bluuebunny/crossref_metadata_2025_split huggingface dataset:
Janus kinase 3 (JAK3) increases phosphorylation of signal transducers and activators of transcription 3 (STAT3) in reproductive cells
Authors: Ndiaye Kalidou , Castonguay Amélie , Benoit Gabriel , Silversides David W , Lussier Jacques G 
Published: November 2023

JAK3 is a member of the membrane‐associated non‐receptor tyrosine kinase protein family and is considered predominantly expressed in hematopoietic cells. We previously identified JAK3 as a differentially expressed gene in granulosa cells (GC) of bovine preovulatory follicles. This study aimed to further investigate JAK3 mRNA and protein regulation and to identify its binding partners in GC of bovine dominant follicles. GC were obtained from small follicles (SF), dominant follicles at day 5 of the estrous cycle (DF), and ovulatory follicles, 24 hours following hCG injection (OF). RT‐PCR analyses showed greatest expression of JAK3 in GC of DF, while the weakest expression was in OF (P < 0.0001). There was a 5‐ and 20‐fold reduction of JAK3 steady‐state mRNA levels in follicular walls, respectively at 12 and 24 hours post‐hCG as compared to 0 hour (P < 0.05). These results were confirmed in western blot analysis showing weakest JAK3 protein levels in OF as compared to DF. Yeast two‐hybrid screening of a DF‐cDNA library resulted in the identification of 23 JAK3 partners in GC. Moreover, the amount of phosphorylated STAT3 in GC was greatest in SF and DF as compared to OF while the opposite was observed for non‐phosphorylated STAT3. In functional studies using bovine endometrial cells, we discovered that phosphorylation of STAT3 (pSTAT3) in a bovine endometrial cell line was increased by JAK3 while inhibition of JAK3 by JANEX‐1 decreased the amount of pSTAT3. In the same study, we showed that JAK3 increased endometrial cell proliferation as well. We found a similar effect of JAK3 on STAT3 phosphorylation using an immortalized murine granulosa cell line. These results support a physiologically relevant role of JAK3 in follicular development and provide insights into the mode of action and function of JAK3 in reproductive tissues.Support or Funding InformationThis work was supported in part by internal funds from Université de Montréal to KN and by a Discovery Grant from Natural Sciences and Engineering Research Council of Canada (NSERC) to JGL.
The Role of Molecular Chaperonin CCT and its Co‐chaperone PhLP1 in the Assembly of mTOR Complexes
Authors: Dhavale Madhura , Plimpton Rebecca , Aoba Takuma , Tensmeyer Nicole 
Published: November 2023

The mechanistic target of rapamycin (mTOR) kinase signaling pathway controls cell proliferation, metabolism and survival. Mutations in components of the mTOR pathway lead to tumerogenesis, angiogenesis and metastasis and therefore are implicated in many types of cancers. mTOR signaling is mediated through two signaling complexes, namely mTORC1 (consisting of mTOR, mLST8, Raptor, Deptor and PRAS40) and mTORC2 (consisting of mTOR, mLST8, Rictor, mSin1, Deptor and Protor1). In these complexes, mTOR is the central kinase whereas the other components are accessory proteins that assist in regulation and recognition of mTOR substrates. The mechanism by which the mTOR components are assembled into functional complexes from nascent polypeptides is a fundamental question yet to be answered. We have evidence indicating that the chaperonin CCT (also called TRiC) and its co‐chaperone PhLP1 assist in mTORC1 assembly by folding two of the mTOR components, mLST8 and Raptor. CCT and PhLP1 form a ternary complex with mLST8, while Raptor binds CCT in the absence of PhLP1. Furthermore, cellular depletion of CCT or PhLP1 disrupts mTORC1 assembly, and CCT depletion inhibits mTORC2‐mediated phosphorylation of Ser473 of AKT. In contrast, the mTORC2 subunit Rictor, which is structurally homologous to Raptor, does not bind CCT. These results suggest that CCT and PhLP1 contribute to mLST8 folding and assembly into mTORC1 and mTORC2, while CCT alone assists in Raptor folding and assembly into mTORC1. Thus, the CCT chaperonin and its co‐chaperones may represent new physiological or therapeutic targets to regulate mTOR signaling.Support or Funding InformationNIH Grants: EY 012287, GM 078550, Brigham Young University Cancer Research Center
The potential link of COMP‐ADAMTS and Aggrecanase‐ADAMTS Genes with Osteoarthrit
Authors: Demircan Kadir , Gürlü Gülsüm , Güven Kübra Nur , Fıstık Tuba , Çakırbay Haşim , Yanık Burcu 
Published: November 2023

