bluuebunny/crossref_metadata_2025_split huggingface dataset:
Age Differences in the Stimulation of Murine Hepatic XBP‐1 Splicing in Response to the Anti‐HIV Drug Efavirenz
Authors: Hamlin Allyson Naugle , Heck Carley J.S. , Bumpus Namandje N. 
Published: June 2021

Efavirenz is a non‐nucleoside reverse transcriptase inhibitor used in combination with other antiretroviral drugs to treat HIV infection. Efavirenz is one of the most commonly prescribed antiretrovirals and is used in both adult and pediatric populations. Efavirenz treatment has been associated clinically with hepatotoxicity; however, the mechanism underlying this is not fully understood and work in this area has largely been focused on adults. With this in mind, using primary mouse hepatocytes, we probed whether the activation status of transcription factors, such as X‐box binding protein‐1 (XBP‐1), that play a crucial role in regulating cell survival are differentially impacted by efavirenz as a function of age. To do so, the ratio of spliced (activated) to unspliced XBP‐1 mRNA was measured following efavirenz treatment. Primary mouse hepatocytes were isolated from pediatric (3–4 week old) and adult (8–12 week old) male C57Bl/6J mice. XBP‐1 splicing following incubation with efavirenz for 4 hours was measured using reverse transcriptase PCR. In 3–4 week old mice, treatment with 10 μM efavirenz for 4 hours resulted in a 3.4‐fold increase in the ratio of spliced to unspliced XBP‐1 as compared to vehicle controls; however, XBP‐1 splicing was not elevated in response to this same treatment in adult mice. In fact, a 30 μM concentration of efavirenz was required in adult mice to stimulate a comparable (3.9‐fold) increase. Because activation of XBP‐1 splicing can affect cell survival, a cell death assay using ethidium bromide and acridine orange was performed and these experiments were confirmed using a TUNEL assay. In primary hepatocytes isolated from 3–4 week old mice, a 4.8‐fold increase in cell death was observed following treatment with 20 μM efavirenz. Similarly, using hepatocytes from adult mice, cell death was stimulated 4‐fold by treatment with 20 μM efavirenz. In the presence of STF080310, a known inhibitor of XBP‐1 splicing, cell death in response to 20 μM efavirenz was reduced to 2.5‐fold and 1.8‐fold over controls in 3–4 week old and adult mice, respectively. These results were commensurate with data obtained using TUNEL staining. In conclusion, hepatocytes from mice at early stages of development may be more sensitive to activation of XBP‐1 splicing than adult mice. Furthermore, cell death was similar in the 3–4 week old mice and adult mice suggesting that age may not result in differential susceptibility to hepatocyte death in response to efavirenz.Support or Funding InformationThis research is funded by NIH R01 GM103853.This abstract is from the Experimental Biology 2018 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
Gender Differences on the Effects of Ibuprofen on Proteasome Function in Mice Heart
Authors: Tiwari Shuchita , Gomes Aldrin V. 
Published: June 2021

Introduction and objectivesIbuprofen, a cyclo‐oxygenase inhibitor, is a commonly used non‐steroidal anti‐inflammatory (NSAID) agent used in the treatment of pain, fever and inflammation. The long‐term use of ibuprofen is associated with increased risks for cardiovascular events and stroke. The effect of ibuprofen on cardiac function as well as if gender differences exist with respect to ibuprofen's effect on cardiac function are not well understood. Accordingly, the aim of this study was to explore the effects of ibuprofen on protein homeostasis in mice cardiac tissue and to determine if ibuprofen affects male and female cardiac proteasome activity in a similar manner.Method/ResultsAged matched male and female C57BL/6J mice were randomly assigned to two groups: (A) control (B) group treated with ibuprofen. Mice were treated with ibuprofen (100 mg/kg) dissolved in drinking water for 6 weeks.Since impaired proteasome function is associated with cardiovascular disorders, β1 caspase‐like activity, β2 trypsin‐like activity, and β5 chymotrypsin‐like proteasome activities were measured using bortezomib as a specific inhibitor of proteasome in cardiac tissue of both male and female mice. In female mice chronic treatment with ibuprofen showed decreased proteasome function as decreased β1, β2, and β5 activity was observed in ibuprofen treated female animals. However, proteasome activity was not affected in male animals treated with ibuprofen. Excess accumulation of misfolded and oxidized proteins alters proteasome function, therefore levels of oxidized proteins in heart lysates was measured by oxyblot method using dinitrophenylhydrazine (DNPH). The levels of oxidized proteins increased in only female animals treated with ibuprofen. The levels of polyubiquitinated proteins was similar in male and female ibuprofen treated hearts compared to control hearts. An increase in free ubiquitin levels was only observed in the hearts of female animals treated with ibuprofen when compared to control animals and ibuprofen treated male animals.ConclusionOur findings suggest, for the first time, that gender differences on proteasome function occur in the heart of ibuprofen treated animals. These data show that chronic treatment of ibuprofen decreased proteasome activities in females as well as increased the levels of oxidized proteins and free ubiquitin levels in only ibuprofen treated female mice hearts.Support or Funding InformationAmerican Heart Association Award #16GRNT31350040This abstract is from the Experimental Biology 2018 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
CCR5 antagonism as a potential therapeutic approach for cardiac fibrosis
Authors: Liang Wenjing , Gorr Matthew W. , Insel Paul A. 
Published: June 2021

