bluuebunny/crossref_metadata_2025 huggingface dataset:
Use of a rapid analytical method for identifying closely related Yersinia species to provide a procedure for food safety and security
Authors: Whittaker Paul
Published: June 2021

Capillary gas chromatography with flame ionization detection (GC‐FID) analysis of chemical components of bacterial cells has provided useful information for rapid detection and identification of bacteria in clinical and diagnostic bacteriology laboratories and currently has increased significance for both food safety and security. GC‐FID was used to determine the cellular fatty acid (CFA) profiles of closely related Yersinia species. For GC‐FID analysis, whole cell fatty acid methyl esters (FAMEs) from cells cultured on brain heart infusion (BHI) agar at 35 (C for 24 h were obtained by saponification, methylation and extraction into hexane/methyl tert‐butyl ether. A data set for each Yersinia species was prepared using fatty acid profiles from five replicates prepared on different days. Major fatty acids of the 26 Yersinia strains evaluated in this study were straight‐chain 12:0, 14:0, 15:0, 16:0 and unsaturated 16:1 ω7c/16:1 ω6c, 18:1 ω7c, and 14:0 3OH/16:1 iso, and 17:0 ωcyclo 7‐8. The Yersinia species Y. enterocolitica, Y. pseudotuberculosis, Y. frederiksenii, Y. intermedii, Y. kristensenii, and Y. pestis have unique fatty acid patterns. The CFA profiles for Y. pestis and Y. pseudotuberculosis are similar, but there are several fatty acids, 16:1 ω5c, 16:0, 17:1 ω7c, 17:0 ωcyclo 7‐8, 19:0 and summed 18:2 ω6c, 9c/18:0 ante that differ significantly between these two species. Analysis of FAMEs from Yersinia strains grown on BHI agar by a rapid GC‐FID method can provide a sensitive procedure for the identification of these organisms, and this analytical method provides a procedure applicable to issues of both food safety and security.
Household food supply is affected by food abundance‐food shortage monthly cycle among food insecure women in Ohio
Authors: Ye Qian , Remley Dan , Zubieta Ana Claudia , Long Cindy , Angell Deb , Mims Sheryl , Melgar‐Quinonez Hugo
Published: June 2021

This study assesses the existence of a monthly cycle of food abundance‐shortage among food insecure Caucasian women in Ohio. The study interviewed 29 low income women at the beginning and the end of two continuous months. Household food supply was assessed using a shelf food‐inventory questionnaire with a list of food items common to Ohioan households. Food insecurity was measured by a Household Food Security Module. Body mass index was calculated using measured heights and weights. Differences in the number of food items between the beginning and the end of the months were determined through paired t‐test. Most of the women were classified as overweight or obese (78% and 66%, respectively), and 93% of them were food insecure. The number of food inventory items at the beginning of each month was significantly higher than at the end of the month (Month 1: 83.6 vs. 65.9; Month 2: 83.7 vs. 68.1; p=0.01). With the exemption of oils, the numbers of food inventory items by food groups (grains, vegetables, fruits, milk, meat & beans, and oils) were significantly higher at the beginning of both months. The biggest difference was observed in the meat & beans group (Month 1: 10.3 vs. 6.9; Month 2: 10.4 vs. 7.5; p=0.01). These findings indicate that food insecure households may suffer monthly cycles with drastic decreases of the food supply by the end of the month. Funded by Ohio Agricultural Research and Development Center (Grant # OHOA1277).

Grant Funding Source

Ohio Agricultural Research and Development Center
Effect of Unbalanced Dietary Habits to the Body Development of Preschool Children
Authors: Shin Kyung Ok , An Chang Hun , Hwang Hyo Jeong , Lee Jun Sik , Choi Kyung Soon , Chung Keun Hee
Published: June 2021

The dietary habit and life style of preschool children will affect nutrient intake for children. Nutrient intake for preschool children is critical because it affects the growth and is an influencing factor for adult health status when they grow up.

The objective of this research is to study how nutrient intake and life style will affect the body development for preschool children. Generally, 87% of children reporting had a good dietary habit, 53% of children had regular meals, 18% of children had balanced meals, and 39% of children had unbalanced meals. Children who eat unbalanced meal had relatively lower intake of meat, fish, egg, soy bean, kimchi, vegetable, fruit, and ate irregular meal compare to the children who eat balanced meal. However, those who eat unbalanced meal had relatively higher sweet food such as ice cream, cookie, and carbonated beverage and had lower intakes of niacin, Ca, and Zn.

According to Kaup Index, 59.8% of reporting children had normal weight(KI=15‐18), 28.9 % are under‐weighted, and 11.3% are over‐weight or obese. In contrast, according to 20% of body fat, 14.9% of reporting children were under‐weighted, 20.6% were normal‐ weighted, 20.6% were obese, and 46.1% were over‐weighted or obese. Among children who eat balanced meal, 64.9% were normal‐weighted and 27.3% were under‐weighted, while among children who eat unbalanced meal, 30.4% were under‐weighted and 58.4% were normal‐weighted (p<0.01)

Consequently, it is imperative to develop the teaching program for nutritionists and teachers who take care the children in the preschool facility.
Effects of partial liver ischemia followed by global liver reperfusion on remote organs: lungs and kidneys
Authors: Capellini Verena Kise , Miranda Luiz Eduardo Correia , Reis Graziela Saraiva , Celotto Andrea Carla , Carlotti Carlos Gilberto , Evora Paulo Roberto Barbosa
Published: June 2021