This project aimed to investigate the role of COMP‐ADAMTS and aggrecanase‐ADAMTS which are responsible for the destruction of cartilage in the ADAMTS (a disintegrin and metalloproteinase with trombospondin motifs) genes that are suspected to play role in the genetic etiology of the osteoarthritis (OA) which is the most common joint disease.88 patients aged between 40–75 years, with the diagnosis of primary knee OA (according to American College of Rheumatology criteria for knee OA) and 88 healthy volunteers aged between 40–75 years, without OA enrolled in the study. Exclusion criteria include, previous knee surgery (total knee prosthesis etc.), inflammatory rheumatic diseases, secondary OA and malignancies. Radiologic staging of the participants is done, pain and functional status are assessed by visual analogue scale, WOMAC, Lequesne index. Blood samples obtained from the participants analyzed for ADAMTS gene family by ELISA and RT‐PCR.Blood samples obtained from the participants analyzed for ADAMTS ‐1,‐4,‐5,‐7,‐8,‐9,‐12,‐15,‐16 ve ‐18 genes by ELISA. Compared OD levels determined by ELISA reader of groups, OD level of ADAMTS‐4,‐8,‐9 and ‐18 was determined significantly high in the patient group compared to the control group. When examined the patients splitting (dividing) into two groups as early–advanced OA, results of ADAMTS‐4,‐5,‐8,‐9,‐15 ve ‐16 according to control group in early OA group, results of ADAMTS‐15 and ‐16 according to advanced OA group in early OA group and results of ADAMTS‐8, ‐9 and ‐18 according to control group in advanced OA group was seen significantly high.Statistical evaluation of RT‐PCR results showed that ADAMTS‐4,‐9 and ‐18 expressions are significantly decreased in the patient group compared to the control group, ADAMTS‐8 expression is not low enough to be meaningful statistically and there isn't any enough difference between two groups in ADAMTS‐5 expression level.Consequently; we observed that in the early stages ADAMTS‐4, ‐5, ‐15 and ‐16, in advanced stages ADAMTS may be the main aggrecanase responsible for the destruction of cartilage in human knee OA. ADAMTS‐8 and ‐9 were determined to play a role in both early and advanced stages of cartilage destruction in OA. We have seen that ADAMTS‐1, ‐7 and ‐12 were not playing an important role in the pathogenesis of OA.Support or Funding InformationTubitak and Turgut Ozal University Hospital Project Management Ofice
Effect of Pathway‐Interconnectors in SEB‐Induced Apoptosis Related Events in Human PBMCs
Authors: Geddes Erin , Hendricks Michelle , Doyle Connor , Borgos Dylan , Burton Danielle , Mendis Chanaka 
Published: November 2023

