bluuebunny/crossref_metadata_2025_split huggingface dataset:
NLRP3 Inflammasomes Contribute to the Impaired Bladder Contraction in Male Diabetic Mice
Authors: Priviero Fernanda , Onuoha Nina , Calmasini Fabiano Beraldi , Ogbi Safia , Webb R. Clinton 
Published: June 2021

BackgroundNLRP3 inflammasomes activation has been demonstrated in both type 1 and type 2 diabetes mellitus and it has been correlated with dysfunctional organs. Although it has been shown that activation of NLRP3 inflammasomes contributes to bladder dysfunction in models of cystitis and bladder outlet obstruction, it is still unknown whether inflammasomes contribute to the development of diabetic bladder dysfunction.AimWe aimed to evaluate the contribution of NLRP3 inflammasomes in the late phase of diabetic bladder dysfunction which is known by hypo responsiveness of the detrusor smooth muscle.MethodsStrips of bladder were excised from male and female wild type (WT) and dbdb−/− mice at 14 – 16 week old and were mounted in a muscle strip myograph system. Contraction of the bladder was evaluated by a single concentration of KCl (120 mM), concentration‐response curves to carbachol (1 nM – 30 μM) and frequency‐response curves to electrical field stimulation (EFS: 1–16 Hz). Carbachol and EFS were repeated in the presence of the NLRP3 inhibitor MCC950 (1 μM).ResultsMale bladder strips presented higher magnitude of contraction than female bladder strips for all contractile agents tested. KCl‐induced contraction was similar in both strains (Female WT: 42 ± 7 mN; dbdb−/−: 40 ± 4 mN; Male WT: 54 ± 6.5 mN; dbdb−/−: 51 ± 17 mN). In males, the potency of carbachol was lower in the bladder of dbdb−/− mice (pEC50: 5.53 ± 0.09) compared to WT (pEC50: 6.11 ± 0.07) and it was not restored in the presence of MCC950 (pEC50: 5.66 ± 0.08). Similarly, the contraction induced by EFS was significantly reduced for all frequencies in the bladder of male dbdb−/− mice (at 16 Hz: 51 ± 10 mN) compared to WT (at 16 Hz: 77 ± 3 mN). However, it was restored in the presence of MCC950 (at 16 Hz: 71 ± 11 mN). In female mice, there was no difference in the contraction of the bladder of dbdb−/− mice when compared to WT and addition of MCC950 did not altered the responses neither in WT nor in dbdb−/−.ConclusionIn conclusion, our data suggest that NLRP3 inflammasomes contribute to the development of hypoactive detrusor response in the late phase of diabetic bladder dysfunction in males.Support or Funding InformationNational Institute of Health (NIH; HL‐134604) São Paulo Research Foundation (FAPESP; 2016/20592‐8)This abstract is from the Experimental Biology 2019 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
Effects of chronic arsenic oral exposure on hepatic and intestinal CYP expression
Authors: Fan Xiaoyu , Liu Pengfei , Han Weiguo , Wu Xiangmeng , Ding Liang , Zhang Donna , Ding Xinxin , Zhang Qing‐Yu 
Published: June 2021

Environmental exposure to arsenic is associated with increased incidences of multiple adverse health consequences in humans, posing a significant health risk to people living near contaminated sites. For example, populations residing near mining sites may be exposed to mine tailings through the inhalation of dust particles or ingestion of contaminated water or food, as the arsenic contaminants may become bio‐accessible. Previous studies in rodents reported the ability of short‐term exposures to arsenic at high doses to increase or decrease the expression of several xenobiotic metabolizing CYPs in liver, lung, heart, or kidney, raising the possibility that exposed human individuals may have altered ability to metabolize drugs and other xenobiotic compounds. The aim of this study is to examine effects of chronic arsenic oral exposure at a relatively low level on expression of various CYPs in the liver. In the first experiment, mice were fed control or sodium arsenite‐containing drinking water (25 ppm) for 20 weeks. Hepatic gene expression was analyzed