects of almonds as a snack or meal accompaniment on appetite, glycemia and body weight (641.9)
Authors: Tan Sze , Mattes Richard
Published: June 2021

Snacking may contribute to overweight and glucose dysregulation because snack foods often elicit weak dietary compensation and promote glycemic swings. Almonds are satiating and moderate post‐prandial glycemia, but it is not known if these properties hold when consumed specifically as a snack. Following an initial OGTT, an acute feeding trial was conducted where participants (49M, 89F) at risk for diabetes consumed 43g of roasted, lightly‐salted almonds with breakfast (N=28) or lunch (N=26), alone as a morning snack (N=28) or afternoon snack (N=28) or no almonds (N=27). Appetitive sensations and blood glucose concentrations were monitored over 515 minutes during which standardized meals were provided. Participants then consumed almonds as per their group assignment for 4 weeks followed by another acute feeding trial and OGTT. Almond consumption as snacks led to significant reductions of appetitive sensations and blood glucose concentrations. They blunted (non‐significantly) glucose rises at meals. There was no change of body weight and MUFA and Vitamin E intake increased during the 4‐wk feeding period. Almonds may be a snack that contributes to weight and glycemic management.

Grant Funding Source: Supported by the Almond Board of California
Role of milk fat globule membrane in the regulation of blood lipids in humans: a randomized trial (642.3)
Authors: Rosqvist Fredrik , Paulsson Marie , Lindmark‐Månsson Helena , Smedman Annika , Risérus Ulf
Published: June 2021

Dairy fat has in general cholesterol raising effects due to its high content of saturated fat. Some studies however suggest differential effects of various dairy products on blood lipids. The presence or absence of milk fat globule membrane (MFGM) might explain such discrepancies, possibly through the phospholipid content of MFGM, or its physical structure, or both. Some recent controlled supplementation studies have indicated that MFGM powder may counteract cholesterol raising effects. However no studies have tested foods that contain MFGM in its native form. We aimed to investigate the role of native MFGM on plasma lipoproteins, using common dairy foods. In a single‐blinded, 8‐week iso‐caloric trial, fifty‐seven overweight (mean BMI 28) but healthy men and women (mean age 58 years) were randomized to consume 40 g dairy fat/day with or without MFGM. All dairy products were provided and subjects were not allowed to consume any other dairy products during the study. Subjects visited the clinic weekly for weighing and food distribution. Based on daily checklists, compliance in both groups was excellent. There were no differences in serum fatty acid composition suggesting similar intake of saturated fat in both groups. Although blood lipids were primary outcome, other aspects of cholesterol metabolism (e.g. PCSK9, lathosterol and C4) will be investigated, as well as cardiometabolic risk factors and gene expression in peripheral blood mononuclear cells. Results will be presented at the meeting. This study could help to understand the divergent associations observed in the literature regarding intake of different dairy foods and cardiovascular risk.

Grant Funding Source: Supported by The Swedish Research Council Formas (222‐2011‐305)
Effects of amino acid intake level in pregnant pigs on plasma metabolome (643.3)
Authors: Moehn Soenke , Han BeomSoo , Krishnamurthy Ramanarayan , Liu Philip , Bouatra Souhaila , Wishart David , Pencharz Paul , Ball Ronald , Plastow Graham
Published: June 2021

The objective of this study was to determine commonalities and differences in the metabolome in non‐pregnant pigs and in early (EG), late (LG) and entire pregnancy during the study of amino acid (AA) requirements. Each pig was fed 6 levels of test AA at maintenance and in both EG and LG. The metabolome of 331 plasma samples from 19 sows in their 2nd to 4th parity was analyzed by nuclear magnetic resonance, gas chromatography‐mass spectroscopy (MS) and direct injection MS. Data were evaluated using partial least square‐discriminant analysis (PLS‐DA) in www.metaboanalyst.ca. Across AA tested, AA limitation affected (P<0.05) the metabolome at maintenance and in EG but not in LG. The AA level, across AA tested, was only significant in EG (P=0.01). For AA level, only urea was an important feature in maintenance, EG and LG. There were no common metabolites for AA limitation. The AA tested (Thr, Ile, Lys, Trp) affected (P<0.001) the metabolome at maintenance and in the entire pregnancy, EG and LG. Within AA tested, no significant results were found for Ile and Trp because of fewer observations (<40). The stage of pregnancy affected (P<0.05) the metabolome when studying Thr or Lys. In EG and LG, threonine and aminoadipate were the only common features for Thr and Lys, respectively. For Lys, AA limitation affected (P=0.036) the metabolome in EG but not in LG. For Thr, AA level affected (P=0.029) the metabolome in LG but not in EG. In this study, PLS‐DA could not identify common key metabolites for different AA and status of pregnancy. Metabolomics to determine key metabolites for the effect of AA intake needs to be applied within AA tested and within stage of pregnancy in pigs.

