bluuebunny/crossref_metadata_2025_split huggingface dataset:
Modulating Human Follicle Stimulating Hormone Signaling Activity Using Negative Allosteric Modulators
Authors: Love Jillian R , Cohen Brian D 
Published: June 2021

Human follicle stimulating hormone (hFSH) is a gonadotropin hormone involved in maturation of ovarian follicles and estrogen production in females and spermatogenesis in males. Its release is regulated via the hypothalamic‐pituitary‐gonadal axis. Proper regulation and function of hFSH in females is necessary for fertility, so its manipulation has been a focus in female fertility treatments and contraception. The hFSH receptor (hFSHR) is a GPCR found in granulosa cells in females and Sertoli cells in males and is responsible for initiating a complex downstream signaling cascade in both cell types. It has recently been discovered that hFSHR is capable of biased signaling, meaning it may be possible to selectively activate some downstream signaling pathways without affecting others. This biased signaling may be achieved through the addition of negative allosteric modulators (NAM's), notably the small molecules ADX68692 and ADX68693. Both NAM's have been shown to decrease FSH‐dependent ovulation in vivo although with differing effects on steroid hormone production. The specific molecular mechanisms by which this occurs is unclear. Utilizing the human granulosa cell line hGrC1 we have observed differences in p44/42 MAPK signaling after treatment with NAM's. In control conditions, activation of p42/44 MAP kinase peaked five minutes following FSH stimulation, but in the presence of NAM's there was a continuous increase in signaling over thirty minutes. Additionally, signaling through protein kinase A (PKA) in the presence of ADX68692 showed increased activation of some substrates but in the presence of ADX68693 basal levels of the same substrates were initially higher and showed a decrease over time. Interestingly, some PKA substrates showed no difference in hFSH activation between untreated and NAM treated samples. The effects of treatment with NAM's show their applicability in the development of novel non‐steroidal contraceptive methods that could be associated with fewer side effects. Studying the complex biased signaling mechanisms of hFSHR could identify ways to manipulate hFSHR for not only improved methods of contraception but also fertility treatments.This abstract is from the Experimental Biology 2019 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
Fyn is recruited to specialized clathrin coated pits and regulates EGF receptor signaling
Authors: Antonescu Costin N , Lucarelli Stefanie , Judge Gurjeet , Abousawan John , DiGiovanni Laura , Vural Dafne , Hong Wanjin , Kim Peter K 
Published: June 2021

The Epidermal Growth Factor (EGF) Receptor (EGFR) is a key receptor tyrosine kinase that controls many facets of cell physiology, including proliferation, survival and migration. EGF stimulation elicits EGFR phosphorylation and/or membrane recruitment of Grb2, Gab1 and phosphatidylinositol‐3‐kinase (PI3K), resulting in production of phosphatidylinositol‐3,4,5‐trisphosphate (PIP3), leading to Akt activation. Concomitantly upon ligand binding, EGFR is simultaneously recruited to clathrin‐coated pits (CCPs), eventually leading to receptor endocytosis. We previously uncovered that perturbation of clathrin, but not of receptor endocytosis, impairs EGF‐stimulated activation of Akt signaling (Garay et al. 2015. Mol Biol Cell.26:3504–19). Moreover, we found that some key EGFR signaling intermediates such as phosphorylated Gab1 are enriched with some CCPs. We thus proposed that in addition to their function as endocytic portals, some CCPs have a direct role in controlling EGFR signaling. How CCPs and clathrin directly control the spatiotemporal properties of EGFR signaling at the plasma membrane prior to receptor endocytosis had remained poorly understood. We uncovered that perturbations of TOM1L1 (a clathrin‐binding regulator of Src‐family kinases) and the TOM1L1‐binding Src‐family kinase Fyn impaired EGF‐stimulated Akt phosphorylation, similarly to the effects of clathrin perturbations. We found that TOM1L1 and Fyn are enriched within a subset of CCPs that also contain EGFR, and that perturbation of TOM1L1 prevented Fyn recruitment to CCPs. Importantly, TOM1L1‐ or Fyn‐positive CCPs exhibit unique properties, such as distinct lifetimes, compared to other CCPs. Thus, our results indicate that a subset of CCPs is specialized by enrichment with key signaling regulators to control EGFR signaling, a phenomenon required for subsequent Akt activation. Given the importance of EGFR in driving growth and survival of certain types of cancer, obtaining a better understanding of how CCPs organize key EGFR signals in space and time within the PM may lead to the development of novel anti‐cancer treatments.Support or Funding InformationFunding for this research was provided by an Operating Grant (no. 125854), a Project Grant and a New Investigator Salary Award from the Canadian Institutes of Health Research, as well as an Early Researcher Award (Ontario Ministry of Research, Innovation and Science) to C.N.A., and an Ontario Graduate Scholarship to S.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.
Cardiac Fibroblast Regulation of Matrix Assembly and Inflammation
Authors: Tallquist Michelle D. 
Published: June 2021

