bluuebunny/crossref_metadata_2025_split huggingface dataset:
Carvedilol protection involves the mitochondrial deacetylase Sirt3 in mitigating Doxorubicin‐induced Cardiomyocyte Injury
Authors: Uche Nnamdi , Dai Qiang , Lai Shuping , Benjamin Ivor 
Published: April 2020

Although anthracyclines such as doxorubicin are highly effective chemotherapeutic agents and widely used for various malignancies, their use in the clinic is accompanied by major cardiovascular events. Cardiotoxicity is a well‐recognized and unpredictable consequence of anthracycline therapy that may be permanent and progressive, leading to heart failure in some patients while severely impacting quality of life. The pathophysiology of anthracycline‐induced cardiotoxicity has not been fully elucidated but the most widely accepted hypothesis is the increased reactive oxygen species (ROS) generated by anthracyclines. Data from our lab as well as other recent studies support an alternative model involving mitochondrial dysfunction perpetuated by anthracycline‐induced dysregulation of mitochondrial protein modifications. The mitochondrial deacetylase Sirt3 has been suggested to play a role in cardio‐protection. Carvedilol, a beta‐blocker, had previously been shown to mitigate anthracycline‐induced mitochondrial dysfunction and although the mechanism is unknown, data from our lab suggest involvement with Sirt3. To explore this further, rat embryonic cardiomyocytes (H9c2 cells) devoid of Sirt3 expression were either treated with doxorubicin or pretreated with carvedilol followed by doxorubicin treatment before downstream assays were performed. Protein expression and/or activity assays were performed to assess proteins involved in apoptosis and mitochondrial function and stability including caspase3 and the Sirt3 target Mn/SOD. MitoSOX assay was done to assess mitochondrial ROS generation. Our results suggest that carvedilol prevents or reverses the anthracycline‐induced inhibition of Sirt3, a protein responsible for activating mitochondrial proteins involved with metabolic processes and maintenance of ROS. Further and ongoing studies in the lab will access the role of Sirt3 in cardio‐protection in vivo.Support or Funding InformationNIH CTSA UL1TR001436
Relative Shedding of Glycosaminoglycans from the Endothelial Glycocalyx During Inflammation
Authors: Lipowsky Herbert H. 
Published: April 2020

The endothelial (EC) surface layer (glycocalyx) has been shown to act as a barrier to transvascular exchange of solutes, and adhesion of leukocytes (WBCs) during the inflammatory process. It is a labile structure whose components are readily shed by the action of proteases and endoglycosidases. Evidence to date suggests that during the inflammatory process, activation of matrix metalloproteases on the EC surface may cleave the principal proteoglycans (syndecans and glypicans), and EC derived lyases may result in shedding of the associated glycosaminoglycans (GAGs) heparan sulfate (HS), chondroitin sulfate (CS) and hyaluronan (HA). Shedding of these components of the glycocalyx serves to increase leukocyte (WBC) access to adhesion receptors on the EC membrane. The objective of the present study was to elucidate the relative role of HS (the dominant GAG) and HA as the major barrier to WBC‐EC adhesion.Shedding of glycans from the EC in post‐capillary venules of the mesentery (rat) in response to topical application of the chemoattractant f‐met‐leu‐phe (fMLP, 10−6 M) was observed by intravital microscopy. The presence and loss of HS, CS and HA was assessed by labeling all components with fluorescently labelled lectin (BS‐1) or HS antibodies, and HA with fluorescently labelled hyaluronan binding protein (HBP). During exposure to fMLP, GAG concentration exhibited an exponential decay. Following a 30 min period about 50% of HBP was lost in contrast to a previously shown loss of 20% of lectin labelled GAGs, and 25% loss of Mab labelled HS. The exponential time‐constant for HBP shedding was 5.8 min, which was one‐third the 14.3 min for BS‐1 labelled GAGs.The relatively rapid shedding of HA compared to HS was consistent with the hypothesis that HA may form the dominant barrier to WBC‐EC adhesion. In situ observations (in rat mesentery) of fMLP induced transient adhesion of circulating beads coated with antibody to ICAM‐1 (1A29) revealed a time constant for adhesion of 10.9 min, which was less than that for BS‐1 labelled GAGs. Prior observations by Fan et al., (Biorheology, 2019, DOI: 10.3233/BIR‐180204) using stochastic optical reconstruction microscopy suggest that HA lies closer to and parallel to the endothelial membrane, compared to HS which lies further away and normal to the EC surface. Thus, a compact layer of HA near the EC membrane may surround WBC adhesion receptors that are much shorter in length than the total thickness of the glycocalyx. Its more rapid shedding, compared to other GAGs, may be the major factor affecting exposure of WBC adhesion receptors on the EC.Support or Funding InformationSupported in part by NIH R01 HL39286
Investigation of type 1 angiotensin II receptor activation induced gene expression changes in rat vascular smooth muscle cells
Authors: Hunyady Laszlo , Balla András , Szalai Laura , Gém Janka , Porkoláb Edit , Kovács Kinga , Szakadáti Gyöngyi 
Published: April 2020

