bluuebunny/crossref_metadata_2025_split huggingface dataset:
Induction of a Non‐Apoptotic Cell Death in Momordica Charantia ‐Treated Human Cancer Cell Lines
Authors: Ehigie Adeola Folasade , Wei Peng , Wei Taotao , Ehigie Leonard Ona , Yan Xiyun , Ojeniyi Fiyinfoluwa Demilade , Olorunsogo Olufunso Olabode 
Published: June 2021

One major mechanism of death in cancer cells is via the induction of apoptosis, the best‐studied form of programmed cell death. Although apoptosis is essential in many normal physiological processes, including tissue sculpting during embryogenesis, development of the immune system and destruction of damaged cells, its alteration is a major cause of cancer. Having confirmed the cytotoxic effect of certain fractions derived from the crude water‐soluble extract (CWSE) of Momordica charantia Linn (MC) in a previous study (Ehigie et al., 2016), we investigated in this study, the mode of death induced by these fractions, both in human breast [MDA‐MB‐436] and lung cancer [A549] cell lines, using PI (Propidium Iodide), DAPI (4,6‐diamidino‐2‐phenylindole dihydrochloride) and Caspase‐3 Activation Determination Assays. Western blotting was also employed to verify the activation of caspase‐3 and the mitochondrial release of cytochrome c. The ethyl acetate (D3), n‐hexane (D4) and dichloromethane (D5) fractions were all obtained from the CWSE via solvent partitioning and then administered at 100 and 125 μg/mL for 24 h. Though we observed that the cancer cells treated with the ethyl acetate fraction of the CWSE of M. charantia were the most killed, we saw neither the evidence of chromatin condensation, cell shrinkage and apoptotic bodies, nor the activation of caspase‐3 or cytochrome c release! Data obtained for caspase‐3 activation by MC show no significant (P<0.05) difference between the negative control and all MC‐treated groups for both cell types with the negative control group for MDA‐MB 436 cells at 90.97±9.03 compared with D3‐treated (89.26±3.17 and 96.29±3.85), D4‐treated (90.99±19.78 and 91.71±4.83) and D5‐treated (93.95±2.95 and 88.91±1.00) cells at 100 and 125 μg/mL respectively, similar to values obtained for A549 cells; negative control (69.69±2.60) compared with D3‐treated (70.39±6.93 and 76.21±2.65), D4‐treated (73.67±1.32 and 74.25±0.00) and D5‐treated (71.80±3.26 and 71.17±14.85) cells at 100 and 125 μg/mL respectively. From the foregoing, we conclude that the cell death induced by the ethyl acetate, n‐hexane and dichloromethane fractions of MC in the selected cancer cell lines is by a non‐classical apoptotic mechanism.Support or Funding InformationOrganization for Women in Science for the Developing World (OWSD) and Swedish International Development Cooperation Agency (SIDA)This abstract is from the Experimental Biology 2019 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
Insights into the Role of the Membrane on PLCβ and Gαq‐Mediated Activation and Adsorption
Authors: Hudson Brianna N. , Lyon Angeline M. 
Published: June 2021

Phospholipase C β (PLCβ) enzymes are peripheral membrane proteins that are required for normal cardiovascular function and whose dysregulation results in cardiovascular disease. PLCβ hydrolyzes phosphatidylinositol‐4,5‐bisphosphate (PIP2) to produce the two potent second messengers inositol‐1,4,5‐triphosphate (IP3) and diacylglycerol (DAG), increasing intracellular Ca2+ and activating protein kinase C. PLCβ has low basal activity, but is activated downstream of G protein‐coupled receptors (GPCRs) through direct interactions with the heterotrimeric G protein subunits Gαq and Gβγ. While G proteins stimulate activity, they are insufficient for full activation, suggesting the membrane contributes to full activation. However, the molecular basis for the role of the membrane in basal and G‐protein stimulated activity are not well understood. We seek to understand how membrane composition helps regulate adsorption of PLCβ to the membrane, and how interfacial activation contributes to basal activity and Gαq‐dependent activation. Using a model membrane system, we are applying an innovative combination of atomic force microscopy, mass spectroscopy, and biochemical assays to begin understanding how the membrane itself, regulatory elements within PLCβ, and Gαq regulate lipase activity. These studies provide the first structure‐based approach to understanding how the cell membrane regulates the activity of this essential effector enzyme.Support or Funding InformationThis work was supported by the Purdue Center for Cancer Research, the Showalter Foundation, American Heart Association Scientist Development Grant 16SDG29920017, and NIH 1R01HL141076‐01 to A. Lyon and a Purdue Doctoral Fellowship to B. Hudson.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 the proteoglycan, serglycin, in platelet exocytosis
Authors: Lykins Joshua , Chanzu Harry , Whiteheart Sidney 
Published: June 2021

