bluuebunny/crossref_metadata_2025_split huggingface dataset:
Potency Adjusted Bovine Heparin is Comparable to Porcine Heparin in Patients Undergoing Open Heart Surgery
Authors: Gomes Walter , Braile Domingo , Leal Joao Carlos , Guizilini Solange , Regenga Marisa , Kouta Ahmed , Walenga Jeanine , Jeske Walter , Hoppensteadt Debra , Ramacciotti Eduardo , Nader Helena , Lima Marcelo , Fareed Jawed 
Published: June 2021

IntroductionHeparin obtained from bovine intestinal mucosa is being considered for reintroduction in cardiovascular, surgical, and interventional procedures. Although the anticoagulant potency of bovine heparin is weaker (130–140 U/mg) than that of porcine heparin (180–200 U/mg), when potencies are adjusted to equivalent USP U/mL activity, both porcine and bovine heparin exhibit comparable anticoagulant activities in in‐vivo and in‐vitro settings.AimThe primary aim of this study was to compare bovine and porcine heparins at equivalent dosing in terms of anticoagulation and potential bleeding effects in patients undergoing coronary artery bypass graft (CABG) surgery.Materials and MethodsCoagulation parameters and biomarkers of inflammation were measured in 32 patients included in a single center, blinded, controlled clinical trial, randomized to CABG surgery with either bovine or porcine heparin. Equivalent dosages of 28,500 units (potency adjusted to 10,000 U/mL) total dose heparin were used. Blood samples were collected at multiple times during and after surgery to measure global clotting activity (ACT, APTT) and amidolytic anti‐Xa levels. TNFa, IL‐6, and TFPI antigen were measured by ELISA; activated protein Cwas measued by a clot‐based assay. A preliminary report of these results has been published (Int J Cardiol 223:611–612, 2016).ResultsAll coagulation parameters (ACT, APTT, anti‐Xa, residual bleeding) demonstrated comparable heparinization (p ≥ 0.05). The only difference between groups was an increased amount of protamine required to neutralize bovine heparin after surgery determined by ACT; expected due to the higher gravimetric quantity of bovine heparin used to adjust the U/mL potency. No differences were found between groups in levels of acute inflammatory markers (p ≥ 0.05).ConclusionThis study shows that bovine and porcine heparins at potency adjusted dosages are equivalent anticoagulant drugs in CABG surgery resulting in comparable perioperative coagulation, bleeding, and anti‐inflammatory effects.Support or Funding InformationNone.This abstract is from the Experimental Biology 2019 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
Mechanistic Differences in the Antiplatelet Effects of DT‐678 and Existing P2Y12 Antagonists
Authors: Henderlong Dawn Shirley , Deters Mackenzie Rae , Christian Barbara DaShawn , Lauver Dale Adam 
Published: June 2021

Arterial thrombosis, leading to tissue ischemia, is the most common cause of death in the developed world. Dual antiplatelet therapy with low dose aspirin and a P2Y12 antagonist is widely used to prevent thrombus formation. The P2Y12 antagonist, clopidogrel, is a prodrug that undergoes a complex metabolism process involving cytochrome P450 enzymes (CYP450) to produce the biologically active metabolite (AM). The AM covalently binds to the P2Y12 receptor on platelets thereby inhibiting receptor activation by ADP resulting in the inhibition of platelet activation. Approximately 60–70% of Asians and 30% of Caucasians carry a genetic polymorphism in the CYP450 that prevents the production of the AM. Our laboratory has previously reported the development of DT‐678, a novel conjugate of the AM that does not require CYP450s. Instead, the drug is nonenzymatically activated through a glutathione exchange reaction.Previous studies have demonstrated that DT‐678 is equally as effective as clopidogrel and ticagrelor at inhibiting platelet activation, while requiring a lower dose than clopidogrel. Additionally, we have shown that while clopidogrel and ticagrelor induce a 1‐ to 2‐fold increase in bleeding time, DT‐678 does not induce a significant increase in bleeding time. The purpose of the present study was to compare the antithrombotic effects of DT‐678 to clopidogrel and ticagrelor after oral administration. To do this, we used a New Zealand white rabbit model of FeCl3‐induced carotid artery injury. All three antiplatelet drugs showed an equal efficacy in prolonging the time to occlusion after FeCl3 application compared to vehicle. Further data suggest that DT‐678 differentially affects calcium mobilization in platelets and other cell types compared to clopidogrel and ticagrelor after application of 2MeSADP. Together, these data suggest mechanistic differences in the action of DT‐678 compared to existing P2Y12 antagonists and may explain differences in the bleeding times observed.Support or Funding InformationNational Institutes of Health National Heart, Lung and Blood Institute [1R43HL139380‐01]This abstract is from the Experimental Biology 2019 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
High Fat, High Sugar Diet Induces Metabolic Changes in Articular Cartilage that are Reversed with Low Intensity Exercise and Genistein Treatment
Authors: Lopez Janelle , Castro Monica , Al‐Nakkash Layla , Broderick Tom L , Plochocki Jeffrey H 
Published: June 2021

