bluuebunny/crossref_metadata_2025_split huggingface dataset:
Protective Effects of Ocimum sanctum Leaf Extract against Lead‐induced Toxicity in Rat Brain
Authors: Kilari Eswar A , Putta Swathi , Qureshi Absar Ahmed 
Published: June 2021

Lead is a known as multi organ toxicant majorly on neuronal system. The present study evaluates the protective role of Ocimum sanctum leaf extract against lead induced oxidative damage to the different regions of the brain. Albino Wistar rats were divided into three groups each consisting of 10 rats. The Ocimum sanctum extract was administered orally at 200 and 400 mg/kg along with 500ppm of lead acetate dissolved water for four weeks to two groups of rats. The control group was administered with sodium acetate dissolved in water for four weeks. At the end of four weeks study period, the rats were sacrificed and brains were isolated and different regions like cerebellum, hippocampus, frontal cortex and brain stem were separated. The delta ALAD levels were estimated in the serum samples. The antioxidant enzymes (SOD, CAT and GST), non enzymatic antioxidants (GSH), lipid peroxidation (TBRS), metal content, tissue nitrite and protein carbonyl content (PCO) were measured in all separated parts of the brain. Finally the histopathology was conducted for all the selected parts of the brain. The treatment with Ocimum sanctum showed significant elevation of serum delta ALAD levels, antioxidant enzymes (SOD, CAT and GST) and compared to that of lead acetate control group. The lipid peroxidation and PCO were found to be lowered with Ocimum sanctum treatment in lead acetate induced brain toxicity. The histopathological studies indicated that the Ocimum sanctum showed marked amelioration of damaged structure of the parts of the brain caused by the lead acetate in rats. The results indicating that the protective effect was found to be more in cerebellum and hippocampus compared to other parts of brain.ConclusionsThe protective effect of Ocimum sanctum might be due to its antioxidant activity.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 Microdialysis Assay for Assessing sGC Stimulator Target Engagement in the Rat Liver
Authors: O'Connell Ryan P. , Hall Katherine C. , Jacobson Sarah , Tobin Jenny V. , Masferrer Jaime L. 
Published: June 2021

Dysfunction of the nitric oxide (NO)‐soluble guanylate cyclase (sGC)‐cyclic guanosine monophosphate (cGMP) pathway has been implicated in numerous cardiovascular, metabolic, and fibrotic diseases. sGC stimulators are small molecules that bind sGC and synergize with NO to increase the enzymatic formation of cGMP and amplify downstream signaling. In preclinical models of liver disease, sGC stimulators have been shown to attenuate liver steatosis, inflammation, and fibrosis. To demonstrate their engagement in the sGC‐cGMP pathway in vivo, a unique microdialysis assay was developed to directly measure changes in cGMP levels in the rat liver.A midline laparotomy was performed on anesthetized male, Sprague Dawley rats (225–250 g) to expose the liver. A 25‐gauge needle was used to place a dialysis probe within the right medial lobe. The probe was perfused with saline at a rate of 2.5 μL/min and dialysate collected in one‐hour periods. Following baseline collection, test compound was administered intravenously, and dialysate collected for the following 2 hours. The NO donor sodium nitroprusside (SNP, 300 nM) was dosed via retrodialysis at study's end as a positive control. cGMP in the dialysate was measured using an enzyme immunoassay.In a study investigating our clinical‐stage sGC stimulator, praliciguat, an oral 3‐mg/kg dose produced a 3.9 ± 1.7‐fold increase (p<.0001) in cGMP from baseline levels of 0.8 ± 0.1 nM. A 1‐mg/kg dose caused a 1.7 ± 0.1‐fold increase in cGMP (p<.001), while the effect of a 0.3‐mg/kg dose was not significant. On average, SNP induced a 4.7 ± 1‐fold increase in cGMP (p<.0001).In summary, our microdialysis assay enabled in vivo monitoring of sGC stimulation in the rat liver. Beyond demonstrating the effects of praliciguat, this method can be used to evaluate and characterize other sGC stimulators for their capacity to amplify liver cGMP production. Further, this assay helps to link previously observed hepatic effects of sGC stimulators in disease models with an enhancement of sGC signaling.This abstract is from the Experimental Biology 2019 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
Ulcerogenic Action of Indomethacin on Gastric and Small Intestinal Mucosa in C57/BL6/J and TRPV1 Knockout Mice
Authors: Filaretova Ludmila , Yarushkina Natalia , Sudalina Maria , Punin Yuriy 
Published: June 2021