Grant Funding Source: Alberta Livestock and Meat Agency
1,25 Dihydroxyvitamin D inhibits glutamine metabolism in ras‐transformed MCF10A breast epithelial cells (644.12)
Authors: Zhou Xuanzhu , Zheng Wei , Tayyari Fariba , Gowda Nagana , Raftery Daniel , Donkin Shawn , Bequette Brian , Teegarden Dorothy
Published: June 2021

Studies were designed to investigate the effect of 1,25 dihydroxyvitamin D (1,25D) on glutamine metabolism during cancer progression employing untransformed (10A) and ras oncogene transformed MCF10A human breast epithelial cells (ras). Glutamine restriction reduced cell number in both cell lines, and 1,25D ameliorated these reductions. Intracellular glutamine and glutamate levels, determined by NMR, were increased 25% in ras cells compared to 10A cells, and 1,25D reduced these levels by 23% in ras cells. Fractional glutamine and glutamate flux, utilizing U13C5‐L‐glutamine, into the TCA cycle was increased in ras cells by 21% and 18%, respectively, compared to 10A cells. 1,25D decreased glutamine flux into the TCA cycle by 18% and 31% in 10A and ras cells, respectively, and decreased glutamate flux into the TCA cycle by 13% and 17% in 10A and ras cells, respectively. The mRNA expression of the glutamine transporter, solute linked carrier family 1 member A5 (SLC1A5), was decreased by 34% in ras compared to 10A cells. 1,25D further decreased SLC1A5 mRNA expression in ras by 45%, but had no effect in 10A cells. These results suggest 1,25D may alter glutamine metabolism in ras cells by inhibiting glutamine uptake and utilization, in part through down‐regulation of SLC1A5.

Grant Funding Source: Supported by NIH R25CA128770 and NCI CCSG CA23168
Decreased coregulation and interaction of mixed dietary carotenoids when delivered through sequential release technology (645.14)
Authors: Venzon Dawna , Kazlova Valentina , Izzy Samantha , Intra Janjira , Gellenbeck Kevin
Published: June 2021

Carotenoids are implicated as beneficial substances in human health yet human dietary intakes are repeatedly shown to be inadequate to current health recommendations. It may be useful to provide a balanced, naturally derived, mixed carotenoid supplement to address the widespread deficits in carotenoid consumption. However, when mixed carotenoids are delivered into the intestinal tract together considerable competition occurs for micelle formation and subsequent intestinal absorption, affecting the bioavailability of the compounds. Previously we have demonstrated a mixed carotenoid formulation that can release individual carotenoids in a sequential structure over the transit time of the human upper gastrointestinal system (6 hours). We hypothesized that temporally separating the individual carotenoids would reduce competition for micelle formation, improve uptake, and maximize efficacy. Here, we test this hypothesis by evaluating the change of carotenoid levels in the plasma chylomicron fraction for 8 hours after oral doses of a sequential release mixed carotenoid formulation in comparison to a matched conventional release formulation, in a randomized, double blind, repeated measure, cross‐over study in adult humans. We find the carotenoid change from baseline area under the curve (AUC) is more favorable following consumption of the sequential release format over that seen following the conventional release. Spearman’s correlation analysis also demonstrates less interaction and regulation between the sequentially spaced carotenoids, suggesting improved bioavailability from a novel sequential release mixed carotenoid formulation.

Grant Funding Source: Supported by Amway Corporation
Liver protein synthesis and glycogen storage with the progression of cancer cachexia (649.2)
Authors: Narsale Aditi , Puppa Melissa , Carson James
Published: June 2021

Cachexia is a multifactorial syndrome characterized by the loss of peripheral tissue and hypertrophy of visceral organs. The ApcMin/+ mouse model develops an IL‐6 dependent cachexia, leading to gradual loss of muscle and fat and hypertrophy of spleen and liver. The purpose of this study was to examine liver function related to protein synthesis and glycogen storage during the progression of cancer cachexia. Livers collected from non‐cachectic and cachectic ApcMin/+ mice were analyzed for marker of protein synthesis (p38) and degradation/angiogenesis marker ‐ MMP2 protein expression. Liver glycogen content was examined by PAS staining intensity. The role of cachexia related inflammation on liver metabolism was studied by 10mg/kg/day administration of PDTC, a STAT3 and NF‐ΚB inhibitor, for 2 weeks. As cachexia progressed there was suppression of Akt and p38 phosphorylation, and a reduction in liver glycogen content. MMP2 expression was also decreased 97% with severe cachexia. PDTC administration during severe cachexia did not rescue suppressed p38 phosphorylation or MMP2 expression. However, glycogen storage was increased in the PDTC treated cachectic Min mice as compared to the untreated mice. These results indicate that signaling regulating liver protein synthesis is suppressed with cachexia progression, and there is a loss of glycogen stores in severely cachectic mice, which is partially rescued by inhibition of STAT and NF‐ΚB signaling, irrespective of p38 suppression.