Hepatic ischemia (I) followed by reperfusion (R) results in mild to severe remote organ injury. Oxidative stress and nitric oxide (NO) seems to be involved in I/R injury. The aim was to investigate the effects of liver I/R on liver function and on lipid peroxidation, leukocyte infiltration and NO synthase (NOS) immunostaining in lung and kidney. 24 male Wistar rats were randomized into 3 groups: 1) sham, 2) 60 min of partial (70%) liver I and 2 h of global liver R, and 3) 60 min of partial (70%) liver I and 6 h of global liver R. The groups 2 and 3 presented a significant increase in plasma levels of alanine and aspartate aminotransferase and in tissue levels of malondialdehyde and myeloperoxidase. In the kidney, the positive endothelial NOS (eNOS) staining was significantly decreased in group 3 compared to 1 and the staining for inducible NOS (iNOS) and neuronal NOS (nNOS) did not differ among groups. In the lung, the staining for eNOS and iNOS did not present statistical differences among groups and no positive nNOS staining was observed in any group. These results suggest that partial liver I followed by global liver R induces liver, kidney and lung injuries characterized by neutrophil sequestration and oxidative stress increase. In addition, we can suppose that the reduction in NO formation via eNOS can be implicated in the moderate impairment of renal function, observed by others, 24 hours after liver I/R. Support: FAPESP; FAEPA.
Selective but Slight Enhancement of Delta Agonist‐Induced Antinociception by Repeated Morphine in Rhesus Monkeys
Authors: Negus S. Stevens , Banks Matthew L. , Schrode Katrina , Morrissey Ember , Rice Kenner C.
Published: June 2021

Repeated treatment with mu opioid receptor agonists (e.g. morphine) may promote insertion of delta receptors into neuronal membranes and enhance the potency/efficacy of delta opioid receptor agonists. The present study compared effects of the delta agonist SNC80 and the mu agonist fentanyl after repeated saline or morphine in rhesus monkeys. In an assay of schedule‐controlled responding (FR30 for food), SNC80 and fentanyl each produced a dose‐dependent decrease in response rates after repeated saline treatment. Repeated morphine treatment (1, 1.6, 2 and 3 mg/kg at 12hr intervals, testing 12hr after last dose) had no significant effect on the potency or efficacy of either drug. In the assay of thermal nociception (tail‐withdrawal from 48 and 54°C water), SNC80 had little effect, whereas fentanyl produced full and partial antinociception at 48 and 54°C, respectively. Repeated morphine produced a significant but only slight enhancement of SNC80 effects and reduction in fentanyl effects at 54°C. Thus, under these conditions, repeated mu agonist treatment produced a selective but slight enhancement of the antinociceptive effects of SNC80 in rhesus monkeys. Supported by NIDA and NIAAA of NIH.
Validating aspects of pica response induced by oxycodone administration
Authors: Batra Vinita , Schrott Lisa M
Published: June 2021

Oxycodone is a semisynthetic opioid analgesic. In our lab prior studies with oxycodone were accompanied by prolonged and intense oral ingestion of non‐food substances. These manifestations resemble pica behavior, an index of nausea and emesis associated with chemotherapeutic drugs and other opioids, but not to date oxycodone. To further explore this behavior, route of administration, time course and receptor mediation were investigated. The results obtained suggested that pica response is associated with oxycodone administration. Proemetic agent‐induced kaolin intake is a well established model to assess pica. To corroborate our results, we assessed kaolin intake following oxycodone administration. Adult female rats were adapted to preweighed kaolin for 2 days followed by baseline testing. Following drug or vehicle treatment, kaolin intake was assessed for 3hr for 2 days. At the end of the experiment, wet weight of stomach was measured and brain was isolated for c‐fos analysis. Oxycodone and cisplatin treatment moderately increased kaolin intake and the wet weight of stomach was elevated 2‐3 fold. Preliminary results indicate positive c‐fos staining in the area postrema and nucleus of solitary tract following oxycodone treatment. These results validate our hypothesis. Future studies will assess the mechanisms underlying the neurobiological basis of this behavior. Supported in part by LEQSF RD‐A‐19, (LA Board of Regents) and USPHS DA018181 (NIDA, NIH).
Heterogeneous Organization of the P450 System in Lipid Microdomains of the Endoplasmic Reticulum
Authors: Brignac‐Huber Lauren M , Reed James R , Backes Wayne L
Published: June 2021

P450 is part of an electron transport chain found in the endoplasmic reticulum (ER), with its catalytic function requiring interactions with other ER‐resident enzymes such as NADPH‐cytochrome P450 Reductase (CPR). The goal of this study is to examine how components of the P450 system are organized, and to determine if they are heterogeneously distributed in ER lipid microdomains. Microsomes were partially solubilized with 1% Brij 98 at 37°C and centrifuged, separating membranes into detergent‐resistant (pellet) and ‐soluble (supernatant) regions. Lipid analysis of these samples illustrated that the pellet was enriched with sphingomyelin and cholesterol when compared to the bulk ER membrane, implicating the presence of lipid raft‐like domains. Detergent‐resistant and ‐soluble membranes were subjected to immune blotting to establish the regional distribution of CPR and P450s within the membrane. Microsomes induced with Phenobarbital and β‐Naphthoflavone, exhibit both CPR (~50%) and CYP1A2 (~50%) residing in a region of the membrane that was resistant to solubilization, whereas CYP2B4 was located in a more detergent‐soluble region. Preliminary studies using sucrose gradient centrifugation has shown similar findings. These results suggest that the proteins of the ER are not randomly distributed in the membrane, but appear to exist in specific lipid microdomains. (Supported by NIEHS ES00434)
The UGT2B15*2 polymorphism is a major determinant of oxazepam clearance in human volunteers.
Authors: He Xi , Hesse Leah , Masse Gina , Harmatz Jerold S , Greenblatt David , Court Michael H
Published: June 2021