Capsaicin‐sensitive afferent neurons are involved in maintenance of gastrointestinal mucosa integrity. Capsaicin action is mediated through TRPV1 receptors. The aim of the present study was to elucidate the role of these receptors in providing of protective or/and pathological mechanisms under circumstances of indomethacin (IM)‐induced injury in gastric and small intestinal mucosa. We compared vulnerability of gastric and small intestine mucosa to IM action in mice with genetically deleted TRPV1 receptor and C57/BL6/J mice. IM‐induced injury was assessed by macroscopically as well as histologically; the somatic pain sensitivity was estimated by tail flick latency (tail flick test); plasma corticosterone levels and body weight were also monitored. A single IM administration (35 mg/kg, sc) into fasting (24 h) mice caused the formation of gastric erosions 4 h after injection and, then, after refeeding, formation of the small intestine injury which was visible 24, 48, 72 h after the injection. Although IM‐induced gastrointestinal injury was detectable in both of C57/BL6/J and TRPV1 knockout mice, area of gastric damage was greater in C57/BL6/J than in TRPV1 knockout mice, whereas the small intestine injury (48 and 72 h after IM injection), on contrary, prevailed in TRPV1 knockout mice compared to C57/BL6/J mice. TRPV1 knockout mice also showed an increased tail flick latencies and lower plasma corticosterone levels than C57/BL6/J mice. The data suggests that in TRPV1 knockout mice the gastric mucosa is less vulnerable to ulcerogenic IM action compared to C57/BL6/J mice, whereas their small intestinal mucosa, on contrary, is more susceptible to IM ulcerogenic action than in C57/BL6/J mice.Support or Funding InformationThe study was supported by grant of Russian Science Foundation (RSF) N 14‐15‐00790 and grant of RFBR N 16‐04‐01196aThis abstract is from the Experimental Biology 2019 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
A Study on the Effect of N453 from Fungi on Inhibiting CFTR Chloride Channels in Intestinal Epithelial Cells
Authors: Chutinun Onjira , Akrimajirachoote Nattaphong , Muanprasat Chatchai 
Published: June 2021

Secretory diarrhea remains a global health problem, especially in developing countries. Cyclic AMP‐stimulated intestinal Cl− secretion plays a crucial role in pathogenesis of diarrheas including cholera and traveler's diarrhea. Inhibition of cAMP‐activated Cl− channel is considered as drug target for secretory diarrhea. The main objective of this study was to identify a novel inhibitor of the cystic fibrosis transmembrane conductance regulator (CFTR), cAMP‐activated Cl− channel, from a screen of fungal metabolites. A fungal derivative compound N453 was identified as an active compound that could inhibit cAMP‐activated Cl− secretion in intact T84 cells with a half maximal inhibitory concentration (IC50) of ~ 0.78 μM in Ussing chamber experiments. Importantly, MTT assays revealed that N453 (up to100 μM) did not affect cell viability, suggesting that inhibition of cAMP‐activated Cl− current was not due to cytotoxic effect. In apical Cl− current analysis, N453 suppressed CFTR‐mediated apical Cl− current in a concentration‐dependent manner with IC50 of ~ 2 μM. On the other hand, the Na+‐K+ ATPase activity was not affected by N453. Of interest, N453 concentration‐dependently inhibited basolateral cAMP‐activated K+ channels with an IC50 of ~ 23 μM. In conclusion, this study demonstrated that N453 inhibits Cl− secretion in intestinal epithelial cells, at least in part, by inhibiting CFTR‐mediated apical Cl− current and basolateral cAMP‐activated K+ channels. N453 may be useful as potential therapy of secretory diarrheas.Support or Funding InformationThis work was supported by the NSTDA Chair Professor grant (the Fourth Grant) of the Crown Property Bureau Foundation, Faculty of Science Mahidol University, Faculty of Graduate Studies Mahidol University and Tonkla Ramathibodi.This abstract is from the Experimental Biology 2019 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
Pentadecapeptide BPC 157 Counteracts the Adverse Effect of Lithium Overdose in Rats
Authors: Strbe Sanja , Filipcic Igor , Seiwerth Sven , Sikiric Predrag 
Published: June 2021