Grant Funding Source: Supported by R01CA121249‐01A2
The effect of TNF on the colonic microbiome and acute colitis (650.11)
Authors: Jones‐Hall Yava , Nakatsu Cindy
Published: June 2021

Inflammatory bowel disease affects approximately 1.4 million people in the United States and consists of two disorders, ulcerative colitis and Crohn’s disease (CD). CD is a chronic condition, primarily mediated by T helper (Th)1 and Th17 effector responses. Tumor necrosis factor (TNF) is a promoter of inflammation in CD and pharmacological blockade of TNF reduces inflammation in many CD patients. Elucidation of the exact mechanism of action of anti‐TNF therapy, as well as the potential effect of TNF on the microbiome is needed. Using the chemical trinitrobenzene sulfonic acid (TNBS) to induce acute CD‐like colitis in wild type (WT) and TNF deficient (Tnf ‐/‐) mice, we found that lack of TNF resulted in diminished colitis, indicating that TNF is pro‐inflammatory in TNBS acute colitis. Microbial community structures were compared using Denaturing Gradient Gel Electrophoresis (DGGE) of 16S rRNA genes amplified by PCR using DNA extracted from fecal samples taken before and 72 hours after TNBS colitis induction. PCR‐DGGE profiles of microbial communities Tnf ‐/‐ and WT mice were distinctly different and clustered separately using Dice similarity comparisons. After TNBS treatment, changes in microbial profiles were greater in the Tnf ‐/‐ mice than WT mice. Currently Mi‐Seq2000 Illumina sequencing of the samples are being performed for a more in‐depth characterization of the microbiome. Our preliminary findings emphasize the potential integrated role of the microbiota and TNF in promoting colitis.
Cortactin regulates intestinal epithelial permeability by stabilizing tight junctions (650.6)
Authors: Citalán‐Madrid Alí , García‐Ponce Alexander , Vargas‐Robles Hilda , Betanzos Abigail , Rottner Klemens , Mennigen Rudolf , Schnoor Michael
Published: June 2021

Chronic inflammatory diseases of the intestines such as inflammatory bowel disease (IBD) are characterized by disturbed epithelial contacts and increased intestinal permeability. The actin cytoskeleton and its regulators are critical for these features. Analyzing cortactin‐KO mice, we found that the actin‐binding molecule cortactin (CTTN) controls vascular permeability. However, it is unknown if CTTN also regulates intestinal epithelial permeability. Thus, we generated a stable CTTN‐depleted Caco‐2 cell line that showed reduced transepithelial resistence (TER) and increased paracellular flux of 4 kDa FITC‐dextran. In these cells, we also observed a less developed cortical actin ring and internalization of occludin and ZO‐1. However, CTTN depletion did not cause increased apoptosis. CTTN‐KO mice also showed increased basal permeability in the colon. In healthy human colon tissue biopsies, we found that CTTN colocalized with actin and the tight junction protein ZO‐1. By contrast, in IBD patients, we observed strong internalization of CTTN resulting in a loss of colocalization with actin and ZO‐1. Our data imply that CTTN regulates intestinal permeability by controlling actin dynamics at cell contacts to stabilize epithelial junctions.

Grant Funding Source: CONACYT 179895
Bile acids and acetaminophen protein adducts in children with acetaminophen overdose (653.1)
Authors: Bhattacharyya Sudeepa , Pence Lisa , Yan Ke , Gill Pritmohinder , Letzig Lynda , Simpson Pippa , Beger Richard , James Laura
Published: June 2021

Disruption of bile acid transport is a recognized mechanism of drug induced liver injury. The relationship of circulating bile acids to acetaminophen protein adducts and to alanine aminotransferase (ALT) levels was examined in healthy controls and children hospitalized for acetaminophen overdose. Mean (range) peak values of ALT and adducts for controls (23) and overdose (64) were ALT: 18 (10‐37 IU/L) vs. 626 (8‐9909 IU/L) and adducts: 0.01 (0‐0.01 nmol/mL) vs. 0.72 (0.03‐7.92 nmol/mL). Mean (range) time to treatment with N‐acetylcysteine for overdose subjects was 15 (1‐85 h). Targeted analysis of nine bile acids using UPLC/TQ‐MS showed that Glycocholic Acid (GCA), Glycodeoxycholic Acid (GDCA), and Taurodeoxycholic Acid (TDCA) were higher in children with APAP overdose (p<0.001) compared to controls. ROC analysis of bile acids showed that GDCA was the best discriminator of APAP overdose (ROC AUC=0.861 [95% CI: 0.758 ‐ 0.964] compared to controls). GDCA > 0.364 uM distinguished APAP overdose from controls (sensitivity 93.8%, specificity 68.4%). Comparison of time to achieve peak values was examined for adducts, bile acids and ALT. Adducts peaked prior to ALT (p=0.057) and bile acids (p<0.001) and ALT peaked prior to GDCA and TDCA (p=0.001). Bile acids are increased in children with APAP overdose compared to controls and GDCA is the best predictor of APAP overdose among the bile acids examined.
Safety of emu oil for intestinal applications (653.9)
Authors: Mashtoub Suzanne , Cheah Ker , Howarth Gordon
Published: June 2021