High fat, high‐sugar diet (HSFD) has been shown to induce obesity, metabolic dysregulation, and inflammation in peripheral tissues. Inflammation within cartilage tissue is associated with metabolic changes that lead to arthrosis. In this study, we examine the effects of HSFD on expression of col10a and MMP‐13 in articular cartilage of the knee joint. Both of these proteins are markers for inflammation in chondral tissue. In the presence of proinflammatory cytokines, col10a expression by chondrocytes decreases, while MMP‐13 increases. We also examined the effects of low intensity exercise and genistein treatment on cartilage metabolism. Both of these treatments have been shown to improve bone health, but their effects on diet‐induced cartilage inflammation are unclear. We randomly assigned 100 mice to five treatment groups of equal sizes: lean controls, HFSD fed, HFSD + exercise, HFSD + genistein, and HFSD + exercise + genistein. HFSD included 60% fat in chow and 42 g/L sugar in the drinking water. Exercise training included treadmill running for 180 min/week. Genistein was given in the diet at 600 mg/kg. Duration of treatment was 12 weeks. Col10a expression was significantly reduced in mice fed HFSD versus lean controls (P < 0.05). Mice treated with exercise and genistein in combination exhibited decreased col10a expression that was similar to levels expressed in cartilage of lean controls. MMP‐13 expression did not differ significantly among the treatment groups (P > 0.05). Examination of articular cartilage stained with H&E revealed the thickness of the uncalcified zones was significantly reduced in mice fed HFSD in comparison to lean control mice (P < 0.05). Staining in this zone also reflected reduced proteoglycan content in the cartilage of mice in the HFSD treatment group. These findings suggest articular cartilage of mice fed HFSD exhibit signs of inflammation and reduced capacity to resist compression. Treatment with genistein and exercise in combination resulted in articular cartilage that more closely resembles cartilage of lean mice fed a standard diet. Low‐intensity exercise and genistein intake may prove beneficial to joint health in the obese condition.This abstract is from the Experimental Biology 2019 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
Lapatinib Metabolism in CYP3A5‐Genotyped Primary Human Hepatocytes
Authors: Bissada Jennifer , Abouda Arsany , Jackson Klarissa 
Published: June 2021