Oxazepam is specifically glucuronidated by UGT2B15. In previous studies we have shown that a common non‐synonymous polymorphism in the UGT2B15 gene (*2, D85Y) results in lower oxazepam glucuronidation by recombinant enzyme and human liver microsomes. The goal of this study was to determine the effect of UGT2B15*2 on the pharmacokinetics of oxazepam in 30 healthy male subjects administered 15 mg oxazepam by mouth. UGT2B15 genotype frequencies were 0.20, 0.67 and 0.13 for *1/*1, *1/*2 and *2/*2, respectively. Mean (+/‐ SD) plasma oxazepam area under the curve values extrapolated to infinity were significantly higher (P<0.001, ANOVA) in *2/*2 subjects (2.50 +/‐ 0.96 ug/mL*hr) compared with *1/*2 (1.37 +/‐ 0.40) and *1/*1 (0.981 +/‐ 0.29) subjects. Furthermore, mean weight‐normalized plasma clearance values were significantly lower (P=0.003, ANOVA) in *2/*2 subjects (1.55 +/‐ 0.61 mL/min/kg) compared with *1/*2 (2.56 +/‐ 0.74) and *1/*1 (3.53 +/‐ 1.14) subjects. These results mirror our previous in vitro results and substantiate the UGT2B15*2 polymorphism as a major determinant of oxazepam clearance in vivo. (Supported by NIH grants GM‐61834, DK‐79300, AI‐058784, and AG‐017880.)
Expression and localization of cholesterol sulfotransferase (SULT) 2B1b in human brain
Authors: Salman Emily Deanna , Runge‐Morris Melissa , Kocarek Thomas , Falany Charles
Published: June 2021

Regulation of cholesterol homeostasis in brain is necessary for neurotransmission, myelination, steroid and oxysterol synthesis, and membrane integrity. SULT2B1b is a cytosolic SULT capable of sulfonating cholesterol, oxysterols, hydroxysteroids, and therapeutic drugs. SULT2B1b mRNA and protein were localized in multiple regions of human brain by RT‐PCR, immunoblot and immunohistochemical (IHC) analysis. SULT2B1b was expressed in all tissues tested, including: superior temporal gyrus, hippocampus, cerebellum, occipital, frontal and temporal lobes. With IHC, SULT2B1b protein was localized to both neurons and glial cells with oligodendrocytes showing the most intense staining. Proteins involved in cholesterol metabolism and regulation, including: LXRα, LXRβ and CYP46, responsible for synthesis of 24(S)‐hydroxycholesterol, have also been localized to these brain regions. SULT2B1b is capable of the sulfation of 24(S)‐hydroxycholesterol, regulating its LXR ligand activity. Therefore, SULT2B1b is apparently the SULT isoform responsible for the conjugation of cholesterol, oxysterols and steroids in the human brain. Supported by PHS grant GM38953 to CNF and ES/CA05823 to MRM.
Prevention of Functional and Pathological Lung Changes in Bleomycin‐Induced Models of Pulmonary Fibrosis with a Pan‐TGF‐b Neutralizing Antibody
Authors: D'Angona Anne Marie , Finn Patrick , Gregory Rick , Philips Lucy , Lonning Scott , Andrews Laura , McVie‐Wylie Alison
Published: June 2021

Idiopathic pulmonary fibrosis (IPF) affects 34/100000 people per year. Currently there is no cure for IPF, partially because the etiology remains unknown. IPF is believed to develop through three phases: inflammation, alveolar epithelial damage, and fibroblast response resulting in excess matrix. TGF‐b is known to play a critical role in the development of fibrosis in IPF as well as other fibrotic diseases. We have employed the use of a bleomycin‐induced model of pulmonary fibrosis to evaluate a pan‐ TGF‐b neutralizing antibody (1D11) as a potential therapeutic. Although bleomycin treatment induces several pro‐inflammatory and pro‐fibrotic responses, TGF‐b is upregulated and modulation of TGF‐b has been suggested to result in decreased fibrosis. Following bleomycin administration, mice developed fibrosis, which was abrogated by 1D11 treatments. In addition, flexiVent analysis revealed significant changes in functional parameters, which 1D11 treatments improved. In initial studies, direct 1D11 administrations prevented the development of alveolar/perivascular inflammation as well as fibrotic lesions. Furthermore, 1D11 treatments prevented reduction of functional parameters, most notably elastance. These studies have demonstrated functional and histologic improvement with 1D11 and provide support for the utilization of therapeutic TGF‐b neutralizing antibodies for the treatment of IPF.
Activation of nitric oxide‐cyclic GMP pathway (NO‐cGMP) with nitrite or sildenafil, but not BAY 41‐2272 (BAY), blunts acute pulmonary embolism (APE)‐induced increases in circulating matrix metalloproteinase‐9 (MMP‐9) and oxidative stress
Authors: Dias‐Junior Carlos Alan , Cau Stefany Bruno Assis , Montenegro Marcelo Freitas , Castro Michele Mazaron , Tanus‐Santos Jose Eduardo
Published: June 2021

Objectives

Oxidative stress is a major upregulator of MMP‐9 and both are induced by APE, thus, we comparatively assessed the effects of NO‐cGMP activators with and without antioxidant properties on APE‐induced increases in MMP‐9.

Methods

Hemodynamic and biochemical evaluations were performed in non‐embolized dogs treated with saline (N= 5), and in embolized dogs (intravenous injections of microspheres) that received saline (n=9), or nitrite (6.75 µmol/kg over 15 min followed by 0.28 µmol/kg/min; n=5), or sildenafil (0.25 mg/kg over 30 min; n=5), or BAY (0.03, 0.1, 0.3, and 1 mg/kg/h; n=5). Plasma thiobarbituric acid reactive substances (TBARS) concentrations and MMP‐2 and MMP‐9 levels were determined using a fluorometric and gelatin zymography methods, respectively.

Results

APE increased pulmonary artery pressure by ~25 mmHg. The infusion of each drug (nitrite, BAY, or sildenafil) reversed this increase by ~40% (all P<0.05) and similar effects were seen on the pulmonary vascular resistance. While both nitrite and sildenafil produced no systemic effects, the infusion of BAY produced systemic hypotension (P<0.05). While nitrite and sildenafil blunted the increases in plasma MMP‐9 levels and TBARS (all P<0.05), BAY produced no such effects.