Platelet granule secretion is essential for the maintenance of the vascular system and influences cardiovascular diseases such as stroke and myocardial infarction. Platelets secrete their granules in response to vascular damage, which promotes clot formation; but, excessive secretion can lead to occlusive thrombosis. Platelets are derived from large progenitor cells called megakaryocytes that reside in the bone marrow. These cells project proplatelet extensions into circulation for platelet production. Platelet secretory granules are synthesized within megakaryocytes and packaged into proplatelets. Serglycin is an intra‐granular, chondroitin sulfate proteoglycan produced by several hematopoietic cells. Serglycin−/− mice exhibit decreased agonist‐dependent secretion of α‐granule cargo (Platelet Factor‐4 (PF4), MIP‐1) despite showing normal exposure of α‐granule membrane protein (P‐selectin), suggesting a defect in granule cargo storage and/or solubility. We hypothesize that the negative charges present on serglycin are vital for the retention and solubility of α‐granule cargo during their synthesis in megakaryocytes and packaging into platelet granules. We examined primary megakaryocytes isolated from bone marrow of wild‐type and serglycin−/− mice. Serglycin−/− megakaryocytes showed an increase in PF4 in the culture media, coupled with a decrease in PF4 associated with cell pellets, as measured by ELISA assays. Fluorescent microscopy also showed a decrease in the retention of fibrinogen, a soluble factor endocytosed by megakaryocytes and sorted into α‐granules, in serglycin−/− megakaryocytes. Additionally, myelofibrosis and splenomegaly were observed in serglycin−/− mice. These studies contribute to an understanding of the physiological function of serglycin in megakaryocytes and to the mechanism of α‐granule biosynthesis.Support or Funding InformationSupported by HL56652, HL138179 and VA Merit AwardThis abstract is from the Experimental Biology 2019 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
Inner mitochondrial translocase Tim50 is central in steroid metabolism in steroidogenic tissues
Authors: Bose Himangshu S , Marshall Brendan , Perry Elizabeth W , Whittal Randy M 
Published: June 2021

Steroid hormones are synthesized in an acute condition but some genes are also transcriptionally active, and they are essential for survival of all mammals. The synthesis of metabolic reaction is initiated in the matrix side of mitochondria. In addition to the presence of different enzymes mitochondria has many translocases, which help in sorting and localization proteins across the subcompartments. The molecular basis of mitochondrial redistribution depends on the different steps of protein folding. In adrenal and gonads, Cytochrome P450 side chain cleavage enzyme (SCC or CYP11A1), catalyzes cholesterol to pregnenolone. We find that gradual mitochondrial import of SCC led to the association with mitochondrial translocase, Tim50. The N‐terminal is cleaved first followed by the C‐terminus amino acids, where deletion of more than ten amino acids from the C‐terminus reduced the activity and interaction with Tim50. Similarly, knocking down Tim50 ablates SCC activity and blocking complete SCC import into the inter mitochondrial membrane ablated activity. These two proteins are colocalized in the mitochondria, where amino acids 208–212 of Tim50 first interact with SCC. A mutation in either of these two proteins reduced their interaction, resulting in a metabolic and developmental disorder. Thus, the metabolism for conversion of pregnenolone depends on SCC and Tim50 together, as the negative feedback is inhibited in the absence of interaction between these two proteins, increasing ACTH levels.Support or Funding InformationNoThis abstract is from the Experimental Biology 2019 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
Avocado Oil Ameliorates Non‐Alcoholic Fatty Liver Disease by Down‐Regulating Inflammatory Cytokines and Improving Mitochondrial Dynamics
Authors: Garcia‐Berumen Claudia Isabel , Olmos‐Orizaba Berenice Eridani , Márquez‐Ramírez Cristian Adrián , Orozco Alain Raimundo Rodríguez , González‐Cortez Aramen , Saavedra‐Molina Alfredo , Montoya‐Pérez Rocio , Cortés‐Rojo Christian 
Published: June 2021