Drug‐induced liver injury is associated with the oral tyrosine kinase inhibitor lapatinib, which is used in metastatic breast cancer therapy. Lapatinib is extensively metabolized by cytochrome P450 (CYP) 3A4 and CYP3A5 to yield an O‐debenzylated metabolite (Lap‐OH), which can undergo further oxidation to a reactive quinoneimine. We hypothesized that polymorphic expression of CYP3A5 may contribute to individual differences in lapatinib bioactivation and influence hepatic exposure to reactive, potentially toxic metabolites. The objective of this study was to assess the metabolism of lapatinib in CYP3A5‐genotyped human hepatocyte cultures, and evaluate whether CYP3A5 genotype and activity lead to differences in lapatinib metabolite formation. Commercially available human hepatocytes were obtained from XenoTech (pooled) and BioIVT (individual). CYP3A5 genotype (*1/*1, *1/*3, and *3/*3) and donor characteristics (sex, age, ethnicity) were provided by the companies. Lapatinib (10 μM) was incubated with genotyped human hepatocytes in suspension for 2 h. Separate incubations were carried out to measure CYP3A and CYP3A5‐selective activity using probe substrates, midazolam (2.5 μM) and T‐5 (5 μM), respectively. Metabolite formation was analyzed by liquid chromatography‐tandem mass spectrometry. The results from experiments with genotyped human hepatocytes from pooled donors showed that formation of Lap‐OH was higher in CYP3A5 expressers (CYP3A5 *1/*1 and CYP3A5*1/*3 donors) compared to CYP3A5 non‐expressers (CYP3A5*3/*3 donors). CYP3A activity, as measured by midazolam 1’‐hydroxylation, and CYP3A5‐selective activity, as measured by T‐5 N‐oxidation, were also higher in CYP3A5 expressers (*1/*1 and *1/*3) compared to non‐expressers (*3/*3). In incubations with single‐donor human hepatocytes (n = 15), the levels of Lap‐OH generated were 2.2‐fold higher in CYP3A5 expressers compared to non‐expressers (p = 0.0497). Moreover, formation of Lap‐OH was 2.5‐fold higher in hepatocytes from female donors compared to males (p = 0.0390). Collectively, these results indicate that CYP3A5 genotype and donor sex influence lapatinib bioactivation in human hepatocyte cultures. Future studies will examine the cytotoxicity of lapatinib in primary human hepatocytes to determine the impact of CYP3A5 genotype, sex, and other individual factors on lapatinib‐induced hepatocellular injury.Support or Funding InformationThis research is funded by the National Cancer Institute of the National Institutes of Health (K01CA190711). Research reported here is solely the responsibility of the authors and does not necessarily represent the official views of the National Institutes of Health.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 Short‐Term and Long‐Term Dexamethasone Action on Expression of Glucocorticoid and Mineralocorticoid Receptors in the Gastric Mucosa
Authors: Filaretova Ludmila , Sudalina Maria 
Published: June 2021

Glucocorticoids may have dual action on the stomach: gastroprotective and proulcerogenic one. Our previous study was designed to investigate how physiological gastroprotective action of glucocorticoids can be transformed to pathological ulcerogenic effect. The results obtained demonstrate that single injection of dexamethasone at the dose of 1 mg/kg may attenuate or aggravate indomethacin‐induced gastric erosions depending on the time of its injection before indomethacin (prolongation of dexamethasone action). At the present study to elucidate the mechanisms of transformation of gastroprotective action of dexamethasone into ulcerogenic one we tested the hypothesis that disbalance between the glucocorticoid and mineralocorticoid receptors (GR and MR) may contribute to this transformation. To test the hypothesis, effect ща dexamethasone at the dose of 1 mg /kg, (i.p.) on the expression of GR and MR in the gastric mucosa was studied 1 h (a short‐term action when the gastroprotective action was observed) and 24 h (long‐term action when the proulcerogenic effect was observed) after its administration. The expression of GR and MR was detected by a immunohistochemical staining technique using α‐glucocorticoid (ab3580; 1:500. Abcam) and mineralоcorticoid (ab41912; 1:500. Abcam) receptor antibodies. The results of the staining were assessed by a morphometric study of microscopic images in five fields of view using the ImajeJ software. A distribution of cytoplasmic staining was estimated with relative stained area (%) in gastric glands and smooth muscle cells of muscularis mucosae. We found that GR are presented in the parietal cells, whereas MR are ubiquitous. Dexamethasone provoked translocation of both GR and MR into nuclei. The short‐term effect of dexamethasone resulted in a decrease of cytoplasmic GR and their translocation into the cell nuclei. The long‐term dexamethasone exposure increased cytoplasmic GR expression not accompanied by nuclear translocation. The short‐term dexamethasone action caused an increase of cytoplasmic MR level, but did not lead to nuclear translocation of these receptors. The long‐term action of dexamethasone induced nuclear translocation of MR, without significant changes of MR expression in cytoplasm. The results suggest that disbalance between GR and MR may contribute to transformation of gastroprotective action of dexamethasone to proulcerogenic effect and support the hypothesis.Support or Funding InformationThe study was supported by grant of Russian Science Foundation (RSF) N 14‐15‐00790.This abstract is from the Experimental Biology 2019 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
Stable Gastric Pentadecapeptide BPC 157 Antagonized Local Anesthetic Effect of Lidocaine
Authors: Lozic Marin , Stambolija Vasilije , Kovacevic Josip , Zivanovic‐Posilovic Gordana , Suran Jelena , Duplancic Bozidar , Mirkovic Ivan , Kokot Antonio , Drmic Domagoj , Seiwerth Sven , Sikiric Predrag 
Published: June 2021