using RNA‐seq, which revealed significant increases in transcript levels for several CYPs, including CYP2A5 (by 10‐fold), an enzyme known for its activities toward nicotine, various environmental toxicants, as well as endogenous compounds. In a follow‐up study, hepatic CYP2A5 transcript level, determined by real‐time PCR, was also found to be increased (by 2.5‐fold) after 4 weeks of exposure to arsenic. Induction of CYP2A protein was confirmed by immunoblot analysis of both sets of samples. Like CYP2A5, hepatic expression of CYP3A11, a major drug‐metabolizing CYP enzyme, was also induced by arsenic exposure for 4 or 20 weeks. CYP expression in the intestine was also examined in the 4‐week exposed groups. CYP3A11, but not CYP2A5, is expressed in the intestine. Induction of CYP3A11 expression was observed in the proximal but not distal intestine, and it was less than 2‐fold. Taken together, these data reveal significant induction of hepatic CYP2A5 and CYP3A11 expression by chronic exposure to a low dose of arsenic in drinking water. The impact of this model of arsenic exposure on hepatic and intestinal xenobiotic metabolism is currently under study.Support or Funding InformationNIH grants ES020867 and GM082978This abstract is from the Experimental Biology 2019 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
A Dual Farnesoid X Receptor Agonist /Soluble Epoxide Hydrolase Inhibitor Prevents Non‐Alcoholic Steatohepatitis in Mice
Authors: Khan Abdul Hye , Stavniichuk Anna , Schmidt Jurema , Merk Daniel , Imig John D 
Published: June 2021

Non‐alcoholic steatohepatitis (NASH) is becoming the most prevalent metabolic liver disorder and is a serious global health burden. Farnesoid X receptor (FXR) agonists are in late stage clinical development to counter the disease and promise therapeutic efficacy. However, the multi‐factorial nature of NASH pathogenesis and progression indicate the need for a therapeutic approach that will target multiple pathophysiological aspects of NASH. We have developed a dual FXR agonist/soluble epoxide hydrolase (sEH) inhibitor (FXRA/sEHI) and investigated its ability to prevent NASH in toxin‐ and diet‐induced mouse NASH models. Toxin‐induced NASH was achieved in male C57BL/6 mice by injecting 200 μg streptozotocin (STZ) solution two days after birth and feeding a high‐fat diet after 4 weeks of age. Diet‐induced NASH was carried out in adult (8–12 weeks old) male C57BL/6J mice fed a methionine‐choline deficient diet (MCD; D518810) and was compared to mice fed a composition‐matched methionine‐choline replete control diet (D518754) for 4 weeks. In toxin‐induced NASH model, mice were orally treated with vehicle or FXRA/sEHI (10 mg/kg, thrice daily) or an FXR agonist obeticholic acid (OCA, 10 mg/kg, once daily) for 4 weeks. The mice from diet‐induced NASH model treated with FXRA/sEHI (10 mg/kg/d) or vehicle given in drinking water for 4 weeks. At the end of the 4‐week treatment period, blood was collected for biochemical analysis, liver tissue was collected for mRNA expression and histopathological analysis. Mice of the toxin‐induced NASH model had higher liver enzyme, hyperlipidemia, liver fibrosis, and steatosis. FXRA/sEHI treatment reduced these parameters (by 20–50%, P<0.05), while OCA displayed weak or no beneficial effect on NASH (P>0.05). In diet‐induced NASH model, the mice exhibited 2‐fold higher hepatic fibrosis, 2–6‐fold higher hepatic mRNA expressions of inflammatory and fibrotic markers, and marked hepatic steatosis (P<0.05). The dual modulator FXRA/sEHI robustly prevented hepatic steatosis, fibrosis, and inflammation (by 50–80%, P<0.05) compared to vehicle. Our results demonstrate strong potential for dual FXRA/sEHI modulators as a novel therapeutic strategy in NASH and advocate multi‐targeted approaches in NASH treatment.This abstract is from the Experimental Biology 2019 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