Conclusion

These findings are consistent with the idea that NO‐cGMP activators with antioxidant effects prevent the release of MMP‐9 during APE. Supported by FAPESP
Modulation of plasma cytokine levels in mice without and with collagen induced arthritis by CO2 and isofluorane.
Authors: Christopher Lawrance , Alexander Heather , Clarke Stephen , Lucas Edralin , Smith Brenda , Kuvibidila Solo
Published: June 2021

Overdose of CO2 or Isofluorane is a common method of euthanasia widely used in mice. Isofluorane has recently been getting much attention due to its immunomodulatory effects. However, the role of both compounds on immune and/or inflammatory markers of mice is limited. In the current study, nineteen‐week old DBA female mice were euthanized with either CO2 (n=55) or isofluorane (n=55) inhalation and plasma was collected for the measurement of soluble ICAM1, tumor necrosis factor‐alpha (TNF‐alpha), and interleukin‐6 (IL‐6) by ELISA. Fifty eight of the mice had collagen‐induced arthritis (CIA) whereas 52 did not. When mice without and with CIA were pooled, compared to CO2, administration of isofluorane was associated with lower production of the pro‐inflammatory cytokines TNF‐alpha (pg/ml, mean ± SD) (26.1 ± 17.6 versus 48.1 ± 55) and IL‐6 (26.8 ± 19 versus 48.1 ± 47) (ANOVA, p <0.05). In contrast to TNF‐alpha and IL‐6, administration of isofluorane induced the secretion of sICAM1 (1177 ± 347 versus 758 ± 177 for CO2) (p <0.00001). When data were analyzed as a function of CIA, the differences between CO2 and isofluorane persisted. Reduced secretion of sICAM by CO2 may be due to hypoxia induced decreased oxygen tension in the vascular endothelium. Since mice are the most common animal model for studying inflammation, researchers should be aware of these iatrogenic experimental variables before interpreting their data. Supp.: Grant #580790706 (Mushroom Council/USDA/NutriCore) and funds from Oklahoma State University.
Cytotoxic Effects of Silver Nanoparticles (Ag‐45) in Coronary Endothelia Cells
Authors: Rosas‐Hernandez Hector , Jimenez‐Badillo S , Gracia‐Espino E , Terrones H , Terrones M , Sadovova N , Zou X , Cheng W , Schlager J J , Hussain S M , Ali S F , Gonzalez C
Published: June 2021

This investigation was designed to determine whether silver nanoparticles (Ag‐45 nm) induce any selective and specific biological effects upon coronary endothelial cells (CEC), such as proliferation, cytotoxicity, and nitric oxide production (NO). CEC were cultured and exposed to increasing concentrations (0.1‐100μg/ml) of Ag‐45 nm for 24 hrs. Ag‐45 induced dual effect upon cellular proliferation, an inhibition at low concentrations and stimulation at high concentrations, and these actions were directly associated with NO production and inversely related with cytotoxicity. Confocal microscopy studies showed that Ag‐45 nm exposure, did not change CEC morphology, and were distributed widely in cellular surface, even with high concentrations of this agglomerate nanomaterial (NM). In addition, physical characterization of these Ag‐45 nm, using transmission electron microscopy, demonstrated that the real range of this NM, compromise a heterogeneous group of silver sizes, from 10‐90 nm. These results clearly demonstrate that Ag‐45 nm show selective and specific effects on this vascular bed, depending on the concentration, and that opposite effect could be result of the heterogeneity of sizes. However, the protective effects we found could be mediated by the production of NO, a free radical produced mostly by endothelium from blood vessels and, mediated angiogenesis and vascular tone.
REDUCED MICROSCOPIC GABAR SENSITIVITY IN RAT HIPPOCAMPAL CA1 NEURONS DURING BENZODIAZEPINE WITHDRAWAL IS REVERSED BY CAMKII INHIBITION
Authors: Das Paromita , Greenfield L. John , Tietz Elizabeth I.
Published: June 2021

Two days after 1‐week oral treatment with the benzodiazepine (BZ), flurazepam (FZP), L‐type voltage‐gated channel (L‐VGCC) whole‐cell Ca2+ current density is doubled and γ‐amino butyric acid type A receptor (GABAR) single‐channel open probability (Po) is reduced in cell‐attached patches from dissociated hippocampal CA1 neurons. Blockade of Ca2+ influx by systemic injection of the L‐VGCC antagonist, nimodipine 1 day after FZP treatment, restored sensitivity of FZP neurons to 5 µM GABA, increasing Po from 0.04 to 0.54 after 2 days to near control levels (0.68). We hypothesized that BZ‐induced elevation of [Ca2+]i may suppress GABAR function by activating Ca2+/calmodulin‐dependent protein kinase II (CaMKII). Single‐channel recordings were obtained after slice pre‐incubation with membrane‐permeant CaMKII inhibitors. At 1 µM GABA, Po in control patches was 0.19, while no openings were observed in FZP patches. After pre‐incubation with KN93, Po in FZP patches was 0.73. No openings were seen with the inactive analog KN92. Pre‐incubation with mAIP, activated 'silent' single channels in FZP patches (Po=0.69) and increased the basal response of control patches (Po=0.57). Membrane‐impermeant AIP and the PKC inhibitor, chelerythrine had no effect. Persistent activation of CaMKII is required to maintain basal GABAR function as well as for BZ‐induced, negative modulation of GABAR function associated with BZ tolerance.
Induction of the antitumorigenic NSAID‐activated gene (NAG‐1) in synthetic hexahydrocannabinol‐induced apoptosis of human colorectal cancer cells
Authors: Cho Mi‐Yeon , Park Min‐A , Babu Dinesh , Thapa Dinesh , Xia Likai , Lee Yong Rok , Kim Jung‐Ae
Published: June 2021

