bluuebunny/crossref_metadata_2025_split huggingface dataset:
CRISPR/Cas9 Knockout‐Based Genome Wide Screening for Pathways Associated with Organochlorine Pesticide Toxicity in Human Dopaminergic Neurons
Authors: Russo Max , Sobh Amin , Tagmount Abderrahmane , Zhang Ping , Vulpe Christopher , Liu Bin 
Published: June 2021

Environmental exposure to organochlorine pesticides (OCP) has been implicated as a risk factor for Parkinson's disease (PD), a devastating movement disorder characterized by massive loss of dopamine (DA) producing neurons in the substantia nigra and clinical symptoms including bradykinesia, rigidity and tremor. PD is the second most prevalent neurodegenerative disease and >95% of PD cases are usually not clinically diagnosed until after the age of sixty. OCPs are highly persistent in the environment and are extremely bio‐accumulative. Altered iron distribution in the brain and accumulation in the substania nigra is also observed in PD and may play a role in PD pathogenesis. To gain insight into the role of OCP exposure and iron dysregulation in PD pathogenesis, we carried out a genome wide (Brunello library, 19,114 genes, 76,411 sgRNA) CRISPR/Cas9 screen in SH‐SY5Y human dopaminergic neuronal cell line exposed to sub toxic doses of dieldrin or iron over one month to identify cellular components which modulate their toxicity. NGS sequencing of sensitive and resistant clones identified multiple genes, which when mutated significantly altered sensitivity to dieldrin or iron. Pathway analysis of the dieldrin data set implicated the protein degradation pathways previously implicated in inherited forms of PD centered on Parkin2 and SCNA. These results suggest a potentially common pathogenesis of idiopathic and inherited forms of PD. Additionally, the dieldrin results also suggested a potential role for the mTOR pathway, which is a signaling pathway that integrates both intracellular and extracellular signals and serves as a central regulator of cell metabolism, growth, proliferation and survival, is significantly involved in dieldrin associated toxicity. Fe toxicity on the other hand was shown to significantly alter dopamine receptor 2 (D2) signaling pathways. Together, these studies provide potentially novel insight into the dopaminergic toxicity associated with PD. Currently, we are conducting targeted validation of candidate genes identified in this study. Our work represents one of the first to apply genome wide CRISPR screens in a neuronal cell line and may reveal how an environmental toxicant and iron dysregulation in the brain contribute to PD. Additionally, this work may allow us to identify molecular mediators of neurodegeneration as potential therapeutic targets and/or diagnostic biomarkers.Support or Funding InformationUniversity of Florida Opportunity Fund.This abstract is from the Experimental Biology 2019 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
Modulatory effects of fibronectin on calcium oxalate crystallization, growth, aggregation, adhesion on renal tubular cells, and invasion through extracellular matrix
Authors: Thongboonkerd Visith , Khamchun Supaporn 
Published: June 2021

Fibronectin, an extracellular matrix (ECM) protein, has been thought to be involved in pathogenic mechanisms of kidney stone disease, especially calcium oxalate (CaOx) type. Nevertheless, its precise roles in modulation of CaOx crystal remained unclear. We thus performed a systematic evaluation of effects of fibronectin on CaOx monohydrate (COM) crystal (the major causative chemical crystal in kidney stone formation) in various stages of kidney stone pathogenesis, including crystallization, crystal growth, aggregation, adhesion onto renal tubular cells, and invasion through ECM in renal interstitium. The data showed that fibronectin significantly decreased crystallization, growth and adhesive capability of COM crystals in a dose‐dependent manner. In contrast, COM crystal aggregation and invasion through ECM migration chamber were significantly enhanced by fibronectin in a dose‐dependent fashion. Sequence analysis revealed two and six calcium‐binding and oxalate‐binding domains, respectively. Saturation of fibronectin with oxalate prior to crystal modulation assays could abolish all the effects of fibronectin, whereas saturation with calcium had minimal interference. These data strongly indicate the dual functions of fibronectin, which serves as an inhibitor for COM crystallization, crystal growth and adhesion onto renal tubular cells, but on the other hand, acts as a promoter for COM crystal aggregation and invasion through ECM. Moreover, its crystal modulatory activities are mainly due to oxalate‐binding capability.Support or Funding InformationThis study was supported by Mahidol University research grant and the Thailand Research Fund (IRN60W0004 and IRG5980006).This abstract is from the Experimental Biology 2019 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
Study the Effect of Irisin on Prostate Cancer Cells
Authors: Ko Fu‐Hsuan , Huang Feng‐Ting 
Published: June 2021

