bluuebunny/crossref_metadata_2025_split huggingface dataset:
Induction of Xenobiotic Metabolizing Enzymes and Transporters in Human Intestinal Cells by Pregnancy‐Related Hormones
Authors: Moscovitz Jamie E , Herrera‐Garcia Guadalupe , Guo Grace L , Aleksunes Lauren M 
Published: November 2023

The intestine expresses many critical xenobiotic metabolizing enzymes and transporters that regulate oral absorption and fecal excretion. Important nutrients and vitamins are absorbed at this interface, and as a result expression of these enzymes and transporters are tightly regulated. While it is known that gastrointestinal motility decreases during pregnancy, additional molecular changes in intestinal cells have yet to be investigated. To determine whether pregnancy alters intestinal disposition pathways, RNA and protein were collected from human colon adenocarcinoma LS174T cells cultured in media supplemented with 20% serum from either non‐pregnant or pregnant women (weeks 31 to 39 of gestation). In intestinal cells treated with pregnant serum, the mRNA expression of the CYP3A4 enzyme, basolateral efflux transporters OSTα and MRP3, as well as the luminal efflux transporters MDR1, MRP2 and BCRP were induced 2‐ to 4‐fold as compared to cells treated with non‐pregnant serum. MRP3 protein exhibited a 2‐fold up‐regulation when treated with pregnant serum. Profiling of differentially expressed genes (induction of CYP3A4 and MDR1 along with down‐regulation of the SHP transcription factor) suggested that serum from pregnant women is activating the Pregnane X Receptor (PXR). Further mechanistic studies revealed that progesterone and its sulfated metabolite PM5S may be responsible, in part, for expression changes observed in cells treated with pregnant serum. Because MDR1, MRP2 and BCRP are important to the luminal efflux of endocrine disrupting chemicals, such as mycotoxins, perfluorinated chemicals, and pesticides, activation of intestinal PXR by hormones in pregnant serum may be an adaptive response to decrease systemic absorption and in turn, fetal exposure.Support or Funding InformationF31HD082965, R01ES020522, T32ES007148, P30ES005022
Sex Specific Differences In Key Signaling Pathways Involved In Sympathetic Nervous System Control Within The Dorsal Medulla Of Adult Sheep With Fetal Betamethasone Exposure
Authors: Hendricks Alexa S , Diz Debra I , Shaltout Hossam A , Chappell Mark C 
Published: November 2023

Glucocorticoids including betamethasone (BM) are a widely accepted and routine therapy administered to women at risk of early preterm labor to facilitate fetal lung development and reduce infant mortality rates. However, fetal steroid exposure may lead to negative long term consequences for autonomic regulation. In a sheep model of fetal programming, BM‐exposed (BMX) adult offspring exhibit elevated mean arterial pressure (MAP), decreased baroreflex sensitivity (BRS) for control of heart rate and insulin resistance accompanied by dysregulation of the brain, renal and circulating renin‐angiotensin system (RAS). In the brain solitary tract nucleus of the dorsal medulla, BMX dysregulation resulting in a shift towards the sympathetic activator angiotensin II (Ang II) actions through the AT1 receptor that oppose the beneficial actions of angiotensin‐(1–7) [Ang‐(1–7)] at the Mas receptor for BRS regulation. Therefore, we examined RAS and insulin/leptin signaling a pathways in the dorsal medulla to understand further the molecular