bluuebunny/crossref_metadata_2025 huggingface dataset:
Relating mood to plasma glutathione and BDNF levels in patients with bipolar disorder
Authors: Singh Nisha , Rosa Adriane R , Whitaker Eleanor , de Britto Marianne , Lewis Alexander , Vieta Edward , Churchill Grant C , Geddes John R , Goodwin Guy M
Published: June 2021

Objective

To correlate self‐reported mood scores (depression and mania) with plasma levels of brain derived neurotrophic factor (BDNF) and glutathione in medicated bipolar patients.

Methods

BDNF (ELISA) and glutathione levels (enzymatic recycling, Ellman's reagent) were measured in plasma from 50 bipolar patients and matched healthy controls. The True Colours database (OXTEXT, University of Oxford) was used to determine mood‐monitoring scores for the bipolar group with the Quick Inventory of Depressive Symptomatology (QIDS) and the Altman Self‐Rating Mania Scale over an 8‐week period.

Results

Bipolar patients had significantly lower levels of reduced glutathione and higher levels of oxidized glutathione, while BDNF levels were not significantly different. 22% and 64% of the patients had scores ≥ 6 on the Altman scale and QIDS, respectively, but mood state did not correlate with antioxidant level.

Discussion

The results suggest that, despite medication, bipolar patients show higher levels of oxidative stress and that the majority still have depressive symptoms. The direction of effect is not established, but these observations appear to be consistent with studies that suggest N‐acetyl cysteine, an antioxidant, reduces depressive symptoms in bipolar patients. Hence, combining an antioxidant with other mood‐stabilizers may be therapeutically beneficial in the treatment of bipolar disorder.
TPA‐induced invasion of HT29 cells is mediated through p67phox‐dependent matrix metalloproteniase‐7 induction
Authors: Banskota Suhrid , Regmi Sushil Chandra , Kang Youra , Kim Jung‐Ae
Published: June 2021

Tumor promoter 12‐O‐tetradecanoylphorbol‐13‐acetate (TPA)‐induced ROS have been implicated in increased expression of matrix metalloproteniase‐7 (Matrilysin, MMP‐7), which is an important enzyme for cancer invasion and also a prognostic marker in colorectal cancer. However, the source of TPA‐induced ROS is still controversial whether it is NADPH oxidase or mitochondria. In the present study, we report that TPA‐induced ROS production and matrigel invasion of HT‐29 were inhibited by apocynin and diphenyleneiodinium (DPI), NADPH oxidase inhibitors, but not by other inhibitors for mitochondria or cytosolic enzymes. TPA increased p67phox expression in a time‐dependent manner without altering the level of other components of NADPH oxidase. In addition, TPA‐induced invasion of HT29 cells was significantly blocked by treatment with p67phox and Nox2 siRNAs but not with Nox1 siRNA. Moreover, TPA decreased mRNA and protein levels of ¥â‐catenin that is an integral part of adherens junction associated with other catenins and cadherins. The TPA‐induced ¥â‐catenin decrease and loss of cell‐cell adherence were also reversed by treatment with apocynin. Taken together, TPA induces HT29 cancer cell invasion by increasing MMP‐7 expression and by downregulating ¥â‐catenin through p67phox‐dependent ROS production.
Tepary bean lectins induces apoptosis on human colon cancer cells and inhibits early tumorogenesis in rats
Authors: Garcia‐Gasca Teresa , Ferriz‐Martinez Roberto , Cervantes‐Jimenez Ricardo , Angeles‐Zaragoza Marco Vinicio , Andrade‐Portillo Veronica , Blanco‐Labra Alejandro , Rodriguez‐Mendez Adriana Jheny , Gallegos‐Corona Marco Alonso , Lopez‐Martinez Francisco Josue , Mendiola‐Olaya Elizabeth
Published: June 2021

The aim of this study was to determine whether a lectin fraction (LF) from Tepary bean seeds (Phaseolus acutifolius) exhibits apoptotic induction on colon cancer cells and its in vivo effect on tumorogenesis in rats. Median lethal concentration (0.402 mg protein/mL) on HT‐29 cells caused a significant increase in caspase 3 activity, an overexpression of caspase 3 (35%) and caspase 9 (78%) and a 36.6% apoptosis increase determined by flow cytometry. Colon cancer was induced on male Sprague Dawley rats treated for 8 weeks with 1,2‐dimethylhydrazine (DMH) (40 mg/body weight kg) and a subsequent administration of 50 mg of LF/body weight kg for 6 weeks. The LF provoked a decrease in food consumption and body weight of rats with a partial recovery at the end of the experiment. Histopathological analysis showed that LF decreased the incidence of early stages tumors in a 53% but no effect was observed on high grade carcinogenesis. The results show that LF induces apoptosis in colon cancer and it has inhibitory effect on early tumorogenesis.

Founding: CONACYT Ciencia Básica (82349)
Rational design and construction of HER2 targeting recombinant human serum albumin
Authors: Dong Daoyuan , Li Zhiyu , Li Zhijun
Published: June 2021

HER2 (Human Epidermal growth factor Receptor 2) is a receptor over expressed in approximately 30% of human breast cancers. Human Serum Albumin (HSA) is the main protein in plasma, its biophysical and biochemical properties make it an ideal candidate for drug delivery. The overall purpose of this study is to develop an HSA‐based drug delivery system capable of targeting HER2 to enhance tumor drug delivery. Several HER2 binding peptides were utilized here. The DNA sequences of these peptides were designed, synthesized and integrated into HSA gene and the recombinant proteins were expressed. The binding affinity of purified proteins to HER2 was tested in SK‐BR‐3 cells and showed specific recognition of HER2. It is hoped that our design can serve as 1) a scaffold to produce highly specific, targeted proteins, or 2) an approach to create bi‐specific proteins capable of promoting a synergistic effect on the inhibition of cancer cell growth when combined with chemotherapeutic agents.
The effect of drugs that alter serotonin homeostasis in rat erectile function
Authors: Linder A. Elizabeth , Poli Anicleto , de Lima e Silva Ana Kelly
Published: June 2021