Angiotensin II (AngII) mainly acts through type 1 angiotensin II receptor (AT1R) to promote a broad variety of biological effects. The vascular smooth muscle cells are one of the main target of AngII and its stimulation activates numerous signaling pathways which could result in gene expression changes in these cells.We carried out Affymetrix GeneChip experiments to analyze the effects of AngII stimulation on gene expression. More than 200 genes were upregulated in response to AngII stimulation of primary rat vascular smooth muscle cells in our experimental setup. We have selected several genes (DUSP5, DUSP6, DUSP10, Lmcd1, HbEGF, and endothelin) to further investigate the kinetics of the gene‐expression changes, and the possible signal transduction processes which lead to the altered expression changes after AngII stimulation. The results of the quantitative PCR measurements confirmed the AngII‐induced upregulation of the investigated genes. The maximal induction of the most genes appeared two hours after the AngII stimulation. Our data show that the upregulated genes can be stimulated through several, possibly parallel signal transduction pathways. We also examined the role of epidermal growth factor receptor (EGFR) transactivation in the upregulation of the investigated genes. According to our data the EGF receptor transactivation is not critical in response to AngII stimulation since silencing of EGF receptor did not significantly alter the AngII induced gene expression changes. Our pharmacological approach revealed that dasatinib, which is a potent inhibitor of ABL and Src family soluble tyrosine kinases, can significantly inhibit several gene expression changes. Interestingly, when we applied ABL kinase inhibitors, which have less potency toward Src tyrosine kinases such as imatinib or nilotinib did not evoke similar effects as dasatinib.Our data can help to understand the details of AngII‐induced long term effects and the pathophysiology of AT1R, moreover it can help to develop potential interventions in symptoms when this receptor over functions such as vascular remodeling or atherosclerosis.Support or Funding InformationThis work was supported by the National Research, Development and Innovation Fund (NKFI K116954 and NVKP_16‐1‐2016‐0039).
Red Blood Cell‐resolved 3D Computational Modeling of Blood Flow in Physiologically Realistic Microvascular Networks
Authors: Bagchi Prosenjit , Ebrahimi Saman 
Published: April 2020

Microvascular networks represent geometrically complex architecture due to constantly bifurcating and merging vessels. Moreover, blood vessels are not necessarily straight, and often highly winding. Mathematical models of blood flow in such networks have treated these vessels as 1D straight segments neglecting the geometric complexity and details. These models also do not consider the deformation and flow of individual red blood cell, and instead use empirical relations to prescribe blood viscosity and red blood cell distribution at a vascular bifurcation. As such, these models cannot predict hemodynamic variations over short time and length scales. To overcome this limitation, we have recently developed a high‐fidelity 3D, first‐principle based, fluid‐‐structure interaction model of blood flow in physiologically realistic microvascular networks. The model retains the exact geometric details of the vasculature, e.g., 3D nature of the blood vessels and bifurcations, and vessel curvature. Deformation and flow of each individual red blood cell as they flow through the vessels are exactly modeled. The model is based on the minimal assumptions: it does not assume any empirical viscosity law or bifurcation law; rather, these laws are predicted by the model. The model provides microscale details of hemodynamic quantities that are not readily available in experiments, and cannot be predicted by the traditional 1D network models, such as wall shear stress gradient and time‐dependent red blood cell partitioning at bifurcations.Of particular interest is the role of red blood cell deformability on hematocrit distribution in the networks. Cell deformability is known to reduce in many diseases, such as sickle cell disease, malaria and diabetes. By using in vivo images of retinal microvasculature, we have performed simulation of network blood flow under varying cell deformability in such networks. The model shows that altered cell deformability significantly changes the cell distribution in the network, and subsequently the flow resistance in vessels. While some vessels show a significant increase in hematocrit, some show a significant decrease. Such a redistribution of hematocrit for diseased red blood cells may have a significant impact on tissue oxygen delivery. The simulation results are used to mechanistically explain the observed redistribution of hematocrit and vascular resistance.Support or Funding InformationFunded by NSF
Use of Mass Spectrometry‐Based Proteomics to Investigate Protein Regulation Associated with COPD‐Related Muscle Dysfunction
Authors: Schiltz Sarah B. , Balnis Joseph , Jaitovich Ariel , Vincent Catherine E. 
Published: April 2020

