bluuebunny/crossref_metadata_2025 huggingface dataset:
Characterization of porcine alternatively activated macrophages
Authors: Dawson Harry Dale , Reece Joshua J. , Solano‐Aguilar Gloria , Kohn Kristen , Vangimalla Vandana , Urban Joseph F.
Published: June 2021

Recent studies indicate that rodent and human alternatively activated macrophages (AAMFs) differ in phenotype. We hypothesized that the phenotype pig and human AAMFs would be more similar than mouse because of more generally comparable immune function. AAMFs were generated in vitro by treating alveolar macrophages with IL‐4, and in vivo by infection with Ascaris suum. AAMF markers were systematically examined by real‐time PCR. CD209, SOCS1, CCL11, CCL17, and CCL26 were the most stable markers in vitro. Like human AAMFs, there was no or low level induction of arginase‐1 (ARG1) and several chitinase family members; however, there was constitutive expression of ARG1 mRNA and enzymatic activity. The mannose receptor (MRC1) and chitinase‐related gene (CHID1) were induced at a low level in vitro and in vivo. In addition, there was a significant induction of several retinol dehydrogenases, RDH10, RDH11, and DHSR9 in vitro and in vivo. Because of this, we examined the effect of all‐trans retinoic acid (ATRA) on development of pig AAMF. Exogenous ATRA increased basal and IL‐4‐induced CCL17 and CCL22 expression, but inhibited basal and IL‐4‐induced CD209 expression. Thus, pig AAMFs express characteristics of both rodent and human AAMF. Additionally, the notable property of dendritic cells to synthesize ATRA that affects local immune responses may also be expressed in pig AAMFs. Research funded through UDSA.
Daily patterns in phagocytosis by zebrafish kidney leukocytes
Authors: Chrenek Ryan , Ruksznis Catarina , Bagley Patrick , Kaplan Jessica , Hannum Lynn
Published: June 2021

The cyclic expression of clock genes creates daily variations in vertebrate behavior and physiology. Oscillations in leukocyte number and activity have been reported in mammalian systems, but little is known about daily rhythms in zebrafish immune function. Expanding the scope of our earlier work on patterns in zebrafish respiratory burst responses, we have examined daily patterns in the phagocytic activity of zebrafish kidney leukocytes. Phagocytosis of bacteria was monitored by flow cytometry at 4 hour intervals throughout 24‐hour periods. The percentage of phagocytes engulfing S. aureus peaked at 2 a.m. (ZT 18) and reached a low at 2 p.m. (ZT 6). Peak relative numbers of bacteria engulfed per cell occurred at 10 a.m. (ZT 2), after which levels fell steadily until dramatically increasing again at 10 a.m. These data provide evidence of rhythmic immune activity in zebrafish, and suggest that zebrafish can be a valuable model in which to study the relationship between circadian gene expression and the activity of immune system cells.

This work was supported by NIH Grant Number P20 RR‐016463 from the INBRE Program of the National Center for Research Resources.
13C and 15N abundance of plasma and saliva protein reflect protein intake from vegetable and animal products
Authors: LIU YUQIU , SHRIVER TIMOTHY C. , SCHOELLER DALE A.
Published: June 2021

Self‐reported dietary intake is often biased. Therefore there is a need for objective biomarkers of diet. We investigated whether isotopic compositions of human protein reflected the proportion of plant and animal protein intake of individuals consuming U.S. foods. We recruited nine each omnivore, lacto‐ova vegetarian, and strict vegan healthy adults. Food frequency questionnaires were administered to assess protein intake. Plasma and saliva protein specimens were analyzed using an isotope ratio mass spectrometry equipped with an elemental analyzer. For plasma protein samples, vegans could be distinguished from omnivores in both 13C (−21.6‰ ± 0.5 vs. −19.3‰ ± 0.5, p < 0. 01) and 15N (8.0‰ ± 0.5 vs. 9.0‰ ± 0.3, p < 0.01) abundances. Lacto‐ova vegetarians were significantly different from vegans for both 13C and 15N abundances. For saliva protein, vegans could be clearly distinguished from omnivores in 13C (−21.5‰ ± 0.6 vs. −19.6‰ ± 0.7, p < 0. 01) and trended in 15N (6.6‰ ± 0.7 vs. 7.3‰ ± 0.7, p = 0.02). The 13C abundances of omnivores were close to values predicted for US foods, but values for vegans contained more 13C than predicted suggesting carbon exchange between macronutrients via intermediary metabolic pathways. The 15N differences displayed the expected trophic level effect. In conclusion, human plasma and saliva 13C and 15N values both reflected the proportion of animal protein in the U.S. diet. Supported by Dairy Management, Inc.
Endotoxin Reduces Muscle Protein Synthesis and Restrains Translation Initiation by Decreasing eIF4G Phosphorylation in Neonatal and Young Pigs
Authors: Orellana Renan , Gazzaneo Maria Carolina , Wilson Fiona , Nguyen Hanh V , Suryawan Agus , Davis Teresa A
Published: June 2021

