bluuebunny/crossref_metadata_2025_split huggingface dataset:
Effects of the Cell Cycle on Vacuole Size in S. cerevisiae yeast
Authors: Sims Jasmine C. 
Published: June 2021

Malfunctions in a cell's ability to degrade unnecessary chemicals or proteins may lead to one of fifty distinctive lysosomal storage disorders in humans. Lysosomal storage diseases such as Tay‐Sach's, Salla, or Sialic Acid Storage diseases often lead to premature death in infants. Lysosomes are vital for degrading subcellular proteins no longer needed by the cell. Our research uses a functional homologue of lysosomes, the vacuole in S. cerevisiae yeast as a model system, which also functions in protein degradation.We study vacuoles in S. cerevisiae yeast to understand protein degradation. We investigate whether changes in vacuole size and degradative function are influenced by the progression of the cell cycle. We hypothesize that changes in vacuolar size and specific phases of the cell cycle play an important role in degradative function. To address this, we designed experiments to approach this by linking vacuole size changes to the progression of the cell cycle. We halted the cells at each cell cycle stage to track vacuolar changes as the cell cycle progresses. We did this by incubating the cells with 4 cell cycle inhibition drugs: Tributyltin (G1 phase), Hydroxyurea (S phase), Epothilone B (G2 phase), and Nocodazole (M phase). Vacuoles were fluorescently labelled with VPH1‐GFP, a vacuolar surface protein. We imaged the samples using Spinning Disk Confocal Microscopy. We then analyzed the images with ImageJ and MatLab algorithms to track, compare, and characterize vacuolar change. Thus far, we have found that the vacuoles change in terms of copy number (the number of vacuolar lobes) as the cell cycle progresses. This preliminary evidence suggests that there is a correlation between changes in vacuolar size along with the progression of the cell cycle. Because of this, we hypothesize that vacuole size is influenced by the progression of the cell cycle.In addition, decreases in degradative ability are correlated with enlarged vacuolar phenotypes, hence a logical future direction would be to test whether degradative ability may also be synchronized with the progression of the cell cycle. Characterizing these changes would allow us to predict degradative efficiency based on the phase of the cell cycle. Further research could determine whether protein degradation and vacuolar morphology depend on one another. Overall, our research is significant because it will provide molecular insight on how vacuolar morphology and degradation are regulated, which may lead to new treatments to help those who suffer from lethal lysosomal storage disorders, and those who struggle with inefficiencies in degradative ability.Support or Funding InformationNSF‐Center for Cellular Construction GrantThis abstract is from the Experimental Biology 2018 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
GlycoSense™: A Rapid Method for Monitoring In Vitro Glycoengineering
Authors: Yang Loretta , Saunders Matthew J. , Samli Kausar N. , Woods Robert J. 
Published: June 2021

The use of enzymes to modify glycans, in order to generate uniform protein glycosylation for optimal bioactivity, or to test glycan‐structure function hypotheses, or in chemo‐enzymatic synthesis, has emerged as a transformative technology. However, there are no convenient cost‐effective methods for monitoring the progress of the enzyme reactions during in vitro glycoengineering.Lectenz® Bio has developed an innovative technology called GlycoSense™ that will permit in vitro glycoenzyme reactions to be monitored in near real‐time using flow cytometry and multiplex microspheres (beads). The GlycoSense™ method detects binding between glycans and glycan‐specific reagents that are conjugated to spectrally‐unique beads. By combining the individual bead‐based reagents into a multiplexed array, a profile of key glycan features can be obtained in less than a minute on a basic cytometer.We illustrate the performance of the GlycoSense™ method using glycans and glycoprotein standards, and demonstrate its utility by monitoring changes in glycosylation patterns upon treatment with glycoenzymes. The GlycoSense™ approach will address the unmet need for a rapid tool to monitor protein glycosylation during in vitro glycoengineering.Support or Funding InformationNational Institutes of Health (U01CA221213, R43GM123863)This abstract is from the Experimental Biology 2018 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
Neuroregeneration improved by ketones
Authors: D'Agostino Csilla Ari , Zippert Monique , D'Agostino Dominic P. 
Published: June 2021