Impact on acetaminophen‐induced hepatotoxicity by long non‐coding RNAs HNF1α‐AS1 and HNF4α‐AS1 in HepaRG cells
Authors: Chen Liming , Manautou Jose E. , Zhong Xiao‐bo 
Published: June 2021

Acetaminophen (APAP) is a commonly used over‐the‐counter drug for its analgesic and antipyretic effects. However, APAP overdose leads to severe hepatotoxicity due to the accumulation of N‐acetyl‐p‐benzoquinone imine (NAPQI), the toxic metabolite of APAP generated by cytochrome P450s (P450s). Long non‐coding RNAs HNF1α‐antisense 1 (HNF1α‐AS1) and HNF4α‐antisense 1 (HNF4α‐AS1) are regulatory RNAs, which are recently identified to be involved in the regulation of hepatic P450 expression. This study aims to determine whether HNF1α‐AS1 and HNF4α‐AS1 have impacts on APAP‐induced hepatotoxicity using the most useful hepatic cell line HepaRG as an in vitro model. Small hairpin RNAs (shRNAs) were used to knock down HNF1α‐AS1 and HNF4α‐AS1 in HepaRG cells. Knocking down of these lncRNAs altered APAP‐induced hepatotoxicity indicated by MTT and LDH cytotoxicity assays. Specifically, HNF1α‐AS1 knockdown decreased APAP toxicity at 10 mM with increased cell viability and decreased LDH release, whereas HNF4α‐AS1 knockdown exacerbated APAP toxicity at 30 mM. Alterations of gene expression by knocking down HNF1α‐AS1 and HNF4α‐AS1 were examined in several APAP‐induced hepatotoxicity pathways, including P450‐mediated APAP biotransformation by CYP1A2, 2E1, and 3A4, APAP detoxification with glucuronidation by UDP‐glucuronosyltransferases, UGT1A1 and 1A9, and sulfation by sulfotransferases SULT1A1, and NAPQI detoxification through glutathione conjugation by glutathione‐S‐transferases, GSTP1 and GSTT1. HNF1α‐AS1 knockdown significantly decreased mRNA levels of CYP1A2, 2E1, and 3A4, while HNF4α‐AS1 knockdown significantly induced mRNA levels of CYP2E1 and 3A4 in HepaRG cells. HNF1α‐AS1 knockdown decreased mRNA expression of SULT1A1 but not UGT1A1 or 1A9, while HNF4α‐AS1 knockdown showed no significant effect on the selected phase II enzymes. Significantly decreased mRNA expression of GSTP1 was observed after either HNF1α‐AS1 or HNF4α‐AS1 knockdown. However, considering the low intrinsic expression of GSTP1 in HepaRG cells, how this decreased expression of GSTP1 affects APAP toxicity is inconclusive. GSTT1 expression showed no change in all groups. Altogether, our study suggests that HNF1α‐AS1 and HNF4α‐AS1 affect APAP‐induced hepatotoxicity mainly through alterations of P450‐mediated APAP biotransformation process in HepaRG cells, indicating an important role of lncRNAs in drug‐induced liver toxicity.Support or Funding InformationThis work was supported by grant NIH/NIGMS R01GM118367.This abstract is from the Experimental Biology 2019 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
Expression and potential involvement of PMAT in mIBG transport in neuroblastoma
Authors: Zhang Yuchen , Quiñones Antonio López , Stevens Jeffrey , Park Julie , Wang Joanne 
Published: June 2021

Neuroblastoma (NB) is the most common extracranial solid tumor in pediatric cancer. Despite recent improvements, the long‐term survival rate for patients with high‐risk NB is still less than 50%. Radiolabeled meta‐iodobenzylguanidine (mIBG), 123I‐mIBG and 131I‐mIBG, are used clinically for both diagnostic and therapeutic purposes, and several clinical trials are ongoing to investigate the potential of 131I‐mIBG as a frontline therapy for high‐risk NB. Previous studies established that mIBG enters NB cells through the norepinephrine transporter (NET). However, not all NET‐expressing NBs respond to 131I‐mIBG treatment and other mechanisms may be involved in the uptake and retention of mIBG in NB. Here, we investigated the expression and potential influence of additional mIBG transporters in NB. By searching through an NCI database with 156 NB patients, mRNA Seq results showed a high and variable mRNA expression of the plasma membrane monoamine transporter (PMAT) in NB tumor samples. Interestingly, Kaplan–Meier estimation