Endotoxin (LPS) reduces muscle protein synthesis by restraining translation in the presence of fed levels of insulin and amino acids. In vivo effects of LPS on translation initiation in muscle during basal fasting insulin and amino acid conditions have not been determined in neonates, whose muscle protein synthesis rates are uniquely high. Overnight fasted 7‐ and 26‐d‐old pigs were infused for 8 h with LPS (0 and 10 μg·kg−1·hr−1) and fractional protein synthesis rates and translational control mechanisms in skeletal muscle were examined (n=5–7/group/age). LPS reduced the basal rate of muscle protein synthesis in 7‐ and 26‐day‐old pigs in the presence of fasting levels of insulin, glucose, and amino acids. LPS decreased eIF4G phosphorylation and the amount of the active eIF4G‐eIF4E complex in muscle of 7‐ and 26‐d‐old pigs, but had no effect on mTOR or eEF2 phosphorylation. Although muscle protein synthesis decreased and eEF2 phosphorylation increased with age, there was no effect of age on translation initiation factor activation in basal fasting conditions. The results suggest that the LPS‐induced decrease in eIF4G phosphorylation in muscle may be a limiting step in translation initiation which contributes to the sepsis‐associated decrease in basal protein synthesis rates in muscle during early postnatal life. NIH AR44474, NIH AR51563, NIH HD41648, and USDA 58‐6250‐6‐001.
Methionine and S‐adenosylmethionine are potential biomarkers to determine safe intake levels for methionine
Authors: Toue Sakino , Chin Keigi , Kodama Riho , Amao Michiko , Kawamata Yasuko , Kimura Takeshi , Sakai Ryosei
Published: June 2021

Although mammals maintain homeostasis in response to various intake levels of nutrients by controlling their metabolic activities, their excessive intakes cause adverse effects. Since intake levels within homeostatic range are considered to be safe, determination of metabolic upper limit for each nutrient could provide information for their safe intake levels. In this study we tried to identify the rate‐limiting step for methionine (Met) in its excess and to find biomarkers for Met excess determination.

F344 male rats were fed a basal diet or diets containing additional Met (0.3, 0.6, 0.9, 1.2 and 2.4%) for 2 weeks. Postprandial plasma and liver samples were collected to measure Met metabolites. More than 0.9% of additional Met increased hepatic Met and S‐adenosylmethionine (SAM) markedly. Met loading at the same doses also decreased the ratio of S‐adenosylhomocysteine (SAH) to SAM, indicating that metabolism from SAM to SAH would be one of rate‐limiting steps for disposal of excessive Met and that both SAM and its upstream Met are possible biomarkers to assess metabolic upper limit of Met. Since NOAEL and LOAEL of additional Met in growing rats were 0.6 and 0.9%, respectively, hepatic SAM, plasma and hepatic Met concentrations discriminated safe intake levels from potentially toxic levels well. These suggest that both Met and SAM are potential biomarkers to determine safe intake level of Met.
Scientific Rigor and Effective Advocacy: The Historical Challenge of Defining Food Insecurity and Hunger in America
Authors: Rose Diego , Chilton Mariana
Published: June 2021

Since the late 1960s, biomedical scientists and others have been documenting the hunger problem in America. These efforts have informed policy debates and legislation on food assistance and welfare programs. Documenting hunger in America represents a dual challenge. On one hand is the importance of getting a scientifically defensible estimate of the magnitude of the problem with only limited amounts of information. On the other hand is the need to provide a policy‐relevant indicator that is easily understood and made available in a timely fashion. Initially, hunger was often equated with malnutrition and estimates relied on knowing the size of the population in poverty. These early estimates were rough, missing the nuances that hunger occurs before obvious symptoms of malnutrition, and that income status is an inadequate proxy for those affected by hunger. Since the mid‐1990s, estimates of hunger have been linked to household food insecurity and derived from direct measures using experiential‐based survey questions in a nationally representative sample. As techniques for estimating food insecurity and hunger have become more refined, there has been increased demand for more precision in the definition and measurement of the hunger condition. This paper traces the history of defining hunger, along with the challenges involved, and suggests new directions for balancing scientific rigor with effective advocacy.
Early growth velocity predicts longitudinal growth failure
Authors: Ross Erin Sundseth , Johnson Susan L , Shroyer A. Laurie , Prochazka Allan , Dickinson L. Miriam , Krebs Nancy , Pharo Susan A
Published: June 2021

Our primary objective was to evaluate whether a decline in growth trajectory, identified as a downward movement in z‐scores between the 4 and 6 month time period, can be used to predict later growth faltering, and whether the time period of 2 – 4 months is as effective of a predictor as the 4 – 6 month period. Longitudinal data from birth to 2 years of age were analyzed for 1978 healthy, term babies seen in a staff model HMO. The outcome variable was a weight‐for‐length ratio z‐score of ≤ −1.67, with measurements collected at 12, 18 and 24 months of age. The odds ratio (OR) for infants with a negative shift in z‐scores > −0.85 across the 4–6 month time period was 2.4 (95% CI 1.5, 3.9), and across the 2–4 month time period was 4.1 (95% CI 1.5, 11.4) compared to those without this shift, holding birthweight constant. Odds were increased as birth weight decreased, with an OR of 2.2 for infants <2.75kg, and an OR of 1.9 for infants ≥2.75kg, <3.0 kg, compared to infants 3.0 kg or heavier at birth. There was no significant difference between the 4–6 month (p=0.0006) or 2–4 month (p=0.0069) periods. Our findings support the hypothesis that a downward shift in growth velocity of this magnitude during early infancy can be used to identify children at‐risk of later poor growth. This information can be used to develop research studies and to support providers in decision‐making. Database development was supported by Kaiser‐Permanente of Colorado.
Effect of different leucine supplementation in low protein diet on Protein Synthesis and Activation of Translation Initiation Factors of Weaned Piglets
Authors: Yin Yulong , Wu Guoyao , Liu Zhaojin , Chu Wuyin , Huang Ruilin , Li Tiejun , Hou Rongqing , Liu Yulan
Published: June 2021