Heart failure is a progressive disease that is often exacerbated by inappropriate cardiac remodeling in response to stress. The excess deposition of extracellular matrix (ECM) by fibroblasts, known as fibrosis, is often coincident with reduced cardiac output . While the detrimental effects of overactive ECM secretion are appreciated, our current understanding of the function of the cardiac fibroblast in the absence of stress is limited. Using a genetic approach in mice to deplete hearts of cardiac fibroblasts, we have investigated how loss of fibroblasts affects heart morphogenesis in the neonate and the adult. In the absence of fibroblasts at postnatal day 5, we observe limited production of ECM especially type I collagen. In the absence of ECM, cardiomyocytes bypass the growth arrest that usually occurs in the first week of postnatal growth and continue to undergo cytokinesis. These data suggest that ECM content may restrict cardiomyocyte growth and loss of ECM may be permissive for continued cardiomyocyte proliferation. To investigate the role of fibroblasts in heart homeostasis and inflammation, we also depleted cardiac fibroblasts from adult hearts. Although subtle alterations in cardiac structure were observed, hearts with 70–80% reduction in fibroblasts were functionally indistinguishable from controls suggesting that type I collagen is more stable than previously thought. Finally, we will describe how the loss of fibroblasts impacts inflammatory responses after injury. While an inflammatory response after acute injury is essential for clearing dead tissue, and some fibrosis may be necessary to maintain the tensile strength of injured myocardium, a sustained inflammatory response can lead to inappropriate fibroblast activation. After injury we observed increased cardiomyocyte hypertrophy and reduced infiltration of neutrophils, monocytes, and T and B cells. Cardiac fibroblast specific translatome analysis revealed that chemokine expression was dynamic after myocardial infarction. In the first four days after injury, Cxcl4, Cxcl10, and Ccl5 were increased, while Ccl17 expression increased and Ccl11 decreased after 8 days of injury. Our results suggest that fibroblasts are essential for ECM production and fibroblast depletion leads to changes in cardiomyocyte biology and inflammation after injury.Support or Funding InformationNHLBI Grant HL144067AHA 17GRNT33660474This abstract is from the Experimental Biology 2019 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
The effects of Se‐allylselenocysteine (ASC), an analogue of garlic compound, on DMBA/TPA‐induced skin tumorigenesis
Authors: Cheng An‐Chin , Lee Ming‐Fen , Pan Min‐Hsiung 
Published: June 2021

Se‐allylselenocysteine (ASC), an analogue of garlic compound, has been shown to inhibit mammary carcinogenesis in vivo and cell growth in vitro. The two‐stage skin carcinogenesis model is an excellent in vivo model which is induced by 7, 12‐dimethylbenz[a]anthracene (DMBA) /12‐O‐tetradecanoylphorbol‐13‐acetate (TPA) in ICR mice, and can well annotate multistep process of carcinogenesis including initiation, promotion and progression. Mice at 6 weeks old were started on 200 nmol DMBA in 200 μL acetone; control mice received 200 μL acetone alone. One week after initiation, the mice were treated topically with 200 μL acetone or promoted with TPA (5 nmol in 200 μL acetone) twice a week for 20 weeks. In the other two groups, the mice were treated with ASC (1 and 5 μmol in 200 μL acetone) 30 min before each TPA treatment. Tumors of at least 1 mm in diameter were counted and recorded every week. The results were expressed as the average number of tumors per mouse, percentage of tumor‐bearing mice, and average tumor weight per mouse. In our finding, topical application of ASC on the dorsal skin of DMBA‐initiated and TPA‐promoted mice could significantly accelerate skin tumorigenesis and raise tumor multiplicity than positive control group (DMBA+TPA). The number of tumors that were 1–3 mm, 3–5 mm and >5 mm in size per mouse was increased in a dose‐dependent manner in the ASC‐pretreated groups. It's probably because ASC needs metabolizing in vivo to have the inhibitory effect of cancer. However, the study is based on the model of applying drug on mouse skin. The pro‐inflammatory activity of ASC can be demonstrated by its effect on COX‐2 expression in TPA‐induced mouse skin.This abstract is from the Experimental Biology 2019 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
Garlic, Ginger and Tumeric Extracts Alter Cell Cycle Arrest through Estrogen Pathway
Authors: Ebuehi Osaretin Albert Taiwo 
Published: June 2021