We investigated wether antidepressants that alter serotonin (5‐hydroxytryptamine; 5‐HT) homeostasis impact rat erection due to 5‐HT uptake and metabolism in the penile dorsal vein (PDV). 5‐HT and its metabolite 5‐hydroxyindole acetic acid (5‐HIAA) were measured by high performance liquid chromatography in rat PDV. Exposure to exogenous 5‐HT increased 5‐HIAA but not 5‐HT content in PDV, suggesting uptake followed by metabolism, and this was supported by increased 5‐HT and decreased 5‐HIAA content induced by pargyline, a monoamine oxidase inhibitor (enzyme that metabolizes 5‐HT). Fluoxetine (5‐HT uptake transporter inhibitor) and imipramine (norepinephrine and 5‐HT uptake transporters inhibitor) did not inhibit 5‐HT uptake in PDV. Concentration effect curve (CEC) induced by 5‐HT was also evaluated in endothelium‐intact rings of PDV in the presence of fluoxetine, imipramine and pargyline. The CEC induced by 5‐HT was shifted to the right by imipramine and unchanged by pargyline. Fluoxetine increased the maximal contraction induced by 5‐HT in PDV. The effect of these drugs was also evaluated in the erections induced by apomorphine in conscious rats. Pargyline decreased, fluoxetine increased and imipramine had no effect in the number of erections induced by apomorphine. As apomorphine, fluoxetine per se induced ~3 erections/rat. Altogether, drugs that alter 5‐HT homeostasis in PDV do impact erection. CAPES; FAPESC.
Doxycycline does not reverse structural observed in SHR aortas
Authors: Gerlach Raquel F , Antonio Raquel Carros , Ceron Carla S , Rizzi Elen , Coelho Eduardo B , dos Santos Jose Eduardo Tanus
Published: June 2021

This study aimed at testing whether doxycycline (dox) would change the vascular structure, gelatinolytic activity and MMP‐2 levels in SHR aortas of 13‐week‐old animals treated for 5 weeks. SHR and Wistar‐Kyoto controls received either dox 30 mg/kg/day by gavage or vehicle. Systolic blood pressure (SBP) was assessed weekly by tail cuff, and at the end the aortas were processed for histological analysis and in situ zymography. Histological sections were used to determine media cross‐sectional area (CSA), media/lumen diameter (M/L), and the number of smooth muscle cells (SMC). Gelatinolytic activity was measured in situ using DQ gelatin. SBP was significantly lower in the SHR + doxy group compared to the SHR vehicle (P< 0.05). No changes were seen in any of the structural parameters (p>;0.05). Gelatinolytic activity was 3 times higher in the SHR aortic media (53.4 ± 15 units) compared with Wkyoto controls (18.2 to 18.9 ± 6 units), and dox treatment decreased the SHR values to 26.0 ±8 units (P <0.0001). Data confirms the high expression and activity of MMP‐2 in aortas of SHRs, and the effect of doxycycline in reversing the increase in SBP. However, at least for the time frame used (5 weeks) these changes were not concurrent with structural changes in the aortas, as shown in other models of hypertension after dox treatment.

Support: State of Sao Paulo Research Foundation (FAPESP) and National (Brazilian) Research Council (CNPq).
Role of phosphatidic acid phosphatase type 2 domain containing 2 (PPAPDC2) in squalestatin 1‐mediated CYP2B1 induction in primary cultured rat hepatocytes
Authors: Pant Asmita , Kocarek Thomas A
Published: June 2021

Inhibition of squalene synthase causes accumulation of farnesyl pyrophosphate (FPP) in primary cultured rat hepatocytes, which increases CYP2B expression through activation of the constitutive androstane receptor (CAR). However, the identity of the CAR‐activating isoprenoid has not been established. Since the lipid phosphatase PPAPDC2 has recently been reported to catalyze FPP dephosphorylation (Miriyala et al., J Biol Chem 285:13918–13929, 2010), we determined the effect of PPAPDC2 overexpression and knockdown on squalestatin 1 (SQ1)‐mediated activation of a CAR‐responsive CYP2B1 luciferase reporter in primary cultured rat hepatocytes. Treatment with 100 nM SQ1 increased reporter expression ~11‐fold in hepatocytes that were co‐transfected with empty expression plasmid. The SQ1‐mediated induction was enhanced ~2‐ or 1.5‐fold when the hepatocytes were co‐transfected with an expression plasmid for rat or human PPAPDC2. SQ1‐inducible reporter expression was not affected by co‐transfection of a non‐targeting shRNA but was reduced ~60% by co‐transfection with a PPAPDC2 shRNA. These results indicate that PPAPDC2 plays an important role in SQ1‐mediated CYP2B1 induction, most probably by catalyzing the conversion of FPP to farnesol. This work was supported by NIH grant R01HL050710. Asmita Pant is supported by a Thomas Rumble Fellowship.
Aging augments apoptosis of endothelial cells in mice
Authors: Campagnaro Bianca Prandi , Tonini Clarissa Loureiro , Porto Marcella Leite , Vasquez Elisardo Corral , Meyrelles Silvana Santos
Published: June 2021

We evaluate mice endothelial cells (EC) apoptosis in aging and atherosclerosis by flow cytometry.

Male apolipoprotein E‐deficient (ApoE, N=10) mice were separated in two groups according to their ages: 2 and 18 months old and compared with C57BL/6 mice (C57, N=10). Animals were euthanized and EC mechanically isolated from aorta. For apoptosis analysis, 105 cells were incubated with Annexin V‐FITC and propidium iodide (PI) at room temperature during 15min. Events were acquired by FACSCanto II and analyzed by FACSDiva software. Data are means±SEM. p<0.05 (two‐way ANOVA).

Flow cytometric analysis showed that atherosclerosis did not affected EC apoptosis between young (C57: 1.9±0.2 vs. ApoE: 1.15±0.12 %) and aged (C57: 23.25±7.7 vs. ApoE: 23.36±0.55 %) groups. However, aging augments apoptosis of EC in ApoE (Young: 1.15±0.12 vs. Aged: 23.36±0.55 %) and C57 (Young: 1.9±0.17 vs. Aged: 23.25±7.71 %) groups.