Acute administration of L‐leucine stimulates protein synthesis in skeletal muscle of young pigs. However, little is known about a role for the chronic enteral feeding of leucine in regulating the activity of translation initiation factors, nutrient utilization, or growth of the neonate. Eighteen 21‐day crossbred pigs with an average live body weight 8 kg were assigned randomly to one of three groups and feed with low protein dietaries (16% crude protein) containing 1.3%, 1.6%, or 1.9% leucine which meets the marginal, adequate and excess leucine requirement of piglets respectively. At 35 d old, protein synthesis was measured and blood samples were obtained at 15, 30, 45min after infusion for metabolite analysis and the abundance and phosphorylation state of mammalian target of rapamycin (mTOR), ribosomal protein S6 kinase 1 (S6K1), and eIF4E‐binding protein‐1 (4E‐BP1) were measured in longissimus muscle and liver. The results shows that excess leucine could increase (P < 0.05) the fractional synthesis rate (FSR) of heart, posterior small intestine, kidney, liver, spleen, stomach and pancreas compared with pigs adequate and marginal group. Data in the present study also show that excess leucine could increase positive translation initiation factors activity in liver and muscle compared with marginal leucine group. These findings suggest that, despite the activation of positive translation initiation factors, adequate availability of essential amino acids may be necessary for leucine to stimulate muscle protein synthesis in young pigs.
Tumor Blood Vessel Dynamics During and After Angiogenesis Inhibition
Authors: McDonald Donald M.
Published: June 2021

Inhibitors of VEGF signaling cause robust and rapid changes in blood vessels of tumors. The inhibitors block angiogenesis, have potent antivascular actions resulting in regression of tumor vessels, and reduce the abnormality of tumor vessels that survive. Within hours, endothelial fenestrations and vascular sprouts disappear, and patency is lost and blood flow ceases in a population of tumor vessels. By the end of the first week, as many as 80% of tumor vessels regress. Surviving tumor vessels acquire more normal structural and functional features, in a process called normalization. Pericytes of tumor vessels are not as severely impacted by VEGF inhibitors as endothelial cells. Empty sleeves of vascular basement membrane persist after tumor vessels regress, providing a historical record of degenerated tumor vessels and a scaffold for vascular regrowth. Withdrawal of treatment leads to regrowth of endothelial sprouts into empty sleeves of basement membrane. Regrowth begins within a day after withdrawal of inhibitors that are rapidly cleared. Within a week, tumors may be fully revascularized with functional blood vessels. These results raise the possibility that empty sleeves of vascular basement membrane and accompanying pericytes provide a scaffold for rapid revascularization of tumors after removal of anti‐VEGF therapy and highlight their importance as potential targets in cancer therapy.
HIFα induces the recruitment of bone marrow‐derived vascular modulatory cells to regulate tumor angiogenesis
Authors: Bergers Gabriele
Published: June 2021

Several studies have shown that neovascularization of tumors is not solely restricted to activation of differentiated endothelial cells but can also involve the recruitment of bone marrow‐derived cells to active sites of vascular remodeling in tumors. These BMDCs include endothelial progenitor cells (EPC) and pericyte progenitor cells (PPC). While EPCs incorporate into the vasculature and differentiate into endothelial cells, PPCs envelope blood vessels and mature into pericytes and vascular smooth muscle cells. In addition, CD45+ cells of the monocytic lineage that function as vascular modulators, but are not physically part of the vasculature present the biggest and most heterogeneous group. Recruitment of BMDC, specifically of EPC, can vary substantially between tumor types and stages but little is known about factors that dictate the mobilization of BMDC from the bone marrow into the blood stream and their recruitment and retention into the tumor. The most prominent factors identified thus far include VEGF, PlGF and PDGF‐B acting as mobilization factors of EPCs and PPCs respectively and SDF1α, which acts to retain CXCR4+ BMDCs within tumors. We hypothesize that hypoxic conditions in tumors may promote neovascularization by mediating BMDC recruitment. We demonstrate that HIF1α, the direct effector of hypoxia, partly through increases in SDF1α, induces recruitment of bone marrow (BM)‐derived CD45+ myeloid cells containing Tie2+, VEGFR1+, CD11b+ and F4/80+ subpopulations, as well as endothelial and pericyte progenitor cells to promote neovascularization in glioblastoma. MMP‐9 activity of BM‐CD45+ cells is essential and sufficient to initiate angiogenesis by increasing VEGF bioavailability. In the absence of HIF1α, SDF1α levels decrease, fewer BM‐derived cells are recruited to the tumors, decreasing MMP‐9 and solubilization of VEGF.
Influence of Lean and Fat mass on Bone Mineral Content in Asian and White Adolescent Girls
Authors: Daida Yee (Yihe) , Novotny Rachel , Vijayadeva Vinutha , Grove John , LeMarchand Loic
Published: June 2021

A high BMI has been associated with increased bone mass. However, BMI does not differentiate between lean and fat mass. The purpose of this study is to examine the influence of dual‐energy x‐ray fat and lean mass on total bone mineral content (TBMC), among Asian and White adolescent girls. Girls were 9–14 y at visit 1, and 11 to 16 y at visit 2. TBMC was regressed on bone area, age, Tanner pubic hair stage, physical activity, dietary energy, protein and calcium intakes, percent Asian ancestry, and fat and lean mass. Age, pubic hair stage, Asian ethnicity and lean mass were positively associated with TBMC, while height and fat mass were negatively associated with TBMC. When fat mass was removed from the model, age, pubic hair stage, Asian ethnicity and lean mass were positively associated with TBMC, height was negatively associated, and dietary calcium intake was marginally associated with TBMC. When lean mass was substituted for fat mass in the model, pubic hair stage and Asian ethnicity were positively associated with TBMC. Age and calcium intake were marginally associated with TBMC while height was negatively associated. The interaction (cross product of lean and fat mass) was added to the first model and was highly significant and positive, meaning that the effect of lean mass was greater among girls with more fat mass. These findings suggest that lean mass, fat mass and their interaction are important influences on bone mass.
Vitamin D status is unaltered by mild weight reduction in obese premenopausal women
Authors: Peterson Catherine Ann , Ring Susan , Loethen Joanne , Rector R. Scott , Thomas Tom R. , Hinton Pamela S.
Published: June 2021