According to the World Health Organization, prostate cancer is the second most common type of cancer and the fifth leading cause of cancer‐related death in men around the world. Epidemiological studies demonstrate that exercise can improve survival and reduce recurrence of prostate cancer. Studies indicate that humans' exercise serum has a growth inhibitory effect on prostate cancer cell proliferation, but which factor plays a major role is not clear. Recently, myokines secreted by skeletal muscle are believed to take part in cancer suppression. Irisin, an exercise‐induced myokine, promotes the “browning” of white adipocytes to brown‐like adipocytes. Several studies have demonstrated that irisin can inhibit cell progression in various cancer cells including those of breast cancer, lung cancer, and osteosarcoma. However, the underlying mechanisms of irisin's inhibitory effects on prostate cancer growth are not yet completely understood.Our present study reveals that human recombinant irisin expressed in the E. coli system can inhibit cell migration and invasion of two prostate cancer cell lines, PC3 and 22Rv1. Irisin suppresses the proliferation of prostate cancer cells in a dose‐dependent manner, in addition to inducing cell cycle arrest and apoptosis in prostate cancer cells. These findings support the possibility of irisin treatment as a new therapeutic strategy against prostate cancer.Support or Funding InformationThis study was supported by the Department of Biochemical Science of Technology of National Taiwan University and the Taiwan Ministry of Science and Technology.This abstract is from the Experimental Biology 2019 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
Ceranib‐2 inhibits HIF1‐α gene expression and induces apoptosis in HepG2 cells
Authors: ERKASAP NILUFER 
Published: June 2021

The aim of this study to investigate the apoptotic effect of novel anti‐cancer drug ceranib‐2 in human liver carcinoma cells (HepG2). HepG2 cells plated at 1 × 105 cell/well in 24 well plates. Cell were treated with ceranib‐2 at 0,1, 1, 5, 10, 25 and 50 μM concentrations for 24 and 48 hours. Cell viability measured using 3‐(4,5‐d‐methylthiazol‐2‐yl)‐2,5‐diphenyltetrazolium bromide thyazolyl blue (MTT). HepG2 cell viability were decreased by 58 % after 24 and 48 hours ceranib‐2 treatment at concentration 10 μM. mRNA abundance of acid ceramidase (ASAH), caspase‐3, caspase‐8, HIF1‐α, and TNF‐ α conducted by Real time PCR (qRT‐PCR). Ceranib‐2 at 10 μM concentration increased mRNA abundance of caspase‐3 and no change were detected on caspase‐8 level vs control group (p<0.05) after 24 hours' treatment. No difference detected on caspase‐3 and caspase‐8 mRNA level after 48 hours' treatment with ceranib‐2 at 10 μM concentration. mRNA abundance of ASAH were reduced after 48 hours treatment with 10 μM ceranib‐2 (p<0.05). HIF1‐α expression was low after 24 hour compared to the control group (p<0,0001). TNF‐α mRNA were increase after 24 and 48 hours (p<0.05). Low abundance of ASAH in treated cells showed that 10 μM ceranib‐2 could inhibit ceramidase after 48 hour and this may trigger elavating ceramide concentration. We also observed that inhibition of ceramidase with ceranib‐2 inhibits HIF1‐α expression. Inhibition of HIF1‐a expression might induce apoptosis by reducing cell viability in HepG‐2 cells via ceranib‐2. In conclusion, ceranib‐2 may have anti‐cancer effect at 10 μM concentration in HepG2 cells. Finally, we have shown that inhibition of ASAH with ceranib‐2, induces apoptosis in HepG2 cells.This abstract is from the Experimental Biology 2019 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
Sex Differences in the Stages of Cocaine Use Disorder and the Influence of the Mesolimbic Dopamine System
Authors: Mauterer Madelyn I , Jones Sara R 
Published: June 2021