Fibrosis of the heart occurs in multiple cardiac disorders and is associated with significant morbidity and mortality. Effective therapy to treat and prevent this pathological process remains a major clinical challenge. G protein‐coupled receptors (GPCRs) play a vital role in altering the function of cardiac cells and are thus major therapeutic targets for cardiovascular disorders. We tested the hypothesis that the chemokine receptor CCR5, which is a co‐receptor for HIV infection of immune cells, may contribute to and thus be a novel therapeutic target in cardiac fibrosis. We found prominent CCR5 expression in cardiac fibroblasts isolated from mice and rats. Expression of CCR5 was significantly increased when fibroblasts were treated with hypoxia to induce a myofibroblast phenotype. Cardiac fibroblasts exposed 72 hours to hypoxia increased CCR5 expression 5.8±1.7‐fold compared to normoxia (p=0.0327) in cells isolated from mice, and 4.2±1.1‐fold in cells isolated from rat (p=0.0178). To assess the role for CCR5 in experimental cardiac fibrosis, we used pharmacological CCR5 activation and inactivation in mouse and rat cardiac fibroblasts. Maraviroc, an FDA‐approved antagonist of CCR5 reduced the expression of fibrotic markers, including α‐smooth muscle actin and collagens I and III in mouse and rat cardiac fibroblasts grown under both normoxic and hypoxic conditions. Treatment with recombinant CCL4, the agonist of CCR5, increased these fibrotic markers in rat cardiac fibroblasts. In addition, Maraviroc reduced cardiac fibroblast proliferation and migration while CCL4 had the opposite effect. We conclude that the CCL4‐CCR5 pathway can regulate activity of cardiac fibroblasts and may be a therapeutic target for cardiac fibrosis.Support or Funding InformationSupported by R21AG52914 and T32HL007444‐34This abstract is from the Experimental Biology 2018 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
Ursodesoxycholic acid Display a Role of Experimental Anti‐hepatic Fibrosis by Inhibiting the Expression of Bnip3L Protein
Authors: Teng Lin‐Hua , Liu Qi‐Lin , Zhang Ji‐Wang , Liu Ze‐Feng , Chen Li , Zhang Guo 
Published: June 2021

Backgound & AimsPrevious reports showed that ursodesoxycholic acid (UDCA) might has an therapeutic effect on liver fibrosis, but the detail pharmacological mechanism remains unclear. This study was aimed to study the relationship between the expression change of mitochondrial Bnip3L protein and inhibitory role of UDCA for experimental hepatic fibrosis.MethodsHepatic stellate cell (HSC)‐LX2 cell line and CCl4‐induced hepatic fibrotic rats model were used in the in vitro or in vivo regimens with UDCA or vehicle control. Western Blot was then performed to analyze the expression of α‐SMA and Bnip3L in LX2 cells. Real‐time fluorescence quantitative PCR (qRT‐PCR) was performed to compare the differential expression of COL1 mRNA in LX2 cells and COL1 mRNA. Bnip3L mRNA in liver tissues among these different groups. The liver fibrosis of rats was evaluated by Masson staining.ResultsOur research showed that the expression of Bnip3L protein was significantly inhibited by UDCA in LX2 cells which were activated by TGF‐β1, and that was positively correlated with the down‐regulated expression of α‐SMA and COL1 mRNA. UDCA could significantly reduce the production of collagen in liver tissues of CCl4‐induced liver fibrosis. In parallel, the expression of Bnip3L mRNA and COL1 mRNA were simultaneously inhibited by UDCA in these liver tissues.ConclusionsOur findings demonstrate that UDCA displays a critical role of anti‐hepatic fibrosis by down‐regulating the expression of Bnip3L protein, which may provide a novel pharmacological strategy for liver fibrosis in the future.Support or Funding InformationSupported by NSFC Grants 81360077, 30630145, 81172260 and GXNSFC Grants 2015GXNSFDA139022 to Guo ZhangThis abstract is from the Experimental Biology 2018 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
POSSIBLE PROTECTIVE EFFICACY OF ETHANOL EXTRACT OF GARCINIA KOLA SEED ON CANDIDIASIS COMPLICATION IN DIABETES MELLITUS
Authors: Adeyeoluwa Temitayo Esther , Okeleye Benjamin Ifeoluwa , McGaw Lyndy , Ashafa Anofi Omotayo 
Published: June 2021