Excess adiposity negatively affects vitamin D status because of its deposition in fat compartments. Limited data are available characterizing weight loss effects on vitamin D status. We hypothesize that body fat loss is associated with increased serum 25‐hydroxyvitamin D (25OHD) concentrations in obese premenopausal women after weight loss. Since increased physical activity is linked with higher serum 25OHD, we also explored the effects of exercise mode on vitamin D status. 37 overweight to class I obese sedentary, non‐smoking women (18–35 y) were randomized to one of three treatments designed to achieve a 5% reduction in body weight in 6 wk: energy restriction (DIET, n=11), energy restriction + non‐weight bearing aerobic exercise (CYCLE, n=13), or energy restriction + weight‐bearing aerobic exercise (RUN, n=13). Body composition was assessed by 3‐site skinfold measurement and 25OHD serum concentrations were measured by RIA before and after weight loss. While weight loss was similar among groups, the RUN group lost more body fat than the other groups (p<0.001). There was no effect of total weight or fat loss, regardless of method, on serum 25OHD concentrations. In conclusion, mild reductions in body weight and percent fat by energy restriction alone or with exercise did not alter vitamin D status in obese premenopausal women. Support: Dept. of Nutr. Sciences Margaret Mangel Research Catalyst & UM Research Council.
Six‐month Changes in Dietary Behaviors, and Physical and Emotional Health of Recent Breast Cancer Survivors
Authors: Tangney Christy C , Ventrelle Jennifer C , Morris Laura R , Oleske Denise M
Published: June 2021

Background: Breast cancer survivors often experience weight gain and clinical depression early in their treatment course. It is uncertain whether weight changes are attributable to changes in lifestyle behaviors or a product of adjuvant therapy.

Methods: Twenty‐six breast cancer survivors (within 1 year of diagnosis) were enrolled in a weight management program at an urban medical center. Anthropometrics (height, weight, waist circumference, and body fat by bio‐impedance and near‐infrared interactance), resting metabolic rate (RMR) measurements, repeated telephone 24‐hour recalls (Nutrition Data System 2006, University of MN), functional performance tests, blood chemistries, and surveys to assess quality of life (FACT‐B) and possible depression (CES‐D) were obtained at baseline and 6 months.

Results: At study entry, all participants were clinically depressed (CES‐D≥ 16), though FACT B scores (103 ± 10) were relatively high. Average age was 46.4 ± 8.1 years, and BMI, 30.4 ± 6.1. Exactly 20% were African Americans. By six months, two‐thirds of participants had either maintained or lost weight. These changes were accompanied by significant improvements in fruit and vegetable consumption (p=0.019), and in CES‐D scores (p=0.026), and reduction in body fat (p=0.051) when compared to those who had gained weight (mean, 3.2 lbs). Weight losers relative to “gainers” had non‐significant decrease in RMR and total sugar consumption with marginally significant increases in walking speed, dietary fiber intakes and energy density.

Conclusion: Weight management support for recent survivors can be successful in improving not only dietary behaviors but also mental health.
Polyphenol content and antioxidant capacity of eggplant skin
Authors: Olson Rachel M , Teo Albert , Singh Ajay P , Luthria Devanand , Banuelos Gary S , Pasakdee Sajeemas M , Vorsa Nicholi , Wilson Ted
Published: June 2021

Eggplant (Solanum melongena) represents one of the best dietary sources of biologically active polyphenolic compounds. Polyphenolic‐rich extracts from darkly colored grape and cranberry fruit skins are known to inhibit the oxidative modification of low density lipoprotein (LDL) lipids and proteins. Dark purple skinned Blackbell and Millionaire eggplant varieties were grown under controlled agricultural conditions after which the skin polyphenolic content was extracted. Skin extracts contained phenolic acids (3‐caffeoylquinic acid, 4‐caffeoylquinic acid,5‐caffeoylqunic, dihydroxy cinnamoyl amide, caffeic acid conjugates), flavonols (myricetin‐3‐galactoside, quercitin‐3‐glucoside, quercitin‐3‐rhamanoside), in addition to anthocyanins, and proanthocyanidins whose specific identity were not determined. Polyphenolic extracts from all sources potently delayed the cupric ion‐mediated lag‐time for LDL lipid oxidation (234 nm), the formation of thiobarbituric acid reactive substances, and protected Apo‐B100 proteins against oxidative modification. Future studies of the biological activities of eggplant polyphenolics may improve eggplant cultivar selection for nutritional health benefits.
Effect of Date Pits on Oxidative Damage and Antioxidant Defense Systems in Rat Tissues
Authors: Habib Hosam H. , Ibrahim Wissam H.
Published: June 2021

The purpose of this study was to determine the effect of date pits on oxidative damage and antioxidant status in vivo. Male Wistar rats (six weeks old) were fed a basal diet with either 0%, 7% or 14 % date pits for 30 days. All three diets were isonitrogenous and isocaloric. At the end of the feeding period, rats were killed and liver homogenate and serum were analyzed for the levels of the lipid peroxidation product malondialdehyde (MDA). Additionally, the levels of the antioxidants glutathione, ascorbic acid, and vitamin E, and the antioxidant enzymes glutathione peroxidase, and catalase were analyzed in the liver. Compared with the control group, rats fed either 7% or 14% date pits had significantly (P < 0.05) lower levels of serum MDA or liver MDA. There was no significant difference in serum MDA or liver MDA between rats fed 7% and those fed 14% dates pits. The levels of vitamin E, GSH, and ascorbic acid and the activities of glutathione peroxidase and catalase were not significantly altered by date pits. The results obtained provide direct experimental evidence of the antioxidant effect of date pits in vivo.
Ingestion of casein and safflower oil, not soy protein and palm oil attenuates renal damage in hyperuricemic rats
Authors: Lo Hui‐Chen , Cheng Chun‐Hsiang , Yang Yu
Published: June 2021

