bluuebunny/crossref_metadata_2025_split huggingface dataset:
Endometriosis‐associated vaginal hyperalgesia is mediated by the balance of reactive aldehyde production and metabolism.
Authors: McAllister Stacy , Sinharoy Pritam , Gross Eric 
Published: June 2021

BackgroundEndometriosis affects ~176 million women worldwide and is characterized by the growth of endometrial tissue outside the uterus (known as endometrial ectopic cysts). How endometriosis becomes associated with painful symptoms is not clear. Here we hypothesize that endometriosis‐associated vaginal hyperalgesia is mediated by an imbalance of reactive aldehyde production and metabolism.MethodsTo test this hypothesis, we used female Sprague‐Dawley rats (n=5/group) and subjected them to a cutting‐edge model of behavioral psychophysics to assess for rat vaginal nociception. Prior to subjecting the rodents to this model, we treated rodents with the reactive aldehyde, acetaldehyde (9 mg/kg intraperitoneal) to mimic an increased production of reactive aldehydes in the intraperitoneal space. To modulate ALDH2 activity, we administered to rodents either the ALDH2 activator, Alda‐1 (1 mg/kg, diluted in DMSO, given intraperitoneal) or the ALDH2 inhibitor disulfiram (15 mg/kg, diluted in DMSO, given intraperitoneal). The ALDH2 activator were delivered 10 minutes before or after acetaldehyde (Alda‐1 + aldehyde or aldehyde +Alda‐1). Disulfiram was given 10 minutes before acetaldehyde (Disulfiram + aldehyde). As controls Alda‐1, disulfiram, saline, and DMSO were delivered alone. Changes in vaginal nociception were expressed as area‐under‐the‐curve (AUC) units and calculated using standard trapezoid methods. For statistical analysis, a one‐way ANOVA with Bonferroni's multiple comparison test was used. Data presented as mean ± SEM.ResultsAcetaldehyde‐induced vaginal hyperalgesia (44±4* vs. saline 28±2, *P<0.05). Alda‐1 markedly attenuated hyperalgesia when given either before (26±3, *P<0.05 vs. acetaldehyde) or after (25±3, *P<0.05 vs. acetaldehyde) acetaldehyde treatment. Conversely, pretreatment with disulfiram before acetaldehyde increased vaginal hyperalgesia (61±5, *P<0.05 vs. acetaldehyde). Alda‐1, DMSO, and disulfiram alone produced no significant changes in vaginal nociception.ConclusionsThese data suggest that the mechanism of endometriosis‐induced hyperalgesia is mediated by changes in reactive aldehyde production and metabolism. Increasing ALDH2 activity in rats, both prevented and alleviated reactive aldehyde‐induced vaginal hyperalgesia. These data suggest a potential novel therapeutic for treating endometriosis by targeting reactive aldehyde metabolism.Support or Funding InformationThis work was supported by the National Institutes of Child Health and Human Development K99HD093858 (SLM), a Stanford Women & Sex Differences in Medicine (WSDM) Seed Grant, and an Endometriosis Foundation of America Research Award.This abstract is from the Experimental Biology 2018 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
EPAC1 Boosts Macrophage Appetite: Implication of EPAC1 in Foam Cell Formation during Atherosclerosis
Authors: Robichaux William Gene , Mei Fang C. , Sun Hua , Wang Hui , Yang Wenli , Teng Ba‐Bie , Cheng Xiaodong 
Published: June 2021