suggested an association of low PMAT expression with poor survival rates in these patients. Using qPCR analysis, we further demonstrated a high expression of PMAT in 32 local NB samples as well as two human NB cell lines SH‐SY5Y and SK‐N‐BE(2). Immunofluorescence imaging of PMAT in NB tumor specimens and cultured cells suggested that unlike its normal expression on plasma membrane, PMAT protein in NB cells is predominantly expressed in intracellular organelles but also with a low expression at the plasma membrane. In HEK293 cells stably expressing human PMAT, mIBG is avidly transported by PMAT with a Km value of 42.0 ± 10.2 μM. In SH‐SY5Y and SK‐N‐BE(2) cells, uptake of mIBG was performed with or without sodium in uptake buffer to distinguish the NET‐mediated uptake. Preliminary data showed that sodium depletion only inhibited 50–60% of mIBG uptake whereas addition of a PMAT inhibitor to sodium‐free buffer completely inhibited mIBG accumulation in both cell lines. In summary, our current study suggests that PMAT is highly expressed in NB tumors and NB‐derived cells lines. PMAT transports mIBG and may be involved in the uptake and retention of mIBG in NB cells, and hence may represent a novel determinant of mIBG tumor targeting and clinical response.Support or Funding InformationThis work is supported by NIH Grants GM066233 and GM066233‐14S1.This abstract is from the Experimental Biology 2019 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
Cell Model for Studying Nucleoside Transporters, a Key Component of the Blood‐Testis Barrier
Authors: Miller Siennah R , Morales Mark N , Frost Kayla L , Cherrington Nathan J , Wright Stephen H 
Published: June 2021

Sertoli cells in the testis represent the principle element of the Blood‐Testis Barrier (BTB). Equilibrative nucleoside transporters (ENTs) are responsible for the transport of nucleosides across rodent BTB and immunohistochemistry suggests that hENT1 is located on the basal membrane and hENT2 is located on the apicolateral membrane of Sertoli cells (Klein et al., 2013). Therefore, the ENTs are of particular interest in studying the disposition of nucleoside reverse transcriptase inhibitors (NRTIs) in the human male genital tract because of their similarity in chemical structure to nucleosides. This study characterized the transport of a specific nucleoside, uridine, to determine the relative roles of ENT1 and ENT2 in nucleoside transport in HeLa S3 cells, a potential model system for the study of ENT selectivity, using the ENT specific inhibitor S‐(4‐nitrobenzyl)‐6‐thioinosine (NBMPR). Uptake of 20 nM [3H]uridine was linear for 7 minutes and 100 nM NBMPR blocked 77% of this uptake, with the remaining uptake blocked by 100 uM NBMPR (to the same level produced by 5 mM unlabeled uridine). ENT1 and ENT2 are known to have different sensitivities for NBMPR and, consistent with this difference, inhibition of [3H]uridine uptake produced by increasing concentration of NBMPR proved to be biphasic, with IC50s of 11.1 nM (ENT1) and 7.2 uM (ENT2). Consequently, uridine uptake measured in the presence of 100 nM NBMPR was taken to represent ENT2‐mediated transport; subtracting that from total uptake represented ENT1‐mediated transport. The kinetics of ENT1‐ and ENT2‐mediated [3H]uridine uptake into HeLa S3 cells showed no difference in Jmax (16.1 and 12.3 fmol cm−2 min−1) and a three‐fold difference in Km (13.3 and 35.2 uM). The resulting 3.5‐fold difference in intrinsic clearance for uridine uptake (i.e., Jmax/Km) for hENT1 and hENT2 mediated transport (1.21 mL cm−2 min−1 and 0.35 mL cm−2 min−1, respectively), accounted for the observed inhibition of uridine transport produced by 100 nM NBMPR, indicating that at low concentrations, hENT1 is primarily responsible for uridine uptake in these cells. Together, these data suggest that HeLa S3 cells are an adequate model for studying the characteristics of nucleoside transporters present in the BTB.Support or Funding InformationSupported by R01GM123643This abstract is from the Experimental Biology 2019 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