Non‐steroidal anti‐inflammatory drug (NSAID)‐activated gene (NAG‐1) is a transforming growth factor (TGF)‐β superfamily member that has pro‐apoptotic and anti‐tumorigenic activities. Cannabinoids have recently been extensively studied for their anti‐tumoral activity, and NAG‐1 has been shown to be up‐regulated by delta 9‐tetrahydrocannabinol. The present study investigated whether a synthetic hexahydrocannabinol, Yong‐8, influences NAG‐1 expression and apoptosis in human colorectal cancer cells, HCT‐116 (wild‐type p53) and HT‐29 (p53 mutant). Yong‐8 induced NAG‐1 protein and mRNA expressions in both cell lines. Concomitantly, Yong‐8 induced apoptosis of both HT‐29 and HCT‐116 in a concentration‐dependent manner. Interestingly, Yong‐8 increased p53 protein expression in both cell lines irrespective of their p53 status. Moreover, inhibition of p53 using antisense oligonucleotide caused a significant decrease in NAG‐1 protein expression and apoptosis in both cell lines under the same conditions. The present study demonstrates for the first time that Yong‐8, a synthetic hexahydrocannabinol, induces apoptosis in colon cancer cells. Taken together, our results suggest that hexahydrocannabinol induces colon cancer apoptosis via up‐regulation of NAG‐1 expression which is regulated by p53.
An endothelin converting enzyme inhibitor (SM‐19712) attenuates dextran sodium sulfate (DSS)‐induced colitis in mice
Authors: Lee Seungjun , Carter Patsy R. , Watts Megan N. , Bao Jianxiong R. , Harris Norman R.
Published: June 2021

Ingestion by mice of dextran sodium sulfate (DSS) induces colonic vasoconstriction and inflammation, with some of the effects potentially mediated by the vasoconstrictor endothelin‐1 (ET‐1). In this study, mice given 5% 40 kD DSS for 5‐6 days had elevated colonic immunostaining for ET‐1 and platelet endothelial cell adhesion molecule‐1 (PECAM‐1). Increased ET‐1 can induce microvascular constriction that could limit blood flow; however, the increase in PECAM‐1 is consistent with previous reports of DSS‐induced angiogenesis that could decrease flow resistance. Intestinal blood flow measured with microspheres revealed a tendency for a DSS‐induced decrease, indicating that the vasoconstriction may be a more dominant factor than angiogenesis in the overall microvascular resistance. DSS mice given daily 15 mg/kg doses of the endothelin converting enzyme inhibitor SM‐19712 had decreased tissue ET‐1, and a tendency for decreased PECAM‐1. SM‐19712 attenuated histologic signs of tissue injury and inflammation, and decreased the extent of loose stools and fecal blood. However, the inhibitor did not significantly decrease DSS‐induced weight loss, colon shortening, or tissue levels of myeloperoxidase (an indicator of neutrophil infiltration). Supported by Project 2 of NIH P01DK043785.
Antecedent Ethanol Attenuates Cerebral Ischemia/Reperfusion‐induced Leukocyte‐Endothelial Adhesive Interactions: Role of Large Conductance, Ca2+‐activated K+ Channels (BKCa)
Authors: Wang Qun , Kalogeris Theodore J , Wang Meifang , Jones Allan W , Korthuis Ronald J
Published: June 2021

Our recent work indicates that ethanol ingestion (EtOH‐PC) 24 hrs prior to induction of cerebral ischemia/reperfusion (I/R) reduces postischemic neuronal injury. In this study, we investigated whether EtOH‐PC would reduce I/R‐induced leukocyte‐endothelial adhesive interactions in pial postcapillary venules, and if so, whether this protective effect was triggered by BKCa channel activation. To address this aim, C57BL6 mice were administered ethanol by gavage [(body weight (in g) ×0.6) + 0.3] or treated with the BKCa channel opener, NS‐1619 (0.1mg/kg, ip) 24 hrs prior to I/R in the absence or presence of the BKCa channel blocker, paxilline (PX; 2.5 mg/kg body wt, i.p., administered 10 min before ethanol or NS1619 administration). 24 hour later, both common carotid arteries (CCA) were occluded for 20 min followed by 2 and 3 hours of reperfusion, then the numbers of rolling (LR) and adherent (LA) leukocytes were quantified in pial venules using intravital microscopy. I/R induced large increases in LR and LA in pial venules compared with sham. The increased LR and LA were prevented by EtOH‐PC or antecedent NS1619 administration. These protective effects were attenuated by PX treatment coincident with ethanol or NS1619 administration. Our results indicate that EtOH‐PC may activate BKCa to prevent cerebral ischemia/reperfusion‐induced leukocyte‐endothelial adhesive interactions. Supported by R01AA014945 from NIH
Low density lipoprotein modulates rat mesenteric lymphatic pumping
Authors: Wang Wei , Muthuchamy Mariappan , Zawieja David C
Published: June 2021

The unique nature & location of mesenteric lymphatics provides a special site for modulating lipoprotein transport, including low density lipoprotein (LDL). Intestinal lymph contains both enterocyte‐synthesized & plasma‐derived lipoproteins. Lipid absorption is associated with alterations in mesenteric lymph pressure & flow, implying that they coordinate lymph transport with intestinal lipid absorption by adjusting lymph pump flow. However, detailed studies of the mechanisms involved haven't been done, so we studied this process in isolated rat mesenteric lymphatics. We found: 1. Prominent expression of LDL receptors in lymphatic cells, endocytosis of LDL into lymphatic endothelium, and endothelial NO synthase (eNOS) expression in lymphatic endothelium; 2. LDL up‐regulates phasic contraction frequency and alters tone, resulting in a significant increase in the lymph pump output. Therefore we hypothesize that: LDL exposure, acting through its receptors and/or modulations of eNOS activity, provides positive feedback to the lymphatic pump via increases in the phasic contraction frequency with accompanying alterations of the tone. These studies may help correlate lymphatic function with lipoprotein metabolism and provide the basis for the prevention and treatment of the disorders related to lipoprotein metabolism. NIH HL70308 & HL80526
Molecular Mechanism of Substance P Mediated Signaling in Lymphatic System
Authors: Chakraborty Sanjukta , Zawieja David C , Muthuchamy Mariappan
Published: June 2021

