bluuebunny/crossref_metadata_2025_split huggingface dataset:
Quantitative Proteomics‐based Bottom‐up PBK Modeling: A Case Study of 3 Statins
Authors: Chan James Chun Yip , Tan Shawn Pei Feng , Chan Eric Chun Yong 
Published: June 2021

In the consumer care, cosmetics and chemical industries, there is a growing need for alternatives to animal testing to derive biokinetic data to evaluate both efficacy and safety of chemicals. One promising alternative is bottom‐up physiologically‐based biokinetic (PBK) modeling, which utilizes in vitro‐to‐in vivo extrapolation (IVIVE) for prediction of biokinetic parameters. The main challenges of IVIVE lie in suboptimal biokinetic predictions, particularly when using in vitro transporter data which often requires empirical fitting to human biokinetic data. As part of an Organization for Economic Cooperation and Development (OECD) case study, the primary objective of this work is to develop and validate quantitative proteomics‐based bottom‐up PBK models using a set of chemicals rich with in vitro and human in vivo data. Three HMG‐CoA reductase inhibitors that undergo differential elimination processes were chosen: rosuvastatin (transporter‐dependent), fluvastatin (metabolism‐dependent) and pitavastatin (mixed‐mode). PBK models were built using the Simcyp® Simulator by incorporating: (1) in vitro transporter and metabolism data (Vmax, Jmax, Km and CLint) and (2) animal tissue distribution data from literature to inform tissue‐to‐plasma equilibrium distribution ratio (Kp). Simulations were performed for single intravenous, single oral and multiple oral dosing of these chemicals. The successful prediction was based on a two‐fold criterion when compared against human biokinetic parameters. Our results showed that predicted systemic exposure (AUC0‐∞h), maximum plasma concentration (Cmax), plasma clearance (CL) and time to reach Cmax (Tmax) were within two‐fold of the observed data, with the exception of parameters associated with multiple oral pitavastatin dosing and Tmax of single oral fluvastatin dosing. The use of animal Kp data improved the predicted plasma‐concentration time profiles but did not significantly alter predicted biokinetic parameters (Figure 1). Our study demonstrated that quantitative proteomics‐based mechanistic IVIVE could account for differences in transporter and metabolic enzyme expression levels between in vitro systems and in vivo organs, allowing the prediction of whole organ clearances without any empirical scaling. We conclude that bottom‐up PBK modeling incorporating mechanistic IVIVE could be a viable alternative to animal testing in obtaining predictions of human biokinetics of chemicals.Support or Funding InformationThis work was supported by the Innovations in Food and Chemical Safety Programme [Grant number H18/01/a0/C14] and NUS Department of Pharmacy [Grant number C‐148‐000‐003‐001]. Travel funding for this conference was provided by the Innovations in Food and Chemical Safety Programme and the Skin Research Institute of Singapore.This abstract is from the Experimental Biology 2019 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
Hyperglycemia increases breast cancer cell and normal breast epithelial cell proliferation through polyamine deregulation
Authors: Capellen Caleb , Ortega Jose , Dunworth Matthew , Casero Robert A. , Chandra Surabhi 
Published: June 2021

Polyamines (spermine, spermidine, and putrescine) are involved in basic cellular processes such as cell growth, replication, and transcription. Upregulation of polyamines can thus develop tumors due to hyper‐proliferation of cells as observed in cancers of the colon, skin, prostate and breast. Inhibitors of polyamine pathway prevent tumor growth as shown in animal models of colon and skin cancer. Diabetic conditions further accelerate the growth and metastasis of cancers such as pancreas, breast, liver, and colorectum, however the role of polyamines in such states has not been investigated. We hypothesized that polyamine pathway rapidly affects proliferation of breast cancer cells and normal breast epithelial cells in diabetic conditions. Breast cancer cells (MDA‐MB‐231) and normal breast epithelial cells (MCF‐10A) were treated with low glucose (LG, 5mM) or high glucose (HG, 25 mM) in the presence/absence of polyamine pathway inhibitors. There was a time dependent increase in cell proliferation with HG from 48–72 hr compared to LG. This was correlated with an increase in polyamines (putrescine, spermidine) though spermine levels were not changed considerably. In addition, gene expression and activity of catabolic enzymes in the polyamine pathway, spermine‐spermidine acetyltransferase (SSAT) and spermine oxidase (SMOX) were decreased with HG treatment, further suggesting a build‐up of polyamines. Simultaneous treatment with an ornithine decarboxylase (polyamine synthesis enzyme) inhibitor, difluoromethylornithine (DFMO) could prevent cell proliferation, restore polyamines, and normalize polyamine catabolic enzymes with high glucose exposure. In conclusion, polyamine pathway is responsible for proliferation of breast cancer cells and normal breast epithelial cells in diabetic states, which can be prevented using inhibitors of polyamine synthesis.This abstract is from the Experimental Biology 2019 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
Heterozygous knockdown insulin‐like growth factor‐1 receptor (IGF‐1R) regulates mitochondrial functions and prevents colitis and colorectal cancer
Authors: Wang Shu Qing , Cui Shu Xiang 
Published: June 2021