Multidrug resistance protein 4 (Mrp4) regulates nutrient and energy metabolism by altering the adipose tissue physiology
Authors: Donepudi Ajay C , Lee Yoojin , Lee Ji‐Young , Schuetz John D , Manautou Jose E 
Published: June 2021

Multidrug resistance protein 4 (Mrp4; encoded by Abcc4) belongs to the ATP‐binding cassette efflux transporter superfamily and transports several xenobiotics and endogenous molecules, such as prostaglandins and cyclic nucleotides. Mrp4 is highly expressed in the kidney and bladder, with minimal expression detected in the liver. We recently determined that the lack of Mrp4 increases adipose tissue and body weights in mice. Interestingly, the expression of Mrp4 and its role in adipose tissue physiology, nutrient metabolism and the pathogenesis of metabolic diseases are unknown. The current study was aimed at characterizing these specific roles of Mrp4 employing wildtype (WT) and knockout (Mrp4−/−) mice. Our studies determined that Mrp4 is expressed in high levels in adipose tissue and that the lack Mrp4 resulted in adipocyte hypertrophy. Along with increases in body and adipose tissue weights, lack of Mrp4 in mice also resulted in increased blood glucose and leptin levels, and impaired glucose tolerance. Additionally, Mrp4−/− mice have increased levels of adipose tissue phospho‐cAMP response element binding protein (p‐Creb), which is known to regulate adipocyte glucose and lipid metabolism. In contrast to altered glucose metabolism, Mrp4−/− mice did not show any significant changes in plasma triglycerides, free fatty acids or total cholesterol levels compared to WT mice. Indirect calorimetry measurements under basal conditions showed that Mrp4−/− mice have decreased volumetric oxygen consumption (VO2), respiratory exchange ratio (RER), energy expenditure (EE), ambulatory activity (AA) and wheel counts compared to WT mice. Decreased indirect calorimetry parameters and impaired glucose metabolism in Mrp4−/− mice can be attributed to increased adipose tissue weights, which complements the observed obese and diabetic phenotype in these mice. Together, our data indicate that Mrp4 regulates nutrient and energy metabolism by altering the adipose tissue physiology. In conclusion, the absence of Mrp4 function in mice results in the development of obesity and diabetes.This abstract is from the Experimental Biology 2019 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
A Novel Embryonic Stem Cell‐based Vaccine for the Prevention of Lung Cancer
Authors: Meng Shuhan , Whitt Aaron G. , Al Rayyan Numan , Richie Jamaal , Tu Allison , Eaton John W. , Yaddanapudi Kavitha , Li Chi 
Published: June 2021

The antigenic similarity between embryos and tumors has raised the idea of using embryonic material as a preventative vaccine against neoplastic disease. Indeed, we have previously reported that a vaccine comprised of allogeneic murine embryonic stem cells (ESCs) and murine fibroblasts expressing immune‐stimulatory granulocyte macrophage‐colony stimulating factor (GM‐CSF) successfully blocks the outgrowth of an implantable lung cancer (Lewis lung carcinoma; LLC) and lung tumors caused by a combination of a mutagen followed by chronic pulmonary inflammation. However, such a vaccine is obviously impractical for application to humans. The use of fibroblasts to produce GM‐CSF is needlessly complicated, and intact live ESCs carry the hazard of generating embryomas/teratomas. Here, we report the successful application of an alternative prophylactic vaccine comprised of exosomes/microvesicles derived from murine ESCs engineered to produce GM‐CSF. The prophylactic potential of the exosomes against implanted (subcutaneous inoculation of LLC) and metastatic (tail vein injection of LLC) lung cancer was tested by immunizing mice with exosomes derived from ESCs. Vaccination of