Ketone bodies serve as alternative fuel for the brain when glucose availability is limited. The ketone body beta‐hydroxybutyrate (BHB) is synthesized in the liver from acetoacetate, the first ketone produced in the fasting state. Several studies show that being in the state of ketosis can be neuroprotective, but the exact mechanism is not known. It is also not known whether ketones would aid regeneration of a nervous system injury. In this study we investigated on cell cultures whether ketone bodies improve neuroregeneration after injury or trauma.The effect of ketone bodies on cell regeneration was tested in 3 weeks old rat (Sprague Dawley) primary neuronal cell cultures. Scratch assay was performed for 24 hours, while the cells were either exposed to 2mM R, S‐sodium‐3‐hydroxibutyrate (BHB) or not (control). One hundred snapshots were taken with the CYTOSMART system to document the cell migration, changes in cell density and the process of cell regeneration into the damaged area over the 24 hours. Immunofluorescence reaction was performed to label beta III tubulin and synapsin I in the regenerating areas.Cell cultures treated with BHB showed more intense cell migration and regeneration of the damaged area over the 24 hour period. Significantly more DAPI stained cell nuclei were found in the damaged area in cell cultures with 2mM BHB at 20× (p=0.01)and at 10× (p=0.0004) magnification photos as well, and the cell coverage was higher than in control cell cultures. Markers that are associated with flagellar movement, cytoskeletal functions and axonogenesis, synaptogenesis were denser in treated neuronal cultures at the regeneration site as revealed by fluorescence microscopy. These above results show potential applications for ketogenic compounds for treating traumatic brain injury or other injury to the central nervous system.Support or Funding InformationUniversity of South Florida FoundationThis abstract is from the Experimental Biology 2018 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
AKR1A1 deficiency is associated with high risk of alcohol‐induced fatty liver syndrome
Authors: Chen Wan‐Ru , Lan Ying‐Wei , Chen Hsiao‐Ling , Chen Chuan‐Mu 
Published: June 2021

Alcoholic liver disease(ALD) is caused by excessive alcohol consumption. The spectrum of alcohol‐related liver injury varies from hepatic steatosis, steatohepatitis, alcoholic fibrosis to irreversible cirrhosis. AKR1A1 is a member of the aldo‐keto reductase (AKR) superfamily, which catalyze the reduction of various aldehydes to their corresponding alcohols in an NADPH‐dependent manner. The enzyme is encoded by the AKR1A1 gene that protects cells against oxidative stress and detoxification. The aim of this study is to identify the role of AKR1A1 deficiency in mouse hepatic diseases after chronic alcohol exposure. In this study, we have established a novel strain of AKR1A1 knockout (KO) mice, which integrated the pCAG‐eGFP transgene and simultaneously deleted 17kb of the AKR1A1 gene locus. The AKR1A1 KO mice (AKR1A1eGFP/eGFP) reduced ascorbic acid biosynthesis, may affect aldehyde reductase metabolism. Male AKR1A1eGFP/eGFP and CD‐1 control mice were divided into four groups(n=6) that fed either the Lieber‐Decarli diet containing 5 % ethanol (Ethanol‐fed) or an isocaloric (Pair‐fed) control diet for 8 weeks. In comparison with ethanol exposure‐CD‐1 control mice, the ethanol exposure‐ AKR1A1eGFP/eGFP mice showed a significant increase in the serum alanine aminotransferase (ALT) and aspartate transaminase (AST) levels (p<0.05), and the levels of pro‐inflammatory cytokines (TNF‐α and IL‐1β), hepatic triglycerides(TG) and cytochrome P4502E1(CYP2E1) were also elevated. However, the levels of antioxidant enzymes, including hepatic glutathione(GSH), malondialdehyde (MDA), and superoxide dismutase (SOD) were significantly reduced in the ethanol exposure‐AKR1A1 KO mice. The histological analysis observed that ethanol exposure‐ AKR1A1eGFP/eGFP mice developed fatty liver syndrome, but not observed in the ethanol exposure‐CD‐1 control mice. In conclusion, we demonstrated that chronic exposure of alcohol lead to lack of hepatic steatosis and inflammation in AKR1A1 KO mice, which may provide a novel insight into the role of AKR1A1 in alcoholic liver disease.This abstract is from the Experimental Biology 2018 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
Honokiol and Pioglitazone Ameliorate Alzheimer's Disease Pathologies in vitro and ex vivo
Authors: Lynd Tyler , Govindarajulu Manoj , Dhanasekaran Murali , Suppiramaniam Vishnu 
Published: June 2021