Nonalcoholic Fatty Liver Disease (NAFLD) is a chronic disease characterized by excessive fat accumulation, inflammation and liver dysfunction in the absence of significant alcohol consumption and without any other liver disease. NAFLD is accompanied by mitochondrial dysfunctions such as decreased activity of the enzymes of the electron transport chain (ETC), impaired β‐oxidation of fatty acids, excessive reactive oxygen species (ROS) production and increased lipid peroxidation. These last two processes have been involved in liver inflammation by augmenting the activity of cytokines like TNF‐α, and IL‐6. Mitochondrial dynamics, a process that modulates both mitochondrial morphology and function by events of fusion and fission, also becomes impaired during NAFLD by exacerbating mitochondria fission. Previously, we have reported that avocado oil, a rich source of C18:1, bioactive sterols and antioxidants, attenuates both mitochondrial dysfunctions and oxidative stress during diabetes and hypertension. Therefore, we aimed to test if avocado oil attenuates NAFLD by counteracting the alterations in both cytokines levels and mitochondrial dynamics in rats feed with a diet with high fat and fructose for 4 months. This diet led to hepatic alterations including inflammation, ballooning, necrosis and increased expression of both TNF‐α and IL‐6. Increased levels of ROS production and lipid peroxidation were observed in mitochondria, along with augmented expression of DRP1 and Fis1, two proteins of fission, and decreased levels of fusion proteins Mfn1/2 and OPA1. All these effects were counteracted when avocado oil was supplemented daily after one month of the beginning of the diet up to the end of the experiment. These data suggest that avocado oil ameliorates NAFLD by decreasing inflammation and improving mitochondrial dynamics. Thus, avocado oil may be a nutritional approach to complement pharmacological treatment of NAFLD.Support or Funding InformationThis work was supported by a Coordinación de la Investigación Científica‐UMSNH grant (to CCR)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 Three Blind Men: How Adhesion Proteins Binding Affects the Retinas of People with Retinoschisis
Authors: Strandberg Stacey , Bauer E. , Cobb E. , DebBaruah C. , Fricker E. , Griffith‐Topps A. , Jessick M. , Kuehn C. , Kulju T. , Lois C. , Sargent C. , Schraufnagel L. , Strandberg S. , Strother C. , Thota A. , Urban S. , Fleischmann Scott 
Published: June 2021

X‐linked retinoschisis, an early‐onset macular degenerative disease occurring primarily in young boys, can result in blindness. As many as two hundred different mutations on the X chromosome can lead to a malformation of retinoschisin, a structural protein which normally holds the photoreceptors and bipolar cells together. The photoreceptors detect light, and bipolar cells process these stimuli for interpretation by the brain. An improperly folded retinoschisin prevents the photoreceptors and bipolar cells from interacting and prevents information from being transmitted to the brain. Retinoschisin is composed of sixteen subunits arranged in a double octomer. Because retinoschisin relies on extensive interactions between subunits, many different mutations can lead to a dysfunctional protein. The C110 and C142 bonds stabilize the spikes protruding from the discoid fold—the part of the protein that allows the retinoschisin to bind to the retinal cells. Mutations in these residues lead to the inability of the protein binding to retinal cells. Since the retinoschisin cannot attach to the retinal cells, the retina begins to split and causes eventual blindness. In order to further understand the structure and function of this protein, the Divine Savior Holy Angels SMART (Students Modeling a Research Topic) team modeled retinoschisin using 3D printing technology. Although no cure exists, current research is focusing on the structure of retinoschisin, and how mutations prevent adhesion between retinal cells with an ultimate goal of developing a gene therapy. By correcting the mutated X chromosome segment, retinoschisis could be prevented.Support or Funding InformationMunzenmaier, Dr. Diane <munzenmaier@msoe.edu>This abstract is from the Experimental Biology 2019 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
Human RGS14 binds NHERF1 and regulates NPT2A‐mediated phosphate uptake: Disruption by genetic variants
Authors: Hepler John R. , Mamonova Tanya , Sneddon W. Bruce , Emlet David , Harbin Nicholas H. , Squires Katherine E. , Friedman Peter A. 
Published: June 2021