The lymphatic system plays a key role in body fluid homeostasis, lipid absorption and immunity, and these vital functions rely on the contractile activity of the lymphatic muscle cells. However the molecular mechanisms governing lymphatic contractility is not completely understood. We have previously shown that substance P (SP), a neuropeptide associated with the sensory innervation of lymphoid tissue, modulates lymphatic vessel function and lymph flow. In this study we tested the hypothesis that SP increases the myosin light chain 20 (MLC20) phosphorylation and modulates lymphatic contractile dynamics. Lymphatic muscle cells (LMCs) were subjected to various agonists of which SP had the most potent effect on MLC20 phosphorylation. SP mediates its various biological responses through the G protein coupled neurokinin (NK) receptors, NK1R, NK2R and NK3R. However, it is unknown which specific receptors or pathways are activated by SP action on the lymphatics. Molecular analyses demonstrate the expression of both NK1R and NK3R in LMCs and lymphatic tissues. Immunofluorescence data show the presence of NK1R in the cytoplasm, whereas, NK3R is predominantly present in the nuclei. Furthermore SP treatment is found to activate the p38 MAPK pathway. We propose that SP modulates lymphatic contractility by regulating MLC20 phosphorylation via p38 MAPK pathway. NIH Support RO1 HL080526, KO2 HL086650 to MM; HL75199 to DCZ.
Plasminogen activator inhibitor‐1 can regulate myoendothelial junction formation
Authors: Heberlein Katherine , Straub Adam C , Best Angela K , Looft‐Wilson Robin Christine , Isakson Brant E
Published: June 2021

Myoendothelial junctions are cellular projections that form between the vascular smooth muscle cell and endothelial cell layers of the resistance vasculature by disruption of the internal elastic lamina. How and why these structures form is unknown, but given the location of the MEJ, it is likely that degradation of the internal elastic lamina is required. We have recently identified plasminogen activator inhibitor ‐1 (PAI‐1), a mediator of the fibrinolytic system, as being preferentially localized at the MEJ both in vivo and in vitro (see abstract by Heberlein, et al.). Using Metamorph software analysis on a vascular cell co‐culture model (Isakson et al., 2005), we show that inhibition of PAI‐1 activity using a monoclonal antibody (1000 ng/mL) has an approximate 20% increase in the number of MEJs formed, as identified by phalloidin labeling of actin. Conversely the addition of PAI‐1 (100 ng/mL) results in an approximate 20% decrease in number of MEJs formed. The formation of MEJs in response to PAI‐1 and PAI‐1 monoclonal antibodies appears to be dose‐dependent. These results were compared to MEJ formation in PAI‐1‐/‐ mice. Based on these data we conclude that PAI‐1 may regulate the formation of MEJs in the vasculature. Supported by NIH RO1 088554 and an AHA SDG.
4‐Phenyl butyrate (4‐PBA) alters ERp29 expression, connexin localization and enhances mesenteric artery agonist‐induced constriction
Authors: Looft‐Wilson Robin Christine , Smith Tekla , Koval Michael , Isakson Brant E
Published: June 2021

4‐PBA, an HDAC inhibitor and chemical chaperone, has therapeutic potential for many diseases. Based upon its known effect on protein processing and transport, we hypothesized that treatment of mesenteric arteries with 4‐PBA would alter the expression/distribution of gap junction proteins (connexins) and vasomotor function. Extra‐luminal 4‐PBA (2‐4 mM) treatment for 2 hr increased expression of ERp29, an endoplasmic reticulum chaperone that facilitates Cx43 transport, by ~50% (p<0.05). This coincided with a trend for increased total Cx40 and Cx43 protein, and increased localization of Cx37 to the endothelial cell membrane. Artery constriction (absolute and % diameter change; pressure myography) to phenylephrine (10−5 M) was significantly greater (p<0.05) after 2 hr extra‐luminal 4‐PBA (2 mM) incubation, and compared to vehicle‐treated arteries, which constricted similarly before and after treatment. There were no significant differences in basal tone or dilation to acetylcholine (10−5 M) before or after 4‐PBA or vehicle treatment, or between groups, indicating that increased vasoconstriction was not due to endothelial impairment. In conclusion, 4‐PBA alters connexin expression and localization. Whether these molecular changes contribute to the enhanced vasoconstriction remains to be determined. Support: NIH‐R15HL082647, R01HL083120, R01HL088554, AHA‐SDG.
Aquaporin5 (AQP5) modulation as a novel mechanism to regulate airway barrier function
Authors: Sidhaye Venkataramana , Shimoda Larissa , King Landon
Published: June 2021

Airflow creates shear forces on the apical surface of airway epithelial cells, the effects of which are largely unknown. In isolated mouse trachea, airflow induced shear stress enhanced epithelial barrier function (permeability of evans blue dye‐4% albumin). Primary human airway epithelial cells differentiated in an air‐interface were exposed to shear stress (3 dynes/cm2) using Krebs solution, leading to increased barrier function (FITC‐dextran permeability) and reduced the abundance of AQP5, an apical water channel, by sequential activation of TRPV4 and VGCC. 16HBE cells, without endogenous AQP5, were infected with either an adeno‐AQP5 expressing or control GFP virus. The presence of AQP5 reduced barrier function and allowed further modulation of barrier function in response to shear stress. Altering barrier function by expressing AQP5 altered receptor‐ligand interactions. Apical ligands (EGF,IFNƒ×) interact with basolateral receptors only when paracellular permeability is increased (induced by AQP5 overexpression). Similarly, Substance P (basolateral ligand) only interacts with apical receptors when AQP5 was overexpressed. We hypothesize that regulation of AQP5 modulates the airway epithelia barrier, which is critical in regulating signaling. This important homeostatic mechanism to regulate epithelial responses may be disrupted in pathologic conditions. Supported by HL085763 and FAMRI.
Spatial Patterns of Transforming Growth Factor Beta Signaling in Mgr‐/‐ Mouse Model of Marfan Syndrome
Authors: Popovic Natasa , Humphrey Jay D. , Wilson Emily
Published: June 2021