Although insulin‐like growth factor‐1 receptor (IGF‐1R) has been accepted as a major determinant of tumorigenesis, the mechanisms of its roles in the pathogenesis of cancers have remained elusive. Herein, we demonstrate that IGF‐1R plays pivotal roles in regulation of mitochondrial functions and resistance to oxidative stress in colitis and colorectal tumorigenesis. Heterozygous knockdown IGF‐1R attenuated azoxymethane (AOM)/dextran sulfate sodium (DSS)‐induced colitis and colitis associated cancer (CAC) in Igf1r+/− mice. Knockdown of IGF‐1R confers the resistance to oxidative stress‐induced damage on colorectal epithelium through protection the mitochondrial dynamics and structures. Knockdown of IGF‐1R develops the function of mitochondrial fusion under oxidative stress. Mechanically, an increase of mitochondrial coupling index RCI and oxidative phosphorylation index ADP/O was seen in the epithelial cells of Igf1r+/− mice. Seahorse XFe‐24 analyzer analysis of mitochondrial bioenergetics showed an increase of oxygen consumption rate (OCR) and a decrease of extracellular acidification rate (ECAR) in Igf1r+/− cells. Further analysis suggests the protection mechanisms of colorectal epithelial cells from oxidative stress through activating mitochondrial respiratory chain and the LKB1/AMPK pathways. These results support the roles of IGF‐1R at the nexus between oxidative damage and mitochondrial function and connect colitis and colorectal tumorigenesis.Support or Funding InformationThis work was supported by Natural Science Foundation of China (81673449, 91629303).This abstract is from the Experimental Biology 2019 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
Sphingosine kinase inhibitor regulates pro‐inflammatory cytokines to subdue the neoplastic lesions in hepatocellular carcinoma
Authors: CHATTEERJEE NABANITA , Das Subhadip , Bose Dipayan , Banerjee Somenath , Saha Krishna Das 
Published: June 2021

Hepatocellular carcinoma (HCC), is the most leading cause of cancer‐related death in all over the world (seems to more than 80% of death cases). It is frequently linked with continuous hepatocytes death, inflammatory cell infiltration, and compensatory liver regeneration with higher angiogenic rate. Considering the different signalling process during this tumorogenesis with the involvement of new ‘lead’ identification of different sources, desire to be less toxic and higher bioavailability. The use of live, attenuated or genetically compensated microbes or its cellular component(s) or metabolites has begun to emerge as a potential new approach in medicinal research to deliver bioactive entities. Thus, advancing our knowledge of such microbes‐mediated therapy may suggest new avenues for therapeutic intervention in many fatal diseases.Sphingosine kinase inhibitor (SKI) II has been reported as a dual inhibitor of sphingosine kinases (SKs) 1 and 2 and has been extensively used to prove the involvement of SKs and sphingosine‐1‐phosphate (S1P) in cellular processes and able to induce apoptosis in human HepG2 (Hepatocellular carcinoma cell, human) cells, as characterized by phosphatidylserine serine exposure, activation of series of caspases and cleaved PARP. SKI also modulates the various transcription factors like NF‐kB‐65, p53 through regulation of cell proliferation exerted the death cause. Moreover, this study also focused that SKI effectively suppressed diethylnitrosamine (DEN) induced liver inflammation and hyperplasia in murine, also regulated the lipid peroxidation and enzymic antioxidants (SOD, CAT, GPx, GR and GST) status with an improvement of their survival rate. It reduced the elevated level of cytokines including tumor necrosis factors, interleukins, interferon (alpha and gamma), and transforming growth factors in hepatocytes from liver fibrosis bearing mice. As evident from an immunochemical analysis, DEN‐induced neoplastic lesions may up‐regulate the various growth factors including EGF, HGF, endothelin‐1 regulation with promoting angiogenic activity via modulation of CD34, VEGF, HIF‐1α expression and we found that the alteration of these changes were balanced by SKI treatment in vivo system. So taken together, these finding indicated that, SKI may recover in hyperproliferating cancer cells with tumor growth regulation in carcinogen‐exposed rodent system. In future direction, we will check the p53−/− and cmyc−/− mice that hoe it could regulate cancer metastasis with this SKI. and the involvement of od tumor microenvironment.Support or Funding InformationCSIR and Department of Science and Technology, IndiaThis abstract is from the Experimental Biology 2019 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
Phloretin ameliorates hyperuricemia‐induced chronic renal dysfunction through inhibiting NLRP3 inflammasome and uric acid reabsorption
Authors: Cui Danli , Liu Shuyun , Zhao Meng , Mao Ruiwen , Wang Chengshi , Li Lan , Chen Younan , Cheng Jingqiu , Lu Yanrong , Liu Jingping 
Published: June 2021