Amyloid beta (AB) plaques are extracellular, neurotoxic protein aggregations. These plaques have historically been considered the hallmark of Alzheimer's disease due to their ability to reduce long term potentiation (LTP) and promote long term depression (LTD). Peroxisome proliferator‐activated receptor gamma (PPARg) agonists have been shown to be a novel and promising class of drugs capable of alleviating AD pathologies. In this study, we observed the neurotoxic effects of amyloid beta on hippocampal field potentials on the Schaffer collateral. In addition, we displayed honokiol, a novel SIRT3 and partial PPARg agonist, and pioglitazone, a full PPARg agonist, is capable of improving mitochondrial function, increasing LTP, and reducing AB aggregation. The hippocampal field potential along the Schaffer collateral was measured by the LTP theta burst protocol, while signaling mechanisms were evaluated using western blot, RT‐qPCR, mitochondrial assays, and imaging techniques. We hypothesize and provide evidence that these compounds regulate mitochondria‐endoplasmic reticulum communication in order to modify AD pathologies providing evidence for the MAM hypothesis. Both compounds performed comparably suggesting honokiol may be a promising compound for the treatment of AD as its blood brain barrier (BBB) permeability is much higher than pioglitazone.This abstract is from the Experimental Biology 2018 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
Cell‐Specific Role of Aryl Hydrocarbon Receptor in Liver Fibrosis
Authors: Yan Jiong , Xie Wen 
Published: June 2021

Background & AimsThe role of aryl hydrocarbon receptor (AhR) in liver fibrosis is controversial, because both AhR ablation and AhR activation lead to liver fibrosis. The liver is an organ of multiple cell types, including hepatocytes, hepatic stellate cells (HSCs) and Kupffer cells. We investigated cell‐type specific role of AhR in liver fibrosis and the specific inhibitory effect of AhR in HSC activation.MethodsPrimary mouse and human stellate cells and LX2 cell line were used to characterize the in vitro function of AhR in the HSC activation. Hepatocytes‐, HSCs‐, and Kupffer cells‐specific AhR knockout and AhR−/− mice were used to study the role of AhR in the spontaneously developed liver fibrosis and carbon tetrachloride (CCl4)‐ and bile duct ligation (BDL)‐induced liver fibrosis in vivo. Activation of HSCs and fibrotic phenotype were measured by quantitative real‐time PCR and Western blot analysis of fibrogenic genes, immunofluorescence of α‐smooth muscle actin (α‐SMA) and Ki67, and histological analysis.ResultsAhR is highly expressed in HSCs and declines with the onset of HSC activation in the primary HSCs. AhR−/− HSCs exhibited accelerated culture‐induced activation. AhR antagonist promoted while agonist inhibited the HSC activation, respectively. Ablation of AhR in HSCs was sufficient to sensitize mice to liver fibrosis.ConclusionsAhR is an intrinsic suppressor for HSC activation and liver fibrosis. Development of non‐toxic AhR agonists or strategies that preferentially activate AhR in the HSCs represent a novel therapeutic for liver fibrosis.Support or Funding InformationNIH DK083952This abstract is from the Experimental Biology 2018 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
Development of novel inhibitors of swelling‐activated LRRC8 anion channels
Authors: Figueroa Eric , Fulton Mark , Kharade Sujay , Kramer Meghan , Weaver David , Lindsley Craig , Denton Jerod 
Published: June 2021