Epidemiological studies indicated that there is a high prevalence of hyperuricemia in Taiwanese, especially in elderly and male aborigines. Using oxonic acid, a uricase inhibitor, and uric acid (UA) induced hyperuricemic rats, we investigated the effects of different dietary protein and lipid on UA excretion and renal damage. Male Wistar rats with hyperuricemia were fed with casein or soy protein as protein source and safflower oil or palm oil as lipid source for 8 weeks. Hyperuricemic rats with or without allopurinol treatment (150 mg/L in drinking water) and normal rats fed with AIN 93M diet, which composed of casein and corn oil, were also included. After 8 weeks, hyperuricemia‐induced decrease in feed efficiency was significantly improved by ingestion of casein and safflower oil; the induced increases in serum UA was significantly attenuated by allopurinol treatment and ingestion of soy protein and safflower oil; whereas the induced increase in blood urea nitrogen was augmented by palm oil ingestion. Moreover, animals ingested with casein and safflower oil, not soy protein and palm oil or allopurinol treatment had significantly attenuated hyperuricemia‐associated tubulointerstitial nephritis, crystals, and fibrosis in the kidneys. Our results suggest that vegetated diet with soy‐based protein and plant‐based saturated fatty acids might not be appropriate for patients to reduce the deleterious effects of hyperuricemia.
Fermented Soy Diet Reduces Weight Gain and Alters Liver Fatty Acid Composition in mice Fed a High Fat Diet
Authors: Cole Rachel M. , Shrode Gayle E. , Koba Kazunori , Tanimoto Hiroyuki , Okabe Yuki , Belury Martha A.
Published: June 2021

Obesity is now a worldwide problem, yet countries with high intake of soy foods have less incidence of obesity. Fermentation of soy leads to accumulation of aglycone isoflavones. In this study, we compared fermented soy (fsoy, n=12) to soy (n=12) and casein (n=11) diet on weight gain and hepatic lipid composition in diet‐induced obese male C57B16/J mice. Food intake was measured semi‐weekly and weights weekly. After the 17 weeks, necropsy was performed, tissue weights were obtained and livers were snap frozen for measurement of triglycerides, fatty acids, and mRNA levels of lipid metabolizing genes. The fsoy and soy diet groups had increased average daily food intakes and lower weight gain compared to casein group. The fsoy group had a higher feed‐to‐gain ratio. The liver weights and triglyceride level did not differ. The hepatic fatty acid composition was different including a higher omega‐3:omega‐6 ratio in the fsoy group. The fsoy group had a lower fold induction of peroxisome proliferator activated receptor alpha (PPAR ), fatty acid binding protein 1(FABP1), and stearoyl Co‐A desaturase 1 (SCD1) mRNA than the casein group. The fsoy group had a higher fold induction of fatty acid synthase (FAS) and PPAR mRNA than the soy group. Fermented soy decreases weight gain and changes the composition of liver fatty acids in C57B16/J mice. Supported by Ajinomoto Corporation
Anti‐oxidant activity increased in human dermal fibroblasts and intact skin by Zingiber officinale CO2 extract
Authors: DaSilva Marissa , Bhagavathula Narasimharao , Barron Adam , Warner Roscoe , Weber Stephanie , Varani James
Published: June 2021

Primary human dermal fibroblasts in monolayer culture exhibit a cytotoxic response to hydrogen peroxide at concentrations greater than 25μM. When pretreated with gingerol enriched Ginger (Zingiber officinale) extract for 72 hours, fibroblasts have an increased capability to cope with peroxide induced injury and cytoxicity. Intact skin was organ cultured for 3 days with and without ginger and exposed to hydrogen peroxide (25mM–50mM). Histology revealed that skin which received pretreatment with gingerol enriched Ginger extract exhibited normal physiology post peroxide stress (37.5mM). Alternatively, non‐treated tissue exhibited necrosis at 37.5mM hydrogen peroxide. Intact skin was organ‐cultured with and without Ginger. The tissue was exposed to peroxide, minced, and explanted. Ginger treated tissue had normal outgrowth of fibroblasts and keratinocytes post peroxide exposure. Conversely, tissue which did not receive pre‐treatment with Ginger had no cellular outgrowth post peroxide exposure. The cell outgrowth assay indicated both keratinocytes and fibroblasts are sensitive to peroxide injury. However, fibroblasts appear to be more sensitive to injury and pretreatment with Ginger. In addition to identifying Ginger as a promoter of antioxidant activity in cells and intact skin, this approach may serve as a useful paradigm in the identification and evaluation of potential antioxidants in skin.
Stem cell marker immunoprofile in glioblastomas and its potential role in tumorigenesis
Authors: Sze Chun‐I , Huang Yu Fen , Hsieh Chia Li , Li Meng Yen
Published: June 2021