ObjectiveHyperuricemia is an important risk factor for renal diseases and contributes to renal fibrosis, and it's a big challenge for clinic. Phloretin, a plant‐derived dihydrochalcone, has shown antioxidant/anti‐inflammatory properties, and could inhibit uric acid (UA) uptake in vitro, but its role on renal injury remains unknown. In this study, we aimed to investigate the therapeutic potential of phloretin on hyperuricemia‐induced renal dysfunction and underlying mechanism in mice.MethodsMice were induced hyperuricemic by oral gavage of adenine/potassium oxonate mixture, and then treated with phloretin. The effects of phloretin on renal function, inflammation and interstitial fibrosis were evaluated in hyperuricemic mice. The changes of renal mitochondrial oxidative stress and UA metabolism were also analyzed.ResultsHyperuricemic mice showed renal dysfunction with elevated renal inflammation and fibrosis. By contrast, phloretin significantly improved renal function and morphologic lesions, as indicated by reduced levels of serum blood urea nitrogen (BUN), creatinine and urinary albumin to creatinine ratio (UACR), as well as decreased levels of kidney tubular necrosis, interstitium extracellular matrix (ECM) deposition and interstitial fibroblasts activation in the kidneys of hyperuricemic mice. Furthermore, phloretin significantly suppressed renal mitochondrial reactive oxygen species (ROS) production and NOD‐like receptor family pyrin domain containing 3 (NLRP3) inflammasome signaling, and inhibited glucose transporter 9 (GLUT9) and UA reabsorption in hyperuricemic mice.ConclusionPhloretin can ameliorate hyperuricemia‐induced chronic renal injury through co‐inhibiting NLRP3 pathway and UA reabsorption, and thus it might be a promising therapy to hyperuricemia‐related renal diseases.Support or Funding InformationThis work was supported by grants from National Natural Science Foundation of China (31200754, 81571808, 31871001) and China Postdoctoral Science Foundation (2012M511931).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 Roles of Qishen Granules Recipes, Qingre Jiedu, Wenyang Yiqi and Huo Xue, in the Treatment of Heart Failure
Authors: Gao Sheng , Wang Yong , Tian Chuan , Li Chun , Wu Dingfeng , Jiao Na , Zhu Lixin , Zhu Ruixin , Wang Wei 
Published: June 2021