The balance between wall stress and structure of a blood vessel is crucial for its optimal function and is maintained by remodeling. Marfan patients, harboring mutations of fibrillin1 (FBN1) gene, exhibit aberrant vascular remodeling, formation of aortic aneurysms and dissections. This is associated with altered signaling through transforming growth factor beta (TGFβ) pathway. Non‐invasive imaging and computational modeling of blood flow predict complex pattern of wall stress in the ascending aorta and arch. We hypothesized that these regions of aorta exhibit altered morphology and signaling patterns. Aortas from 15 wild type and 15 FBN1 deficient (FBN1‐/‐) mice were isolated and divided into 4 segments: ascending aorta, arch, proximal and distal thoracic aorta. VVG staining of aortic cross‐sections showed that aortas of FBN1‐/‐ mice have diffuse fragmentation of elastic lamella and decreased elastic lamellar thickness (p=0.005). Increased wall thickness (p=0.002), inner radius (p=0.000), and wall cross‐sectional area (p=0.000) was present in the ascending aorta and arch of FBN1‐/‐ mice. The same area was associated with increased levels of pSMAD3 protein and decreased relative expression of SMAD3 mRNA. More research is needed to determine relationships between TGFβ signaling, distributions of wall stresses, and increased wall stiffness due to elastin fragmentation in aortas of Marfan patients.
The effect of HIV‐1 Tat on the aortic response to vasoconstrictors
Authors: McDonough Kathleen H. , Giaimo Mary , Prakash Om
Published: June 2021

The human immunodeficiency virus (HIV‐1) has long term effects on the immune and cardiovascular systems. The HIV‐1 protein tat alters synthesis of proteins such as nitric oxide synthase and cytokines. The combination of alcohol (Alc) and Tat may exacerbate the effects of either Alc or Tat alone. We tested the hypothesis that Tat, expressed in transgenic mice, would depress aortic responses to constrictor agents and Alc would exacerbate this effect. To test this hypothesis, transgenic and control mice were given water or water with Alc (20% v/v) for 6‐8 weeks and then aortic ring contraction to depolarization with KCl and alpha mediated contraction with phenylephrine (PE) was determined. Some mice were given endotoxin (Endo) to simulate inflammation. Isolated rings from control mice increased tension in response to KCl and PE by approximately 75% and 92%, respectively. Similar changes were noted in the Tat control group. Alc increased the response of rings from control mice to KCl but not to PE. Alc increased the response to constrictors in the Tat mice in the opposite way ‐ increased response to PE but no change in response to KCl. 24 hr after Endo, the per cent change in tension was increased by PE but not by KCl. Results suggest that the major influence on constrictor tone was recovery from Endo rather than Tat or Alc provision. The Endo effect was primarily demonstrated by the increased response to PE, a receptor mediated process.

(Supported by NIH AA013555)
Alpha1‐Adrenoceptor mediated aortic contraction involves transactivation of EGF receptor
Authors: Ulu Nadir , Landheer Sjoerd W , Roggeveld Jan , Duin Marry , Henning Robert H
Published: June 2021

Prolonged elevation of plasma catecholamines is a risk factor for vascular diseases such as hypertension and atherosclerosis. Catecholamines induce hypertrophy of vascular smooth muscle through alpha1‐adrenoceptors (α1‐ARs). In cell culture this involves the transactivation of epidermal growth factor receptor (EGFR). We hypothesized EGFR transactivation to be involved also in the α1‐AR mediated aortic contraction. Thoracic aorta was isolated from 12‐14 weeks old male Wistar rats. In vitro aortic contractile responses to cumulative doses of phenylephrine (PE) were investigated in the absence and presence of EGFR antagonists AG1478 and DAPH in intact and endothelium‐denuded rings. Involvement of signal transduction pathways was investigated by using heparin and inhibitors of Src, MMP, ERK1/2 and IP3K. AG1478 concentration dependently decreased PE‐induced contractile responses in both intact (pA2=5,52) and denuded (pA2=5,47) rings, as did DAPH. In contrast, AG1478 did not affect contraction response to the thromboxane analog U46619. Blockade of IP3K by wortmannin abrogated the effect of AG1478 on the PE response, whereas heparin and inhibitors of Src, MMP and ERK1/2 did not affect the AG1478 action. α1‐AR mediated contraction of aortic ring involves transactivation of EGFR, most likely via activation of the IP3K pathway and not by shedding of heparin‐binding EGF. This study was supported by GUIDE.
The CaV1.2 α1 subunit N‐terminus regulates cerebral artery contractility
Authors: Bannister John Phillip , Adebiyi Adebowale , Cheng Xiaoyang , Zhao Guiling , Jaggar Jonathan H
Published: June 2021

Voltage‐dependent Ca2+ (Cav1.2) channels are the major Ca2+ entry pathway in arterial myocytes and are essential for myogenic constriction. Although Cav1.2 α1 subunits undergo splicing, physiological functions of vascular splice variants are unclear. We have shown that arterial myocytes express two different Cav1.2 α1 subunit N‐terminal variants (J Biol Chem, 282: 29211‐21, 2007); a novel isoform from exon 1c (e1c) and the previously described exon 1b (e1b). Here, we examined protein expression and physiological functions of Cav1.2 α1 subunit N‐termini in resistance‐size rat cerebral arteries. Custom antibodies recognizing peptide sequences encoded by e1b and e1c detected Cav1.2 α1 subunits in cerebral arteries. ShRNA targeting e1b and e1c both reduced Cav1.2 α1 subunit expression, with e1c shRNA causing a larger reduction than did e1b shRNA. E1b or e1c overexpression reduced pressure (60 mmHg)‐induced myogenic tone, with e1c causing a greater reduction than did e1b. Acute application of a membrane‐permeant peptide corresponding to the sequence encoded by e1c inhibited myocyte voltage‐dependent Ba2+ currents and reversibly dilated pressurized arteries. In contrast, a control peptide had no effect. In summary, data suggest that Cav1.2 α1 subunits containing e1b and e1c are expressed in cerebral artery myocytes and that e1c regulates voltage‐dependent Ca2+ currents and myogenic tone.
Dihydrotestosterone (DHT) increases cyclooxygenase‐2 (COX‐2) in human coronary artery smooth muscle cells following IL‐1β‐induced inflammation
Authors: Osterlund Kristen , Gutierrez Anthony , Gonzales Rayna J
Published: June 2021