Chronic Obstructive Pulmonary Disease (COPD) is an umbrella term used to encompass several progressive lung diseases. A common comorbidity of COPD is skeletal muscle dysfunction with muscle mass loss, a condition shown to decrease quality of life and increase mortality rates independently of the level of pulmonary disease. Interestingly, rehabilitation exercises for the recovery of skeletal muscle integrity has been associated with improved disease outcomes. There are two mouse models that have been developed to mimic the COPD phenotype and demonstrate skeletal muscle wasting that recapitulates the disease seen in humans. A comprehensive mass spectrometry‐based proteomics study in one of these models (a mouse conditionally over‐expressing interleukin‐13, IL13TG) yielded a set of potential protein biomarkers for the degree of muscle dysfunction in skeletal muscle, including haptoglobin, hemopexin, and ceruloplasmin. Our present study will use mass spectrometry‐based proteomics to interrogate protein regulation in the EDL muscle of the other mouse model (a mouse that has lung‐specific expression of human matrix metalloproteinase‐1 (MMP‐1), an interstitial collagenase). We hypothesize that we will find a set of regulated proteins in common between the two studies, providing a set of conclusive biomarkers for COPD‐related skeletal muscle dysfunction that are independent of genetic modification. If our hypothesis is confirmed, we will then have a list of potential targets that can help facilitate early detection and/or potential interventions for COPD in humans to achieve improved patient outcomes.Support or Funding InformationPart of the results reported herein have been funded Hartwick College and by NHLBI of the National Institutes of Health under the award number K01‐HL130704 (AJ), and by the Collins Family Foundation Endowment (AJ); NIH/NHLBI 5R01HL049426 (HAS); NIH/NHLBI PO1 HL114501(JAE); R01 HL115813 (CGL); NIH/NIGMS grant 1R01GM124133 (APA)
Melatonin plus Folic Acid Treatment Ameliorates Reserpine‐Induced Fibromyalgia: Evaluation of Pain, Oxidative Stress and Inflammation
Authors: Fusco Roberta , Siracusa Rosalba , D’Amico Ramona , Peritore Alessio Filippo , Cordaro Marika , Gugliandolo Enrico , Crupi Rosalia , Impellizzeri Daniela , Cuzzocrea Salvatore , Di Paola Rosanna 
Published: April 2020

BackgroundFibromyalgia is a chronic condition characterized by increased sensory perception of pain, neuropathic/neurodegenerative modifications, oxidative and nitrosative stress. An appropriate therapy is hard to find, and the currently used treatments are able to target only one of these aspects.MethodsThe aim of this study was to investigate the beneficial effects of a melatonin plus folic acid administration in a rat model of reserpine‐induced fibromyalgia. Sprague–Dawley male rats were injected with 1 mg/kg of reserpine for three consecutive days and later administered with melatonin, folic acid or both for twenty‐one days.ResultsAdministration of reserpine led to a significant decrease in nociceptive threshold as well as significant increase in depressive‐like symptoms. These behavioral changes were accompanied with increased oxidative and nitrosative stress. Lipid peroxidation was significantly increased, as well as nitrotyrosine and PARP expression, while superoxide dismutase, nonprotein thiols and catalase were significantly decreased. Endogenously produced oxidants species are responsible of mast cells infiltration, increased expression pro‐inflammatory mediators and microglia activation.ConclusionMelatonin plus acid folic administration was able to ameliorate the behavioral defects, oxidative and nitrosative stress, mast cells infiltration, inflammatory mediators overexpression and microglia activation induced by reserpine injection with more efficacy than them separate administration.
Structural and Functional Characterization of Phosphoglycosyl Transferases from Human Pathogens
Authors: Dodge Gregory , O’Toole Katherine , Bernstein Hannah , Seebald Leah , Allen Karen N. , Imperiali Barbara 
Published: April 2020