Traditional drug abuse research has primarily focused on male behavior and neurobiology; however, recent work in the field has revealed distinct differences between male and female cocaine responsivity. Specifically, females appear to be more vulnerable to the effects of cocaine, a phenomenon that has been shown to fluctuate with the reproductive cycle in both clinical and preclinical studies. For example, in human studies, women report greater subjective effects of cocaine during the follicular phase of the menstrual cycle, while in rodent studies, mice and rats exhibit greater cocaine taking behavior and preference during the estrus phase of the estrous cycle. Increased sensitivity to cocaine reward has been shown using many behavioral assays including those used to measure initiation of cocaine use, motivation for drug, extinction, and reinstatement. Increased cocaine taking behavior in females has been manipulated further to study the effects of social interaction, natural reward, exercise, and stress on intravenous cocaine self‐administration. Altogether, sex differences, both between males and females and across the stages of estrous, appear to be mediated by differences in the mesolimbic dopamine system according to recent work implicating dopamine projections from the ventral tegmental area to the nucleus accumbens core (NAc).Here, intravenous cocaine self‐administration is used to assess drug taking behavior during the stages of drug addiction including initiation of use (acquisition), escalation, motivation, extinction, and reinstatement. Females in estrus showed faster times to acquisition of cocaine self‐administration and greater motivation to self‐administer cocaine using the behavioral economics protocol Threshold. Female cocaine abuse vulnerability further appears to be mediated by underlying neurobiological differences in cocaine taking between sexes and estrous stage. Females in estrus exhibit less synaptogyrin 3 expression in the NAc compared to their non‐estrus counterparts, a significant finding as decreased synaptogyrin expression in the NAc is associated with higher breakpoints in progressive ratio in males and is associated with dopamine transporter function. Furthermore, using ex vivo fast scan cyclic voltammetry (FSCV) in the NAc, females also exhibit greater dopamine transporter sensitivity to cocaine than males. Overall, these results add to the growing body of literature suggesting that females exhibit greater vulnerability to cocaine use and abuse across the stages of addiction and that these effects are due to an inherent neurobiological difference that this linked to the reproductive cycle and the mesolimbic dopamine system.Support or Funding InformationT32AA007565, P50DA006634This abstract is from the Experimental Biology 2019 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
Activation of protein kinase C impairs endothelium‐dependent vasorelaxation in isolated human omental resistance arteries
Authors: Grunewald Zachary Ian , Ghiarone Thaysa , Quinones‐Morales Mariana , Ramirez‐Perez Francisco , Martinez‐Lemus Luis , Padilla Jaume 
Published: June 2021