Our data showed that atherosclerosis per se was not able to induce apoptosis. However, aging was directly responsible for augmentation of EC apoptosis.
Physical exercise and taurine supplementation restore endothelial dysfunction and antioxidant defense in obese rats.
Authors: de Moraes Camila , Delbin Maria Andréia , de Freitas Ellen Cristini , Maia Aline Rosa , Davel Ana Paula
Published: June 2021

This study aimed to evaluate the effects of physical exercise (PE) and taurine (Tau) supplementation on endothelium‐dependent relaxation and oxidative stress biomarkers in obese rats. Thirty male Wistar rats were divided: lean control; obese sedentary; obese sedentary taking Tau or trained; and obese trained and taking Tau. Exercise training (treadmill: 20 m/min, 60 min, 5 days/week) and Tau supplementation (2%) started after 4 weeks of cafeteria diet consumption and was carried out for further 7 weeks. Data were analyzed using ANOVA and Tukey post‐hoc (p<0.05). Obese rats showed an impaired endothelium‐dependent relaxation to acetylcholine (about 26%) and higher plasma levels of TBARS (138%) and lower NOx (62%), compared to lean rats. Besides, Cu/Zn SOD and EC SOD aortic protein expression were reduced in obese rats (52% and 28%, for Cu/Zn and EC respectively). Either exercise or Tau restored the impaired endothelium‐dependent relaxation observed in obese sedentary rats. Both treatments increased NOx concentration (88% in Tau and 108% in exercised rats) and lowered TBARS (18% and 46% to Tau and exercised rats). Increase in aortic expression of Cu/Zn and EC SOD was also seen in Tau or exercised rats. Concomitant intervention with exercise+Tau did not produce additive effects. Physical exercise, as well Tau supplementation, improves endothelium‐dependent relaxation related to a reduction in oxidative stress due to improvement on antioxidant defense. Financial support: FAPESP
Evidence that chronic inhibition of the mammalian Target of Rapamycin (mTOR) reduces skin blood flow in humans
Authors: Kellogg Dean Lundt , Zhao Joan , Wu Yubo , Roman Linda , Johnson John Marshall
Published: June 2021

Chronic mTOR inhibition causes vascular dysfunction in rodents. We hypothesized that a similar effect would be observed in humans. 6 middle‐aged persons participated. mTOR was antagonized by topical application of 8% rapamycin (RAPA) ointment to skin. RAPA was applied twice daily for 7 days to an 8×8 cm area of skin on the abdomen while a second site was similarly treated with ointment vehicle only. We examined the effect of these treatments on skin blood flow (SkBF) as monitored by laser‐Doppler flowmetry (LDF). Local skin temperature (Tloc) was controlled with combined LDF/local heating probe holders. Tloc was maintained initially at 34°C and then was raised to 43°C to cause maximal local vasodilation (maxLDF).

Results

At Tloc=34°C, SkBF was 72±4.3% maxLDF at RAPA treated sites and was 80±4.9% maxLDF at sites that received vehicle only (p<0.03 between sites). maxLDF (in mV) when Tloc was 43°C did not differ between RAPA and vehicle treated sites (p>;0.05). We conclude that chronic RAPA treatment reduces SkBF levels at normothermic skin temperatures in middle‐aged persons without an effect on maximal vasodilation. (supported by NIH Grant AG041365)
High‐Fructose Consumption and Cardiac Substrate Utilization
Authors: Kertowidjojo Elizabeth Calista , Eisenberg Carol , Wolin Michael , Hintze Thomas H
Published: June 2021

Many studies have shown a correlation between high‐fructose consumption and the metabolic syndrome, a known harbinger of cardiovascular disease. In this study, we explored the effects of high‐fructose diet on the metabolism of the heart in chronically instrumented conscious dogs. One of the hallmarks of heart failure is a shift from the utilization of free fatty acid (FFA) to glucose and lactate. We hypothesize that high‐fructose diet causes disruption of normal myocardial metabolism, leading to an increase of carbohydrate utilization. Male mongrel dogs were fed an isocaloric 60% fructose diet for 49 days (FF, N=6). Comparison of aortic and coronary sinus blood samples revealed a significant increase in glucose (by 98 ± 14%) and lactate uptake (by 58 ± 12%) by the hearts of FF dogs. On the other hand, uptake of FFA was significantly decreased (4.1 ± 1.0% and 1.7 ± 0.3% of arterial FFA in control and FF dogs, respectively). There was no significant difference in total calculated cardiac ATP production. However, percent of ATP produced from FFA metabolism dropped significantly from 87 ± 4% in control to 53 ± 4% in FF group. Accordingly, ATP produced from glucose and lactate was significantly increased in FF dog hearts as compared to control. Finally, analysis of total blood O2 content revealed significantly increased cardiac O2 consumption in FF dogs (12.9 ± 1.5 mL in FF compared to 8.7 ± 1.1 mL in control). Supported by PO43023.
Knockout of type VI collagen preserves mitochondrial structure and function following myocardial infarction
Authors: Luther Daniel J. , Patel Hemal , Niesman Ingrid R. , Kang Patrick T. , Adapala Ravi K. , Thoppil Roslin , Bonaldo Paolo , Chilian William M. , Chenn Yong‐Renn , Thodeti Charles K. , Meszaros J. Gary
Published: June 2021