With the increasing age of the global population, cardiovascular diseases (CVD) is the leading contributor of morbidity and mortality in growing patient populations. Thus, further elucidating the pathophysiology and developing novel treatments for CVD are critical to combat this urgent unmet medical need. The cAMP second messenger, is a major stress‐response signal found to play an important role in cardiovascular functions. Moreover, the downstream contribution of the exchange protein directly activated by cAMP 1 (EPAC1) in the development of atherosclerosis has yet to be explored. Genetic ablation of EPAC1 in atherosclerosis prone LDb (Ldlr−/−/Apobec1−/−) mice were developed as a model to investigate the functional consequences attributable to loss of EPAC1 in the pathogenesis of atherosclerosis. Remarkably, absence of EPAC1 protected against atherosclerotic lesion development compared to mice expressing EPAC1. One prevalent source attributing to atherosclerotic pathologies is chronic arterial inflammation, implicating innate immunological responders, such as macrophages. Within the lesion, macrophages are subject to a multitude of inflammatory cytokines and initiate engulfment of oxidized lipids that trigger lipid‐laden foam cell formation. Although the presence of foam cells is observed to further perpetuate macrophage aggregation and subsequent lesion advancement, the precise molecular mechanisms governing formation of foam cells and initial progression of atherosclerosis are still not fully understood. Recent evidence demonstrates EPAC1 participating in functions downstream of cAMP during phagocytosis, and thus may be involved in macrophage accumulation of oxidized‐LDL. Therefore, we hypothesize that loss of EPAC1 could attenuate atherosclerotic lesion development, in part by disrupting physiological endocytosis of oxidized‐LDL in macrophages. Indeed, further investigation demonstrates reduced phagocytosis and lipid‐burden in bone‐marrow derived macrophages not expressing EPAC1 when exposed to oxidized‐LDL. These results depict an unidentified functional role for EPAC1 in endocytosis of oxidized lipids, and suggest targeting EPAC1 may be beneficial in attenuating atherosclerotic lesion development in patients.Support or Funding InformationThis work was supported by the U.S. National Institutes of Health (NIH) Grants R01‐GM066170 and R35‐GM122536.This abstract is from the Experimental Biology 2018 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
Interaction of the Plasma Membrane Monoamine Transporter and Organic Cation Transporters with Meta‐iodobenzylguanidine (mIBG)
Authors: Lopez Quinones Antonio J. , Wagner Dave , Wang Joanne 
Published: June 2021

Meta‐iodobenzylguanidine (mIBG) is a radiopharmaceutical used as both a diagnostic imaging agent (123I‐mIBG) and a targeted frontline radiotherapy (131I‐mIBG) for neuroblastoma. Scintigraphic imaging of mIBG in human and biodistribution studies in xenografted animal models have shown that alongside its uptake into neuroblastoma tumor tissues, mIBG is also significantly transported into normal organs and tissues including the liver, salivary glands, and kidney. 123I‐or 131I‐mIBG accumulation in normal tissues can interfere with its tumor imaging quality and contribute to peripheral toxicities. While mIBG uptake into the neuroblastoma tumors has been shown to be facilitated predominantly by the human norepinephrine transporter (hNET), the molecular mechanisms involved in its uptake into the peripheral tissues have been poorly understood. We hypothesized that the organic cation transporters 1–3 (hOCT1‐3) and the human plasma membrane monoamine transporter (hPMAT) are involved in mIBG transport and accumulation in normal tissues. Thus, the purpose of this project is to characterize the interaction and transport kinetics of mIBG with these transporters and compare them with hNET. This was done by executing uptake assays in human embryonic kidney cells stably expressing hPMAT, hNET, or hOCT1‐3, followed with mIBG quantification by liquid chromatography‐tandem mass spectrometry. Time‐dependent studies showed that mIBG is an excellent substrate of hPMAT, hNET, and hOCT1‐3. Concentration‐dependent studies were fitted to the Michaelis‐Menten equation, resulting in Km values for hPMAT, hNET, and hOCT2 of 42.0 ± 10.7, 35.7 ± 8.91, and 42.9 ± 12.5 μM, respectively, suggesting that mIBG has a similar apparent affinity towards these transporters. Our data demonstrated that mIBG is a novel substrate for hPMAT and hOCTs, providing a molecular basis for these transporters as potential facilitators for mIBG uptake into normal tissues.Support or Funding InformationThis work is supported by NIH Grants GM066233 and T32‐GM007750.This abstract is from the Experimental Biology 2018 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
Multidrug resistance protein 4 (Mrp4), a drug transporter is a novel genetic determinant for adipogenesis
Authors: Donepudi Ajay C. , Manautou Jose E. 
Published: June 2021