Staphylococcus enterotoxin B (SEB) is produced by the bacterium Staphylococcus aureus. SEB is a common cause of apoptosis within human peripheral blood mononuclear cells (PBMCs). The purpose of this study is to further investigate human PBMCs in order to better understand the apoptosis related events induced by SEB. C‐Jun N‐Terminal Kinase (JNK) has been previously identified to be induced by SEB, and has demonstrated that it inter‐connects multiple signaling cascades. As a crucial pathway inter‐connector in SEB‐induced human PBMCs, we believe that inhibiting JNK may possess inhibitory effects on SEB‐induced apoptosis. In this study, JNK specific inhibitor (SP600125) was used to inhibit JNK. We will further evaluate the gene expression profile using RT and PCR. Protein expression patterns of some genes will be examined through the use of enzyme‐linked immunosorbent assays (ELISAs). We believe that our work will allow us to better understand the complex interactions of multiple signal transduction pathways induced by SEB.Support or Funding InformationWe want to thank James Waldschmidt, Tricia Mason, Aaron Hopkins, Emily Freehauf, Amanda Smet, and Nicole Krausert who worked on different aspects of the project. We also wanted to thank our Collaborator Dr. Marti Jett, MEDCOM, Frederick, MD.The project was funded by Centerpoint and PURF, SAIF grants (University of Wisconsin ‐Platteville).
Myocardin Regulated Genetic Pathway Modulates Mitochondrial Permeability Transition Pore Closure to Prevent Cell Death during Cardiac Differentiation
Authors: Mughal Wajihah , da Silva Rosa Simone , Chapman Donald , Hai Yan , Diehl‐Jones William , Gordon Joseph 
Published: November 2023

Myocardin is a transcriptional co‐activator required for cardiovascular development and cardiomyocyte differentiation. Recent studies have shown that genetic inhibition of myocardin results in congenital heart defects associated with increased programmed cell death (PCD). In addition to its established role in regulating PCD, the mitochondrial permeability transition pore (PTP) has been implicated in cardiac calcium homeostasis and myocyte maturation during development. Utilizing fluorescent staining and biochemical assays, we demonstrate that myocardin, a modulator of cardiac gene function, regulates mitochondrial PTP closure during myocyte differentiation to oppose pathways of PCD induced by protein kinase‐A (PKA) activating agents. We show that a loss of myocardin function reduces microRNA‐133a (miR‐133a) mRNA levels and induces mitochondrial dysfunction by PTP opening and reducing mitochondrial membrane potential. Furthermore, myocardin knockdown increased mitochondrial oxidative stress and mitochondrial calcium uptake in differentiated myocytes. Conversely, restoring myocardin expression rescued mitochondrial function, concurrent with a decreased expression of a mitochondrial death protein, Nix. Furthermore, miR‐133a protects against PKA‐activating agents while inhibitory molecules opposing miR‐133a increase Nix expression. With the use of genetically encoded and organelle targeted calcium sensors, our data demonstrates that miR‐133a reverses endoplasmic reticulum calcium release and prevents mitochondrial calcium uptake, in a manner dependent on Nix repression. Finally, we provide mechanistic evidence that myocardin‐dependent miR‐133a expression is regulated by histone deactylase‐5 (HDAC5) and the PKA‐regulated salt inducible kinase‐1 (SIK1). These findings support the notion that myocardin opposes PCD through mitochondrial permeability transition by regulating calcium homeostasis; however, this genetic pathway may be attenuated by pathological PKA signaling.Support or Funding InformationSupported by Children's Hospital Foundation of Manitoba, Research Manitoba and NSERC Canada.
Isolated Extracellular Domain of DR5 is a Potential inhibitor of Apoptosis
Authors: Vunnam Nagamani , Sachs Jonathan N 
Published: November 2023