Cardiac remodeling is a dynamic process that is accelerated following myocardial infarction (MI) injury. Previously, fibrillar collagens type I and III have been the focus of the remodeling field however, our lab has uncovered a potentially novel role for non‐fibrillar type VI collagen (Col6) in this process. Mutations in Col6 lead to an age‐related skeletal muscle disorder known as Bethlem's Myopathy due to altered mitochondrial function and myocyte apoptosis. However, recently we were the first to report that Col6 knockout leads to a “paradoxical” improvement in post‐MI cardiac remodeling and function in Col6−/− mice. To determine the mechanisms by which the absence of Col6 confers protection against ischemic injury, we assessed structural and functional differences of mitochondria between WT and Col6−/− mice pre‐ and post‐MI. Electron micrographs and western blot analysis revealed differences in mitochondrial fusion/fission protein flux as early as 24 hrs. post‐MI. The basal respiratory control index (RCI) was lower in Col6−/− mice compared to WT. However, the RCI of Col6−/− mice did not significantly decline following MI, where as this parameter was attenuated in WT mice. Together, these data indicate that the preservation of mitochondrial morphology and respiration following MI may be an important mechanism underlying cardioprotective effects of col6 knockout in response to MI.
Involvement of exercise‐induced macrophage migration inhibitory factor in the prevention of fatty liver disease
Authors: Moon Hyo Youl , Suh Pann‐Ghill
Published: June 2021

Physical inactivity can lead to obesity and fat accumulation in various tissues. Critical complications of obesity include type II diabetes and nonalcoholic fatty liver disease (NAFLD). Exercise has been reported to have ameliorating effects on obesity and NAFLD. However, the underlying mechanism is not fully understood. We showed that liver expression of macrophage migration inhibitory factor (MIF) was increased after 4 weeks of treadmill exercise. Phosphorylation of AMP‐activated protein kinase and acetyl‐CoA carboxylase in primary human hepatocyte cell lines was enhanced after MIF treatment. These responses were accompanied by increases in lipid oxidation. Moreover, inhibition of either AMPK or CD74 resulted in inhibition of MIF‐induced lipid oxidation. Furthermore, the administration of MIF to a human hepatocyte cell line reduced LXR agonist‐induced lipid accumulation. Taken together, these results indicate that MIF is expressed and secreted in the liver during physical exercise and prevents hepatic steatosis by activating the AMPK pathway.
Voluntary exercise at young stage prevents glucose intolerance in type 2 diabetic rats
Authors: Tsuzuki Takamasa , Yoshihara Toshinori , Ichinoseki‐Sekine Noriko , Kobayashi Hiroyuki , Naito Hisashi
Published: June 2021

Purpose

To investigate the effects of voluntary exercise at young stage on whole‐body metabolism and glucose tolerance in type 2 diabetic rats.

Methods

Otsuka Long‐Evans Tokushima Fatty (OLETF) rats as an animal model of type 2 diabetes and their counterparts (Long‐Evans Tokushima Otsuka; LETO) were used. Twenty male OLETF and LETO rats were divided into three groups, respectively: Sedentary (SED, n=8), Wheel Running (WR, n=6), or Wheel Running‐Sedentary (WS, n=6) group. WR and WS groups were allowed to access a running wheel freely from 5 to 25 week‐old. In WS group, however, a running wheel was locked from 15 to 25 week‐old and rats could not run in the wheel. At 25 week‐old, resting oxygen consumption (VO2 rest) of each rat was measured in a metabolic chamber. Subsequently an intraperitoneal glucose tolerance test (IPGTT) was performed.

Results

There were no significant differences in the areas under the curve of IPGTT (AUC) and VO2 rest (ml/kg/min) among three LETO groups. In OLETF rats, AUC in WR and WS groups were significantly lower than SED group, with AUC being lower for WR group. Moreover, VO2 rest of OLETF in WR and WS groups were significantly higher than SED group.

Conclusion

Our data suggest that voluntary exercise at young stage improves resting metabolism and prevents glucose intolerance in grown‐up type 2 diabetic rats, and these effects remained following exercise cessation.
Differential HSP expression between liver and quadriceps in response to chronic exertional heat stress
Authors: Haak Jodie L , Bloomer Steven A , Stauss Harald M , Liaboe Frederick O , Kregel Kevin C
Published: June 2021

Heat shock proteins (HSP) are upregulated as a key component of the cellular stress response to physiological challenges such as heat and exercise. In this experiment, we investigated the HSP response to the combined stressor of exercise plus hyperthermia in a chronic exertional heat stress (ExHS) model. In the ExHS experiments, male Sprague‐Dawley rats were run at 7–11 m/min in an environmental chamber (39°C) until core temperature (Tc) reached 41.8°C. Animals were exposed to 2 bouts of exertional heat stress per day for 5 consecutive days. In the passive heat stress experiment (HS), animals were placed in the chamber (45°C) until Tc reached 41.8°C. Nonheated animals served as controls. Three days after the final bout of ExHS or HS, rats were euthanized, and livers and quadriceps were harvested. HSPs were evaluated on whole‐tissue homogenates by immunoblotting. In the quadriceps and liver, HSP70 was barely detectable in the nonheated condition; however it was elevated in both tissues in response to HS and ExHS. Interestingly, liver in the the ExHS group displayed a 77% greater expression of HSP70 and a 35% greater expression of HSP60 compared to the HS group. However, in the quadriceps expression of HSP60 and HSP70 did not differ between HS and ExHS. Compared to the quadriceps, this augmented response of the liver to exertional heat stress suggests greater perturbation to homeostasis in the liver after ExHS.
The effect of plasma pH on proton kinetics of wrist flexor muscles following forearm and whole body exercise
Authors: Volianitis Stefanos , Secher Frederik , Secher Niels H , Quistorff Bjørn
Published: June 2021