disturbances that lead to the observed alterations in sympathetic outflow in adult male and female adult sheep (n = 16 sheep; 4 each control or BMX, male or female).A protein array for kinases (PathScan® Intracellular Signaling Array Kit, Cell Signaling) and subsequent verification by Western blot hybridization revealed that female BMX sheep exhibit lower expression of proteins in the PI3 kinase pathway (pGSK3β: 33±0.10%, pAkt: 22±0.33%; p = 0.03) which are known to be related to regulation of metabolic function and blood pressure. In contrast, in male BMX sheep the PI3 kinase pathway components are not different from control, but there is a trend (p = 0.11) for higher expression of phosphorylated MAP kinase pathway protein ERK (37%) and p38 (68%), known to increase sympathetic activation and facilitate insulin actions. Future studies will explorethe potential mechanism for the lower PI3 Kinase related pathway in female sheep. We conclude that in utero BMX promotes an imbalance in adult brain medullary RAS and insulin signaling pathways known to influence sympathetic regulation with different patterns observed in male and female sheep.Support or Funding InformationSupport: HD47584
Modulation by NADPH Oxidase/Rho‐Kinase Oxidative Pathway of Chronic Cyclosporine‐ Diclofenac Hemodynamic Interaction in Female Rats
Authors: El‐Mas Mahmoud M , Ibrahim Karim S , El‐Yazbi Ahmed F , El‐Deeb Nevine M , El‐Gowelli Hanan M 
Published: November 2023

We recently reported contrasting effects for nonsteroidal antiinflammatory drugs (NSAIDs) on the hypertensive effect of acutely administered cyclosporine (CSA) in female rats. CSA hypertension was ameliorated or exaggerated after selective cyclooxygenase‐2 (COX2) and nonselective COX1/COX2 inhibition, respectively. Here we investigated (i) whether a similar interaction could be seen when drugs were administered chronically, (ii) effect of interacting drugs on left ventricular (LV) and cardiac autonomic and histologic profiles, and (iii) role of the NADPH oxidase/Rho‐kinase pathway in the interaction. Rats were pre‐instrumented with femoral indwelling catheters and treated for 10 days with CSA (25 mg/kg/day), diclofenac (nonselective NSAIDs, 1 mg/kg/day), celecoxib (COX2 inhibitor, 10 mg/kg/day), or their combinations. CSA hypertension was maintained upon concurrent administration of either NSAID whereas the concomitant reductions in the time‐ and frequency‐domain indices of heart rate variability were accentuated in presence of diclofenac but not celecoxib. The isovolumic relaxation constant (Tau, τ), a measure of cardiac diastolic function, was similarly reduced by single or combined drug regimens whereas LV contractility (dP/dtmax) remained unaffected. Histopathologically, compared with CSA or CSA/celecoxib treatment, the CSA/diclofenac regimen caused more extensive cardiac structural damage (ischemic necrosis and edema). The co‐administration of diphenyleneiodonium (NADPH‐ox inhibitor), but not fasudil (Rho‐kinase inhibitor), reversed the CSA/diclofenac‐evoked (i) increases in MAP, (ii) decreases in LF/HF and Tau, and (iii) cardiac structural damage. Overall, female rats treated chronically with CSA plus diclofenac exhibit exacerbated hemodynamic, autonomic, histopathologic, and LV functions via Rho‐kinase‐independent upregulation of the NADPH oxidase pathway.Support or Funding InformationSupported by the ALEX‐REP Grant Fund, Alexandria University, Egypt (Grant HLTH‐13‐01).