In recent years, there has been a great interest in pre‐ligand assembly of tumor necrosis factor receptors (TNFRs). TNF‐receptor superfamily plays a major role in regulation of apoptosis. Recent studies suggest that disrupting the pre‐ligand assembly of TNFRs maybe a useful approach for the treatment of TNF‐disorders like rheumatoid arthritis. Thus, strategies for the manipulation of pre‐ligand assembly of TNFRs maybe valuable in prevention and mediation of TNF‐receptor related ailments. In the current study, we targeted the pre‐ligand assembly of death receptor 5 (DR5) to inhibit the apoptosis induced by its cognate ligand. DR5 is a pro‐apoptotic TNF‐receptor protein with a N‐terminal extracellular domain (ECD), a transmembrane domain, and a cytoplasmic death domain, which is crucial for initiation of death signaling upon binding of TNF‐related apoptosis inducing ligand (TRAIL). We hypothesized that ECD, which does not have the death domain, of DR5 could prevent the TRAIL‐induced apoptosis by disrupting the pre‐ligand assembly of endogenous‐DR5 (eDR5) and forming the death‐inhibitory complex with it. To test this hypothesis we first determined the interactions between isolated ECD, eDR5 and TRAIL by using electrophoresis, spectroscopy, FRET and Coimmunoprecipitation (CoIP) studies. These studies show that isolated ECD interacts with both TRAIL and eDR5 and does not significantly influence the eDR5‐TRAIL interactions. We further investigated the effect of ECD on TRAIL‐induced apoptosis by using cell culture studies. Here, we show that ECD inhibits the TRAIL‐induced apoptosis by disrupting the pre‐ligand assembly of eDR5, which is required for induction of death signaling, and forming the anti‐apoptotic complex with eDR5. In summary, our biophysical and cell culture data suggest that the ECD is a potential inhibitor of apoptosis and may have a prospective therapeutic value for the treatment of TNF‐receptor related disorders.Support or Funding InformationThis work was supported by National Institutes of Health Grant GM107175‐02
Modulation of PON2 and Proinflammatory Cytokine Genes in Rat Tissue Exposed to Combined oral Contraceptive Ethinylestradiol and Levonorgestrel
Authors: Wusu Adedoja Dorcas , Rotimi Solomin Oladapo , Saibu Gbemisola Morounke , Rotimi Oluwakemi Anuoluwapo , Giwa Rahman Ayodele 
Published: November 2023

Paraoxonase (PON2) was identified as a genetic risk factor for cardiovascular disease (CVD) and usage of oral contraceptive (OC) is associated with increased cervical cancer and cardiovascular risk. PON2 protect against atherosclerosis development at the cellular level and this phenomenon could be related to their antioxidative properties. Therefore, the aim of the present study was to investigate the effect of OC on the expression of PON2, pro‐inflammatory cytokines interleukin one alpha (IL1α) and tumor necrosis factor alpha (TNFα) in the liver, kidney and brain of rats. Different dosage groups of eight female rats were treated with oral contraceptive (0.15mg levonorgestrel 0.03mg ethinylestradiol(A); 0.3mg levonorgestrel 0.06 mg ethinylestradiol (B) and 0.075 mg levonorgestrel 0.015 mg ethinylestradiol (C))/kg body‐weight(bw)). Two groups of eight rats were included in the study for a control group (D) and ≤0.1% DMSO (drug vehicle) group (E), which were not subject to drug administration for 21days. The levels of expression of the gene were assessed using quantitative reverse polymerase chain reaction technique. Combined oral contraceptive treatment produced a significant increase(p<0.001) in the level expression of renal IL1α and TNFα in all the groups compared to control in a dose‐dependent manner but has no significant effect on PON2.Meanwhile, OC resulted in significantly (p<0.0001) reduced level of expression of hepatic IL1α with no significant effect on hepatic PON2 and TNFα level. In the brain, OC resulted in significantly (p<0.0001) reduced level of expression of TNFα in all dose groups and IL1α level at 0.015mg/bw. Although OC treatment did increase the expression of brain PON2 significantly (p<0.05) at the lowest dose. Therefore, pharmacological modulation of the expression of genes could constitute a useful approach for preventing atherosclerosis
Epidermal Growth Factor Substrate 8 is an essential partnering factor of the Forkhead box transcription factor FOXM1 in the regulation of cell proliferation in cervical cancer cells
Authors: Ngan Wan Ling , Yao Kwok‐Ming , Leung Wai Ying , Chan David Wai 
Published: November 2023