Multidrug resistance protein 4 (Mrp4; encoded by Abcc4) is a basolateral; efflux pump that transports several xenobiotic and endogenous molecules such as prostaglandins and cyclic nucleotides. Mrp4 is highly expressed in kidney and bladder, with minimal expression in liver. Recently, our laboratory determined that mice lacking Mrp4 develop hepatic steatosis during compensatory tissue regeneration following partial hepatectomy. These studies indicate that Mrp4 may play a significant role in lipid homeostasis. Recent studies also showed that Mrp4 is expressed in adipose tissue, yet the role of Mrp4 in adipose tissue physiology and lipid metabolism is unknown. To begin addressing this, WT and Mrp4 knockout (Mrp4−/−) mice were subjected to 24‐hr fasting. The mouse embryonic fibroblast cell line 3T3‐L1 was also used to investigate the role of Mrp4 in adipogenesis. The results showed that both male and female Mrp4−/− mice have increased adipose tissue to body weight ratio and adipocyte cell size in both fed and fasted conditions. Additionally, the lack of Mrp4 is associated with increased serum glucose levels and impaired glucose tolerance in mice challenged with the glucose tolerance test. In parallel to increased adipose tissue weights, Mrp4−/− mice also showed increase in leptin gene expression in adipose tissue. In vitro studies using 3T3‐L1 cells showed that pharmacological inhibition of Mrp4 function using either MK571 or C1 increases adipogenesis and expression of adipose tissue markers such as fatty acid binding protein 4 (Fabp4), lipoprotein lipase (Lpl), glucose transporter type 4 (Glut4) and peroxisome proliferator‐activated receptor gamma (Ppar‐γ). In conclusion, our studies show that the absence of Mrp4 or chemical inhibition promotes adipogenesis, in addition to its well‐established role as a drug transporter. The implications of these findings are that alterations in Mrp4 expression and function can now be considered as a potential risk factor in the pathogenesis of metabolic diseases such as fatty liver disease, obesity and diabetes.This abstract is from the Experimental Biology 2018 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
A Blockade of IL‐6 Released from Human Breast Carcinoma MDA‐MB‐231 in Presence of NK‐92 Inhibits an Invasion of Breast Carcinoma
Authors: Lee HwanHee , Cho Hyosun 
Published: June 2021

A variety of cytokines are existed in tumor microenvironment, which might affect tumorigenesis and metastasis. Recent studies have demonstrated that interleukin(IL)‐6, a pro‐inflammatory cytokine, is significantly present in breast tumor microenvironment. It is suggested to have a critical role in an epithelial to mesenchymal transmission(EMT) process in breast cancer cells. Natural killer (NK) cells are a large innate lymphocyte that has a cytotoxic effect to cancer cells or virus‐infected cells. However, in breast tumor microenvironment, NK cells are shown to have pro‐tumor activity in presence of various inflammatory cytokines and growth factors. We found that IL‐6 is highly produced in coculture of invasive breast cancer cell MDA‐MB‐231 and natural killer(NK)‐92 cell, but is little produced in coculture of non‐invasive breast cancer cell MCF‐7 and NK cells. Subsequently, we found that the expression of p‐stat3, p‐erk, akt and pi3k proteins is statistically up‐regulated in a coculture of MDA‐MB‐231 and NK‐92 compared to in coculture with MCF‐7. A blockade of IL‐6 by anti‐IL‐6 (1ng/ml) or shRNA in invasive breast cancer cell MDA‐MB‐231 reduced a migratory and invasive capability as well as down‐regulated an expression of p‐stat3 protein. Further we examined the inhibitory effect of apigenin on invasion or migration of invasive breast cancer cell MDA‐MB‐231. We found that a blockade of IL‐6 by apigenin in MDA‐MB‐231 mediates the inhibitory effect on migration and invasion of MDA‐MB‐231.Support or Funding Informationthis research was supported by basic science research program through the national research foundation of korea(nrf) funded by the ministry of education(2016R1A6A1A03007648)This abstract is from the Experimental Biology 2018 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
Transient Increase in Sympathetic Constriction of Mesenteric Arteries from High Fat Diet‐Fed Dahl Salt‐sensitive Female Rats
Authors: Alula Kibrom Meles , Xu Hui , Galligan James 
Published: June 2021