MSC Exosomes Deliver Cardioprotective miR‐21
Authors: Luther Kristin , McGuinness Michael , Haar Lauren , Xu Huan , Chen Jenny , Medvedovic Mario , Jones W. Keith 
Published: November 2023

IntroductionMesenchymal stem cell (MSC) transplantation has been shown to improve outcomes including infarct size in animal models of myocardial infarction (MI), but the mechanism of the beneficial effects is not well understood. Much data supports that it involves direct protection of the ischemic myocardium via paracrine transfer of factors from the MSC to cardiac myocytes. If these factors can be identified, they can provide new opportunities for therapeutic development. Exosomes are small vesicles that contain miRNA, which can be exchanged between numerous different types of cells. The objective of this study was to examine the role of miRNA in paracrine effects mediated by MSC exosomes.HypothesisOur study addresses the hypothesis that exosomal miR‐21 is a mediator of protection in MSC therapy.Methods and ResultsIn vitro, exosomes dose dependently decreased cell death (39% reduction at maximum) in an oxygen‐glucose deprivation model of ischemia‐reperfusion injury. To determine whether cardioprotective miRNAs are found within MSC exosomes, their RNA was isolated, and miRNA‐Seq was performed. The most highly expressed exosomal miRNA was found to be miR‐21, which is known to be upregulated by numerous cardioprotective stimuli. To determine the contribution of miR‐21 in exosome‐mediated protection, we cultured MSCs from miR‐21− mice, and compared their exosomes to those of wild‐type MSCs in vitro and in a mouse model of MI. We found that the miR‐21− exosomes were significantly less protective than wild‐type exosomes. To determine the gene regulatory mechanism behind the observed protection, we investigated the expression of miR‐21 target genes in H9c2 cells, and found significant downregulation of Programmed Cell Death 4 and Fas ligand.ConclusionsMSC exosomes are protective against ischemia‐reperfusion injury in vitro and in vivo, and they contain miRNAs that influence expression of proteins in cell death pathways. The data support a key role for miR‐21 in these protective effects and aid in the understanding of how this mechanism could be translated into cardioprotective therapies.Support or Funding InformationThis research was supported by the National Heart, Lung, and Blood Institute grant # 5R01HL091478‐07.
Human Perivascular Adipose Tissue Contains an Adrenergic System
Authors: Ayala‐Lopez Nadia , Pereira‐Hicks Cristiane , Burnett Robert , Fink Greg D. , Watts Stephanie W. 
Published: November 2023

Perivascular adipose tissue (PVAT), fat tissue that wraps most blood vessels of the body, can alter vascular tone through various mechanisms. Our team discovered a source of a releasable pool of catecholamines in PVAT from the rat but the relevance to human physiology is unknown. Increased adipose tissue mass is observed in obesity. Sympathetic activity is also increased in obesity‐associated hypertension. Thus, the link between obesity and hypertension could be the PVAT adrenergic system. PVAT may affect vascular tone negatively in disease leading to hypertensive states. We hypothesized that similar to the rat, human PVAT would contain an adrenergic system that could synthesize and store catecholamines. Tissue was collected from patients (body mass index= 43.58±3.23, age range 29–58; mean age= 41.56±3.62, 8 females and 1 male) during elective bariatric surgery and placed into physiological saline solution (PSS). Splanchnic blood vessels with associated PVAT were dissected from the tissue and snap frozen for HPLC analysis or saved for immununohistochemistry (IHC). Dopamine (DA) and norepinephrine (NE) were quantified by HPLC in human mesenteric artery PVAT. NE was the most abundant catecholamine with concentrations of 14.63±1.39 ng/g NE vs. 0.80±0.09 ng/g DA (N=9). Two key enzymes in catecholamine synthesis; tyrosine hydroxylase and dopamine β‐hydroxylase, were located to PVAT adipocytes by IHC (N=9). In order for catecholamines to be released, a storage reservoir that holds catecholamines may be present in PVAT. IHC for the vesicular monoamine transporter 2 (VMAT2) indicated the presence of the transporter in the adipocyte cytoplasm (N=9). Negative controls were performed without the inclusion of the primary antibody and these were not positive. These data indicate that human PVAT contains elements of an adrenergic system complete with capabilities of synthesis and storage of catecholamines. Sympathetic activity of an adrenergic system located within PVAT may effect vascular tone in a balance that could be altered in disease to promote contraction. Future studies aimed to understand the differences in the PVAT adrenergic system of obese hypertensives compared to obese non‐hypertensives are necessary to address the question as to why a subset and not all obese people develop hypertension.Support or Funding InformationNIH NHLBI P01HL70687IPSTP 5 T32 GM 92715‐4NIH NRSA F31 HL128035‐01
Effect of Ang III on JNK mitogen activated protein (MAP) kinase in Wistar rat VSMCs
Authors: Alanazi Ahmed Z. , Clark Michelle A. 