It has been suggested that the concept of gliomas originate from tumor stem cell (TSC) may provide clues to our understanding of gliomas pathogenesis. Here, we are using immunohistochemistry (IHC) and immunofluorescence (IF) methods to examine 16 glioblastomas (GBM), 2 anaplastic astrocytomas, 3 astrocytomas, and 4 normal brains with stem cell markers to test the hypothesis that TSCs and stem cell markers are differentially expressed in the gliomas. IHC revealed that there were relatively low number of CD133 and nestin antibodies positive cells in normal brain, grade II and grade III astrocytomas. CD133 and nestin were strongly expressed and co‐expressed in the smooth muscle cells of proliferating glomeruloid vessels in GBMs but not in the small cortical or larger meningeal vessels. CD133+ cells either weakly or not co‐express with vimentin. Nestin+ cells co‐express vimentin and GFAP. The IF density and number of CD133+or nestin+ cells decreased inversely to the distance of the glomeruloid vessels in GBMs. IF also revealed that nestin co‐express with vimentin, and GFAP. MAP2 and tubulin III antibodies highlight entrapped neurofilaments in the tumors. Given these three intermediate filaments present in different stage of glial cell development, and nestin and vimentin differentially co‐expressed in CD133+ cells, our findings suggest that intermediate filaments may play important role in stem cell derived GBMs.
Phosphorylation of Myocardin by Mitogen Activated Protein Kinase.
Authors: Taurin Sebastien , Dulin Nickolai O.
Published: June 2021

Myocardin (MyoCD) is a transcription factor that drives smooth muscle (SM) cell differentiation by activating serum response factor (SRF) and by inducing transcription of SM‐specific genes. Here we report that MyoCD is heavily phosphorylated by mitogen activated protein kinase (MAPK) both in vitro and in intact cells. We identified four phosphorylation sites on MyoCD and confirmed those by mutation analysis. Phosphorylation‐deficient mutant of MyoCD showed a reduced ability to transactivate the SRF‐dependent gene transcription, as assessed by using luciferase reporters for SRF and for promoters of SM genes. Furthermore, compared to wild type MyoCD, the phosphorylation‐deficient mutant was less efficient in promoting the binding of SRF to its DNA target, CArG box, as assessed by electrophoretic mobility shift assay (EMSA). By contrast, phosphorylation did not affect MyoCD localization, dimerization and binding to SRF or to an acetyltransferase, CREB‐binding protein (CBP). In conclusion, we demonstrate a novel crosstalk between MAPK and SRF signaling through a direct phosphorylation of MyoCD.
Characterization of the regulatory mechanism controlling Regulator of R‐Protein Signaling‐5 (RGS5) expression
Authors: Mahoney William M. , Schwartz Stephen M.
Published: June 2021

We are interested in examining the role of G‐protein coupled receptor (GPCR) signaling with respect to vascular remodeling as a model for cardiovascular disease. The regulator of G‐protein signaling (RGS) protein family primarily functions to control the duration of cellular signals mediated through GPCRs. RGS proteins act as GTPase activating proteins (GAPs) by facilitating the exchange of GDP for GTP upon Gα subunits of the heterotrimeric G‐protein complex, thereby returning the complex to its inactive, yet primed state. RGS5, a member of the RGS‐R4 subfamily of RGS proteins, is the most predominant RGS‐R4 subfamily member expressed in vascular smooth muscle cells (SMCs), and it uniquely identifies arterial SMCs relative to venous SMCs. The putative RGS5 promoter was isolated and characterized by promoter‐luciferase assays in rat aortic SMCs. Chromatin immunoprecipitation (ChIP) assays were performed to confirm the binding of specific transcription factors in vivo. By understanding how RGS5 is regulated, we hope to understand a basic mechanism in determining arterial and venous identity, and to further characterize a key protein in understanding hypertension.
CB2 cannabinoid receptor stimulation attenuates TNF‐alpha‐induced human endothelial cell activation, transendothelial migration of monocytes, and monocyte‐endothelial adhesion.
Authors: Rajesh Mohanraj , Mukhopadhyay Partha , Bátkai Sandor , Huffman John , Csiszar Anna , Ungvári Zoltán , Chatterjee Subroto , Pacher Pal
Published: June 2021

Targeting cannabinoid‐2 (CB2) receptors with selective agonists may represent a novel therapeutic avenue in various inflammatory diseases, but the mechanisms by which CB2 activation exerts its anti‐inflammatory effects and the cellular targets are elusive. Here, we investigated the effects of CB2‐receptor activation on TNF‐alpha‐induced signal transduction in human coronary artery endothelial cells in vitro and on endotoxin‐induced vascular inflammatory response in vivo. TNF‐alpha induced NF‐kappaB and RhoA activation and upregulation of adhesion molecules ICAM‐1 and VCAM‐1, increased expression of monocyte chemoattractant protein, enhanced transendothelial migration of monocytes, and augmented monocyte‐endothelial adhesion. Remarkably, all of the above‐mentioned effects of TNF‐alpha were attenuated by CB2 agonists. CB2 agonists also decreased the TNF‐alpha‐ and/or endotoxin‐induced ICAM‐1 and VCAM‐1 expression in isolated aortas and the adhesion of monocytes to aortic vascular endothelium. CB1 and CB2 receptors were detectable in human coronary artery endothelial cells by Western blotting, RT‐PCR, real‐time PCR, and immunofluorescence staining. Because the above‐mentioned TNF‐alpha‐induced phenotypic changes are critical in the initiation and progression of atherosclerosis and restenosis, our findings suggest that targeting CB2 receptors on endothelial cells may offer a novel approach in the treatment of these pathologies.
AIP1 functions as a negative regulator in flow‐mediated vascular remodeling
Authors: Min Wang , Yu Luyang , Zhang Haifeng , He Yun
Published: June 2021