Bacterial virulence and viability are strongly dependent on extracellular glycoconjugates including lipopolysaccharide (LPS), capsular polysaccharide (CPS), wall teichoic acid (WTA) and glycoproteins. While these glycoconjugates are diverse in structure and function, they share a common biosynthetic logic. For example, in Gram‐negative bacteria the biosynthesis of a variety of glycoconjugates are initiated on the cytoplasmic face of the bacterial inner membrane, with a phosphoglycosyl transferase (PGT) catalyzing the first membrane‐committed step of the pathway. PGTs transfer a phosphosugar from a nucleotide‐activated donor substrate onto a polyprenol phosphate such as undecaprenol phosphate (UndP). PGTs comprise two distinct superfamilies – polytopic, with multiple membrane spanning helices, and the more recently described monotopic, which contain an unusual re‐entrant membrane intercalating helix. To date, monotopic PGTs have only been observed in bacteria, and remain underexplored antimicrobial targets. In an effort to understand NDP‐sugar specificity and inform inhibitor design, a library of monotopic PGTs from human pathogens was expressed, purified, and subjected to structural and functional characterization. Both detergent and styrene maleic acid (SMA) copolymers were assessed for solubilization towards the purification and stabilization of these challenging protein targets. These efforts have led to purified samples of diverse monotopic PGTs for activity analyses using the UMP‐Glo ™ assay and crystallization screens, which have afforded reproduceable crystals for further investigation.Support or Funding InformationR01GM131627, F32GM134576
Gut microbiome induces leaky gut and inflammation by activating miRNAs which in turn reduces tight junction proteins
Authors: Mishra Sidharth P. , Nagpal Ravinder , Razazan Atefeh P. , Wang Bo , Yadav Hariom 
Published: April 2020

In obesity and diabetes, abnormal gut microbiota and increased gut permeability (leaky gut) are common risk factors which in turn contributes in increasing chronic low grade inflammation. However, precise mechanism(s) whether and how gut microbiota contributes in induction of leaky gut remain largely unknown. Here we, demonstrated that obese/ diabetic gut microbiota not only associated with leaky gut and inflammation, but it plays a causative role to induce leaky gut and inflammation. Using fecal microbiota transplantation, intestinal organoids and human epithelial Caco2 cell systems, we demonstrated that obese microbiota dramatically reduces intestinal barriers such as tight junction proteins (TJPs), that are associated with increased leaky gut and inflammation. Intestinal epithelial TJPs are important structural proteins that holds epithelial cells tight enough to regulate non‐specific permeability of microbial/ food and other antigens from intestinal lumen, thereby, keep a check on inflammatory reactions in the mucosal immune system. We also demonstrated that gut microbiota metabolites that are enriched in obese/ diabetic gut contributes in reducing the expression of TJPs and increasing leaky gut. To decipher the mechanism, we found that obese/diabetic microbiota derived metabolites increases an array of miRNAs (including miR‐10a and miR101a‐3p) that downregulates TJPs expression, which in turn results leaky gut and chronic inflammation in diabetes and obesity. Results demonstrating that obese/diabetic microbiota and its metabolites are causative to increase leaky gut and inflammation by activating miRNAs that disrupts TJPs, are not only of great significance to understand the underlying biology of leaky gut and inflammation in obesity and diabetes, but will also pave the ways to develop novel strategies to prevent/ treat these debilitating public health diseases.Support or Funding InformationWe thank funding from the Department of Defense, National Institutes of Health and Wake Forest School of Medicine, and wonderful support from other team members.
Employing Eye Tracking Technology to Understand How Novices Learn Neuroanatomy
Authors: Nguyen Angela , Cheung Beata , Leclair Rebecca , Saini Jessica , Lyons Jim , Wainman Bruce , Heisz Jennifer , Brewer-Deluce Danielle 
Published: April 2020