Effects of androgens on normal physiological and pathophysiological vascular function remain unclear. Both protective and non‐protective effects of androgens on the cardiovascular system have been documented. Our previous studies show that chronic androgen treatment augments vascular tone under normal physiological conditions and exacerbates endotoxin‐induced inflammation in cerebral arteries isolated from rodents. Here we investigated whether dihydrotestosterone (DHT; 100nM), the more potent androgen receptor agonist, modulates COX‐2 in primary human coronary vascular smooth muscle cells following stimulation with either IL‐1β (5ng/ml) or lipopolysaccharide (LPS 100 ug/ml). A time course for cytokine‐ or endotoxin‐induced COX‐2 levels assessed via Western blot showed that COX‐2 levels were greatest at 6 and 9 hours post cytokine or endotoxin exposure. Similar to IL‐1β or LPS, DHT treatment increased COX‐2 protein compared to vehicle. Interestingly, LPS induced increases in COX‐2 levels were blunted when administered concomitantly with DHT. In conclusion, androgens may have a differential impact on inflammation during pathophysiological compared to normal physiological conditions. Support: AHA SDG RG
Phosphodiesterase type 5 inhibitors relax female rat internal pudendal arteries: potential treatment for female sexual dysfunction.
Authors: Allahdadi Kyan J , Tostes Rita C , Webb R.Clinton
Published: June 2021

The beneficial effects of phosphodiesterase 5 inhibitors (PDE5I) are well documented in erectile dysfunction treatment. The PDE5I use in the treatment of female sexual dysfunction has not been fully explored. Here we compared the effects of PDE5I: sildenafil®, vardenafil® and tadalafil® between male and female rat internal pudendal arteries. We hypothesize that PDE5I present similar affects between male and female rat pudendal arteries. Male and female pudendal arteries (Sprague Dawley; 11 weeks) were mounted in microvessel chambers. Segments were contracted with phenylephrine (50%) then submitted to increasing concentrations of one of the PDE5 inhibitors (10‐10 to 10‐5 mol/L). Values are expressed as % vasodilation from constricted levels and pD2. All PDE5I relaxed the female and male pudendal arteries concentration‐dependently [(sildenafil®: female 105.00±7.73, 6.60±1.7; male: 103±0.9, 5.67±0.9) (vardenafil®: female, 115.6±11.63, 6.67±0.24; male: 105.1±89.88, 6.4±0.23) (tadalafil®: female: 80.13±5.39, 7.98±0.24; male: 102.3±23.65, 5.4±0.23)]. These results indicate that both male and female pudendal artery are sensitive to PDE5 inhibition. Female pudendal arteries were more selective to sildenafil® compared to male arteries, suggesting that this PDE5I indeed may have a different mechanism of action. PDE5I may be useful in female sexual dysfunction treatment.
Peripheral vasodilation and aerobic exercise equally affect skeletal muscle substrate utilization in older and younger adults
Authors: Casperson Shanon , Dillon Edgar L , Paddon‐Jones Douglas , Durham William , Sheffield‐Moore Melinda
Published: June 2021

We examined leg glucose uptake (GU), net phenylalanine uptake (PheU) and Phe net balance (PheNB) in sixteen older (O) and seventeen younger (Y) adults who received either a vasodilator (sodium nitroprusside, SNP: N=8 O and N=8 Y) or performed aerobic exercise (EX: N=8 O and N=9 Y) to induce similar increases in microvascular blood flow and capillary recruitment. We hypothesized that muscle contraction and exercise per se, in the presence of amino acids, would result in greater increases in skeletal muscle substrate utilization than peripheral vasodilation (SNP) without contraction. Indocyanine green (ICG) was used to measure limb blood flow (LBF) and contrast‐enhanced ultrasound to measure microvascular blood volume (MBV, an indicator of capillary recruitment), microvascular flow velocity (MFV) and microvascular blood flow (MBV x MFV). GU, PheU, PheNB and all blood flow measures were obtained 60 min after SNP or EX. LBF, MBV, MFV, microvascular blood flow, GU, PheU, and PheNB were all similarly increased in response to SNP and EX in both age groups. Thus, vasodilation in the absence of muscle contraction is capable of increasing skeletal muscle substrate utilization to a similar extent as aerobic exercise; likely by increasing microvascular blood flow and capillary recruitment. These results indicate that common vasodilators, not unlike exercise, may promote favorable metabolic responses in skeletal muscle.
Hypoxic regulation of early vascular and hematopoietic development in the chick embryo
Authors: Sheng Guojun
Published: June 2021

During embryogenesis, blood and vascular systems provide essential supply of oxygen to developing organs with increasing complexity. It is not well‐understood, however, how changes in oxygen supply affect normal differentiation of hematopoietic and vascular tissues. In chickens, oxygen supply to developing organs is largely achieved via extra‐embryonically generated blood and vascular cells. The effect of hypoxia on extra‐embryonic hematopoietic and vascular differentiation is investigated with a focus on two critical periods (during primitive red cell generation and during early phase of definitive red cell generation). The roles of HIFs, aquaporins, heme and iron supply in this context will also be discussed.