BACKGROUND. Previously, we have reported that Emu Oil (EO) lengthens intestinal crypts, a feature of intestinal growth, in experimental colitis (2012) and mucositis (2013). However, it remains unclear whether the crypt lengthening represents normal or aberrant intestinal growth. AIM. We aimed to determine if crypt depth measurements in EO‐treated rats returned to normal levels following withdrawal of EO therapy. METHODS. Dark Agouti rats (n=8/group) were gavaged daily for 10 days with Water, Olive Oil (OO) or EO (1ml) or 0.5ml EO (0.5EO). Rats were euthanized on day 10 or day 17. Intestinal weights, lengths, villus height (VH) and crypt depth (CD) were quantified. p<0.05 was considered significant. RESULTS. On day 10, jejuno‐ileum (JI) weight was increased by OO (26%) and EO (0.5ml: 15%; 1ml: 29%; p<0.01), which was normalised by day 17. On days 10 and 17, JI length was greater in OO‐ (12%) and EO‐treated rats (0.5ml: 8%; 1ml: 12%; p<0.05), relative to water controls. On day 10, OO and EO increased ileal VH (OO: 32%; 0.5EO: 22%; EO: 35%; p<0.01) and CD (OO: 17%; 0.5EO: 13%; EO: 22%); importantly however, after withdrawal of all oils, VH and CD returned to normal levels. Moreover, the VH:CD ratio (a feature of dysplasia) was unaffected in all oil‐treated rats compared to normal controls. CONCLUSIONS. The restoration of normal intestinal growth following cessation of Emu Oil therapy supports its safety for intestinal conditions.

Grant Funding Source: Supported by The Rural Industries Research and Development Corporation Australia
Identification and characterization of a novel small molecule agonist of lipoprotein lipase by random screening (654.12)
Authors: Sadana Prabodh , Aring Danielle , Burke Jamie , Geldenhuys Werner
Published: June 2021

Elevated triglycerides and decreased HDL cholesterol are hallmarks of metabolic syndrome. Lipoprotein lipase (LPL) found lining the endothelium tissue in the adipose tissue, heart and muscle is an enzyme that catalyzes the hydrolysis of triglyceride rich lipoproteins and chylomicrons into free fatty acids. Regulation of LPL is a key physiological mechanism for the control of lipid levels and atherosclerosis risk. Angiopoietin‐like 4 (ANGPTL4) which inhibits the activity of LPL is a critical regulatory protein in LPL physiology and triglyceride metabolism. Currently, the most well characterized LPL agonist is NO‐1886. We screened an in‐house chemical library at a fixed concentration of 40µM in an in vitro LPL assay. Our screening identified a compound designated C10, showing LPL agonist activity equipotent to NO‐1886. Further, we determined that this novel LPL agonist is able to rescue the LPL inhibition by ANGPTL4, whereas NO‐1886 doesn’t rescue against ANGPTL4 inhibition. Additionally, we performed structure activity relationship study for C10 and identified a more potent LPL agonist (C10d) exhibiting at least 3 fold greater increase in LPL activity than NO‐1886. Our studies have identified a novel LPL agonist that can find therapeutic use in dyslipidemias associated with diabetes and cardio‐metabolic conditions like atherosclerosis.
The study of anti‐aging of Zishen Yigan Wan in vivo and vitro (657.2)
Authors: Liu Ying , Liu Ming , Wang Xuanbin , Li Hongliang , Zhang Qiufang , Zhang Xiaoyan , Wang Linhai
Published: June 2021

The aging‐associate diseases result in greater and greater burden for the families and community in China. A traditional folk formulation, Zishen Yigan Wan comprises 11 traditional Chinese medicinal herbs. This study aimed to observe the effects of Zishen Yigan Wan on anti‐aging and the underlying mechanisms in vivo and in vitro. Compared with the model group of ICR mice, the escape latency of each group treated by Zishen Yigan Wan with low, medium and high dosage decreased significantly (P<0.01), and the number of passing though the original platform increased significantly (P<0.01); the thymus, spleen index, GSH‐PX activity of liver, SOD activity of plasma, liver and brain tissue increased significantly, respectively (P<0.01); while the MDA level of the liver and brain, the level of NO in plasma dropped significantly (P<0.01); The viability of PC12 cells pretreated with Zishen Yigan Wan at low, medium and high dosage were increased significantly (P<0.01). The studies showed that Zishen Yigan Wan could delay ICR mice aging induced by D‐galactose, and improve the ability of learning and memory and immune function of aging mice. The mechanism may be that Zishen Yigan Wan could clear away free radicals and improve the antioxidant system’s activity of the body to slow down the oxidative stress damage of the body induced by D‐galactose.