Cells critically regulate their volume in response to hypoosmotic swelling by transporting chloride and small organic osmolytes out of the cell through so‐called volume‐regulated anion channels (VRACs). The 5 genes encoding VRACs were recently identified in two independent genome‐wide siRNA screens and are named LRRC8 (for Leucine Rich Repeat Containing) A‐E. LRRC8‐containing VRACs have recently been implicated in drug uptake, adipocyte size, and insulin signaling. Unfortunately, the pharmacological tools available to study the physiology and therapeutic potential of LRRC8 channels is limited to low‐affinity small‐molecule inhibitors that exhibit promiscuous activity toward other anion channels, cation channels, and myriad other off‐targets. To circumvent this barrier, we are employing high‐throughput screening (HTS) of small‐molecule libraries and medicinal chemistry to develop more potent and specific inhibitors of LRRC8 channels. HTS will be performed using a fluorescence assay based on halide quenching of a YFP mutant (i.e. F46L, H148Q, I152L) termed Ozzy. Briefly, HEK‐293 cells stably transfected with Ozzy will be plated in black‐walled, clear‐bottomed 384‐well plates, treated with hypotonic solution containing compounds from the Vanderbilt Institute of Chemical Biology library, and then treated with sodium iodide (NaI). Cell swelling opens endogenously expressed VRAC channels, through which iodide enters the cell and quenches Ozzy fluorescence. Small‐molecule inhibitors should block the iodide‐induced quenching of Ozzy. Checkerboard analysis using the best‐in‐class VRAC inhibitor, DCPIB, yielded a Z′ value of 0.74, indicating the assay is reproducible and robust enough to ensure high‐confidence hit‐picking in a large‐scale screen. The assay also reports dose‐dependent inhibition of iodide influx by DCPIB with an IC50 of 11 micromolar, which is close to values derived from electrophysiological experiments. We are currently using this assay to interrogate an analog library based on the DCPIB scaffold, with the goal of identifying more potent and specific inhibitors of VRAC.Support or Funding InformationDepartment of Anesthesiology Faculty Development FundsThis abstract is from the Experimental Biology 2018 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
LRRC26 is functional as an Auxiliary Subunit of Large‐Conductance Ca2+‐Activated K+ (BK) Channel and regulates BK channel activity in Bronchial Smooth Muscle Cells
Authors: Noda Sayuri , Suzuki Yoshiaki , Yamamura Hisao , Imaizumi Yuji 
Published: June 2021

Large‐conductance Ca2+‐activated K+ (BK) channel activity is known to be an essential factor for the control of tone and/or motility in various types of tissues containing smooth muscle cells (SMCs) as one of main components. Recently leucine‐rich‐repeat‐containing proteins (LRRC) have been identified as novel auxiliary (γ) subunits of BK channel. However, the distribution and contribution of BKγ subunits in SMCs is unknown. We found that LRRC26 (BKγ1) is highly and specifically expressed in mouse bronchial SMCs (mBSMCs). In the present study, we examined functional roles of BKγ subunits in mouse bronchial SMCs (mBSMCs). Co‐immunoprecipitation and single molecule image analyses showed the direct interaction between BKα and BKγ1 in BSMCs. Whole‐cell patch‐clamp analyses revealed that BK channel sensitivity to intracellular Ca2+ concentration was much higher in mBSMCs than that in mouse aortic SMCs (mASMCs). Mallotoxin, which activates BK channels only in the absence of BKγ1, dose‐dependently activated BK channel currents in mASMCs, but not in mBSMCs. Knock‐down of BKγ1 in mBSMCs significantly reduced BK channel Ca2+ sensitivity and contraction induced by a BK channel blocker. Taken together, BKγ1 in mBSMCs substantially contributes to keep BK channel activity high and the resting membrane potential deep under resting conditions. Thus, BKγ1 in mBSMCs may be responsible to keep the airway resistance stably low under physiological conditions.This abstract is from the Experimental Biology 2018 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
Changes in the participation and expression of angiotensin II receptor At1 in heart and kidney of rats 6 weeks old with neonatal administration of de dexametasone and hyperbaric oxygen therapy
Authors: Duron‐Gil Alejandra , Guevara‐Balcazar Gustavo , Martinez‐Godinez Maria Angeles , Rubio‐Gayosso Ivan , Franco‐Vadillo Antonio , Castillo‐Hernandez Maria Carmen 
Published: June 2021