Obesity (BMI ≥ 30) is a risk factor for hypertension. Obesity‐associated hypertension is a risk factor for cardiovascular disease, stroke, type 2 diabetes and kidney failure. Hypertension is less common in premenopausal women compared to age‐matched men. This disparity fades away after the onset of menopause. Increased sympathoexcitation in the neuroeffector junction leads to increase in constriction of blood vessels. Differences in the mechanism/s neurogenic arterial constriction in male and female are not well understood. We hypothesized that sympathetic nerve activity in high fat diet‐induced hypertension is greater in male than female rat mesenteric arteries (MA). Rats were fed either control (C) or high fat (HF) diet beginning at 3 weeks of age, and used at 10, 17 and 24 weeks on diet. Body weight and systolic blood pressure (SBP) were measured weekly. HF males gained more weight than HF females at all time points. SBP was higher in HF rats compared to C rats in both sexes and across all time points. The estrus cycle stage was determined cytologically by vaginal lavage prior to euthanasia. Mesenteric arteries (Inner diameter; male=275–350 μm, female=245–315 μm) were harvested and mounted in a pressure (60 mmHg) myograph and inner diameter was measured continuously. MA contractile responses to electrical field stimulation (30 stimuli at 0.2–30 Hz) and drug applications were measured. Neurogenic constriction of MA from HF females was greater than HF males at 5 and 10 Hz at 10 wks, and at 5–30 Hz at 17 wks. At 24 wks the difference disappears. Constrictions caused by norepinephrine (NE) and ATP were similar in MA from HF male and female rats. Immunostaining for tyrosine hydroxylase (sympathetic nerve marker) and vesicular nucleotide transporter (ATP marker) revealed greater sympathetic nerve intensity in HF male than female rats at 10 and 24 wks. There was also greater intensity in MA from HF males than HF females at 10 and 17 wks. Moreover, HF males have higher NE and ATP nerve density than HF females at 10 and 17 but not for ATP at 24 wks. MA NE levels were similar in male and female rats. Prejunctional α2‐AR and NE transporter functions were not impaired in MA from HF Dahl salt‐sensitive rats. Taken together, despite sex differences in sympathetic nerve density in MA, we detected only transient sex differences in sympathetic neurogenic arterial constrictions. Contrary to our hypothesis, we found that HF females transiently have an upregulation of neurogenic sympathetic activity and this warrants further investigation into the mechanism.Support or Funding InformationResearch supported by PPG NIH Grant P01HL070687This abstract is from the Experimental Biology 2018 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
Neuroperptides, their interactions and gastric mucosal protection in the rat.
Authors: Gyires Klara , Zádori Zoltán 
Published: June 2021

In the last decades several neuropeptides, such as thyreotropin‐releasing hormone (TRH), amylin, neurotensin, ghrelin, melatonin, opioid peptides, substance P (SP), angiotensin II (Ang II) have been shown inhibit the gastric mucosal lesions induced by different ulcerogenic agents given centrally in the dose range of fmol to nmol. The site of action is likely to be the dorsal vagal complex. However, the different neuropeptides can interact with each other at different levels and can affect each other's biological actions. Among others nociceptin ‐ opioid; angiotensin ‐ cannabinoid, opioid; CRF ‐ beta‐endorphin, somatostatin interactions have been demonstrated. Aim of the present study was to elucidate potential interactions between neuropeptides and cannabinoids in gastric mucosal defense.MethodsGastric damage was induced by acidified ethanol in the rat (150–170 g, males); the gastric mucosal lesions were detected 60 min after ethanol administration. The dugs were given intracerebroventricularly (i.c.v.) 10 min before ethanol challenge. Gastric mucosal CGRP, somatostatin levels were determined by RIA.ResultsThe gastroprotective effect of SP, nociceptin or cannabinoids was reversed by endomorphin2‐antiserum or the opioid receptor antagonist naloxone, while the Ang II‐induced protection was decreased by the CB1 receptor antagonists, AM‐251.ConclusionNeuropeptides have prominent role in central regulation of gastric mucosal integrity. In addition, interactions between neuropeptides result in redundancy in centrally‐initiated gastric mucosal defense.This abstract is from the Experimental Biology 2018 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
Muscarinic Receptor‐Mediated Regulation of Epineurial Blood Vessels of the Sciatic Nerve
Authors: Killey Chelsa , Cleary Shane , Orr Julie , Frisbee Jefferson C. , Jackson Dwayne N. , Twynstra Jasna 
Published: June 2021