Grant Funding Source: Supported by 2011 Project of Hubei Province, NO. 4
Role of reactive oxygen species as signal transducers in β2 adrenergic receptor mediated β‐arrestin signaling (662.2)
Authors: Singh Monalisa , Burns Rebecca , Moniri Nader
Published: June 2021

Reactive oxygen species (ROS) have previously been linked to deleterious effects caused by oxidative stress as well as to inflammatory disorders, degenerative disorders and cancers. More recently, ROS have been recognized as important signaling molecules, and have been shown to be involved in G protein‐coupled receptor (GPCR) signal transduction. Our laboratory has previously shown that agonism of the prototypical GPCR, the β2 adrenergic receptor (β2AR), generates intracellular ROS and that β2AR‐associated G protein signaling is dependent on ROS. Here, we demonstrate that ROS are critical mediators of β2AR signaling to β‐arrestins. Using coimmunoprecipitation and BRET, we reveal that the physical interaction of β2AR and β‐arrestins are dependent on ROS. Furthermore, using phosphorylation of ERK1/2 as an endpoint for both G‐protein and β‐arrestin signaling, we demonstrate that ROS are required for both G protein and β‐arrestin dependent ERK1/2 phosphorylation. These results are presented in light of data, which demonstrate that β2AR agonism can lead to receptor S‐Sulfenation, a process that is dependent on ROS, and which suggests that ROS may serve a purposeful role in stabilizing β2AR in signal‐capable conformations.

Grant Funding Source: Internal funding
Discovery and characterization of a G protein‐biased agonist of the D2 dopamine receptor (662.7)
Authors: Chun Lani , Free R. , Moritz Amy , Conroy Jennie , Meade Julie , Xiao Jingbo , Dulcey Andrés , Vekariya Rakesh , Ferrer Marc , Javitch Jonathan , Beuming Thijs , Shi Lei , Southall Noel , Marugan Juan , Aubé Jeff , Frankowski Kevin , Sibley David
Published: June 2021

The D2 dopamine receptor (D2R) can activate a spectrum of signaling cascades primarily through G proteins and β‐arrestin recruitment, making it an attractive target for the development of signaling biased ligands. Unlike dopamine, which simultaneously activates G proteins and recruits β‐arrestin, a biased ligand affects only one pathway, and the development of such ligands can allow for more fine‐tuned study of receptor signaling. Our lab has identified a compound (MLS1547) that is a highly efficacious agonist at D2R‐mediated G protein‐linked signaling, but it does not recruit β‐arrestin. Rather, this compound is an antagonist of D2R‐stimulated β‐arrestin‐mediated signaling. A number of structural analogs of MLS1547 were characterized for their signaling properties, which ranged from fully biased, partially biased, to unbiased. These results provided the basis to use pharmacophore modeling and molecular docking analyses to build a preliminary structure‐activity relationship of the functionally‐selective properties of these compounds. This, along with medicinal chemistry approaches, will allow for the identification of more potent G protein‐biased analogs of MLS1547 that will enable a highly targeted approach for investigating D2R signaling in vivo, and may eventually allow for selective treatment of disease symptoms associated with D2R dysfunction.

Grant Funding Source: Supported by the NIH IRP.
Endothelial cell phenotype varies by sex and tissue of origin (664.3)
Authors: Bingaman Susan , Stockard Kevin , Huxley Virginia
Published: June 2021

Endothelial cell (EC) dysfunction is a hallmark of several vascular diseases (CVD) that also show female (F) vs. male (M) differences in incidence and severity. As these differences change with age, i.e., pre‐menopausal F < M, post‐menopausal F 蠅 M, it has been assumed that the reproductive sex hormones, particularly E2, account for the dichotomy. In spite of the correlations between hormonal changes and CVD incidence, data exist demonstrating sex‐hormone independent variations. Before the mechanisms regulating health and disease can be determined, the phenotypes of the F and M EC require description and comparison in the absence of changes in reproductive hormones. To this end we isolated EC from aorta (AEC), skeletal muscle (SkmEC) and mesenteric microvasculature (MesMEC) of young adult F and M Sprague Dawley rats. The EC evaluated under basal conditions for a variety of attributes, including morphology (size, shape, topology, EC‐specific characteristics), growth, metabolism, and receptor and signaling protein expression displayed multiple sex‐ and tissue‐specific differences. This knowledge facilitates the development of appropriate models for future investigations, both in vitro and in vivo.