We documented that the stable gastric pentadecapeptide BPC 157 antagonized local anaesthetic effect of lidocaine. 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). Stable gastric pentadecapeptide BPC 157 was previously shown as a cardioprotective compound in a model of arrhythmia induced by bupivacaine toxicity where it counteracts arrhythmias and prevents lethal outcome (Eur J Pharmacol. 2016 Dec 15;793:56–65) much like in other cardiotoxicity mainly related to potassium disturbances, both hyperkalemia and hypokalemia, in vivo and in vitro (HEK293 cells) (Curr Pharm Des. 2018;24(18):1990–2001). We used Wistar Albino male rats, underwent regional blocks with lidocaine (spinal intrathecal block (lidocaine 6 mg/kg, 0.1 ml/rat, 550 g b.w.) or axillary block (lidocaine 15 mg/kg, 0.3 ml/rat, 220 g b.w.). Rats received BPC 157 (10 μg, 10 ng, 10 pg/kg intraperitoneally or intragastrically) or an equivolume of saline (5 ml/kg), either immediately or at 10 min when local anesthesia was fully established. Consistently, while lidocaine application produced a prolong function failure, all BPC 157 regimens significantly shortened time to full function recovery, either given immediately after lidocaine application or later, in the conditions of full local anesthesia. In other experiments, using a hot plate (55°C for 3 minutes) when rat hind paws were infiltrated with 2% lidocaine (0.1 ml/paw), a subsequent infiltration with BPC 157 (10 μg, 10 ng, 10 pg/kg) results in the faster feet lifting and much less edema (Figure 1). Finally, ECG recording documented that the mentioned regimens of BPC 157 markedly counteracted the lidocaine‐induced arrhythmias as well. Therefore, it may be possible that pentadecapeptide BPC 157 acts as the missing antidote to local anesthethics, and potentially deleterious and even life threatening adverse effects of toxic doses of local anesthethics would be markedly attenuated or even abolished.Support or Funding InformationThis work was supported by the University of Zagreb scientific support fund [grant number BM099].This abstract is from the Experimental Biology 2019 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
Small GTPase Rab7 Mediates Aquaporin‐2 Recycling and Apical Trafficking
Authors: WANG WEI‐LING , YANG CHAN‐WEI , WONG KIT YEE , YU MING‐JIUN 
Published: June 2021

Water permeability of the kidney collecting ducts is regulated by the abundance of the molecular water channel protein aquaporin‐2 (AQP2) in the apical plasma membrane of the collecting duct cells. The apical AQP2 abundance in turn is regulated by trafficking of AQP2‐containing intracellular vesicles to and from the apical membrane by the peptide hormone vasopressin. Here we used confocal microscopy to follow how AQP2 trafficked among biotin‐labeled apical membrane, Rab5+ early endosome, Rab7+ late endosome, and Rab11+ recycling endosome in response to vasopressin. Before vasopressin stimulation, most AQP2 stayed in Rab5+ early endosome and Rab11+ recycling endosome. In response to vasopressin, AQP2 in the Rab11+ recycling endosome trafficked first to the apical membrane followed by AQP2 in the Rab5+ early endosome. Upon vasopressin withdrawal, apical AQP2 returned to the Rab5+ early endosome before joining the Rab11+ recycling endosome. Rab5 knockdown slowed APQ2 internalization upon vasopressin withdrawal. Rab7 knockdown resulted in AQP2 accumulation in the Rab5+ early endosome upon vasopressin withdrawal, consistent with a role of Rab7 in AQP2 sorting to the late endosome. However, Rab7 knockdown also reduced AQP2 in the Rab11+ recycling endosome upon vasopressin withdrawal, suggesting that Rab7 participates in AQP2 sorting from the early endosome to the recycling endosome. In fact, in the Rab7 knockdown cells vasopressin‐stimulated apical AQP2 trafficking was greatly impaired. These results indicate that internalizing AQP2 first arrives at the early endosome where Rab7 sorts it to the recycling endosome where it waits for the next vasopressin signal for apical trafficking. Rab11 knockdown did not affect vasopressin‐induced apical AQP2 trafficking as reported previously. In contrast, Vps35 (vacuolar protein sorting‐associated protein 35) knockdown trapped AQP2 in the Rab11+ recycling endosome and reduced apical AQP2 trafficking. We conclude that Rab7 plays an important role in AQP2 sorting from the early endosome to the recycling endosome where vasopressin excites its apical trafficking mediated by Vps35.Support or Funding InformationThis work was supported by the Ministry of Science and Technology, TAIWAN (MOST 107‐2320‐B‐002‐057‐MY3).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 and characterization of novel small‐molecule inhibitors of SLC26A3
Authors: Park Jinhong , Namkung Wan 
Published: June 2021