IntroductionNeurophobia is the fear of neuroscience experienced by students who are learning and applying content. Although the cause remains unclear, neurophobia presents a major barrier to student success in neuroanatomy. A preliminary study suggested that individuals with low working memory capacity (WMC) benefit from using high‐contrast anatomical images when learning neuroanatomy. However, the reason for the improvement is unknown.AimThis study uses eye tracking technology to better understand how individuals of varying WMC and expertise study neuroanatomical images. We aim to: 1) assess differences in gaze patterns between students of high and low WMC when viewing high and low contrast brain slices and 2) compare novices and experts to uncover potential competency‐related differences.MethodsUndergraduate students with no prior anatomical education (n=120) and neuroanatomical experts were recruited. During an eye tracking session, participants were given 5 minutes to study twelve structures on digital images of 4 brain slices (either coronal or transverse slices in high or low contrast) and were then tested on their ability to identify the learned structures on similar low‐contrast images. This procedure was repeated such that all participants were exposed to a set of coronal/transverse and high/low contrast images in a randomized fashion. All participants’ eye tracking data and test accuracy were recorded. After testing, participants completed the Automated Operation Span Task (OSPAN) to quantify WMC.ResultsPreliminary results show that students with high WMC are more accurate in identifying neuroanatomical structures when compared to low WMC students [F(1, 32), p =.088]. Dwell time was a significant predicting factor for accuracy [r(32)=40, p =.02]. High WMC students fixated longer on neuroanatomical structures when compared to low WMC students [F(1, 32)=6.50, p =.02]. Data collection from expert participants is ongoing.Discussion/ConclusionOverall, these results offer a quantitative measure of how neuroanatomical information is viewed and learned to gain insight into neurophobia and influence future teaching practices in neuroanatomy.
Perfluorooctanoic Acid (in the Presence of Fetal Bovine Serum) Induces Proliferation in ERα Positive and ERα Negative Breast Cancer Cell Lines
Authors: Gaw Victoria , Glenn Manderrious , Cannon Jennifer 
Published: April 2020

Perfluorooctanoic acid (PFOA) is a synthetic chemical belonging to a larger group of fluorotelomers. These compounds have been used in the production of both industrial and consumer products as surfactants and due to their chemical nature are environmentally persistent pollutants. Individuals can be exposed to PFOA through ingesting PFOA‐contaminated water or food. PFOA has been shown to have a half‐life of approximately 4 years in humans. While the long‐term effects of PFOA are largely unknown, there is increasing evidence suggesting it to be an endocrine disruptor. Studies have shown that PFOA binds to and activates peroxisome‐proliferator‐activated receptor α (PPARα), which can regulate the expression of other genes and receptors such as the other PPAR isoforms as well as estrogen receptor α (ERα).Previous experiments in our lab demonstrated that PFOA treatment of MCF‐7 breast cancer cells (an ERα positive cell line) decreased expression of ERα mRNA and protein, and decreased cell viability by ~20% within 48h of treatment compared to DMSO controls. However, these cells were treated in the absence of fetal bovine serum (FBS). When we repeated these experiments without serum withdrawal, we initially noted a tendency towards increased proliferation in MCF‐7 cells treated with 50μM and 100μM PFOA at both 24h and 48h compared to control. To further examine the role of ERα in this PFOA‐induced proliferation, we carried out additional experiments in MCF‐7 cells along with experiments in another ERα‐positive cell line, T47D, as well as an ERα‐negative cell line, MDA‐MB‐231. There was a tendency for increased viability in all three cell lines when treated with PFOA, although only the MDA‐MB‐231 cells and the T47D cells showed a statistically significant (~20%) increase at 48h with 50μM PFOA and 100μM PFOA, respectively. These data suggest that the PFOA‐induced increase in cell viability in these cell lines is not dependent on ERα expression. In addition, the opposing effects of PFOA on proliferation in MCF‐7 cells in the presence and absence of FBS demonstrates the importance of accurately and completely reporting cell culture and treatment conditions.Support or Funding InformationAugusta University Department of Biological Sciences, Augusta University Center for Undergraduate Research and Scholarship