Grant Funding Source: Supported by DK095501, HCO6RR017353 & the MU Pulmonary/MPP Partnership
Adenosine prevents TNFα‐induced decrease in endothelial mitochondrial mass via activation of eNOS‐PGC‐1α regulatory axis (664.5)
Authors: Kalogeris Theodore , Baines Christopher , Korthuis Ronald
Published: June 2021

To gain insight into the potential role of endothelial mitochondrial function and biogenesis in the preconditioning effects of adenosine (Ado), we incubated human microvascular endothelial cells (HMEC‐1) with TNFα in the presence or absence of Ado. TNFα produced time and dose‐dependent decreases in mitochondrial membrane potential, cellular ATP levels, and mitochondrial mass, which preceded an increase in apoptosis. These effects were prevented by co‐incubation with Ado, an NO donor (detaNO), a guanylate cyclase (GC) activator (YC‐1), or a cell‐permeant cGMP analog (8‐Br‐cGMP). The effects of Ado were blocked by a NO synthase inhibitor (L‐NIO), a soluble guanylate cyclase inhibitor (ODQ), a morpholino antisense oligonucleotide to eNOS, or siRNA knockdown of PGC‐1α. DetaNO reversed the the effect of eNOS knockdown, as did YC‐1 and 8‐Br‐cGMP, while the effect of detaNO was blocked by ODQ. Both detaNO’s and 8‐Br‐cGMP’s effects were prevented by siRNA to PGC‐1α. TNFα also decreased expression of eNOS and PGC‐1α, which was reversed by Ado. DetaNO, but not Ado, rescued expression of PGC‐1α in cells in which eNOS expression was knocked down by eNOS antisense treatment. These results support the existence of an adenosine‐triggered, mito‐and cytoprotective mechanism dependent upon an eNOS‐PGC‐1α regulatory pathway, which acts to preserve endothelial mitochondrial function and mass in the face of inflammatory challenge.

Grant Funding Source: Supported by NIH grants: AA014945 & HL095486
Modulating Ca2+ entry into endothelial tubes by controlling membrane potential during muscarinic receptor activation (664.7)
Authors: Behringer Erik , Segal Steven
Published: June 2021

Coupling of intracellular Ca2+ [Ca2+]i to hyperpolarization of endothelial cells underlies smooth muscle cell relaxation in resistance vessels. Whether membrane potential (Vm) influences [Ca2+]i levels in the endothelium remains controversial. To resolve the impact of Vm on Ca2+ influx, Fura‐2 photometry was evaluated during intracellular current injection (‐5 to +5 nA; 20 s pulses) to control Vm of endothelial tubes (width, 60 µm; length, 蠄 1 mm) freshly isolated from superior epigastric arteries of adult (4‐6 mo) male C57BL/6 mice (n蠅4). Relative to resting Vm (‐40±2 mV), current injection hyperpolarized (Vm in mV; ‐1 nA: ‐52±2, ‐3 nA: ‐64±2, ‐5 nA: ‐79±2) and depolarized (+1 nA: ‐33±1, +3 nA: ‐13±2, +5 nA: 8±3) cells. Under resting conditions, changing Vm did not affect [Ca2+]i (e.g., F340/F380 = 0.68±0.03 at rest and ‐5 nA). However, during exposure to acetylcholine (ACh; 100 nM, ~EC50), the plateau phase of the [Ca2+]i response was either enhanced [∆F340/F380 & ∆Vm (mV); ‐1 nA: 0.07±0.02 & ‐10±1; ‐3 nA: 0.16±0.02 & ‐23±2; ‐5 nA: 0.21±0.02 & ‐38±1] or reduced (+1 nA: ‐0.08±0.02 & 11±2; +3 nA: ‐0.18±0.03 & 28±3; +5 nA: ‐0.34±0.03 & 49±4) according to voltage amplitude and polarity (P<0.05). These effects of ACh and ∆Vm were abolished in the absence of extracellular Ca2+ (2 mM). Thus Vm can modulate Ca2+ entry during submaximal activation of ion channels that enable Ca2+ influx in accord with the electrochemical driving force.

Grant Funding Source: Supported by R37‐HL041026 & 5T32‐AR048523
Interaction between endotoxin and histones in thrombosis (671.5)
Authors: Lam Fong , Parikh Kathan , Cruz Miguel , Rumbaut Rolando
Published: June 2021

Histones, the protein component of nucleosomes, are released from neutrophils during inflammation and have a pro‐aggregatory effect on platelets. Recent reports suggest that histones interact with toll‐like receptor‐4; therefore, we studied the effect of endotoxin (LPS) on nucleosome formation and histone‐induced platelet aggregation. We measured plasma nucleosome levels in mice exposed to either LPS or control, and the effects of exogenous histones of photochemical‐induced thombus formation in cremaster venules. Finally, we compared the effects of LPS and histones on platelet aggregation. We observed that 4 hours after LPS, mice had a 4‐fold higher level of plasma nucleosomes compared to saline‐exposed mice (p<0.01). Furthermore, exogenous histones resulted in a decreased platelet counts (from 1.1 x 106 to 0.86 x 106/µL; p< 0.05), increased thrombin‐anti‐thrombin complexes (from 0.99 to 6.1 ng/mL; p<0.01), and accelerated rates of thrombosis (time to thrombotic occlusion was reduced from 6.5 to 4.4 min; p<0.05). Finally, whereas LPS alone did not induce platelet aggregation, it enhanced the aggregatory response after histone administration in a dose‐dependent manner (22, 28, and 38% aggregation with LPS 0, 10, and 100 ng/mL, respectively; p<0.01). Taken together, these data suggest a synergistic interaction between endotoxin and histones in producing a prothrombotic state.