The proliferation‐associated Forkhead box transcription factor FOXM1 (named Molecule of the Year in 2010) is increasingly recognized as marker and therapeutic target in cancers of multiple tissues of origin. FOXM1 plays critical role in maintaining the proliferation of cancer cells and its function is activated by mitogenic signals. In a yeast two‐hybrid screen for FOXM1‐interacting proteins, we identified Epidermal Growth Factor (EGF) Substrate 8 (EPS8), the phosphorylation of which is stimulated by EGF signaling. FOXM1 co‐localized with EPS8 at the G2/M phase of the cell cycle and both proteins can be reciprocally immunoprecipitated from lysates prepared from cervical cancer cells. Interestingly, the nuclear translocation of EPS8 was dramatically enhanced by treatment with the exportin 1 inhibitor leptomycin B, suggesting that EPS8 is actively regulated by nuclear export. Site‐directed mutagenesis defined a functional nuclear export signal within EPS8. To test the relevance of FOXM1‐EPS8 interaction in the regulation of FOXM1 function, we depleted EPS8 expression in cervical cancer cells by RNA interference. Importantly, depletion of EPS8 led to G2/M arrest and suppression of expression of FOXM1 and its known target genes including CCNB1 and CDK1. Moreover, chromatin immunoprecipitation analysis revealed recruitment of both FOXM1 and EPS8 to the promoters of CCNB1 and CDK1, consistent with EPS8 being a partnering factor of FOXM1 in transcriptional regulation. Taken together, our findings provide the first evidence to support a novel role of EPS8 in modulating the function of FOXM1, which is relevant to the development of cancer diagnostic strategy and targeted therapy.Support or Funding InformationThis work was supported by a grant from the Hong Kong Research Grants Council (17125214) awarded to Kwok‐Ming Yao
Role of ERK1/2 and p38 MAPKs in Tributyltin‐stimulated Interleukin 1 Beta Secretion and Production from Human Immune Cells.
Authors: Boules Mariam , Brown Shyretha , Whalen Margaret 
Published: November 2023

Tributyltin (TBT) is an organotin compound that contaminates the environment due to its use as a biocide in a wide variety of applications. TBT has been found in human blood samples at levels as high as 265 nM. Previously, we have demonstrated that TBT interferes with the lytic function of human natural killer lymphocytes and alters the secretion of several pro‐inflammatory cytokines from lymphocytes, including interleukin 1 beta (IL‐1β). IL‐1β regulates cellular growth and immune responsiveness. Elevated levels of IL‐1β have been shown to increase the invasiveness of tumors. Our recent studies showed that TBT exposures of 5–25 nM (levels that are seen in human blood samples) caused increased secretion of IL‐1β from monocyte‐depleted peripheral blood mononuclear cells (MD‐PBMCs). In this study we examine the roles of the ERK1/2 and p38 MAPK pathways in TBT‐induced increases in IL‐1β secretion, protein production, and mRNA levels. MD‐PBMCs were treated with the MEK1/2 inhibitor PD98059 or the p38 inhibitor SB202190 for 1 h prior to exposure to TBT concentrations of 25, 10, and 5 nM for 24 h. Secretion of IL‐1β was measured using ELISA, intracellular protein levels were monitored using western blot, and mRNA expression was determined by RT‐qPCR. Results indicated that the TBT‐induced secretion of IL‐1β requires the ERK1/2 pathway. The increases in secretion are accompanied by increased protein levels in TBT‐exposed cells and this increase in protein production also utilized the ERK1/2 pathway. These data suggest that increased protein production accompanies the increases in secretion of IL‐1β seen in human immune cells exposed to TBT and these increases are dependent on MAPK pathway activation.Support or Funding InformationSupported by NIH grant 5U54CA163066.
Effects of Pentachlorophenol (PCP) on Secretion of Interferon Gamma (IFNγ) from Human Immune Cells
Authors: Massawe Reda , Whalen Margaret 
Published: November 2023