BackgroundRecipes (Qingre Jiedu (QJ), Wenyang Yiqi (WYYQ) and HuoXue (HX)) in Qishen granules (QSG) are believed to synergistically exert cardio‐protective effects and improve heart function. Further studies, however, are necessary to interpret the underlying pattern of each decomposed recipe in QSG and their synergistic effect in the treatment of heart failure.MethodsIn this study, the efficacy of QJ, WYYQ and HX was evaluated by using heart failure model Sprague‐Dawley rats. Cardiac tissues were harvested for RNA Sequencing. Differentially expressed genes (DEGs) analysis and pathway enrichment analysis were performed with NOIseq and clusterProfiler approach, respectively. Furthermore, an optimized quantitative pathway analysis method that maximally acquires gene expression information was introduced to perform differential pathways analysis between different groups.ResultsQJ, WYYQ and HX are shown to improve left ventricular ejection fraction, indicating their common contribution within QSG to the treatment of heart failure. Compared to model group, there are 858, 124, 1195, 1733 DEGs in QSG, QJ, WYYQ and HX group respectively, which are enriched in all or part of pathways including energy metabolism, inflammation reactions and ventricular remodelling. Quantitative pathway analysis show that decomposed recipes QJ, WYYQ and HX have common but differentiated contributions in the treatment of heart failure. Especially, ATP1A3 gene in thyroid hormone signalling pathway is up‐regulated significantly in both QSG and HX groups. Besides, ADCY1 gene in relaxin signalling pathway is up‐regulated in both QSG, WYYQ and HX groups. Therefore, WYYQ and HX might have complementary effects on energy supply in the treatment of heart failure through activation of thyroid hormone signalling and relaxin signalling pathways.ConclusionOur results have indicated that QJ, WYYQ and HX recipes in QSG achieve synergistic therapeutic effects in myocardial energy metabolism. This provides theoretical foundations and evidence for the optimization of QSG formulas. Quantitative pathway analysis is proved as an effective tool to decompose the recipes of Traditional Chinese Medicine.Support or Funding InformationThis work was supported by National Natural Science Foundation of China 81770571 (to LZ), 81774152 (to RZ), 41530105 (to RZ).This abstract is from the Experimental Biology 2019 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
Efficacy of Tramadol in Thermoreversibles Gels in a Model of Postoperative Pain
Authors: Tofoli Giovana , Papini Juliana , Cereda Cintia , Napimoga Juliana , Macedo Cristina , de Araujo Daniele 
Published: June 2021

Tramadol (TR) is an opioid analgesic with short duration of action, still it may be consider valuable when compared to other opioids, as it produces mild adverse effects. Due to these properties, trmadol was incorporated in thermoreversible poloxamers and the effectiveness of these new drug delivery formulations was evaluated in a rat model of postoperative pain. The poloxamer (PL)‐tramadol systems were prepared by direct dispersion of the drug in solutions with PL 407 (20%) or the association of PL 407 (20%) and PL 188 (10%). Male Wistar rats were submitted to hindpaw incision and after 24 hours treated by subcutaneous route with one of the two formulations or plain TR 2% (10 μg.kg−1), saline or the poloxamers vehicles. Mechanical hypersensitivity was measured using von Frey filaments after 15, 30, 60, 90, 120,180, 240, 300 and 360 minutes from the injections. The protein levels of TNF‐α, IL‐1β, Substance P and CGRP were obtained through ELISA assays after 1, 3 and 6 hours after tramadol injections. All animals presented postincisional hypersensitivity showed by increased withdrawal response when compared to the preoperative period (p<0.05). Poloxamers formulations presented greater and longer analgesic effect than plain tramadol (p<0.05). Also, the new formulations were able to reduce the levels of TNF‐α after 6 hours (p<0.05). One hour far ahead of the formulations' injection CGRP levels with the PL 407 +PL 188 +TR 2% formulation was smaller than this vehicle without TR (p<0.05). The association of PL 407 and PL188 in an injectable formulation of tramadol promoted prolonged and effective postoperative pain relief in a rat model.Support or Funding InformationFAPESP (#2015/20744‐0)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 Glucocorticoids in Gastroprotection Induced by Capsaicin
Authors: Filaretova Ludmila , Podvigina Tatiana , Morozova Olga 
Published: June 2021