Grant Funding Source: Supported by a BLR&D Merit Review Award from the Department of Veterans Affairs
Niacin‐induced enhancement of lysosomal cholesterol efflux in macrophages through CD38 ‐ NAADP signaling pathway: implication in reduced foam cell formation (671.6)
Authors: Zhang Fan , Zhang Qiufang , Li Ningjun , Li Pin‐Lan
Published: June 2021

Niacin (Nicotinic acid), a hyperlipidemia‐lowering agent, acts as a substrate in the enzymatic synthesis of lysosomal Ca2+ messenger, NAADP by CD38. Our recent studies have demonstrated that the deficiency of CD38 led to lysosomal lipid accumulation in macrophages, foam cell formation and atherosclerosis, as observed in CD38‐/‐ mice. The present study was designed to examine the effects of niacin on the foam cell formation and explore the underlying mechanisms. Using fluorescent microplate reader and confocal microscope, we found that niacin dose‐dependently (in mM: 0, 0.05, 0.1, 0.3, 1 and 2) decreased lysosomal lipid accumulation in bone marrow‐derived wild type macrophages treated with oxLDL (40 µg/ml) as stained by lysosomal lipid specific dye of Bodipy 493/503, but it had no effects in CD38‐/‐ macrophages. This niacin‐induced lysosomal lipid lowering effects were almost completely abolished by NAADP antagonist of PPADS (100 µM). In the niacin‐treated macrophages, the cholesterol level in the purified lysosomal fractions was markedly decreased. Correspondingly, oil red o staining and electron microscope assay showed that the foam cell formation was significantly reduced. These results for the first time reveal that niacin promotes lysosomal cholesterol efflux and thereby attenuates macrophage foam cell formation, which is associated with activation of CD38‐NAADP signaling pathway.

Grant Funding Source: NIH R01HL115068 and R01HL091464
Changing shear stress‐induced ATP release from erythrocytes increases endothelial calcium and vascular permeability independent from nitric oxide production in intact rat venules (672.2)
Authors: Xu Sulei , He Pingnian
Published: June 2021

Disturbed shear stress (SS) promotes atherosclerosis, thrombosis, and inflammation of the vessel wall. Endothelial cell (EC) barrier dysfunction has been implicated to play important roles in the pathogenesis of many cardiovascular diseases. However, how the disturbed blood flow affects EC barrier function in vivo and the involved cellular mechanisms remain to be identified. Our study investigated changing SS‐induced endothelial signaling and permeability in intact rat venules. EC [Ca2+]i and NO were measured in Fura‐2 AM and DAF‐2 DA loaded venules. EC eNOS activation was evaluated by immunofluorescence staining with confocal imaging. ATP released from red blood cells (RBCs) in response to SS is measured by bioluminescent assay. In response to a sudden change of SS, transient shear magnitude‐dependent increases in EC [Ca2+]i occurred only in vessels perfused with whole blood or perfusate containing RBCs, which was correlated with EC gap formation illustrated by fluorescent microsphere accumulation. Carbenoxolone, a Pannexin 1 inhibitor, significantly reduced shear magnitude‐dependent ATP release from RBCs and also abolished SS‐induced increases in EC [Ca2+]i. However, both plasma and whole blood perfusion induced shear magnitude‐dependent NO production and eNOS‐ser1177 phosphorylation. These results indicated that in intact venules ATP released from RBCs in response to the sudden change of SS is accountable for the increases in EC [Ca2+]i and vascular permeability, while SS‐induced NO production can be independent of increased EC [Ca2+]i.

Grant Funding Source: Supported by HL56237, HL084338 and AHA 12PRE11470010
Evidence for a Rac1 independent mechanism for sphingosine‐1‐phosphate mediated endothelial barrier enhancement (672.7)
Authors: Zhang Xun , Adderley Shaquria , Breslin Jerome
Published: June 2021

S1P is a bioactive lipid that tightens intercellular junctions between endothelial cells (EC) through coordinated spreading of local lamellipodia. We tested the hypothesis that S1P causes protrusions through activation of the small GTPase Rac1. Cultured human umbilical vein EC or human dermal microvascular EC monolayers were used to model the endothelial barrier. Trans‐endothelial electrical resistance (TER) served as an index of barrier function and time‐lapse imaging of EC was employed to study local lamellipodia. Rac1 expression/activity was modulated by transfection of siRNA or plasmids encoding wild‐type (WT) or dominant‐negative (DN) GFP‐Rac1, and confirmed by Western blot or fluorescence microscopy, respectively. Non‐targeting RNA sequences and GFP transfection served as respective controls. The results show that S1P (2 μM) caused rapid, coordinated protrusions of local lamellipodia from individual cells that coincided with increased TER. Neither Rac1 knockdown, nor overexpression with WT or DN Rac1 significantly changed TER, compared to controls, at 72h. Additionally, S1P significantly increased TER in Rac1 knockdown and overexpression groups, at an equivalent magnitude as controls. These data suggest that a Rac1 independent mechanism controls S1P‐induced EC protrusions and endothelial barrier enhancement. Supported by NIH R01HL098215.