RGS14 is a multifunctional scaffolding protein that integrates G protein, MAPK and Ca++/CaM signaling pathways. Most of what we know about RGS14 comes from studies in rodent brain where it acts as a natural suppressor of synaptic plasticity and hippocampal‐based learning, though other studies implicate a role for RGS14 in myocardial remodeling. Recent genome‐wide association studies have implicated RGS14 in disordered phosphate metabolism, an effect of chronic kidney disease. The RGS14 gene is adjacent to SLC34A1 on human chromosome 5 that encodes the NPT2A sodium‐phosphate cotransporter, which mediates hormone‐sensitive phosphate resorption in renal proximal tubules. Therefore, the goal of these studies was to define a possible role for RGS14 in hormone‐sensitive phosphate transport. Human/primate RGS14 differ from rodent proteins by possessing a carboxy‐terminal extension encoding a Type I PDZ ligand (DSAL). Of note, several rare human mutations have been reported in this region of RGS14 located at residues 563 and 565, corresponding to positions −1 (A565S, A565V) and −3 (D563N, D563G) of the PDZ ligand. The family of PDZ adapter proteins assemble a variety of membrane‐associated proteins, including transporters, receptors, and ion channels, and signaling molecules in short‐lived functional units. The PDZ adapter NHERF1 is expressed in renal proximal tubules, and contains two PDZ domains (PDZ1 and PDZ2). NHERF1 binds the PDZ ligand carboxy tail of NPT2A to promote its phosphate uptake, and this NPT2A:NHERF1 complex is inhibited by parathyroid hormone (PTH) acting at the PTH1 receptor. Here we report that RGS14 is expressed in proximal and distal renal tubules of humans, and that RGS14 binds to NHERF1 via the PDZ2 domain. PTH inhibits NPT2A uptake of phosphate, and RGS14 blocks this action. Of the RGS14 genetic variants tested, D563N disrupted binding to NHERF1 and RGS14 actions on PTH‐sensitive phosphate uptake, whereas D563G, A565S and A565V were innocuous. Computational analysis and molecular dynamics modeling of NHERF1 PDZ binding to the RGS14 C‐terminal PDZ ligand define the structural determinants of this interaction. Taken together, our findings are consistent with the idea that RGS14 contributes to PTH‐sensitive phosphate transport in humans, and that RGS14 coding variants may cause disordered phosphate metabolism.Support or Funding InformationNational Institutes of Health (NIH) awards DK105811, DK111427 (PAF); NS037112, NS102652 (JRH).This abstract is from the Experimental Biology 2019 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
Deletion of Telomere‐Rap1 aggravates cardiac aging by impairing fatty acid oxidation via PPARα signaling
Authors: Liu Hao , Cai Yin , Ying Fan , Liu Shiming , Xia Zhengyuan 
Published: June 2021