Garlic (Allium sativum), ginger (Zingiber officinale)) and turmeric (Curcuma longa) possess diverse nutritional, gastronomic and therapeutic functions. The effects of administration of aqueous garlic and ginger extracts, and acetone extract of tumeric at different concentrations from 4.88–1,250μg/ml were tested against the human (MCF‐7) adenocarcinoma breast cancer cell line. The phytochemical examination of these plants showed the presence of alkaloids, flavonoids, and phenol in the plant extracts, while tannins were present in garlic and ginger extracts. Terpenoids was present in garlic and turmeric extracts, cardiac glycoside was present in ginger and turmeric, steroid and saponin were exclusively present in turmeric, while anthroquinone was not detected. The radical scavenging properties of extracts were examined against 2,2‐Diphenyl‐1‐Picrylhydrazyl, Nitric Oxide (NO) and reducing power assay show significant antioxidant activity in all plant extracts along the concentration gradient. Gas chromatography‐mass spectrometry screening of the extracts identifies various biomolecules that might be responsible for anti‐carcinogenicity. The percentage viability of human adenocarcinoma breast cancer cell line (MCF‐7) was determined using MTT test, the percentage viability of cell increase across all concentrations (4.88–1250μg/ml) above 100% original viability. Cisplatin (standard drug) used as a control increase in the same pattern parallel to the garlic extract increase from 0.98 to 250 μg/ml. This result suggests that the effects of garlic, ginger and turmeric extracts do not significantly inhibit carcinogenesis / pathogenesis of human breast cancer rather promote proliferation of breast cancer cell probably due to it lower concentration of the extract that cannot effectively act on the oncogene/tumour suppressor, cyclins E and D1, and p27 of the cell. Data of study indicate that garlic, gingrer and turmeric extracts allow free promotion of the estrogen stimulation that leads to increase cellular proliferation via upregulation of cyclin D1 levels and CDK4/6 activity, as well as c‐myc, and cyclin E/CDK2 levels.This abstract is from the Experimental Biology 2019 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
Glia Maturation Factor‐Antibody Injection Reduces Behavioral Impairment, Neuro Inflammation and Amyloid Pathology in 5XFAD Mice Brains
Authors: AHMED MOHAMMAD EJAZ , Thangavel Ramasamy , Selvakumar Govindhasamy Pushpavathi , Kempuraj Duraisamy , Dubova Iuliia , Zahoor Haris , Raikwar Sudhanshu P , Mentor Shireen , Zaheer Smita , Iyer Shankar S , Zaheer Asgar 
Published: June 2021

Alzheimer's disease (AD) is a progressive neurodegenerative disorder characterized by the accumulation of beta‐amyloid (Aβ) plaques and neurofibrillary tangles (NFTs) in the brain. Emerging evidence suggested that in neurodegenerative disease, aggregation of these proteins can promotes neuroinflammation and neurodegeneration. Glia maturation factor (GMF), a neuroinflammatory protein isolated and cloned in our laboratory play an important role in the pathogenesis of AD. To study the relationship between overexpression of GMF, glial activation and neuroinflammation, we used 9‐month‐old 5XFAD mice, a transgenic model of AD. We intravenously (iv) injected single dose of GMF antibody and found that antibody injection leads to: improvements in behavioral deficits, decreased activation of glial cells and reduced expression of pro‐inflammatory cytokines and amyloid pathology. We report that aberrant expression of GMF, GFAP, IBA1, and co‐localization of GMF with GFAP, IBA1 and Aβ significantly more in 5XFAD mice as compared with wild type mice. However, following GMF antibody injection, we found significant reduced expression of GMF, GFAP, IBA1 and Aβ in the cortex region of 5XFAD mice. We conclude that GMF‐antibody may be used to reduce neuroinflammation in neurodegenerative diseases including Alzheimer's disease.Support or Funding InformationNational Institutes of Health Grants NS073670, AG048205 and Veterans Affairs Merit Award I01BX002477 to AZ.This abstract is from the Experimental Biology 2019 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
Na,K‐ATPase β2‐subunit/AMOG as a regulator of Merlin/NF2 signaling in the cerebellum
Authors: Langhans Sigrid A , Litan Alisa , Li Zhiqin , Tokhtaeva Elmira , Kelly Patience , Vagin Olga 
Published: June 2021