One therapy that has demostrated that it normalize the cardiovascular responses of angiotensin receptor in one experimental model, is the hyperbaric oxygen therapy. It has been described the wide use of dexametasone during the neonatal period of time, having different and controversial responses. Dexametasone has been used commonly due to the improvement in the lung function into the respiratory difficulty syndrome. Some studies reports that the use of steroid during the neonatal period could have some beneficial effects but also it could induce cardiovascular and renal disorder during the development. As it has been described one of the main pathway involved is the renin‐aldosterone‐angiotensin system. Some researchers describe that steroids regulate the angiotensin II receptor and function.Objetivethe main idea is to analize the modification of the expression of angiotensin II receptor in rat with administration neonatal of dexametasone with/without hyperbaric oxygen therapy.Methodologyneonatal rat (2 days) were administrated with dexametasone and were divided in 2 groups one with hyperbaric oxygen therapy and the other without therapy. At the age of 6 weeks the animals were sacrificed, and the expression of the receptor At1 were measured in the heart and the kidney by PCR and inmunohistochemistry.ResultsThe group only with dexametasone showed has major expression of At1 receptor, however the group with dexametasone and hyperbaric oxygen therapy has a reduction in the expression of this receptor.ConclusionThis results could suggest that the hyperbaric oxygen therapy could normalize the expresion in this rats with dexametasone treatment, being beneficial because it could reduce the possibility of one cardiovascular pathology like and infarct.Support or Funding InformationSIP, COFAA, IPNThis abstract is from the Experimental Biology 2018 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
Arginase inhibition improves vascular function and restores NO production in diabetic conditions
Authors: Shatanawi Alia , Momani Munther S. 
Published: June 2021

Cardiovascular complications of diabetes are a leading cause of morbidity and mortality. Vascular endothelial dysfunction (VED) is strongly implicated in the pathogenesis of diabetic vascular complications. Diabetes has been linked to elevated arginase and associated with vascular endothelial dysfunction. The pathological process is characterized by impaired endothelial cell production of nitric oxide (NO) and/or decreased NO bioavailability. NO is produced by activity of endothelial NO synthase (eNOS) on its substrate L‐arginine. Arginase in ECs also uses L‐arginine as a substrate to produce urea and ornithine. Thus, elevated levels of arginase will limit availability of L‐arginine to eNOS and decrease NO production.We performed analysis of arginase activity in type 2 diabetes mellitus (T2DM). Arginase activity was elevated in diabetes patients as compared to age‐matched healthy volunteers. Levels of arginase activity had a positive correlation with HbA1c levels in diabetic patients (R2=0.8 Pearson r=0.87). Cell studies also agreed with these findings, as high glucose (25 mmol/L, 72 hrs) treatment to ECs resulted in a significant increase in arginase activity. This increase in arginase activity was concomitant with a 27% drop in NO produced by EC. Inhibition of arginase by ABH (100 uM) restored NO level to normal.The effects of L‐citrulline, a natural arginase inhibitor, on inhibiting arginase activity in T2DM patients was assessed. Twenty five patients received L‐citrulline supplements (2000 mg/day) for 1 month. Arginase activity was decreased 30% in T2DM patients after taking L‐citrulline supplements. There was a modest improvement on HbA1c levels in these patients. The effect on L‐citrulline on arginase activity was also measured in BAECs grown in high glucose conditions. L‐citrulline 100μM completely prevented the increase in arginase activity and restored NO production levels. These data indicate that inhibiting arginase activity can have therapeutic benefits in diabetic patients by preventing vascular dysfunction and maintaining NO levels.Our results indicate that arginase can be regarded as a novel marker for the vascular complications of diabetes. Drugs targeting arginase or its signaling pathway may show benefits in delaying or preventing these vascular complications.Support or Funding InformationThe University of Jordan deanship of researchThis abstract is from the Experimental Biology 2018 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.