ObjectiveThe objective of this study was to determine the role of muscarinic receptors on the diameter of the epineurial vessels of the sciatic nerve in situ.MethodsAn anesthetized rat model (318.4±7.1 g) was used to expose the sciatic nerve and directly visualize its microvasculature using intravital video microscopy. Rats were tracheotomized and catheters were inserted into the jugular vein for supplemental anesthesia, and the carotid artery for measures of mean arteriolar pressure. Alexa Fluor 488 Isolectin GS‐IB4 conjugate (178.6 μm/ml) was topically applied for 10 minutes to the exposed sciatic nerve to stain the endothelial cells of the epineurial vessels of the nerve. Following a 30‐min equilibration period, acetylcholine (10−4 M, n=8) or atropine (10−5 M, n=6) was topically applied for 4 and 5 minutes, respectively. Images were captured at baseline (immediately prior to drug application) and every minute during the application period. The images were analyzed offline using ImageJ to measure the diameter of the vessels at each time point. All data are expressed as mean ± standard error.ResultsAcetylcholine caused a vasodilation of the epineurial vessels (baseline: 22.9±2.4 μm, acetylcholine: 29.2±2.9 μm, p<0.01). There was no difference in mean arteriolar pressure from baseline (103.8±1.5 mmHg) to maximum vasodilation (102.3±2.7 mmHg). Atropine caused a vasoconstriction of the epineurial vessels (baseline: 26.7±3.2 μm, atropine: 15.5±3.6 μm, p<0.05). There was no difference in mean arteriolar pressure from baseline (104.0±1.9 mmHg) to maximum vasoconstriction (105.2±2.2 mmHg).ConclusionApplication of acetylcholine, a muscarinic receptor agonist, resulted in a vasodilation while application of atropine, a muscarinic receptor antagonist, caused vasoconstriction. This finding suggests that muscarinic receptors are involved in blood flow regulation of peripheral nerves. Further, since atropine constricted the vessels at baseline conditions, muscarinic receptors may be involved in maintaining baseline diameter.Support or Funding InformationThis study was supported by the SEED Research Grant from The University of Western Ontario.This abstract is from the Experimental Biology 2018 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
In Vivo Measurement of Mouse Endothelial Surface Layer in Surface Cortical Microvasculature
Authors: Perez Mario James , Ford Joshay , Schmidt Eric 
Published: June 2021

Sepsis, a systemic inflammatory response to infection, is the most common cause of in‐hospital mortality in the United States. Sepsis survivors often demonstrate chronic neurocognitive dysfunction. However, little is known about the mechanisms underlying the development of neurocognitive dysfunction during sepsis. We have previously demonstrated that septic lung and kidney injury is mediated by degradation of the endothelial surface layer (ESL), a thick glycosaminoglycan‐rich layer lining the pulmonary and glomerular microcirculation, respectively. We now postulate that degradation of the neurovascular ESL may contribute to septic neurocognitive dysfunction. To pursue this hypothesis, we have adapted a surgical approach to in vivo brain confocal microscopy that allows for direct visualization of the ESL in surface cortical microvasculature. We placed a cranial window in anesthetized mice. After allowing for 4 days of recovery, we re‐anesthetized mice and administered an intravenous bolus of 200 μl Fluorescein isothiocyanate (FITC)‐labeled 150 kDa dextran (excluded by the ESL), followed by a bolus of 200 μl Tetramethylrhodamine isothiocyanate (TRITC)‐labeled 40 kDa dextran (which can penetrate the ESL). Using an in‐focus frame, we identified surface cortical microvessels (< 20 μm diameter); at least 3 microvessels are typically found on a single frame. We performed in vivo confocal microscopy through the cranial window, simultaneously measuring TRITC (inclusive of the ESL) and FITC (exclusive of the ESL) microvessel widths. Assuming equal ESL thickness at both edges of the vessel, the ESL size is defined by one‐half the difference between TRICT‐ and FITC‐dextran vascular widths. After measurement of baseline ESL thickness, mice were injected with lipopolysaccharide (LPS, 20 μg/g body weight in 100 μl saline) to model sepsis. ESL thickness was followed every 30 min thereafter. The baseline ESL thickness of surface cortical microvessels was 0.51 μm, a size less than that which we have previously observed in lungs (1.67 μm) but similar to that observed in systemic vessels (0.6 – 0.7 μm). Endotoxemia led to a rapid loss of ESL thickness (0.08 μm vs. 0.71 μm 30 min after LPS or saline, respectively). In conclusion, by use of surgical placement of a cranial window, we have developed an experimental model that allows for the detailed observation of the neurovascular ESL. Using this model, we have demonstrated that the neurovascular ESL thickness is approximately 0.51 μm in thickness and may be degraded during endotoxemia, similar to other vascular beds of clinical relevance in sepsis.Support or Funding InformationR01 HL125371 K08 HL105538This abstract is from the Experimental Biology 2018 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.