The pivotal role of capsaicin‐sensitive peptidergic sensory fibers in maintaining of gastric mucosal integrity against injurious interventions has been well established in rats and then in human. Clinical finding suggests that capsaicin‐induced gastric mucosal protection against the injurious effects of ethanol or indomethacin is due to the acute stimulatory effect of the compound. It was concluded that patients taking non‐steroidal anti‐inflammatory drugs (NSAID) regularly can decrease the incidence and severity of gastric ulceration, if they have moderate spicy foods. Opposite effect has been observed in rats desensitized by capsaicin. Functional blockade of gastric sensory nerve endings elicited by systemic capsaicin application was resulted in impaired mucosal protection against various ulcerogenic agents. Our previous findings suggest a pivotal compensatory role of glucocorticoids in maintenance of gastric mucosal integrity in rats desensitized by capsaicin. In this study, we tested if glucocorticoids can participate in capsaicin‐caused gastroprotection during NSAID‐caused gastropathy accompanied by prostaglandin deficiency in rats. Gastric injury was induced by indomethacin (IM). A single IM administration (35 mg/kg, sc) into fasting (24 h) rats caused the formation of gastric erosions 4 h after injection. The effects of small doses (0.1, 0.5, 1.0, 2.0, 3.0, 5.0, 10.0 mg/kg, sc) of capsaicin on the blood corticosterone and glucose levels and IM‐induced gastric injury were tested. A single capsaicin administration was performed one hour before IM administration. To test the participation of corticosterone in the gastroprotective effect of capsaicin, the pretreatment by the inhibitor of glucocorticoid synthesis, metyrapone (30 mg/kg, i.p., 30 min before capsaicin) was used. We found that the pretreatment with capsaicin (0.5–10.0 mg/kg, sc) reduced dose dependently the area of IM‐induced erosions. This protection was accompanied by a gradual increase in blood corticosterone and glucose levels. Metyrapone injected shortly before capsaicin administration (1.0 mg/kg) caused a fast inhibition of capsaicin‐induced corticosterone response and reversed the protective effect of capsaicin on the gastric mucosa against the IM‐produced injury. The present study confirms evidence for the powerful gastroprotective potency of capsaicin during NSAID‐caused gastropathy. The findings obtained demonstrate that capsaicin administration resulted in an increase in plasma corticosterone levels which in turn plays a pivotal role in maintaining of gastric mucosal integrity against injurious intervention of IM.Support or Funding InformationThe study was supported by grants of RSF N 19‐15‐00430 (NSAID‐caused gastropathy with prostaglandin deficiency) and RFBR N 19‐015‐00514a (capsaicin‐induced gastroprotection)This abstract is from the Experimental Biology 2019 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
Mechanosensitive Ion Channels in the Development of Ileus
Authors: Uray Karen , Sipos Ádám , Vo Duong Nhu Y , Thi Hai Yen Nguyen , Docsa Tibor 
Published: June 2021

IntroductionApproximately 30% of trauma patients develop ileus, a decrease or cessation of intestinal motility resulting in pathological distension of the intestinal wall. Mechanosensitive ion channels play a role in both the physiological and pathological response to mechanical stimuli.ObjectiveThe objective of this study was to determine if mechanosensitive ion channels play a role in the development of ileus and the response of the intestine to pathological stretch.MethodsIn a rodent model, we induced ileus by a combination of hemodilution and mesenteric venous hypertension. We measured intestinal function using an ex vivo organ bath system. We also measured smooth muscle myosin light chain phosphorylation and intestinal wall fluid content. We collected tissue for microarray analysis.ResultsIntestinal wall fluid content increased significantly, indicating the development of intestinal edema. Edema development resulted in distension of the intestinal wall, decreased intestinal smooth muscle contractile activity, and decreased smooth muscle myosin light chain phosphorylation. We used microarray analysis to compare normal (unstretched) intestinal smooth muscle to edematous (stretched) intestinal smooth muscle to characterize changes in ion channel expression. We identified hyperpolarization‐activated cyclic nucleotide‐gated 2 (HCN2), a cyclic nucleotide gated ion channel, which is also stretch‐activated. HCN2 gene expression decreased in edematous intestinal tissue. In the ex vivo organ bath system, we tested the effects of HCN2 inhibitors on intestinal function. In preliminary experiments, HCN2 inhibition with ZD7288 resulted in decreased response to carbachol, with little change in spontaneous contractile activity or frequency.ConclusionsOur data suggest that HCN2 downregulation/inhibition can suppress intestinal contractile activity. Thus, HCN2 is a potential therapeutic target for improving intestinal motility. In future experiments, we will determine the role of HCN2 in the intestinal response to stretch.Support or Funding InformationHungarian National Research Development and Innovation Office (K120669, EFOP‐3.6.2‐16‐2017‐00006)This abstract is from the Experimental Biology 2019 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.