Published: November 2023

ObjectiveWe investigated whether angiotensin (Ang) III induces JNK mitogen activated protein (MAP) kinase protein phosphorylation in isolated rat vascular smooth muscle cells (VSMCs). Moreover, we compared its effect with Ang II.BackgroundThe molecular mechanisms by which Ang III induces various biological effects have not been fully investigated. Most studies have shown that MAP kinases mediate Ang II apoptosis and growth promoting effects in VSMCs. In addition, Ang II induces vascular remodeling in these cells leading to increases in blood pressure. MAP kinases regulate or induce two crucial actions in VSMCs, proliferation and migration, which are associated with atherosclerosis and restenosis.MethodsPrimary cultures of VSMCs were isolated from the thoracic aorta of adult Wistar rats by the explant technique. VSMCs were treated with Ang II and Ang III ranging in concentration from 0.1 nM to 1000 nM for 10 minutes or with 100 nM Ang II and Ang III for 1 minute to 60 minutes. Western blotting technique was used to determine the effects of both peptides on JNK MAP kinase protein phosphorylation.ResultsConcentration studies showed that Ang II and Ang III caused a dose‐dependent increase in JNK MAP kinase protein phosphorylation. The effects of the peptides on this MAP kinase phosphorylation were maximal between 10 nM and 100 nM. The peptides’ effects were rapid and significant, occurring within minutes of treatment and the maximal effects on MAP kinases phosphorylation was observed by 10 minutes.ConclusionThese findings provide insight into the molecular nature of the actions of Ang III and offer possible mechanism by which Ang III physiological actions occur in VSMCs.Support or Funding InformationFunding for this project was provided through: Nova Southeastern University Health Professions Division Grant & Saudi Arabian Cultural Mission.
TBX20 Regulates Endothelial Angiogenesis and Migration
Authors: Meng Shu , Wong Wing Tak , Cooke John P 
Published: November 2023

BackgroundT‐box 20 (TBX20) is a crucial transcription factor (TF) for heart development. It is expressed in myocardium and endocardium. In previous work, our lab has identified TBX20 as a key transcription factor for endothelial differentiation. In this study, we investigated the role of TBX20 in mature endothelial function.MethodsGain‐ and loss‐of‐function studies were performed in human umbilical vein endothelial cells (HUVEC) with overexpression (OE) and knock down (KD) of TBX20 respectively. Lentiviral shRNA targeting TBX20 gene was used to KD studies while lentiviral constructs encoding open reading frame of TBX20 were used for overexpression. Gene expression and protein level were examined by RT‐PCR and western blot. Endothelial angiogenesis was examined by tube formation assay. Angiogenesis PCR array of 84 genes was used to screen the potential angiogenesis related molecular targets. ELISA assay was used to measure secreted protein level. Migration assay and BrdU incorporation assay were used for migration and proliferation analysis.ResultsTBX20 KD in HUVECs resulted in 50% reduction in both TBX20 mRNA and protein level. KD of TBX20 significantly impaired tube formation compared with ECs transfected with control shRNA. After screening of 84 potential angiogenesis related molecular targets, hepatocyte growth factor (HGF) gene expression was found downregulated by TBX20 KD. The secreted HGF level of TBX20 KD cells was found only half of the level of CT KD cells detected by ELISA. Further, supplement of exogenous HGF reversed the impaired angiogenesis of TBX20 KD cells. TBX20 KD also impairs endothelial migration (mean migration distance 442 μm vs CT 569 μm) through downregulation of PROK2 gene. TBX20 OE upregulates PROK2 to 20 fold and significantly promote migration (mean migration distance 729.6 μm vs CT 441 μm).ConclusionTBX20 is a crucial transcription factor to maintain endothelial angiogenesis and migration. TBX20 regulates angiogenesis through HGF secretion. TBX20 promotes endothelial migration partially through PROK2.