The heart undergoes multiple functional and structural declining with aging, including impaired fatty acid metabolism, cardiac dysfunction and hypertrophy. Repressor activator protein 1 (Rap1) as a component of shelterin complex is essential for the maintenance of telomere function. Moreover, Rap1 also regulates gene transcription by binding to non‐telomeric sites such as the nuclear hormone receptor ‐ peroxisome proliferator‐activated receptor α (PPARα) which plays a central role in the cardiac fatty acid metabolism and mitochondrial function. However, it's still not clear whether or not the heart of Rap1−/− mice also suffers more aging associated disorders. Thus, the present study aims at the role of Rap1 in cardiac aging and its possible mechanism. Echocardiography showed enlarged left ventricular internal dimension (LVID) and LV mass, which suggested dilated hypertrophy, and elevated myocardial performance index (MPI) in Rap1−/− mice at age of 85weeks, indicating systolic and diastolic dysfunction (all P<0.05 vs. Rap1+/+). The greater adverse cardiac remodeling in aged Rap1−/− mice was paralleled by elevated cardiomyocyte area showed by Wheat germ agglutinin (WGA) staining (P<0.05 vs. Rap1+/+). p53 as a senescence marker was also increased indicating more severe cellular senescence in cardiomyocyte of Rap1−/− mice (P<0.05 vs. Rap1+/+). From apex cordis of Rap1 wildtype and knockout mice, electron microscopy showed more enlarged pleomorphic mitochondria containing cristae fragmentation, vacuolization and disrupted external membranes in aged Rap1 knockout mice compared to age‐matched wildtype counterparts. Decreased Acetyl‐CoA carboxylase (ACC) (catalyzes the production of malonyl‐CoA) and Elevated Carnitine palmitoyl transferase I (CPT‐1) (transports fatty acyl‐CoA into mitochondria) and Acyl‐CoA dehydrogenase long chain (ACADL, catalyzes most of fatty acid β‐oxidation) were detected in aged Rap1 knockout mice(all P<0.05 vs. Rap1+/+), indicating Rap1 deficiency may lead to reduced fatty acid oxidation during aging. In the heart of aged Rap1−/− mice, PPARα was reduced compared with age‐matched wildtype counterparts (P<0.05 vs. Rap1+/+), suggesting that with aging, Rap1 may down‐regulate PPARα signaling in the heart and control fatty acid metabolism resulting in cardiac dysfunction and hypertrophy. In conclusion, Rap1 deficiency may impair myocardial fatty acid oxidation via PPARα signaling, leading to more severe cardiac aging as cardiac dysfunction and hypertrophy. These findings may identify a new extratelomeric role of Rap1 in cardiac metabolism during aging.Support or Funding InformationThe authors' work was supported by the Health and Medical Research Fund (04151816, Hong Kong) and General Research Fund (17121315M, 17123915M and 17117217M, Research Grants Council of Hong Kong).This abstract is from the Experimental Biology 2019 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
A model of steady state ultrafiltration through an endothelial glycocalyx bi‐layer formed by a porous outer layer and a more selective membrane‐associated inner layer
Authors: Curry FitzRoy E , Michel Charles 
Published: June 2021

Ultrastructural investigations of the endothelial glycocalyx reveal a layer adjacent to the cell surface with a structure consistent with the primary ultrafilter of vascular walls. Theory predicts this inner layer can be no greater than 200–300 nm thick, a result to be reconciled with observations that red cells and large macromolecules are excluded from a region up to 1 micron or more from the cell membrane. We tested whether this apparent inconsistency might be accounted for by a model of steady state water and protein transport through a glycocalyx bi‐layer. Expressions for coupled water and albumin fluxes through the two layers were used to describe steady state ultrafiltration. The model demonstrated that albumin may accumulate at the interface between the porous layer and the selective inner layer. The osmotic pressure of accumulated albumin would significantly modify the observed permeability properties of the microvessel wall by an effective unstirred layer effect. Our results place constraints on the outer layer permeability properties. For example, with an perfusate albumin concentration of 50 mg/ml, Adamson et al ( Am. J. Physiol. 557: 889–907,2004) reported that the hydraulic conductivity measured in rat mesenteric microvessels under conditions of steady state ultrafiltration were similar to that measured before steady ultrafiltration was established. The only properties of the outer layer consistent with these observations are albumin permeability coefficients and hydraulic conductivities close to an order of magnitude larger than that of the inner layer. A high filtration coefficient for the outer layer is also consistent with estimates of the hydraulic conductance of the outer layer needed to account for red cell movement over the endothelial surface up to 1 micron thick.This abstract is from the Experimental Biology 2019 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.