Management of the complications of diabetes mellitus (DM) like candidiasis, a prevalent condition in DM patients, is important in the survival and longevity of life of the patient. In this study, Garcinia Kola nut, a medicinal seed with established anti‐diabetic and anti‐inflammatory activity, was investigated for its anti‐candida properties. Three (3) different extracts of G. kola, viz; the aqueous, hydro‐ethanol and ethanol extracts were prepared and in vitro assays were carried out to determine the anti‐ candida activity and minimum inhibitory concentration (MIC) of the seed extracts using agar well and microdilution techniques, while routine, in vitro analyses of the phytochemical constituents, anti‐oxidative, anti‐diabetic and anti‐inflammatory assays were also carried out on the seed extracts. Ethanol extract showed high anti‐oxidative and anti‐diabetic activities compared to the aqueous and hydro‐ethanol extracts. It also had higher phenolic, flavonoid and flavonol contents. The ethanol extract also showed low α‐amylase and high α‐glucosidase inhibitory activity. Similarly, the ethanol extract had a zone of inhibition of 12–15 mm in the agar‐gel diffusion assay against all the strains of Candida albicans. It had an MIC of 0.39 mg/ml and MFC of 1.563 mg/mL compared with the MIC and MFC of 0.781 mg/mL and 3.125 mg/mL for both aqueous and hydro‐ethanol extracts respectively. The cytotoxicity assay also showed that the ethanol extract has LC 50 of 90 μg/mL. The results of this study show that Garcinia kola seed can be an effective anti‐candida agent suitable for use in diabetic patients.Support or Funding InformationDirectorate of research development. University of the Free state, South Africa. Internal fundThis abstract is from the Experimental Biology 2018 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
Low‐Density Lipoprotein Receptor‐Related Protein 1 (LRP1) Is Degraded Primarily by The Lysosomal Degradation System in Hepatocellular Carcinoma (HepG2) Cells
Authors: Tenglin Karin , Aragòn Brian Aranibar , Aguirre Stefany Huaynasi , Rondon‐Ortiz Alejandro Nico , Pino‐Figueroa Alejandro 
Published: June 2021

LRP1 is a transmembrane protein responsible for binding and internalizing at least 50 different ligands, one of which is amyloid beta (Aβ). LRP1 has a role in clearance of brain and systemic Aβ. Understanding LRP1's role in Aβ degradation could have important implications for understanding Alzheimer's Disease (AD) pathogenesis. Since LRP1 in the liver is primarily responsible for systemic clearance of Aβ, its degradation could have important implications for AD pathogenesis.Normal degradation of proteins typically occurs through the ubiquitin proteasome system (UPS) and/or the lysosomal degradation system. To study LRP1 degradation through either of these two processes, one system was inhibited, then subsequent LRP1 output was analyzed. By inhibiting one degradation system at a time, we can analyze the protein output to quantify how much protein is degraded by each system. Bafilomycin A1 was used to inhibit the lysosomal degradation system, and MG132 was used to inhibit UPS. Finally, cycloheximide, a protein synthesis inhibitor, was used to determine LRP1 half‐life and to determine whether protein output is significantly affected by protein synthesis during inhibition of the protein degradation system (cycloheximide chase analysis).HepG2 cells were used to study LRP1 degradation in these experiments. The cells were treated with Bafilomycin A1, MG132, or dimethyl sulfoxide (DMSO; control group) and lysed at time points 0, 8, 16, and 24 hours to analyze LRP1 protein levels over time. Experiments with Bafilomycin A1 showed increasing levels of LRP1 over time, whereas experiments with MG132 showed unchanged, then decreasing levels of LRP1 over time. Cycloheximide was tested to determine the optimal concentration for HepG2 cells, which was found to be 30 μM. Then, experiments were performed with either cycloheximide and DMSO (as control), cycloheximide and Bafilomycin A1, or cycloheximide and MG132 at time points 0, 8, 16, and 24 hours to determine the half‐life of LRP1. Our results showed an extension of LRP1 half‐life by inhibition of lysosomal degradation system when using Bafilomycin A1, but half‐life was not extended when UPS was inhibited by MG132. Due to its significant increase in half‐life with Bafilomycin A1, but not with MG132, it's concluded that LRP1's degradation is primarily produced through the lysosomal degradation system.Support or Funding InformationMCPHS UniversityThis abstract is from the Experimental Biology 2018 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
Silencing of MAP Kinase Phosphatase 1 Blocks the Renal Inflammatory Response Induced by Chronic Stress
Authors: Roland Miranda M. , Langreck Cory , Duric Vanja , Babich Victor , Di Sole Francesca 
Published: June 2021