mice with the exosomes bearing GM‐CSF significantly slowed or blocked the outgrowth of LLC while control exosomes lacking GM‐CSF were ineffective. Examination of tumor‐infiltrating immune cells showed robust tumor‐reactive immune responses in mice vaccinated with the exosomes bearing GM‐CSF, including an increase in cytotoxic T cells and a decrease in immunosuppressive T regulatory cells. Overall, vaccination of mice with ESCs‐exo bearing GM‐CSF is very effective in preventing both implantable and metastatic lung tumors with no detectable toxicity. Therefore, we conclude that a similar vaccine derived from GM‐CSF‐expressing human ESCs may be applicable to humans with increased risk of developing lung cancer.Support or Funding InformationNIH grant CA198249 (K.Y.); Free to Breathe Research Grant (K.Y.); NIH grant AA018016‐01 (J.W.E); Commonwealth of Kentucky Research Challenge Trust Fund (J.W.E.); NIH grants CA106599 and CA175003 (C.L.); 1P30GM106396 Phase III CoBRE grant (C.L.).This abstract is from the Experimental Biology 2019 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
Excessive salt consumption increases susceptibility to cerebrovascular dysfunction and cognitive impairments in the elderly of both sexes
Authors: Wang Shaoxun , Jiao Feng , Guo Ya , Zhang Huawei , He Xiaochen , Maranon Rodrigo O , Alexandra Barbara , Pabbidi Mallikarjuna , Roman Richard J , Fan Fan 
Published: June 2021

Epidemiological study indicates that the prevalence of dementia is higher in women than men. Excessive salt consumption increases the incidence of cardiovascular disorders, which is one of the major risk factors for dementia. However, whether high‐salt intake that can but not always elevate blood pressure contributes to the development of dementia is still not well understood. Moreover, if there is a correlation between excessive salt consumption, cerebral vascular dysfunction, and dementia, especially if there is a relationship between these factors and sex and age is unclear. In the present study, we examined the myogenic response of middle cerebral artery, autoregulation of cerebral blood flow (CBF), functional hyperemia, and cognitive function in young (3‐month) and old (18‐month) SD rats in both males and females. Blood pressure was not altered in all groups of rats fed 8% high salt (HS) diet. Yong and old female rats all displayed more spontaneous tone, less pressure‐induced myogenic response compared to age‐matched males. HS fed rats developed the impaired myogenic response to pressure and to Ach‐induced vasodilation in all age and sex. CBF measured by laser Doppler flowmetry (LDF) rose by 27.7 ± 4.8% (male, young), 57.4 ± 4.4% (female, young), 35.9 ± 3.3% (male, old), and 60.3 ± 12.3% (female, old), respectively, when MAP increases from 100–180 mmHg indicating that female rats have impaired CBF autoregulation in both ages. HS intake diminished and aggravated CBF autoregulation in male and female rats, respectively, in all age. Elderly female rats exhibited impaired functional hyperemia measured by LDF during whisker stimulation compared to age‐matched males. Excessive salt intake also exacerbated the declined functional hyperemia and resulted in cognitive impairment examined by eight‐arm water maze in old male and female rats. These results demonstrated that excessive salt consumption increases susceptibility to the development of cerebrovascular dysfunction and cognitive impairments in both male and female elderly independent of blood pressure. This study provides novel insight to understand the effects of metabolic stressors on sex and age in the development of cerebral vascular function and dementia.Support or Funding InformationThis study was supported by grants AG050049, P20GM104357, DK104184, and HL138685, from the National Institutes of Health; 16GRNT31200036 from the American Heart Association.This abstract is from the Experimental Biology 2019 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.