Pentachlorophenol ( PCP) is an environmental pollutant due to its use as an insecticide, fungicide, herbicide, and disinfectant. PCP has been found in human blood and urine samples. Exposure to PCP has been linked with cancers of the blood (lymphoma and myeloma) and kidney. Previous studies in our lab, have demonstrated that PCP interferes with the lytic function of human natural killer (NK) lymphocytes and reduces the levels of several cell surface proteins in NK cells. Interferon gamma (IFNγ) is an inflammatory cytokine that is produced by activated T cells and NK cells and to some extent by monocytes. IFNγ is a regulator of immune system function and inhibits viral infections. The objective of this study is to examine whether PCP is able to affect the secretion of IFNγ from several human immune cell preparations, which include: enriched NK cells; monocyte depleted (MD) peripheral blood mononuclear cells (PBMCs); and PBMCs. PBMCs were prepared by back flushing leukocyte filters followed by density gradient separation from red blood cells and granulocytes, with additional enrichment to yield MD‐PBMCs. Enriched NK cells were prepared using a rosetting procedure. IFNγ secretion was detected using an enzyme linked immunosorbent assay (ELISA). The results indicate that the secretion of IFNγ from all three types of cell preparations was altered when cells were exposed to concentrations of PCP ranging from 0.05 – 5 μM for 24 h, 48 h, and 6 days. The data showed that at least one concentration of PCP at one or more length of exposure significantly increased secretion of IFNγ from NK cells, MD‐PBMCs, and PBMCs in from all donors examined. These data suggest that exposure to PCP may significantly disrupt the immune regulation mediated by IFNγ. Additionally, the cell signaling pathways involved in the PCP‐induced increases in IFNγ secretion were examined.Support or Funding InformationSupported by NIH grant 5U54CA163066.
Dibutyltin Exposures Alter Secretion of Interleukin 6 from Human Immune Cells
Authors: Brown Shyretha , Wilburn Wendy , Whalen Margaret 
Published: November 2023

Dibutyltin (DBT) is an organotin compound that is used as a stabilizer in polyvinyl chloride (PVC) plastics (including pipes that distribute drinking water and bottles) and as a de‐worming agent in poultry. DBT is found in human blood samples, and DBT exposures alter the secretion of tumor necrosis factor alpha (TNFα) from lymphocytes. IL‐6 is an anti‐inflammatory agent, as well as a pro‐inflammatory mediator, that is produced by T lymphocytes and monocytes. It is responsible for immune response regulation as well as tissue repair and cellular growth. The aim of the current study is to determine whether exposure to DBT alters the secretion of IL‐6 from increasingly reconstituted preparations of human immune cells. We examined whether exposure to DBT concentrations of 0.05 to 5 μM after 24h, 48h, or 6 days alters IL‐6 secretion in a preparation of highly enriched human NK cells, a monocyte‐depleted preparation of human peripheral blood mononuclear cells (PBMCs) (MD‐PBMCs), PBMCs, granulocytes, and a preparation combining both PBMCs and granulocytes. The levels of IL‐6 were monitored using an enzyme‐linked immunosorbent assay (ELISA). The results indicate that DBT alters IL‐6 secretion from all cell preparations. The concentrations ranging from 0.5 μM to 5 μM caused diminished IL‐6 secretion, while the lowest concentration caused elevation of IL‐6 secretion. The concentrations and lengths of exposure to DBT that caused statistically significant increases in IL‐6 secretion from human immune cells varied from one donor to the next. Therefore, the data indicates that DBT‐induced alterations of IL‐6 secretion from immune cells may be a significant consequence of DBT exposures that may potentially affect immune competence.Support or Funding InformationSupported by NIH grant 5U54CA163066