Attenuation of the Deleterious Course and Gastric Lesions after Bilateral Nephrectomy in Rats, NO‐system relation. BPC 157, L‐arginine, L‐NAME
Authors: Drmic Domagoj , Sucic Mario , Sever Anita Zenko , Kolenc Danijela , Andrijasevic Vedran , Suran Jelena , Seiwerth Sven , Sikiric Predrag 
Published: November 2023

AimChronic renal insufficiency might be associated with increased peptic ulcer presentation. Recently, stable gastric pentadecapeptide BPC 157 (an antiulcer peptide tested in clinical trials, for inflammatory bowel disease and multiple sclerosis, effective against a variety of different ulcerogens), counteracted gastric lesions and other disturbances, after bilateral nephrectomy in rats. Now, we demonstrate that this effect might be NO‐system related.Materials and methodsRats were subjected to bilateral nephrectomy, sacrificed either at 24h or 48h. Medication (/kg i.p.) BPC 157 (10μg, 10ng), L‐arginine (100mg), L‐NAME (5mg) alone and/or combined, was given immediately after nephrectomy. Assessment included gastric lesions, gross clinical presentation, urea, creatinine, AST, ALT, bilirubin, K, Na, Mg, Ca, inorganic phosphate, values in serum as described (Sikiric et al, Curr Pharm Des. 2014;20(7):1126–35) as well as brain lesion presentation, as described (Sikiric et al., Curr Neuropharm. 2015, in press).ResultsComparison of gastric lesions (means±SD, sum of the longest lesions diameters, mm) in the nephrectomized rats (10.5±1.5 (24h), 15.0±3.0 (48h)), revealed that the gastric lesions were either consistently attenuated (vs. *P<0.05, at least) in BPC 157‐rats (1.8±0.5 (24h), 1.0±0.5 (48h)), L‐arginine‐rats (5.8±0.8 (24h), 5.0±0.7 (48h)), BPC 157+L‐arginine‐rats (0.8±0.5 (24h), 1.3±0.4 (48h)), L‐arginine+L‐NAME‐rats (4.9±0.4 (24h), 5.3±0.5 (48h)), BPC 157+L‐NAME‐rats (1.3±0.4 (24h), 1.2±0.5 (48h)), BPC 157+L‐arginine+L‐NAME‐rats (1.6±0.4 (24h), 1.4±0.3 (48h)) or aggravated (vs. *P<0.05, at least) in L‐NAME‐rats (20.5±1.7(24h), 25.0±2.0 (48h)). The influence (decreasing/increasing) on the otherwise increased values of the urea, creatinine, AST, inorganic phosphates as well as on rat edema formation (controls rats were edematous (scored 0–3) (Min/Med/Max, scored 3/3/3)) (attenuation/aggravation) was in accordance with the influence on gastric lesions(attenutation/aggravation) while K, Na, Mg, Ca values in serum were not affected.ConclusionsApplied after nephrectomy, the otherwise deleterious course was attenuated after BPC 157, and/or L‐arginine administration, while they were aggravated by L‐NAME admisitration, an effect also counteracted by BPC 157 and L‐arginine, portraying it as NO‐related. Gastric lesions seem to be a particular indicator of the severity of the course of events which ensue after nephrectomy.Support or Funding InformationThis research was supported by Grant 108‐1083570‐3635, Ministry of Science, Education and Sports, Republic of Croatiaand by Grant BM099, University of Zagreb, Republic of Croatia.