The Na,K‐ATPase, consisting of a catalytic α‐subunit and a regulatory β‐subunit, is a ubiquitously expressed ion pump that carries out the transport of Na+ and K+ across the plasma membranes of most animal cells. In addition to its pump function, Na,K‐ATPase serves as a signaling scaffold and a cell adhesion molecule. Of three b‐subunit isoforms, β1 is found almost in all tissues, while β2 expression is mostly restricted to brain and muscle. In cerebellar granule cells, the b2‐subunit, also known as Adhesion Molecule on Glia (AMOG), has been linked to neuron‐astrocyte adhesion and granule cell migration, suggesting its role in cerebellar development. Nevertheless, little is known about molecular pathways that link the b2‐subunit to its cellular functions. Using cerebellar granule precursor cells, we found that the β2‐subunit, but not the b1‐subunit, negatively regulates expression of a key activator of the Hippo/YAP signaling pathway, Merlin/neurofibromin‐2 (NF2). The knockout of the β2‐subunit in mice or its knockdown in cerebellar granule cell precursors resulted in increased Merlin/NF2 expression and affected down‐stream targets of Hippo signaling, i.e. increased YAP phosphorylation and decreased expression of N‐Ras. Further, the β2‐subunit knockdown altered the kinetics of Epidermal Growth Factor Receptor (EGFR) signaling in a Merlin‐dependent mode and impaired EGF‐induced reorganization of the actin cytoskeleton. Therefore, our studies for the first time provide a functional link between the Na,K‐ATPase β2‐subunit and Merlin/NF2 and suggest a role for the β2‐subunit in regulating cytoskeletal dynamics and Hippo/YAP signaling during neuronal differentiation.Support or Funding InformationThis work was supported by funds from NIGMS‐8P20GM103464, the Nemours Foundation and the DO Believe Foundation. The Biomolecular Core Lab is supported by grants NIGMS P30GM114736 (COBRE) and NIGMS‐P20GM103446 (INBRE).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 flavor chemicals commonly found in E‐vaping liquids on IL‐8 increase and barrier function in human epithelial cells
Authors: Ware Blue , Waters Alexander , Roecklein‐Canfield Jennifer 
Published: June 2021

The number of individuals using e‐cigarettes or e‐vaporizers has significantly increased, within the last decade. Many use these products as a way stop smoking cigarettes, as they are advertised as a safe alternative. Recent studies, however, have shown that e‐liquids can contain numerous flavor chemicals at various quantities, since there is currently no government regulation determining how much of these chemicals can be added to e‐liquids. While recent studies have analyzed the contents of e‐liquids, almost nothing is known about the harmful effects of these flavor chemicals on lung cell health. The goal of this study was to evaluate barrier function and the release of pro‐inflammatory cytokines, specifically interleukin‐8 (IL‐8), in human epithelial cells in response to three different e‐vaping flavoring chemicals. The barrier function is highly regulated and acts as an obstacle against harmful gases and vapors. In response to cellular stresses, the function of the barrier is negatively impacted, which increases pollutants access to the submucosal membrane, resulting in an inflammatory response. Inflammation can often lead to irreversible scarring that can damage lung epithelial cells.. This research analyzes how various concentrations of cinnamaldehyde, benzaldehyde and vanillan, which have been found in e‐liquids at high concentrations, affect epithelial cell health. After exposure the relative expression of IL‐8 were measured using qPCR and ELISA. The barrier function was analyzed using Multiplex in order to look at tight junctions (TJs) in relation to epithelial barrier function. These findings will be used to elucidate the harmful effects flavor chemicals, in e‐liquids, have on lung cell 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.
Attenuated Innate Immunity in Mouse Embryonic Stem Cells as a Self‐Protective Mechanism for Early Embryogenesis
Authors: Chen Bohan , Guo Yan‐Lin 
Published: June 2021