Chronic stress can lead to disruption of the body systemic equilibrium which may correspond to dysfunction of both nervous and peripheral organ systems. A relationship between chronic stress, mood disorders (e.g. major depression) and systemic disease (e.g. kidney disease) is proposed via an immune system reaction and increased level of proinflammatory cytokines. This study aims to determine whether: 1. chronic stress induced by exposure of rats to corticosterone (CORT) in drinking water for 3 weeks (i.e., 50 μg/mL CORT ad libitum for 2 weeks; 25 μg/mL for 3 days and, then, 12.5 μg/mL for another 3 days) associates with a renal immune reaction and, possibly, kidney dysfunction and 2. silencing of MAP Kinase Phosphatase 1 (MKP‐1) affects the renal immune reaction mediated by CORT. MKP‐1 is a negative regulator of the MAP kinase signaling that is overexpressed within multiple human hippocampal sub‐regions of the brain of depressed subjects and is fundamental for the development of depressive‐like behaviors in rodent model of chronic stress. The neutrophil gelatinase‐associated lipocalin (NGAL) and IL‐18 are early diagnostic biomarkers. They accumulate in the kidney in response to inflammation, kidney injury, and decreased kidney function. Protein levels of NGAL and IL18 were analyzed by immunocytochemistry. NGAL and IL18 protein levels were significantly increased in rats treated with CORT compared to their controls (over 70% increase in NGAL and IL18 protein levels in the glomeruli, 40% increase in NGAL and 6% increase in IL18 protein levels in the tubules). The increase in NGAL and IL18 protein levels was blocked by micro‐infusion of recombinant adeno‐associated virus encoding Cas9 with guide RNA targeting MKP‐1 used to silence MKP‐1 protein expression prior to exposure to CORT. In summary, renal inflammation induced by chronic stress was blocked by MKP1 silencing. This study supports the therapeutic potential of MKP‐1 as target for signaling pathways between chronic stress, mood disorders and kidney dysfunction.Support or Funding InformationIowa Osteopathic Education and Research FundsThis abstract is from the Experimental Biology 2018 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
Angiotensin AT2 Receptor Stimulation is Protective in Lipopolysaccharide‐Induced Inflammation and Renal Injury
Authors: Hussain Tahir , Dhande Isha , Patel Sanket Niranjanbhai , Ali Quaisar 
Published: June 2021

Recently angiotensin type‐2 receptor (AT2R) activation has been shown to exert anti‐inflammatory and reno‐protective response against chronic injury. Our prior in vitro studies using the proximal tubule epithelial cells (PTECs) and macrophages revealed that prior activation of angiotensin‐II type 2 receptor (AT2R) reduces pro‐inflammatory cytokine production in response to lipopolysaccharide (LPS). However, the in vivo effects of AT2R stimulation on LPS‐mediated inflammatory renal injury are not known. The present study tests the hypothesis that AT2R activation attenuates inflammation and preserves renal structure and function in the early stages of LPS‐mediated renal injury. Mice were pretreated with vehicle or AT2R agonist C21 (300 μg/kg, i.p.) prior to LPS (5 mg/kg, i.p) challenge. Pretreatment with C21 significantly prevented the LPS‐induced increase in the levels of renal pro‐inflammatory cytokines (TNF‐α, IL‐6), TLR4 expression, and markers of renal injury (blood urea nitrogen, plasma creatinine and albuminuria). Moreover, C21 treatment also protected against LPS‐induced tubular vacuolization and tubular degeneration. Renal and circulating anti‐inflammatory IL‐10 levels were higher with C21 pretreatment at time of LPS challenge. In vitro studies in HK‐2 cells (human PTECs) demonstrate that AT2R activation directly stimulates IL‐10 production. Further, conditioned‐media derived from the C21‐treated HK‐2 cells reduced LPS‐induced TNF‐α production via IL‐10 originating from HK‐2 cells. Our findings suggest that prior activation of AT2R prevents LPS‐mediated inflammatory renal injury, possibly via an increase in renal IL‐10 production.Support or Funding InformationThis study was supported by NIH R01 grant DK61578This abstract is from the Experimental Biology 2018 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
Gonadectomy Affects Water‐Deprivation‐Induced Urinary Concentration in Female and Male Mice
Authors: Delaney Dalton , Waturuocha Chiedozie , Fan Liming , Rouch Al 
Published: June 2021