Chronic diseases such as obesity and diabetes are associated with endothelial dysfunction and increased cardiovascular disease risk. Notably, protein kinase C (PKC) has been identified as a major molecular contributor to chronic disease‐associated microvascular impairments. However, the molecular mechanisms by which PKC activation causes microvascular endothelial dysfunction are not fully understood. Herein, we tested the hypothesis that PKC activation impairs microvascular vasorelaxation through an endothelial nitric oxide synthase (eNOS)‐dependent mechanism in human isolated resistance arteries and cultured endothelial cells. Accordingly, resistance arteries were isolated from the omentum of 32 patients (52±3 years of age, 43.3±1.7 BMI, 76% women) that underwent abdominal surgery and analyzed for vasomotor function ex vivo using wire myography. We found that pharmacological inhibition of PKC (ruboxistaurin, 1μM) for 30 minutes enhanced endothelium‐dependent vasorelaxation in response to bradykinin relative to untreated arteries (p<0.05). This effect of ruboxistaurin was abrogated by NOS inhibition (L‐NAME, 300μM), suggesting that improved endothelium‐dependent relaxation with PKC antagonism is nitric oxide‐dependent (p<0.05). Next, we found that treatment with the PKC activator, phorbol 12‐myristate 13‐acetate (PMA, 0.5μM), for 30 minutes impaired endothelium‐dependent and ‐independent vasorelaxation, which was abolished with concomitant incubation of ruboxistaurin (p<0.05). Furthermore, we found that PMA treatment reduced Akt and eNOS activation in cultured endothelial cells (HUVEC; p<0.05). In conclusion, these findings suggest that in isolated human abdominal adipose tissue resistance arteries, PKC activation impairs microvascular endothelial function likely through an eNOS‐dependent mechanism. Follow‐up experiments are ongoing to determine the involvement of specific PKC isoforms and upstream molecular regulators contributing to PKC‐mediated impairments in endothelial function.Support or Funding InformationResearch support: National Institutes of Health grants: R01 HL137769 (JP), R01 HL088105 (LM‐L).This abstract is from the Experimental Biology 2019 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
Secreted Klotho Protects Against Aging‐associated Coronary Endothelial Dysfunction via SIRT1 Activation
Authors: Chen Kai , Sun Zhongjie 
Published: June 2021

Background and ObjectiveCoronary heart disease (CHD) is the leading cause of death in the world. Endothelial dysfunction is an initial step in the development of CHD and may be a critical early target for the prevention of CHD. Klotho, originally identified as an aging‐suppressor gene, is predominately expressed in kidneys. Klotho protein is secreted into the circulation. The objective of this study is to investigate whether secreted Klotho protein protects against aging‐associated coronary endothelial dysfunction and CHD.Methods and ResultsWe found that aging‐associated coronary endothelial dysfunction assessed by endothelium‐dependent vasodilation and coronary blood flow was accompanied by a reduction in serum level of secreted Klotho (sKL) protein. Using klotho‐hypomorphic (KL (−/−)) mice, we confirmed that Klotho deficiency caused coronary endothelial dysfunction. Interestingly, the impaired endothelium‐dependent vasorelaxation and coronary flow were remarkably improved by in vivo delivery of exogenous sKL protein in aged mice and KL (−/−) mice, suggesting that sKL rescued aging‐induced coronary artery endothelial dysfunction. In addition, sKL treatment protected against aging‐associated and Klotho deficiency‐induced cardiac dysfunction, as evidenced by restoration of stroke volume and cardiac output in aged and KL (−/−) mice. sKL treatment also enhanced coronary endothelial nitric oxide bioavailability and attenuated oxidative stress and apoptosis. Mechanistically, aging via Klotho deficiency suppressed SIRT1‐AMPKα‐eNOS pathway in the coronary endothelial cell through activation of transcription repressor HIC‐1. Furthermore, pharmacological activation of SIRT1 by SRT 1720 protected against Klotho deficiency‐induced coronary artery endothelial dysfunction and cardiac dysfunction.ConclusionKlotho regulates coronary artery endothelial function via SIRT1 activation. Supplement of exogenous Klotho protein represents a promising therapeutic strategy for the treatment of aging‐associated coronary heart disease.Support or Funding InformationAG049780, HL118558This abstract is from the Experimental Biology 2019 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
Building a Data Collection Architecture for Exercise Training and Health Monitoring
Authors: Lee Hsiu An , Hsu Chien‐Yeh 
Published: June 2021