Embryonic stem cells (ESCs) have been intensively studied as a promising cell source for regenerative medicine. However, their immunological property is an important yet poorly understood issue in the therapeutic application of ESCs and in developmental biology. Innate immune responses are critical defense mechanisms that are well‐developed in somatic cells, however, a series of our recent studies demonstrated that ESCs are deficient in innate immune responses to viral/bacterial infections and inflammatory cytokines. These findings raised an intriguing question about the rationale for ESCs, the progenitors of all tissue cells, to not have an innate immunity that has adapted so well in somatic cells. Immune and inflammatory responses have been viewed as a double‐edged sword. On the one hand, they defend the organism against pathogens; on the other hand, they can cause collateral damage to the tissue cells, such as cell cycle inhibition and even cell death. From this perspective, we hypothesize that the attenuated innate immunity could be a self‐protective mechanism for ESCs to avoid the cytotoxic effect of immune/inflammatory responses. To test this hypothesis, we differentiated ESCs to fibroblast‐like cells (ESC‐FBs), which have gained partial innate immunity. In comparative analyses, we exposed ESCs and ESC‐FBs to several inflammatory conditions, including inflammatory cytokines (TNFα and IFNγ), lipopolysaccharide (LPS), the conditioned medium collected from LPS‐stimulated macrophages, and heat killed bacteria. The effects were assessed by cell viability analysis and the induction of inflammatory genes. In all conditions tested, ESCs were less susceptible to the cytotoxic effects of the inflammatory insults and showed diminished induction of inflammatory genes than ESC‐FBs. At the molecular level, the signaling pathways that mediate the effects of LPS, TNFα and IFNγ are mostly defective in ESCs. We conclude that the attenuated innate immune responses could be an adaptive mechanism for ESCs to avoid the cytotoxicity caused by immune reactions resulting from embryo implantation or pathogen infection at the early stage of embryogenesis.Support or Funding InformationThis work was supported by National Institutes of Health Grants R15GM109299 and R15GM128196 (to Y‐L. G.). We thank Mississippi‐INBRE for the use of the imaging facility, funded by an Institutional Development Award (IDeA) from the National Institute of General Medical Sciences of the National Institutes of Health under grant number P20GM103476This abstract is from the Experimental Biology 2019 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
Dissecting the Role of ArhGAP12 in Natural Killer Cell‐Mediated Cytotoxicity
Authors: Heyblom Raini A. , Overlee Brittany L. , Billadeau Daniel B. 
Published: June 2021

The immune system consists of two broad branches: innate and adaptive. In the innate branch of the immune system, one will find natural killer (NK) cells, which are responsible for recognizing and killing virally infected or cancerous cells within the human body. Numerous immunodeficiency syndromes arise from mutations in genes that encode proteins essential to NK cells including those involved in NK cell development and effector functions. Among the most important NK cell effector functions is cell‐mediated cytotoxicity. Our lab has been interested in understanding the molecular machinery that regulates this process, in particular those regulated by the small GTP‐binding proteins of the Rho family. With this in mind, the ArhGAP12 protein, which functions as a GTPase activating protein for members of the Rho family, was tested in NK cells to see if it had an effect on cytotoxicity. Significantly, knockdown of ArhGAP12 in NK cells resulted in diminished NK cell‐mediated cytotoxicity. In addition, we found that ArhGAP12 is tyrosine phosphorylated following stimulation through the NKG2D+2B4 activating receptors, indicating that it is linked to this important signaling pathway. As ArhGAP12 is a multi‐modular signaling protein containing a src‐homology 3 (SH3), WW, pleckstrin homology (PH), and GAP domains, we performed proteomics to identify potential interacting partners using the SH3 domain as bait. We found that ArhGAP12 interacts strongly with multiple regulators of F‐actin dynamics including WAVE2, WASP, Evl and VASP. Taken together, these results suggest that ArhGAP12 is linked to activating receptors on NK cells, associates with F‐actin regulators, and is involved in NK cell‐mediated cytotoxicity.Support or Funding InformationMayo Clinic Graduate School of Biomedical Sciences SURF Program and the R01 grant AI120949 to D.D.B.This abstract is from the Experimental Biology 2019 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.