This study tested the hypothesis that clearance of protons from the muscles depends not only on the muscle to plasma proton gradient but also on plasma pH. Seven rowers (6 males; 23 ± 3 years; mean ± SD) performed, in random order, a maximal “2000 m” trial on a rowing ergometer and maximal rhythmic isometric handgrip contractions for 5 min. The variation of intracellular and extracellular pH of the wrist flexor muscle was evaluated using magnetic resonance spectroscopy (31P‐MRS) for 20 min of the recovery. Exercise elicited a decrease in intracellular pH from 7.0 ± 0.0 at rest to 6.3 ± 0.2 and 6.5 ± 0.1 (P < 0.05) during rowing and handgrip exercise, respectively. Also, rowing lowered arterial pH from 7.4 ± 0.0 to 7.1 ± 0.0 and forearm venous pH from 7.36 ± 0.03 to 6.95 ± 0.06 (all P < 0.05), while during handgrip exercise arterial pH was not different from rest (7.43 ± 0.01 to 7.39 ± 0.02) and venous pH decreased only from 7.39 ± 0.02 to 7.20 ± 0.04 (P < 0.05). Even though the rate of intracellular pH change was correlated with the intracellular to plasma pH gradient, the slope of this correlation was different (−0.6872 vs −0.0391 μM/min, handgrip vs rowing, respectively; P < 0.05). Thus, muscle proton elimination depends not only on the muscle to plasma gradient but also is attenuated by a low plasma pH, suggesting that proton clearance from working skeletal muscle cannot be explained by passive transport.
Central apelin contributes to stress‐induced impaired gastric motility in rats
Authors: Bülbül Mehmet , Uysal Nimet , Birsen Ilknur , Gemici Burcu , Tanriöver Gamze
Published: June 2021

Stress plays a major role in functional gastrointestinal (GI) disorders. GI symptoms may develop as a result of accumulation stressors in some individuals while others can adapt to stressors. Exposure to repeated experience with the same stressor can produce an adaptation of behavioral and hypothalamic‐pituitary‐adrenal (HPA) axis response. However, the adaptation mechanism remains unclear. Acute restraint stress (ARS) increases corticotrophin releasing factor (CRF) in hypothalamic paraventricular nucleus (PVN) leads delayed gastric emptying (GE) via autonomic neural pathways in rats. ARS‐induced delayed GE is completely restored following chronic single stress (CSS) while restoration is no longer observed following chronic complicated stress (CCS). Apelin is upregulated in PVN in response to stress. Central effects of apelinergic system are attributed to regulation of stress response. We performed GE measurement and brain microdialysis from PVN under non‐stressed, ARS, CSS and CCS conditions in rats. ARS significantly increased CRF and apelin concentrations. CSS attenuated this response, whilst concentrations remained significantly high following CCS. ARS delayed GE which was completely restored following CSS, but not CCS. Central administration of apelin antagonist F13A restored delayed GE following ARS and CCS. Thus, apelin may be a candidate for treatment of stress‐induced GI disorders.
Lipid production in Chlorella protothecoides (1000.2)
Authors: Sizemore Brislyn , Waikel Rebekah
Published: June 2021

Depleting fossil fuel supply has prompted increased demand for the discovery of an alternative energy resource. Our study seeks to identify the genes that control lipid production, as well as, the regulation of these genes in Chlorella Protothecoides. To define gene expression profiles in lipid producing algae, Next Generation Sequencing was performed on total RNA extracted from algae grown in growth conditions (nitrogen sufficient) verses algae grown in lipid producing conditions (nitrogen deficient). Lipid production in nitrogen deficient was confirmed using Bodipy 505/515 staining, analyzed by flow cytometry. Next Generation Sequencing data yielded distinct algal gene expression profiles for both lipid producing and growth conditions. Blast analysis of differentially regulated mRNA sequences revealed several known genes likely to be involved in lipid production and 8 yet to be identified genes. Candidate genes were further analyzed by TaqMan to determine relative expression levels. We have established expression profiles of 10 of these candidate genes, which can be used as markers to identify different environmental condition which optimize lipid production.
Epigenetic regulation of LPS‐induced lung injury by sphingosine‐1‐phosphate lyase (1001.10)
Authors: Ebenezer David , Zhao Yutong , Ackerman Steven , Natarajan Viswanathan
Published: June 2021

Sphingosine‐1‐Phosphate Lyase (SGPL1) irreversibly hydrolyses Sphingosine‐1‐Phosphate (S1P) to ethanolamine phosphate and hexadecenal in mammalian cells. We have earlier demonstrated that inhibition or down‐regulation of Sphingose‐1‐Phosphate Lyase (SGPL1) ameliorates lipopolysaccharide (LPS)‐induced lung inflammation in mice. Therefore, challenge causes lung injury by increasing Sphingosine‐1‐Phosphate Lyase (SGPL1) expression, and inhibition of SGPL1 activity could produce therapeutic levels of immunosuppression. Our study is aimed to define SGPL1 mediated epigenetic regulation of LPS‐induced inflammatory responses in lung endothelium. Our preliminary data showed that down regulation of S1PL with siRNA blocked LPS‐induced IL‐6 secretion and partially blocked LPS‐induced acetylation of Histone H3 and Histone H4. Also, knock down of SGPL1 decreased MYST2 and IL‐6 promoter activity and potentiated Histone Deacetylase (HDAC) 5 expression. Moreover, Blocking HDAC activity with Trischostatin A potentiated LPS‐induced IL‐6 secretion while Histone Acetyl transferase (HAT) inhibitor Anacardic acid attenuated IL‐6 production. Taken together, these data reveal a potential epigenetic regulation mediated by SGPL1 in LPS‐induced lung inflammation.

Grant Funding Source: Supported by National Institute of health P01HL098050
Steroids contents of wound healing preparations from the skin of the Arabian Gulf catfish (Arius bilineatus, Val.) (1001.7)
Authors: Al‐Hassan Jassim , George Sosamma , Afzal Mohammad
Published: June 2021

The Arabian Gulf catfish secretes copious amounts of epidermal gel‐like material when threatened or injured. The freeze dried gel contained 85% proteins, 14.4% lipids 2.6% nucleic acid components and carbohydrates.