The selection of athletes for a sports competition is based on his/her achievements and experience. According to research, in the professional sports program, the selection criteria should be based on the characteristics of the competition, the size, physiology, test value, psychological and physiological development and characteristics of the athletes. However, the qualifications of athletes on cross‐disciplinary and skill have not been fully implemented.This study is to build a data collection architecture for exercise training and health status monitoring for athletes. The data items which will be stored including sports training data, biochemical indicators, dietary intake, psychological and physiological data, fatigue index etc. There is a data analysis platform build for future data processing.The design of the collection architecture is divided into five phases, including definition, capture, contribution, share, and analysis. The definition phase and the capture phase are happened in each data source unit and data are transferred and stored in a central data repository. Each data source plays as a contribution role which will follow a prearranged research protocol of the central repository. The information acquisition and storage mechanism are also designed for the repository. The sharing stage is to design a transferring and sharing mechanism for athlete training and competition data to build an exercise research database which is used as a base in the analysis platform in the repository.The system includes five core modules: Data Contributor, Data Dictionary, Data Access, User Management, and Repository Management. The Data Contributor module provides users with a research case on the web page so that participants can upload data. A screening process is also designed to verify the correctness of the sports data and research database. The Data Dictionary module is to provide the definitions of the Basic Data Elements (BDE) which is the most commonly used fields defined by defined by the project, and Extended Data Elements (EDE) which is the user defined data fields and can be added to the BDE.Data Access module is the control module for data access permission. The owner of the research case can share the information, once shared, all members of the team will have the right to view and obtain all data, and can query the data in the database in the system. The User Management module is the research team member and its data usage rights. The research owner can set the usage rights of each member. The Repository Management module manages the database structure and provides views of the contents of the database. Using the data collection architecture, the researcher can manage, share, distribute, and process the information and training schedules of participating athletes.This abstract is from the Experimental Biology 2019 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
Endurance Exercise Capacity and Substrate Metabolism in Male and Female Mice
Authors: Rowe Patrick , O'Neill Caitlin , DeWitt Elizabeth , Kolwicz Stephen C 
Published: June 2021

The investigation of sex‐specific differences in physiological and pathophysiological processes is of increased importance in scientific research. Previous studies, in both rodents and humans, observed that females generally have a higher endurance exercise capacity (EEC) than males. Additional research suggests that the increased EEC in females is mediated, in part, by differences in substrate metabolism during exercise. The objective of this study was to determine whether changes in exercise substrate metabolism contributed to sex‐specific differences in EEC in mice. To evaluate EEC, male and female, C57BL/6‐NCrl mice (n=9 each group) were subjected to a graded exercise test on a motorized treadmill. The treadmill was set at a 10% incline and the speed gradually increased from 10.2 m/min to 22.2 m/min at fixed intervals for up to 2.5 hrs. At exhaustion, blood glucose (BG) was measured from a tail clip using a hand‐held glucometer. Blood and tissues (heart, liver, skeletal muscle, and adipose) were harvested from all mice. Serum levels of non‐esterified fatty acids (NEFA), triglycerides (TG), cholesterol (CHOL), lactate (LAC), and ketone bodies (KET) were measured using commercially available kits. Endurance exercise times were ~25% higher in females compared to males (121.2 ± 4.0 min vs. 97.3 ± 4.3 min, P<0.05). At exhaustion, BG was significantly lower in females (66.7 ± 3.0 mg/dl) vs. males (75.8 ± 2.4 mg/dl); however, there was no significant difference in LAC. Although post‐exercise serum TG levels were similar, CHOL was significantly lower in females (40.3 ± 2.5 mg/dl vs. 50.6 ± 3.0 mg/dl). Interestingly, serum levels of both NEFA and KET were ~50% higher in females at exhaustion. Although body weight was significantly greater in males (25.6 ± 0.8g vs. 20.1 ± 0.6g), the quadriceps to body weight ratio was similar, suggesting that body or muscle mass was not a factor in the differences. In conclusion, female mice have higher EEC than male mice, which may be due to the supply of exogenous substrates. In particular, female mice have elevated serum levels of fatty acids and ketone bodies at exhaustion, which may reflect an increased ability to mobilize these substrates to prolong endurance exercise. Additional studies will investigate whether endogenous cellular metabolism, especially glycogen and triglycerides in skeletal muscle, contribute to the enhanced endurance capacity phenotype.Support or Funding InformationThis work was supported by funding from the American Heart Association and Ursinus College.This abstract is from the Experimental Biology 2019 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.