We documented that stable gastric pentadecapeptide BPC 157 mitigates lithium‐induced intoxication in rats. BPC 157, LD1 not achieved, was implemented as an anti‐ulcer peptide in inflammatory bowel disease trials and now in a multiple sclerosis trial (Curr Pharm Des. 2018;24(18):1990–2001). Previously, BPC 157 counteracts consequences of dopamine (D), receptors blockade (neuroleptics‐induced catalepsy, prolonged QT intervals, sphincters dysfunction and gastric lesions), much like over‐stimulation (amphetamine acute and chronic disturbances; much like D‐receptors supersensitivity (amphetamine after haloperidol)), nigrostriatal lesions (MPTP Parkinsogenic neurotoxin), D‐vesicles depletion (reserpine) (see Sikiric et al., Brain‐gut axis and pentadecapeptide BPC 157: Theoretical and practical implications. Curr Neuropharmacol. 2016;14(8):857–865. Review.). Similarly, BPC 157 counteracts immobility more than imipramine in depression‐models (Porsolt's and chronic unpredictable stress‐open field) and induces 5‐HT release in particular brain areas (nigrostriatum) when given peripherally, acute and chronically (see Sikiric et al., Brain‐gut axis and pentadecapeptide BPC 157: Theoretical and practical implications. Curr Neuropharmacol. 2016;14(8):857–865. Review.). In this study, we applied lithium (500 mg/kg/day, intraperitoneally, once daily through 3 subsequent days). Medication (mg/kg) includes BPC 157(0.01; 0,00001), L‐NAME (5.0), L‐arginine (100.0), applied alone and /or together) while control rats received an equivolume of saline (5 mL/kg). Thereafter, at 20 minutes after medication, we assessed muscular weakness (score 1–5) during next 8 minutes. Then, for next 5 minutes, we recorded ECG. At 3 hours thereafter, the brain, heart, quadriceps muscle and diaphragm muscle were used for histopathological analysis. Regularly, lithium immediately produced severe syndrome (severe muscular weakness and prostration, reduced quadriceps muscle fibers and diaphragm, myocardial infarction, and edema of various brain areas, the most prominent in the cerebral cortex). The worsening appeared with subsequent applications. L‐NAME and L‐arginine, given separately, both induced severe aggravation. This aggravation disappeared when L‐NAME and L‐arginine were given together. Contrarily, given alone or together with NO‐agents, BPC 157 reduced muscular weakness and prostration and muscle damage and mitigated lithium induced myocardial damage. BPC 157 reduced nerve damage and brain edema.ConclusionBPC 157 can be a therapy of lithium intoxication.Support or Funding InformationThis work was supported by the Ministry of Science, Education and Sports, Republic of Croatia [grant number 108‐1083570‐3635].This abstract is from the Experimental Biology 2019 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
Identification of BKca channel activation effect of Kushen extract containing Kurarinone to ease urinary incontinence or overactive bladder
Authors: Lee Narasaem , Han Jisun , Jo Heeji , Jang Sung Joo , Ryu Seunghwan , Moh Sang Hyun , Park Chul‐Seung 
Published: June 2021

The large‐conductance Ca2+‐activated K+ channel (or BKCa channel) requires membrane depolarization or increased intracellular Ca2+ for its activation. This channel makes one of the most effective ways to generate membrane hyperpolarization and regulates several key physiological processes. In the bladder, BKCa channel evokes the relaxation of urinary bladder smooth muscle and thus represents a therapeutic target for urinary incontinence (UI) or overactive bladder (OAB). In our previous study, we reported that kurarinone, a prenylated flavanone from Sophora flavescens, potently activates BKCa channel and established its therapeutic potential for treating OAB (Lee et al., Mol Pharmacol. 90:140, 2016). Subsequently, we identified five more flavonoids from Kushen, the dry root of S. flavescens. including kuraridinol, a prenylated chalcone, significantly potentiating BKCa channel activity (Lee et al., Biol Pharm Bull. 41:1295, 2018). In this study, we analyzed the components of Kushen extracts isolated with differential ethanol concentrations and tested their efficacy for BKCa channel activation. We found that the potentiation effect of various extracts for BKCa channel correlates well with the concentration of kurainone in the extracts. In addition, the Kushen extracts potently relaxed acetylcholine‐induced contraction of rat bladder smooth muscle and thus decreased the micturition frequency of rats with overactive bladder symptoms. Our results strongly support the idea that the Kushen extract containing kurarinone as the main ingredient can be utilized as nutraceutical or functional food against lower urinary tract symptom such as UI or OAB.Support or Funding InformationThis work was supported by Korea Institute of Planning and Evaluation for Technology in Food, Agriculture, Forestry (IPET) through Agri‐Bio industry Technology Development Program, funded by Ministry of Agriculture, Food and Rural Affairs (MAFRA) (317070‐4)This abstract is from the Experimental Biology 2019 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
The TRPP2 (PC2)‐dependent ion channel of renal primary cilia also requires TRPM3
Authors: Bryant Gillian , Kleene Nancy , Siroky Brian , Landero‐Figueroa Julio , Dixon Brad , Pachciarz Nolan , Lu Lu , Kleene Steve 
Published: June 2021