Amygdaloid Corticotropin Releasing Factor as the Key Mediator of Chronic Visceral Pain in Females with a History of Early Life Stress
Authors: Prusator Dawn K , Meerveld Beverley Greenwood‐Van 
Published: November 2023

In females. early life stress (ELS) has been identified as a risk factor for the development of functional pain disorders such as irritable bowel syndrome (IBS) in adults. Previously, our laboratory has shown that unpredictable ELS results in female specific heightened visceral pain behaviors in adulthood. The central amygdala (CeA) has been identified as a key brain nucleus in the processing of stress and pain, however, the role of the CeA in modulating pain following ELS has not been investigated. Therefore, we hypothesized that ELS increases visceral sensitivity in adulthood via altered CeA function, specifically through altered corticotropin releasing factor (CRF) expression.Neonatal rats underwent classical conditioning using unpredictable odor‐shock pairings or odor only as a control. In adulthood, CRF expression in the CeA was quantified via qRT‐PCR. CRF knockdown occurred via cannulae implanted into the CeA for infusion of antisense or random sense oligodeoxynucleotides (ASO/RSO n=6–7/group). Visceral sensitivity was assessed via visceromotor response (VMR) quantified as the number of abdominal contractions in response to graded pressures (0–60mmHg) of isobaric colorectal distension (CRD). The role of CRF receptor 1 was assessed via administration of a selective CRF receptor 1 antagonist CP 376395 5μg/0.5μl administered bilaterally (n=6/group).Expression of CRF was upregulated in the CeA following unpredictable ELS (p<0.01) compared to odor only controls. Administration of the RSO non‐targeting control sequence did not alter visceral hypersensitivity previously observed in the unpredictable group (60mmHg: 35.3±0.6) compared to odor only controls (26.2± 0.5). Knockdown of CRF in the CeA via ASO infusion significantly decreased the number of pain responses in unpredictable females as quantified by VMR to CRD (27.6±0.9, p< 0.01), while ASO treatment in the control group did not significantly alter pain responses (23.5±1.4). Administration of CRF1R antagonist mirrored the effects of CRF ASO infusion by significantly decreasing the VMR to CRD in adult females experiencing unpredictable ELS (26.8±1.1, p<0.01) to that of odor only control (27.0±1.1)In summary, this study suggests a novel consequence of ELS on visceral pain responses in adulthood that are centrally driven via CRF and directly related to the context, predictable or unpredictable, of the ELS experienced. Furthermore, this study identifies for the first time a mechanism by which females develop chronic visceral pain following ELS.Support or Funding InformationThis work was supported by a department of Veterans Affairs Merit Grant to BG‐VM #01BX001195
A new carbon monoxide (CO)‐releasing molecule (CORM401) suppresses PMN migratory potential through an F‐actin‐dependent mechanism
Authors: Inoue Ken , Patterson Eric , Chung Kyukwang , Potter Richard F , Capretta Alfredo , Fraser Douglas D , Cepinskas Gediminas 
Published: November 2023

BackgroundOur previous work suggested that a newly synthesized Mn2+‐based water‐soluble carbon monoxide (CO)‐releasing molecules (CORM401) interfered with PMN recruitment by inhibiting transendothelial migration across the endothelium. In this study, we further investigated the effects of CORM401 on PMN chemotaxis by an F‐actin‐dependent mechanism.MethodsFreshly isolated human PMN were placed into a μ‐Slide chemotaxis chamber and allowed to migrate in response to an fMLP (10μM) gradient for 50 min in the absence or presence of 100 μM CORM401 or inactive CORM401(iCORM401). PMN migration was recorded and analyzed for migration directionality and velocity. To assess the effect of CORM401 on fMLP‐stimulated PMN function, F‐actin polymerization was measured by flow cytometry. In addition, the phosphorylation of MAPK and p21‐activated kinases (PAK) in PMN were examined by immunoblotting.ResultsPre‐treating PMN with CORM401 significantly reduced their directionality and mean velocity (p<0.01) in response to fMLP compared to controls. CORM401 significantly reduced F‐actin polymerization in fMLP‐stimulated PMN (p<0.01). Pre‐treating PMN with CORM401 did not result in decreased MAPK phosphorylation, but resulted in decreased PAK phosphorylation (p<0.05).ConclusionCORM401 suppresses neutrophil migratory potential through an F‐actin‐dependent mechanisms. This study showed for the first time that the use of CORM401 has an effect on F‐actin‐dependent mechanisms of PMN.Support or Funding InformationHSFO 393, IRF‐25‐12