We have previously shown that AIP1 mediates apoptotic signaling in vascular endothelial cells (EC). However, the in vivo function of AIP1 in vasculature is not known. Here we investigate the role of AIP in flow‐mediated vascular remodeling using AIP1‐knockout mice (AIP1−/−). AIP1+/+ and AIP1−/− were subjected to a complete common carotid ligation, which is characterized by both neointimal hyperplasia and inward vessel remodeling. Compared to AIP+/+ littermates, AIP−/− mice had much larger media area, thicker vessel wall and increased neointima formation Moreover, the neointima in AIP−/− mice exhibited high levels of iNOS expression and phospho‐STAT3, suggesting that an augmented interferon‐gamma (IFN‐?) signaling in AIP1−/− mice. Since IFN‐? is critical for neointima formation, we compare the effects of AIP1 on IFN‐? signaling in isolated mouse EC and vascular smooth muscle cells (VSMC) where AIP1 is highly expressed. IFN‐?‐induced activation of JAK1‐STAT1 signaling in AIP−/− VSMC was significantly higher than that in AIP+/+ cells. In contrast, AIP1 deficiency had no effects on IFN‐? signaling in EC. Based on these results, we propose that AIP may function as a negative regulator in flow‐mediated vascular remodeling, in part, by downregulating IFN‐?‐JAK‐STAT‐dependent proliferative signaling in VSMC. The molecular mechanism by which AIP1 inhibits IFN‐ ?‐JAK‐STAT pathway will be discussed.
The transcription factor LMO2 is a robust marker of vascular endothelium and vascular neoplasms with rare exceptions
Authors: Gratzinger Dita , Zhao Shuchun , Vogel Hannes , Gil Elena Cubedo , Levy Ronald , Lossos Izidore , Natkunam Yasodha
Published: June 2021

The transcription factor LMO2 is involved in vascular and hematopoietic development. Here we demonstrate that LMO2 is expressed nearly ubiquitously in native vascular endothelium including lymphatic endothelium. Rare exceptions include the vasculature of adult and fetal heart and the brain vasculature of older individuals, as well as hepatic sinusoidal endothelium. While LMO2 is expressed in the vasculature of a variety of carcinomas and non‐epithelial neoplasms, expression was rare in hepatocellular carcinomas (1/10) and was lacking in a subset of clear cell renal cell carcinomas (14/22). LMO2 was uniformly expressed in a variety of benign vascular (11/11) and lymphatic (5/5) neoplasms. LMO2 was also expressed in most vascular endothelium‐derived malignancies, including 11/11 non‐epithelioid angiosarcomas and 6/6 Kaposi sarcomas. Among epithelioid vascular malignancies, LMO2 expression was lacking in primary epithelioid vascular neoplasms of bone (1/4 positive) and pleura (0/3 positive), but was uniformly present in epithelioid vascular malignancies originating at other sites (9/9, p<0.002 and chi value 12.8 for the three way comparison). In the context of diagnostic immunohistochemistry, LMO2 may be added to the arsenal of markers of vascular differentiation; in epithelioid vascular malignancies that lack LMO2, assessment for a primary site in bone or pleura may be indicated.
Differential solubility analysis of fractionated heart proteins in laboratory animals
Authors: Suzuki Osamu , Koura Minako , Takano Kaoru , Noguchi Yoko , Uchio‐Yamada Kozue , Matsuda Junichiro
Published: June 2021

At the EB2007 meeting, we reported an abnormality of collagen type VI in our transgenic mice with dilated cardiomyopathy based on an analysis of heart proteins fractionated using differential solubilization. To examine if this protein fractionation technique can be applied to proteomic analysis of the hearts of other laboratory animals, we compared fractionated heart proteins among four normal laboratory animals: mice, rats, guinea pigs, and rabbits. Heart proteins were extracted sequentially using differential solubilization with SB3‐10 (ReadyPrep Sequential Extraction Kit; Bio‐Rad) and the three fractions were separated independently using SDS‐PAGE. As in our previous study using mice, we obtained similar band patterns involving three fractions from rat, guinea pig, and rabbit heart proteins. In particular, bands for collagen type VI alpha‐3 subunits and cardiac myosin heavy chains in the most insoluble fractions, both of which were suggested by results in mice, were also visualized for rat, guinea pig and rabbit proteins. Our results suggest that the fractionation of heart proteins would be useful for proteomic analysis of impaired hearts in various animals, especially for analyzing the collagen type VI content. This work was supported by a Health and Labor Science Research Grant from MHLW, Japan.
Nitric Oxide‐Mediated Growth Inhibition of an Endothelialized Tissue Engineered Aortic Valve
Authors: Hageraats‐Boucher Jami‐Lee , Boughner Derek R. , Rogers Kem A.
Published: June 2021

The tissue engineering (TE) of aortic valves may provide an alternative to the use of non‐viable bioprosthetic replacements. Endothelialization of a TE construct is necessary to establish a non‐thrombogenic surface and has been shown to inhibit the proliferation of vascular smooth muscle cells within the construct. Previously it has been shown that nitric oxide (NO) production by endothelial cells may be responsible for this growth inhibition. To test this hypothesis, cells scaffold constructs (CSCs) were produced consisting of radial artery cells seeded onto small intestine submucosa. After one week in culture, an analog inhibitor of nitric oxide synthase, L‐iminoethyl‐L‐orthinine (L‐NIO, 100 μM) was added to either CSCs or endothelialiazed CSCs (ECSCs). Constructs were cultured for an additional week, followed by fixation and tissue thickness analysis by confocal microscopy. ECSC and CSC control groups were cultured for two weeks in the absence of L‐NIO. The presence of an endothelium (ECSCs) significantly inhibited growth of the construct, however, in the presence of L‐NIO, growth of the construct continued in a manner similar to CSCs (75.6μ m vs 55.3μm). Results suggest that the production of NO by the endothelium may be responsible for growth inhibition of the ECSCs providing a valuable tool for controlling TE valve construct thickness in vitro. Funding provided by Heart and Stroke foundation of Ontario.
The atypical antipsychotic drug, aripiprazole, reverses amphetamine‐induced focused stereotypy in rats
Authors: Fowler Stephen C. , Pinkston Jonathan , Vorontsova Elena
Published: June 2021

