bluuebunny/crossref_metadata_2025_split huggingface dataset:
Characterization of Urinary Exosomes from Diabetic Hypertensive db/db Mice
Authors: Schramm Whitney , Tuna Kubra , Chacko Kevin , Sanchez Victor , Ahmed Hamzah , Liu Lauren , Alli Abdel 
Published: June 2021

Hypertension can occur before or after the onset of type 2 diabetes mellitus. The db/db mouse model is commonly used to study type 2 diabetes mellitus and its comorbidities including obesity and hypertension. Diabetic db/db mice develop salt‐sensitive hypertension after salt loading. Urinary exosomes are nano‐sized vesicles derived from cell types within the kidney that contain lipids, proteins, and nucleic acids. Recent studies showed urinary exosomes are useful biomarkers for kidney associated diseases including hypertension. We hypothesized the production of urinary exosomes is augmented and that their cargo is altered during the development of salt‐sensitive hypertension. Metabolic cage studies were performed to collect urine for the isolation of urinary exosomes from diabetic db/db mice and wild‐type littermate mice before and after salt‐loading. Western blot analysis was performed using validated antibodies to probe for the exosomal markers flotillin‐1, Hsp70, and annexin V in urinary exosomal lysates. NanoSight analysis showed an increase in the concentration of urinary exosomes from salt‐loaded, diabetic, hypertensive db/db mice and Western blot analysis revealed the enrichment of ENaC and MARCKS in these exosomes compared to non‐salt loaded diabetic db/db mice or healthy wild‐type littermate mice after salt‐loading. Future studies will investigate the physiological role of exosomal MARCKS in the development of diabetes associated hypertension.Support or Funding InformationThis research was supported by a National Institutes of Health grant K01 DK099617 (to A. A. Alli).This abstract is from the Experimental Biology 2018 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
A Mathematical Approach for Gas Exchange in the Lungs; Balance Between Ventilation and Perfusion
Authors: PIZANO ALEJANDRO , CALVACHI PAOLA , GIRÓN FELIPE , CORDOVEZ JUAN MANUEL 
Published: June 2021

IntroductionChronic pulmonary diseases and respiratory infections remain as one of the most morbid and deadliest diseases in the world. Airway and environment play an important role in the etiology of these pathologies. The balance between oxygen and carbon dioxide exchange is the pillar of the respiratory system. The aim of this project is to understand the gas exchange in the lungs using a mathematical model that describes the dynamics in each alveoli, in order to evaluate the relationship between ventilation and perfusion.MethodsA mathematical model was design based on the alveoli gas exchange and its relationship between ventilation and perfusion (V/Q). So, the lungs were divided into 9 segments, each one with different ratio according to their relative exchange area and the anatomical and physiological characteristics. The model assumed mass balance for O2 and CO2 and ideal gas law. Parameters for the model were determined according to data obtained in the literature. The model was implemented in MATLAB (The MathWorks®, Inc.) and the equations were solved to steady state.ResultsThe model predicted gas concentrations and flows in different lung segments. There was an evident strong association between V/Q, and its mechanisms to compensate. The model showed the behavior of lungs with and without disease. One of the most important outcomes was the evidence of enviromental influence (i.e. altitude, polution) in the imparment of the small airways and its association with the cardiovascular system.ConclusionThe mathematical model was helpful to understand the gas exchange dynamics. The model can be used to predict the interaction between ventilation and perfusion under different conditions, giving the possibility to estimate the efficacy of medical intervention. It can also shed some light on how to prevent and decrease the incidence of lung diseases.Support or Funding InformationResearch Group in Mathematical and Computational Biology (BIOMAC), Department of Biomedical Engineering, Universidad de los Andes, Colombia.This abstract is from the Experimental Biology 2018 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
Developmental Differences in the Contribution of PKCβ Signaling to Chronic Hypoxia‐Induced Pulmonary Arterial Tone
Authors: Yan Simin , Sheak Joshua R. , Jernigan Nikki L. , Walker Benjimen R. , Resta Thomas C. 
Published: June 2021

Vasoconstriction contributes to the pathogenesis of chronic hypoxia (CH)‐induced pulmonary hypertension in both neonates and adults. Although reactive oxygen species (ROS) have been implicated in this response in both age groups, it is not clear whether the subcellular source of ROS and signaling mechanisms involved are similar between adults and neonates. Based on preliminary data from our laboratory in neonatal rats, we hypothesized that PKCβ and mitochondrial ROS mediate enhanced basal pulmonary arterial tone following CH in both neonatal and adult rats. To test our hypothesis, we measured basal tone by videomicroscopy in isolated, pressurized small pulmonary arteries [~150 μm inner diameter (i.d.)] from normoxic and CH (12 d, 0.5 atm for neonates; 4 wk, 0.5 atm for adults,) rats following administration of the PKCβ inhibitor, LY‐333,531 (10 nM), the mitochondrial‐targeted antioxidant, MitoQ (1 μM), or vehicle. All experiments were performed in the presence of the nitric oxide (NO) synthase inhibitor Nω‐nitro‐L‐arginine (300 μM) to evaluate vascular smooth muscle tone independent of influences of endogenous NO. Pulmonary arterial tone was calculated as the percent difference in i.d. between Ca2+‐free and Ca2+‐containing conditions. Our results indicate that exposure to CH significantly (P < 0.05) enhanced basal tone in both neonatal and adult rats as compared to normoxic animals. Whereas LY‐333,531 significantly attenuated basal tone development in arteries from neonatal CH rats, PKCβ inhibition was without effect in arteries from either CH or normoxic adult animals. In contrast, MitoQ eliminated CH‐dependent increases in basal tone in arteries from both neonatal and adult rats. These findings suggest the contribution of PKCβ to regulation of CH‐induced basal pulmonary arterial tone is developmentally regulated, being important in neonates but not adults. Furthermore, mitochondrial ROS appear to be critically involved in basal tone development in pulmonary arteries from both neonatal and adult CH rats, with potential significance to the etiology of CH‐induced pulmonary hypertension.Support or Funding InformationThis work was supported by NIH grants R01 HL132883 (to T.C. Resta) and F31 HL131334 (to J.R. Sheak).This abstract is from the Experimental Biology 2018 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
Implementing a Dissection‐based Human Anatomy Laboratory Manual: An Assessment of Students' Use, Perceived Usefulness, and Resulting Laboratory Preparedness
Authors: Forster Victoria N. , Marrelli Kristina M. , Jadeski Lorraine C. 
Published: June 2021