Preparations from the skin of the catfish were found to accelerate the rate of wound healing, as well as being potent anti‐inflammatory. The lipids were composed of non‐polar lipids, glycolipids and phospholipids. Analysis of the lipids for steroids using TLC, FTIR, 1HNMR, 13CNMR, GC/MS, and GC/MS/MS showed them to contain 14 steroids, including 12 oxysterols. The following compounds were characterized: cholesterol, 7α‐hydroxy cholesterol, 7β‐hydroxycholesterol, 5,6 epoxycholesterol, 3β‐hydroxycholest‐5‐ene‐7‐one, cholest‐3,5‐diene and cholesta‐3,5‐dien‐7‐one. Progesterone, cholest‐3,5,6‐triol and 4‐cholesten‐3‐one were found as minor components, and were identified by comparing their MS, 1HNMR and FTIR spectral data with those of standards and library data. Cholest‐3,6‐dione, cholesta‐4,6‐diene‐3‐one, cholesta‐2,4‐diene, and cholesta‐5,20(22)‐dien‐3‐ol were found only in trace amounts and were identified by GC/MS and MS and their spectral data were compared with library data. The bile of the catfish was found to contain 6 bile acids and salts. The presence of the oxysteroids together with eicosanoids, platelet activating factors, furan fatty acids and unsaturated fatty acids in the skin secretion may help understand the pharmacological and therapeutic activities of the preparations from the skin of the catfish.

Grant Funding Source: This project is supported by Research Administration, Kuwait University Grant No. SL04/09
The hypertriglyceridemic waist, waist‐to‐height ratios and cardiometabolic risk in university students (1001.8)
Authors: Villalobos Raffael D , Villalobos Daniel , Murguía‐Romero Miguel , Jiménez‐Flores Raffael , Méndez‐Cruz René , Sigrist‐Flores Santiago , Medrano Gabriel , Villalobos‐Molina Rafael
Published: June 2021

We used hypertriglyceridemic waist (HW) and waist‐to‐height ratios (WHTR) as simple clinical approaches to identify young adults at risk of cardiometabolic disorders. This cross‐sectional design study was assessed in 4619 students (3142 women) aged 17‐24 years from first grade university, México. Body mass index (BMI), waist circumference (WC) and biochemical variables were determined. The HW phenotype was defined as a waist circumference > 90th percentile for age and sex, and triglyceride concentrations > 1.24 mM, and a high WHTR was defined as > 0.5. We found 42.2% students with increased WC, 18% hypertriglyceridemic and 11.6% of HW phenotype among the sample. Waist circumference and the HW phenotype seem to predict better those young adults at increased risk for cardiometabolic disorders, i.e., those who bear components of metabolic syndrome/insulin resistance.

Grant Funding Source: RV‐M is supported by PASPA, DGAPA, UNAM
Human dendritic cells induce B‐cell responses to polysaccharide antigens (1004.2)
Authors: Mendoza Daniel , Xu Yi , Rodriguez‐Barradas Maria , Tweardy David , Orson Frank , Corry David
Published: June 2021

Background: Carbohydrates mediate host‐pathogen interactions and play prominent roles in cancer and inflammatory diseases. The large majority of carbohydrates directly activate B cells without T cell help to produce antibody responses. For a long time, these responses were assumed to be independent of antigen‐presenting cells (APC). However, recent studies in mice indicate a role of APC in T‐cell independent responses. To determine whether human APC play a role in immune responses to carbohydrates, we characterized dendritic cell (DC) responses to pneumococcal polysaccharides (PS) and PS‐based vaccines (PCV13 and PPV23).

Methods: we incubated human monocyte‐derived DC (MDDC) with a fluorochrome‐labeled PS at various time points to measure antigen internalization in DC by deconvolution microscopy. In different experiments, we exposed PBMC of unvaccinated healthy donors to PS‐based vaccines for 6 hours ex vivo to measure IL‐8 production in DC by flow cytometry. Finally, we incubated human B cells with PS with and without autologous MDDC to measure IgG production by ELISA 6 days later.

Results: Human DC internalized PS within 1 hour of exposure. IL‐8 was the main cytokine produced in PBMC in response to PS. Percent of DC producing IL‐8 in response to PS‐based vaccines were: medium: 5.15%, PCV13: 23%, PPV23 10%, (p<0.0001). DC induced IgG responses to PS: B cells alone: 33.3; B cells+PS: 25.5; DC+B cell alone: 60.2; DC+B cell+PS: 114 ug/ml, p=0.04. DC induced IgM production to PS as well.

Conclusions: human DC interact with PS and induce B‐cell responses to PS. Overall, we have identified a novel and essential immune role for human DC in immune responses to carbohydrate antigens with important implications for vaccine design and immunogenicity.
Reciprocal actions of Wnt9A and BMP4 on chondroitin‐4‐sulfotransferase (CHST11) expression in relation to arylsulfatase B activity (1005.2)
Authors: Bhattacharyya Sumit , Feferman Leo , Tobacman Joanne
Published: June 2021

Initial experiments in human colonic epithelial cell lines and in the arylsulfatase B (N‐acetylgalactosamine‐4‐sulfatase; ARSB)‐deficient mouse showed that reduction in ARSB and the resultant increase in chondroitin‐4‐sulfate (C4S) were associated with reduction in chondroitin‐4‐sulfotransferase expression and ~50% reduction in sulfotransferase activity with overall increase in total sulfated glycosaminoglycans and C4S. Our experiments have also shown that decline in ARSB leads to increase in Wnt9A expression in human colonic epithelial cells and mouse intestine. Since modeling of the extracellular matrix by Wnt and BMP4 involves their interactions with sulfated glycosaminoglycans, including chondroitin sulfate, we evaluated the interactions among ARSB, Wnt9A, BMP4, and chondroitin‐4‐sulfotransferase (CHST11). Following silencing of Wnt9A by siRNA, the CHST11 expression increased to ~3 times baseline and the sulfotransferase activity increased to ~1.6 times the baseline. In contrast, when BMP4 was neutralized by blocking antibody, the CHST11 expression declined to ~50% of the baseline level. Exogenous administration of BMP4 increased the CHST11 expression to 2‐3 times the baseline level. In ARSB‐deficient mice, the BMP4 expression was ~50% of the value in the wild‐type control, and consistent with a decline in CHST11 expression. Both BMP4 and Wnt exert crucial effects in modeling of the extracellular environment, and these observed opposing effects on CHST11 expression and in relation to ARSB activity may be fundamental to their interaction and to their structural and functional effects.
The role of VH CDR3 in determining Tn‐antigen specificities of MLS128 and 83D4 monoclonal antibodies (1007.1)
Authors: Sato Hajime
Published: June 2021