Although the dopamine D2 partial agonist and atypical antipsychotic drug (AAPD), aripiprazole (ARPZ), has been shown to antagonize apomorphine‐induced behaviors in rats, it is unknown whether ARPZ antagonizes amphetamine (AMPH)‐induced behaviors in this species. Accordingly, this study examined effects of ARPZ on AMPH‐induced focused stereotypy in rats. Clozapine was included as a comparison AAPD. Rats were first sensitized to AMPH by 5 treatments (5.0 mg/kg, ip.) each separated by 72 hr. Subsequently, AMPH treatments continued but were followed 30 min later by ARPZ (6, 12, and 18 mg/kg, po.) or clozapine (5, 10, 20 mg/kg, po.). Focused stereotypy and locomotor activity were quantified with force‐plate actometers in 4‐hr recording sessions. Clozapine prolonged AMPH‐induced focused stereotypy and slowed the rats’ rhythmic head movements. On the other hand, ARPZ significantly reversed the expression of focused stereotypy and left intact intermediate amounts of locomotion. Thus, ARPZ behaviorally antagonized the effects of the indirect‐acting dopamine agonist AMPH, and this effect is consistent with ARPZ's D2 partial agonist properties. Supported by MH043429 & HD002528.
The role of CRF in individual differences in response to social defeat: neuroendocrine, behavior, and viscera
Authors: Wood Susan K , Iyer Vikram , Valentino Rita J , Bhatnagar Seema
Published: June 2021

Individual differences in response to stress may determine vulnerability to adverse psychological consequences. During a 30 minute social defeat paradigm, Sprague Dawley intruders exhibited individual differences in average latency to be defeated (exhibiting a supine posture) by an aggressive resident Long Evans during 7 daily defeats. Rats responding submissively exhibited short latencies (SL; 197±15 sec) whereas those displaying defensive behaviors exhibited greater latencies (LL; 438±12 sec). All defeated rats exhibited blunted weight gain and adrenal hypertrophy, however when challenged with restraint, LL rats exhibited facilitated adrenocorticotropin hormone response whereas SL exhibited blunted release vs control. SL rats also displayed increased immobility during Porsolt forced swim vs control. As previously reported in mice, SL rats also developed bladder hypertrophy. Corticotropin‐releasing factor (CRF), in addition to its initiating role in the HPA axis and anxiogenic effects, inhibits micturition via barrington's nucleus, thereby inhibiting urination and, in extreme cases, promotes bladder hypertrophy. Immunohistochemical analysis revealed increased CRF‐expressing cells in barrington's of SL. The results implicate a role of individual differences in coping strategy in determining neuroendocrine, neuropeptide, behavioral and visceral repercussions of repeated defeat.
α2‐Adrenoceptors increase glycogen synthesis in chick astrocytes: possible mechanism to increase memory formation
Authors: Hutchinson Dana Sabine , Gibbs Marie E , Summers Roger J
Published: June 2021

From experiments using a discriminated bead task in chicks, we have defined when and where adrenoceptors (ARs) are involved in memory modulation. All 3 AR subtypes (α1‐, α2‐ and β‐ARs) are found in the chick brain and in regions associated with memory formation. Glucose and glycogen are important in memory formation since an increase in glucose levels improves while inhibiting glucose transporters (GLUTs) or glycogen breakdown inhibits memory consolidation. α2‐ARs are expressed on chick astrocytes (Bmax 66.3 ± 17.7 fmol/mg protein) and stimulation with clonidine increases glycogen synthesis in a concentration‐dependent manner, whereas stimulation of β‐ or α1‐ARs has no effect on increases in glycogen synthesis. The effect of clonidine was sensitive to inhibition by both pertussis toxin (Gi) and inhibitors of PI3K. With respect to memory, injection of α2‐AR agonists into the medial striatum (MSt), but not the intermediate medial mesopallium (IMM) of chicks, increased memory consolidation. Injection of the postsynaptic α2‐AR antagonist ARC239 decreased memory consolidation when injected into either the MSt or IMM. These results show that α2‐ARs increase glycogen synthesis in chick astrocytes and are involved in memory consolidation in the chick.

This research was supported by National Health and Medical Research Council Project Grants 236884 and 491169.
Ouabain‐induced hypertension and rho kinase
Authors: Özdemir Aysun , Aypar Eda , Þahan Gürkan , Gözübüyük Gürkan , Demirtaþ Ayþegül , Turan Nilüfer Nermin , Ark Mustafa
Published: June 2021

Rho/Rho kinase pathway has been implicated various models of hypertension. However, there is no report whether this pathway involves in ouabain‐induced hypertension. In our current work, we evaluated possible involvement of rho kinase in ouabain‐induced hypertension in rats. Ouabain was injected to rats 20μg/kg per day for six weeks. After ouabain treatment, Rho kinase‐1 and Rho kinase‐2 expressions of aortas were evaluated by Western Blotting. It was shown that expressions of neither 1 nor 2 isoform of Rho kinase were altered in ouabain group. Dose dependent relaxations Y27632 were determined in KCl and Phenylephrine precontracted aortas of both control and ouabain treated groups. Although there was no statistically significant difference between KCl precontracted tissues of control and hypertensive groups, it was shown that Y27632 relaxations were enhanced in Phenylephrine precontracted aortas of ouabain treated group compared to control. As a conclusion, the results of this study indicate that although Rho kinase expressions did not contribute to ouabain induced hypertension, Rho kinase activation may have a role in this hypertension model.