The urinary concentrating mechanism is critical for maintaining whole‐body fluid balance and plasma osmolarity. Estrogen has been shown to downregulate vasopressin‐sensitive aquaporin‐2 (Aqp2) expression (Cheema, MU, et al. Am J Physiol, Renal Physiol, 309, F305–F317, 2015). While sex steroids have been reported to influence urinary concentration, additional studies are clearly needed. The purpose of this study was to determine if gonadectomy affects urine osmolarity (Uosm) during water deprivation (WD) in mice. Intact female and male (IF, IM), ovariectomized (OVX), and castrated (CA) CD‐1 mice at four weeks of age were obtained from Envigo, Inc. (Indianapolis, IN). Two studies were conducted: the first served as a control study and the second involved WD. The control study consisted of a four‐day period where IF, OVX, IM, and CA mice (3/group), in metabolic cages, consumed, ad libitum, standard rodent chow (0.25% salt) and water for four days. The WD study also consisted of four days where water was removed from 7 pm to 7 am in the final three days. Daily measurements included food and water intake, urine flow rate, Uosm, and sodium and potassium excretion. Urine samples were collected after 7 am just before replacing water for the mice. Uosm was measured using a vapor‐pressure osmometer. In the control study no differences between any groups were measured. ANOVA revealed that in the WD study, the IF mice had significantly lower Uosm (mOsm/L), averaged from the three WD days, compared to OVX mice (5,924 ± 431 vs 8,400 ± 691, respectively, p < 0.01). The IM mice had significantly higher Uosm compared to CA mice (8,683 ± 691 vs 3,590 ± 382, respectively, p<0.0001). Other Uosm comparisons: IM > IF (p<0.005) and IF > CA (p<0.01). We conclude that gonadectomy augments urinary concentration in female mice while it reduces urinary concentration in male mice during WD. Results suggest significant roles for the sex steroids, estrogen and testosterone, in urinary concentration. RT‐qPCR studies are underway to determine potential expression differences in Aqp2 in all groups from this study.Support or Funding InformationOSU‐CHSThis abstract is from the Experimental Biology 2018 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
Submaximal Level Single Twitch Kinetics Dependent on Disease State in Duchenne Muscular Dystrophy Mouse Model
Authors: Peczkowski Kyra K. , Rastogi Neha , Schultz Eric J. , Rafael‐Fortney Jill A. , Janssen Paul M. L. 
Published: June 2021

In muscular dystrophy research, it is pertinent to analyze the force of the muscles affected by the disease to assess pathology and to assess potential effectiveness of interventions. Maximal tetanic contractions are the most commonly used protocol to assess muscle function. However, the vast majority of in vivo muscle function is at submaximal contraction. Moreover, although force of contraction is assessed, the kinetics of the contraction are rarely reported. To investigate the effect of muscular dystrophy on twitch contraction kinetics, isolated diaphragm and extensor digitorum longus (EDL) of 20‐week untreated “het” (dystrophin deficient and utrophin haploinsufficient) and wild‐type C57 mice were analyzed. In isolated diaphragm strips, Time‐to‐Peak (TTP) is slower in “het” mice (14.2 ± 0.3 ms) than in C57 (11.5 ± 0.3 ms) mice, while no significant difference was identified for Time for 50% Relaxation (RT50) of the twitch contraction. In isolated EDL muscles, TTP was also slower in “het” mice (24.0 ± 0.4 ms) compared to C57 mice (20.7 ± 0.3 ms). Unlike the diaphragm, it was found that RT50 of EDL muscles is also disease‐dependent. The RT50 of “het” mice (15.3 ± 0.6 ms) was significantly slower than the C57 model (11.8 ± 0.2 ms). These findings suggest that not only is the maximal force of the muscles important, but also the kinetics of the muscles are impacted by disease.This abstract is from the Experimental Biology 2018 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.