Tn‐antigen (GalNAcα‐Ser/Thr) is associated with carcinomas. MLS128 and 83D4 are Tn‐antigen specific monoclonal antibodies. They bind carbohydrate epitopes consisting of three or two consecutive Tn‐antigens (Tn3 or Tn2) with different affinities. Since VH sequences of both mAbs are nearly identical except a few amino acids in the CDR3, the aim of this study was to evaluate the role of the VH CDR3 in determining their Tn‐antigen specificities.The VH domain of 83D4 was cloned into pCI‐MLS128 VL‐Fc to construct pCI/MLS128/83D4 VH CDR3 scFv‐Fc vector. CHO cells transfected with the vector were cultured in a selective medium to screen clones expressing MLS128/83D4 VH CDR3 scFv‐Fc protein. Fc‐levels in media of stable clones were determined by ELISA. Of several stable clones producing the MLS128/83D4 VH CDR3 scFv‐Fc protein at 2 µg/ml levels established, one clone was chosen for purification and characterization of the MLS128/83D4 VH CDR3 scFv‐Fc protein. From 330 ml of culture medium, 70 μg of the scFv‐Fc protein were purified to near homogeneity by Protein A‐Sepharose chromatography. Binding activity of three scFv‐Fc proteins (MLS128/83D4 VH CDR3, MLS128, 83D4) was compared by ELISA using Tn2‐ and Tn3 peptides.
Angiotensin‐II‐induced expression of the early growth response protein 1 is mediated by CaMKII‐dependent pathway in vascular smooth muscle cells (1011.7)
Authors: Simo Cheyou Estelle , Srivastava Ashok
Published: June 2021

Egr‐1 is a transcription factor implicated in the regulation of several growth‐related cascades. It has been suggested to participate in the development of vascular disease processes such as intimal thickening following vascular injury, atherosclerosis and cardiac hypertrophy. Upregulation of Egr‐1 was recently shown to be a key contributing factor in stimuli‐induced VSMC proliferation and hypertrophy. Ang‐II, a potent vasoactive peptide with pathogenic role in vascular disease, has been demonstrated to contribute to aberrant remodeling of the vessel wall. This is mainly due to the ability of Ang‐II to activate hypertrophic and proliferative pathways such as the mitogen‐activated protein kinase and the phosphatidylinositol‐3‐kinase pathways in VSMC. It has been shown earlier that Ang‐II requires H2O2 generation to activate these signaling pathways and we have shown that Ca2+, calmodulin (CaM) and Ca2+/CaM‐dependent protein kinase II(CaMKII), play a critical role to trigger H2O2‐induced effects in VSMC. We have also shown that endothelin‐1, another mitogenic vasoactive peptide, requires CaMKII to upregulate Egr‐1 expression and to mediate its physiological responses. However, Ang‐II‐induced modulation of Egr‐1 expression in VSMC and the implication of CaMKII in this process remain unexplored. Therefore these studies were undertaken to characterize the effect of Ang‐II on Egr‐1 expression in A10 VSMC and to examine the role of Ca2+, CaM and CaMKII on this response. Ang‐II, at 100nM, induced an increase in Egr‐1 expression in a time‐dependent fashion in the nucleus of A10 VSMC. Pharmacological blockade of CaM and CaMKII by calmidazolium and KN‐93 respectively, significantly attenuated Ang‐II‐induced Egr‐1 expression. These results demonstrate that Egr‐1 expression is upregulated in response to Ang‐II via CaM/CaMKII‐dependent pathways in VSMC.

Grant Funding Source: Supported by the Canadian Institutes of Health Research
PP2A activation in vivo is sufficient to disrupt interactions among vascular Akt‐Hsp90‐eNOS (1013.16)
Authors: Panneerseelan Leenalitha , Ruan Ting , Li You You , Wan Xin , Kunz David , Walker Matthew , Nhan Jennifer , Ravindran Anindita , Babu P. V. A. , Zhang Quan , Abel E. Dale , Symons J. David
Published: June 2021

We have shown in bovine aortic endothelial cells (BAECs) treated with palmitate (P) and in arteries from obese mice that protein phosphatase 2A (PP2A) associates directly with eNOS. When PP2A co‐localizes with eNOS, interactions among Akt‐Hsp90‐eNOS are disrupted, p‐eNOS/eNOS is decreased, and NO bioavailability is impaired. We hypothesized that PP2A activation is sufficient to disrupt interactions among Akt‐Hsp90‐eNOS in arteries from fat‐fed mice. To test efficacy of the in vivo PP2A inhibitor Lixte Biotechnology 1 (LB1, Setauket, NY), BAECs were treated (3h) ± 500 uM P ± 4 uM LB1. P‐induced increases (p<0.05) in PP2A activity, and reductions (p<0.05) in p‐eNOS/eNOS and NO production (ELISA and EPR), were prevented by LB1. To determine the safety / efficacy of LB1, mice were treated (1 mg/kg/day IP) ± LB1 for 21 days. PP2A activity was reduced (p<0.05) and p‐PP2A Y307 to PP2A was elevated (p<0.05) in arteries from LB1 vs. vehicle‐treated mice after 3 and 21 days. To confirm the importance of PP2A vs. other phosphatases, BAECs incubated ± PP1 (3 uM tautomycin), PP2B (100 nM cyclosporine), or PP2A (4 uM LB1) inhibition. P‐induced reductions (p<0.05) in p‐eNOS/eNOS were prevented only by LB1. Finally, 7‐week old, male C57B16 mice consumed standard (CON, n=20) or high‐fat (HF, n=20) chow for 12‐weeks. Subgroups (n=10) of CON and HF mice received IP injections of saline (vehicle; V) or LB1 for the last 14‐days. HF mice gained weight and developed peripheral glucose intolerance vs. CON mice regardless of LB1 treatment. p‐eNOS/eNOS was impaired (53±6%; p<0.05) in vascular homogenates from HF‐V vs. all groups. After eNOS immunoprecipitation (IP), the association between: (i) PP2A:eNOS increased 95±11%, and the co‐localization between (ii) Akt:eNOS decreased 53±8%, and (iii) Hsp90:eNOS decreased 56±12%, in HF‐V vs. CON mice (all p<0.05). These findings suggest that PP2A activation is sufficient to disrupt interactions among Akt‐Hsp90‐eNOS in arteries from fat‐fed mice.