INTRODUCTIONPrimary cilia are found in numerous places throughout the body and serve many different functions (e.g., cell migration, intercellular signaling, and sensation). Mutations in primary cilia cause many human diseases. Polycystic kidney disease (PKD) is a ciliopathy that results from mutations in the ciliary proteins polycystin 1, TRPP2/polycystin 2 (PC2), or polyductin/fibrocystin. Although the mechanism is not completely understood, abnormally low intracellular Ca2+ levels are thought to be an early step in cystogenesis.AIMWe study renal ciliary ion channels in search of a pharmacological means of compensating for the lower cytoplasmic Ca2+. One channel present in renal tissue is TRPM3. When testing cilia with a TRPM3 agonist, we did not detect a channel with traits characteristic of TRPM3, but, to our surprise, the agonist activated the TRPP2‐dependent ciliary channel.HYPOTHESISThe TRPP2‐dependent channel also requires TRPM3.METHODSWe used electrophysiology, qPCR, western blotting, and immunocytochemistry with a murine immortalized renal epithelial cell line.RESULTSThe TRPP2‐dependent channel was activated by a known agonist of TRPM3 and inhibited by a known antagonist of TRPM3. The TRPP2‐dependent channel current was not detectable when TRPM3 was knocked out with CRISPR/Cas9 genome editing. Knocking out TRPP2 did not reduce TRPM3 mRNA expression, and knocking out TRPM3 did not markedly decrease TRPP2 mRNA expression. Knocking out TRPM3 did not change TRPP2 protein expression. Immunocytochemistry indicated that removal of TRPM3 expression reduced the amount of TRPP2 trafficking to the cilium.CONCLUSIONThe TRPP2‐dependent channel also requires TRPM3. Further studies will be required to determine the nature of the interaction between TRPM3 and TRPP2 and if the interaction can be manipulated to reduce cystogenesis in PKD.Support or Funding InformationThis work was done with financial support from The University of Alabama Hepato/Renal Fibrocystic Disease Core Center, Cincinnati Children's Hospital Medical Center (CCHMC), and the University of Cincinnati.This abstract is from the Experimental Biology 2019 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
Rififylin (RFFL), a Novel Regulator of Transient Outward (Ito) Potassium Channels
Authors: Roder Karim , Kabakov Anatoli , Moshal Karni S , Xie An , Turan Nilüfer N , Lu Yichun , Koren Gideon 
Published: June 2021

The expression of sarcolemmal ion channels is a net result of the rate of coordinated biosynthesis, proper folding, forward trafficking, recycling, and degradation. The importance of these processes was highlighted by several genome‐wide association studies that reported novel loci that modify the QT interval and are located in or near genes involved in protein trafficking and degradation (Roder et al., 2014; Roder et al. 2018). One of the genes identified, the ring finger and FYVE‐like domain containing E3 ubiquitin protein ligase rififylin (RFFL), has also been shown to regulate cardiac excitation and growth in a congenic rat strain (Gopalakrishnan et al. 2011). As transient outward K+ currents (Ito) are essential during the whole repolarization phase of the action potential in rodents, we hypothesized that RFFL could regulate cardiac excitation through Ito‐encoding channels. Adult rabbit cardiomyocytes with adenovirus‐expressed RFFL exhibited reduced Ito,f. In HEK cells, transient RFFL overexpression caused polyubiquitination and downregulation of total and surface levels of Kv4.2 and Kv4.3 as well as the corresponding current Ito,f. In contrast, in transiently co‐transfected HEK cells, overexpressed RFFL monoubiquitinated and upregulated total and membrane levels of Kv1.4 and the corresponding current Ito,s in a RING domain‐dependent manner. We conclude that RFFL, through its interactions with and ubiquitination of Ito‐encoding channels Kv4.2, Kv4.3 and Kv1.4 regulate their forward trafficking and degradation, which may affect cardiac action potential shape and duration.Support or Funding InformationNHLBI grant 5R01HL134706‐02 (GK).This abstract is from the Experimental Biology 2019 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
LITAF regulates action potential duration by modulating NEDD4‐1‐mediated degradation of L‐type calcium channels
Authors: Moshal Karni S , Roder Karim , Kabakov Anatoli , Turan Nilüfer N , Xie An , Werdich Andreas A , Kim Tae Yun , Cooper Leroy L , Lu Yichun , Terentyev Dmitry , Choi Bum‐Rak , MacRae Calum A , Koren Gideon 
Published: June 2021