Grant Funding Source: Supported by NIH R01HL098215
Effects of color temperature (Kelvin) of LED bulbs on blood physiological variables of broilers grown to heavy weights (674.1)
Authors: Olanrewaju Hammed , Purswell Joseph , Collier Stephanie , Branton Scott
Published: June 2021

Light‐emitting diode (LED) has been used in the poultry industry to reduce energy usage in broiler production facilities. However, limited data are available comparing efficacy of different wavelength of LED bulbs. The present study evaluated the effects of color temperature (Kelvin) of LED bulbs on blood physiological variables of broilers grown to heavy weights. The experimental was a randomized complete block design. Four treatments consisted of 3 LED light bulbs (2700k, Warm [LED‐1], 3500k, [LED‐2,], 5000k, Cool [LED‐3]) and incandescent light (ICD, control) from 0 to 56 d of age along with 5 lx intensity starting from 22 to 56 d of age. Chicks were equally and randomly distributed into 16 environmentally‐controlled rooms at 50% RH. Each of the 4 treatments were represented by 4 rooms with 4 replicates. Feed and water were provided ad libitum. Venous blood samples were collected on d 21 (base line) and d 56 of age. Treatments and sex had significant (P 蠄 0.05) effect on BW, pH, pCO2, pO2, and sO2. All these acid‐base changes are still within the normal acid‐base homeostasis and physiological ranges. Plasma corticosterone was not affected by treatments. This study shows the positive impact on profits to commercial poultry industry that are using LED light sources in comparison to ICD. It was concluded that LED light sources are better potential light source for broiler production without inducing stress on broiler health.
Integration of microvascular reactivity, structure and hemodynamics for skeletal muscle function in metabolic syndrome (674.8)
Authors: Butcher Joshua , Chantler Paul , Wu Fan , Brooks Steven , Stanley Shyla , Frisbee Jefferson
Published: June 2021

With metabolic syndrome (MS) in obese Zucker rats (OZR), the ability of in situ skeletal muscle to resist fatigue is compromised well before muscle function; implicating microvascular or perfusion‐based impairments as playing a causal role. However, our results suggest that bulk flow to muscle is not sufficiently constrained to explain the poor performance, and indices such as dilator/constrictor reactivity, vessel wall mechanics and capillary density are not strong predictors of functional outcomes. We have determined that altered RBC distribution at arteriolar bifurcations (γ) is increasingly heterogeneous in OZR muscle, iterating to produce a wide heterogeneity of pre‐capillary flow distribution vs. controls. This increased spatial heterogeneity of perfusion at bifurcations is not compensated for via temporal switching, rather it is exacerbated owing to blunted temporal activity. The combined effect of these behaviors is that microvascular hematocrit becomes increasingly heterogeneous and fixed, compromising perfusion:demand matching and muscle performance. The magnitude of the deviation of γ from 0.5, and its temporal stability are the strongest predictors of muscle performance to date.

Grant Funding Source: Supported by National Institutes of Health and American Heart Association
Altered skin vasomotion in hypertensive humans (677.1)
Authors: Bruning Rebecca , Kenney W. , Alexander Lacy
Published: June 2021

Endothelium‐dependent cutaneous vasodilation (VD) is attenuated in essential hypertensive (HTN) humans. Using spectral analysis (fast Fourier transformation) we aimed to characterize vasomotion in laser Doppler flowmetry (LDF) recordings during local heating (LH)‐induced VD before and after inhibition of nitric oxide (NO)‐dependent signaling (NOS‐I: L‐NAME) in HTN and normotensive (NTN) humans. We hypothesized that HTN would have lower total power spectral densities (PSD), and in the frequency intervals (FI) associated with endothelial and neurogenic vascular control. LDF recordings during LH experiments from 18 HTN (MAP: 108±2mmHg) and 18 NTN (MAP: 88±2mmHg) men and women were analyzed. LH‐induced VD increased total PSD for all FI (all p<0.001). HTN had a lower total PSD (HTN:30±6AU2, NTN:14±3AU2, p=0.03) and neurogenic FI (HTN:6±1AU2, NTN:17±3AU2, p<0.01) versus NTN. As a percentage of total PSD, HTN had reduced neurogenic (HTN:41±3%, NTN:51±1%, p<0.001) and higher myogenic contributions (HTN:29±3%, NTN:24±1%, p=0.04) to the total spectrum. NOS‐I decreased total PSD (p<0.001) for both groups, but HTN exhibited lower endothelial (p<0.01), neurogenic (p<0.05), and total PSD (p<0.001) compared to NTN. These data suggest that HTN results in altered neurogenic and NO‐dependent control of skin vasomotion and support the use of spectral analysis as a non‐invasive technique to study vasoreactivity.

Grant Funding Source: Supported by R01‐HL093238‐02