Grant Funding Source: ADA 7‐08‐RA‐164, ADA1‐12‐BS‐208, 2R15HL091493
An empirical study on accountability for promoting healthy food environments in England through the Public Health Responsibility Deal Food Network (1014.2)
Authors: Kraak Vivica , Swinburn Boyd , Lawrence Mark , Harrison Paul
Published: June 2021

Objective: In 2011, the United Kingdom launched five Public Health Responsibility Deal Networks inspired by ‘nudge theory’ to facilitate healthy‐lifestyle behaviors. This study used Q methodology to examine stakeholders’ views about responsibility and accountability for healthy food environments to reduce obesity and diet‐related chronic diseases.

Design: A purposive sample of policy elites (n=31) from government, academia, food industry and civil society sorted 48 statements grounded in three theoretical perspectives (i.e., legitimacy, nudge and public health law). Factor analysis identified intra‐individual statement sorting differences.

Results: A three‐factor solution explained 64 percent of the variance across three distinct viewpoints: food environment protectors (n=17) underscored government responsibility to address unhealthy food environments; partnership pioneers (n=12) recognized government‐industry partnerships as legitimate; and the commercial market defenders (n=1) emphasized individual responsibility for food choices and rejected any government intervention.

Conclusions: Building trust and strengthening accountability structures may help stakeholders navigate differences to engage in constructive actions. This research may inform efforts in other countries where voluntary industry partnerships are pursued to address unhealthy food environments.

Grant Funding Source: World Health Collaborating Centre for Obesity Prevention, Deakin University, Australia
Analysis of hospital feeding practices at Bach Mai Hospital, Hanoi Vietnam (1015.7)
Authors: Young Lorraine , Huong Pham , Lam Nguyen , Thu Nghiem , Van Ha , Lien Dinh , Hoc Tran , Anh Nguyen , Henry Elizabeth , Lenders Carine , Gura Kathy , Apovian Caroline , Ziegler Thomas
Published: June 2021

Background: Limited data on nutrition support practices in the hospital setting are available in the country of Vietnam.

Methods: From October 2011‐December 2011, a collaboration between investigators from the U.S. and the Vietnamese National Institute of Nutrition enrolled 72 subjects admitted for elective GI surgery for this observational study at Bach Mai Hospital, a major Hanoi teaching hospital. Baseline height, weight, BMI, mid upper arm circumference (MUAC), Subjective Global Assessment (SGA), body weight status, and daily kcal and protein/amino acid intake from oral diet, tube feeding, and IV feedings from admission until discharge were recorded.

Results: Mean age of subjects was 56±15 years and length of stay 12±5 days. 50% of subjects scored a B or C (moderate to severe malnutrition) on the SGA. BMI range was 13‐28; 48% had a BMI < 18.5. MUAC was low normal (24±4 cm). All subjects lost weight during admission. Almost all patients (98%) were fed using parenteral nutrition (PN) postoperatively with oral feeding starting on postoperative day 4. Tube feedings were used in only one patient. Mean daily total calorie intake was 15 kcal/kg/day and protein intake was 0.61g/kg/day. Minimal or no micronutrient supplementation was given to most patients.

Conclusions: Study findings support the development of a nutrition intervention with feeding protocols to increase postoperative energy and micronutrient intake.
Determinants of exclusive breastfeeding cessation in a peer counseling breastfeeding support program for low‐income women (1016.4)
Authors: Rozga Mary , Benton Pat , Kerver Jean , Olson Beth
Published: June 2021

Breastfeeding peer counseling programs have demonstrated efficacy in improving exclusive breastfeeding (EBF) rates, but the impact of specific program protocols on this outcome is unknown.

OBJECTIVE: To describe associations between individual and combinations of program components and EBF cessation in a peer counseling breastfeeding support program.

METHODS: Data was collected for 5,886 low‐income women prenatally enrolled in Michigan’s Breastfeeding Initiative Program from 2005‐2011. Cox proportional hazards models and chi‐square analyses were used to determine the impact of program components on EBF cessation. “Optimal” quantities of contacts were based on program guidelines.

RESULTS: Each home, phone and other contact had a similar impact on risk of EBF cessation at three months postpartum (HR (95% CI): 0.92 (0.89, 0.94); 0.90 (0.88, 0.91); and 0.94 (0.91, 0.98) respectively). Women who received a more in‐person and less phone contacts than were considered optimum had the lowest risk of EBF cessation (HR (95% CI): 0.3 (0.2, 0.3)).

CONCLUSIONS: The current study demonstrated the importance of in‐person contacts between participants and trained peer counselors compared to phone contacts and further demonstrates that the combination of in‐person and phone contacts may have an appreciable impact on EBF cessation.
Clinical trials for foods: satiety panels (1018.10)
Authors: Kessler Jessica , Harris Matthew , Frestedt Joy
Published: June 2021

The purpose of a satiety panel is to explore feelings of fullness and hunger after consuming a test product (food, food ingredient, dietary supplement, etc). We considered 5,264 people for a satiety panel breakfast meal. Recruitment methods included email, phone calls, internet posts, surveys, and flyers posted in high traffic areas. The trial involved 5 visits to our research site to consume 5 different breakfast meals composed of orange juice, a breakfast bar and yogurt. Subjects were required to respond to questionnaires within specified time limits. Of the 63 enrolled, 16 were withdrawn due to non‐compliance with study protocol. A total of 47 people completed all five visits with questionnaires completed within the time allotted. Recruitment difficulties were related to lack of a treatment benefit (this was a food not a therapeutic offering), travel time and distance from the research site, schedule conflicts, unresponsiveness, and ineligibility per exclusion criteria. The high retention rate added to the success of the satiety panel and was affected by the efforts of study staff to contact subjects to ensure completion of questionnaires. This study was supported by a consumer products company.