SLC26A3 (downregulated in adenoma, DRA), an anion exchanger, is highly expressed in the luminal membrane of intestinal epithelial cells, and loss‐of‐function mutations in this gene result in congenital chloride diarrhea caused by detect of chloride absorption and bicarbonate secretion in the intestine. Here, we report development of the selective small‐molecule inhibitors of DRA for the investigation of the effect of DRA inhibition on constipation and metabolic acidosis. A screen of ~3,000 compounds using a cell‐based high‐throughput screening (HTS) assay revealed four novel compounds of DRA inhibitors that fully blocked Cl−/I− exchange activity of DRA with IC50 < 10 μM. The hit compounds strongly inhibited both Cl−/I− and Cl−/HCO3− exchange activity of DRA. The most potent inhibitor significantly blocked Cl−/I− exchange activity of DRA (IC50 of ~3 μM) but it did not affect the activity of SLC26A4 (pendrin), SLC26A7, SLC26A9, CFTR and ANO1. The small‐molecule DRA inhibitors identified here may be useful tools for pharmacological dissection of DRA and drug discovery for treating constipation and metabolic acidosis.This abstract is from the Experimental Biology 2019 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
ROLE OF CHLORIDE AND ANO1 FOR THE CONTRACTILITY OF MESENTERIC SMALL ARTERIES IN RATS WITH STREPTOZOTOCIN INDUCED DIABETES
Authors: Aalkjaer Christian , Kamaev Dmitry , Jensen Anders Bisgaard , Matchkov Vladimir 
Published: June 2021

Diabetes is associated with altered contractile responses of small arteries. In the streptozotocin (STZ) induced rat model of diabetes enhanced contractility several groups have reported an increased contractility. In this study we tested the hypothesis that chloride is important for the enhanced contractility in the STZ rats.Diabetes was induced in 6 weeks old rats by a single intraperitoneal injection of 60 mg/kg STZ, control rats received vehicle injection. Eight weeks later blood glucose was significantly increased in the STZ rats compared to control rats (24.6±1.2 mmol/L and 6.7±1.1 mmol/L, STZ (n=9) and control (n=9), respectively). The rats were killed and mesenteric small arteries were isolated. The contractile response to increasing concentrations of norepinephrine of mesenteric small arteries was assessed in isometric myographs in physiological salt solution and in physiological salt solution where Cl− was substituted with aspartate. Expression of the Ca2+ activated Cl− channel ANO1 was assessed with qPCR and Western blotting in mesenteric small arteries.The maximal contraction to norepinephrine in physiological salt solution was higher in arteries from STZ rats compared to control rats (5.12±0.33 N/m and 4.07±0.24 N/m, STZ (n=14) and control (n=13), respectively). In Cl− free solutions the maximal response to norepinephrine was reduced and no difference between STZ and control rats was found (3.76±0.26 N/m and 3.23±0.22 N/m, STZ (n=14) and control (n=13), respectively). Furthermore the effect of Cl− free solution on the area under the NE concentration response curve was bigger in arteries from STZ rats (2.47±0.42 and 1.39±0.26 STZ (n=14) and control (n=13), respectively). Expression of ANO1 mRNA was higher in arteries from STZ rats (181±10% and 100±18%, STZ (n=4) and control (n=4), respectively). Expression of ANO1 protein was higher in arteries from STZ rats (195±24% and 100±18%, STZ (n=9) and control (n=8), respectively).This study provides evidence that the enhanced contractility of mesenteric arteries from STZ rats is dependent on the chloride gradient over the cell membrane, and suggests that this may be consequent to enhanced expression of the Ca2+ activated Cl− channel ANO1.Support or Funding InformationDanish Council for Independent Research (7016‐00013B)This abstract is from the Experimental Biology 2019 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.