RationaleThe QT interval is a predictor of arrhythmias and sudden cardiac death. A genetic variant upstream of the gene encoding LITAF (lipopolysaccharide‐induced tumor necrosis factor) is associated with a prolonged QT interval.ObjectiveWe sought to determine the role of LITAF in modulating cardiac excitation at the cellular and organ levels.Methods and ResultsMorpholino knockdown of LITAF in zebrafish embryos resulted in a robust increase in calcium transients in optically mapped hearts, which primarily depends on calcium influx. Adenovirally expressed LITAF shortened action potential duration and lowered Cava1c expression in neonatal rabbit cardiomyocytes, whereas a shRNA‐mediated LITAF knockdown had the opposite effect. Using whole‐cell patch clamping and western blotting of 3‐week old rabbit cardiomyocytes, overexpressed LITAF resulted in a decrease in ICa,L and Cava1c expression, whereas a LITAF knockdown increased ICa,L and Cava1c protein levels. Live‐cell confocal calcium imaging detected a decrease in calcium transients in LITAF‐overexpressing adult rabbit cardiomyocytes, which was accompanied by lower Cava1c expression. In tsA201 cells, overexpressed LITAF downregulated the total and surface pools of Cava1c via increased Cava1c ubiquitination and subsequent lysosomal degradation. Co‐immunoprecipitation and immunofluorescence experiments showed complex formation and co‐localization of LITAF and L‐type calcium channels (LTCC) in tsA201 cells and adult rabbit cardiomyocytes, respectively. In tsA201 cells, NEDD4‐1, but not its catalytically inactive form NEDD4‐1‐C867A, increased Cava1c ubiquitination compared to control. Cava1c ubiquitination was further increased by co‐expressed LITAF and NEDD4‐1 but not NEDD4‐1‐C867A. Finally, adenovirally co‐expressed NEDD4‐1 shRNA abolished the negative effect of LITAF on ICa,L and Cava1c expression in 3‐week old rabbit cardiomyocytes.ConclusionsThe data suggest that LITAF promotes NEDD4‐1‐mediated ubiquitination and subsequent degradation of cardiac LTCC, thereby controlling LTCC membrane expression and function and thus APD and the QT interval.Support or Funding InformationNHLBI grant 5R01HL134706‐02 (GK).This abstract is from the Experimental Biology 2019 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
Discovery of Pranlukast in a High‐Throughput Screen for Novel Inhibitors of LRRC8 Volume Regulated Anion Channels
Authors: Figueroa Eric E , Kramer Meghan , Denton Jerod S 
Published: June 2021

Cells critically regulate their volume in response to hypotonic cell swelling by transporting chloride and small organic osmolytes out of the cell through volume regulated anion channels (VRACs) encoded by members of the recently defined LRRC8 gene family. VRACs are ubiquitously expressed in mammalian cells and have been implicated in diverse cellular functions in addition to cell volume regulation, including endothelial cell calcium signaling and pancreatic beta cell insulin secretion. The lack of potent and specific pharmacological tool compounds represents a critical barrier to evaluating the integrative physiology and therapeutic potential of VRACs. We therefore carried out a high‐throughput screen (HTS) of 1,184 of FDA approved drugs using a fluorescence‐based assay for novel inhibitors of native VRACs expressed in HEK‐293 cells. The assay measures the quenching of intracellular YFP by iodide influx through VRACs activated by hypotonic‐induced cell swelling. In this screen, the cysteinyl leukotriene receptor 1 (CysLT1R) antagonist, Pranlukast, was identified as a novel inhibitor of VRAC. In YFP quenching assays and patch clamp electrophysiology experiments, Pranlukast inhibits HEK‐293 cell VRAC reversibly and dose‐dependently with an IC50 of ~6 μM and ~239 nM, respectively. Interestingly, the CysLT1R pathway has been implicated in activation of VRAC in other cell types, raising the possibility that Pranlukast inhibits VRAC indirectly via the CysLT1R pathway. Using a series of fluorescence assays and qPCR we show that CysLT1R is not functionally expressed in HEK‐293 cells. Additionally, using a structurally distinct CysLT1R inhibitor, Zafirlukast, did not inhibit VRAC. These studies provide clarifying insights into the putative role of CysLT1 receptor signaling in regulation of VRACs and identify Pranlukast as a structurally novel small‐molecule inhibitor of LRRC8 channels.Support or Funding InformationRuth L. Kirschstein National Research Service Award F31 (1F31DK120225‐01)This abstract is from the Experimental Biology 2019 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.