The University of Guelph provides year‐round dissection‐ and prosection‐based human anatomy courses to undergraduate students; the combined enrolment in laboratory‐based courses exceeds 1000 students per academic year. Given such large enrolment numbers, an ongoing challenge is to continue to provide a very high level of human anatomy education to students with current resources. Consequently, various educational tools are being developed to enhance the current human anatomy program. To specifically target the course and laboratory learning objectives, a dissection‐based human anatomy laboratory manual has been created and is under further development, with the goal of promoting student preparedness and self‐sufficiency in the laboratory. Detailed cadaveric, dissection‐based images with clear and simple dissection instructions are the fundamental qualities of the manual. This resource also incorporates active learning exercises including drawing, colouring, and labelling components. A pilot edition of the manual was released to students enrolled in the third‐year dissection‐ and prosection‐based courses in the fall semester of 2017 (HK*3401 and HK*3501). Data pertaining to the students' use and perceived usefulness of the manual, as well as their perceived preparedness for laboratory periods have been collected. Analysis of these data is ongoing, and will be used to gain insight for the future development of subsequent editions of the manual.This abstract is from the Experimental Biology 2018 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
Treatment with Resveratrol Reduces Bone Fracture Risk Associated with Neurodegenerative Disease
Authors: Alkhouli Mustafa F. , Hung Jun , Squire Michaela , Anderson Miranda , Castro Monica , Al‐Nakkash Layla , Broderick Tom L. , Plochocki Jeffrey H. 
Published: June 2021

Neurodegenerative disorders like Alzheimer's disease are associated with significant decreases in bone mineral density (BMD) and elevated fracture risk in comparison to healthy individuals of the same age. Low BMD is largely attributable to decreased physical activity and systemic inflammation related to neurodegenerative processes. Here, we test the hypothesis that treatment with moderate exercise and resveratrol, a natural polyphenolic phytostilbene with anti‐inflammatory effects, will improve fracture resistance in a mouse model for Alzheimer's disease. Our 4‐month study examined bone mechanical and histological properties in 7‐month‐old male wild type mice (C57BL/6, n=10) and (age‐matched) neurologically compromised mice that express regulatory genes affecting amyloid‐beta plaque formation (3xTg‐AD, n=28). 3xTg‐AD mice were divided into groups treated with treadmill running 1 hour 5 days/week, 3g/100g dietary resveratrol, both, or neither (n=7 per group). We find treatment with exercise and resveratrol, in combination and alone, significantly improved ultimate force and post‐yield displacement in three‐point bending tests of the tibia (P < 0.05). We also find combined exercise and resveratrol treatment improved cross‐sectional geometric resistance to loading and maintenance of BMD in the tibia (P < 0.05). Bone stiffness was correlated with corrected total tissue fluorescence, an indicator of bone quality that measures advanced glycation end product accumulation in the tissue (r = 0.41, P < 0.01). Our findings suggest exercise and resveratrol may provide benefits to bone health in individuals with neurodegenerative‐related bone loss.This abstract is from the Experimental Biology 2018 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
Forelimb Myology of the Kinkajou (Potos flavus)
Authors: Ahrens Heather E. , Walker Allison M. 
Published: June 2021

The highly arboreal kinkajou (Carnivora: Procyonidae) exhibits many characteristics representative of its locomotor function, including hind foot reversal, diagonal sequence gait, and a prehensile tail. Detailed examination of the myology of P. flavus will aid in the understanding of the terrestrial‐arboreal locomotor gradient and can provide potential phylogenetic characters clarifying musteloid relationships. Here, we present the dissection of the forelimb of a fresh‐frozen male kinkajou and document the musculature via description and muscle maps. Noteworthy findings include the absence of m. biceps brachii caput breve, fused m. extensor carpi radialis, and a distinct humeral origin of the m. flexor digitorum superficialis. The latter finding contradicts a more recent suggestion that identification of a humeral origin was due to oversight and outdated nomenclature. Overall, the kinkajou resembles our primary comparative taxon, the closely‐allied and arboreal red panda (Ailurus fulgens). Similarities include the two insertion sites of the m. latissimus dorsi and the arrangement of the deep extensor compartment. This new description of the kinkajou is an important step in evaluating phylogenetic and functional patterns in the myology of carnivorans. However, further dissection of representatives of additional taxonomic and locomotor groups is necessary to differentiate what is typical of arboreal taxa from ancestral musteloid characteristics.This abstract is from the Experimental Biology 2018 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
Molecular Mechanism of Transcriptional Regulation by Chromatin Reader Transcription Factor 19 (TCF19) during Metabolic Stress
Authors: Sen Sabyasachi , Das Chandrima 
Published: June 2021

Changing nutritional environment confers epigenetic memories that often pre‐dispose the human body to certain metabolic disorders. Histone post‐translational modifications (PTMs), and their selective recognition by reader domain containing proteins constitute a transcription regulatory network that helps in maintaining cellular metabolic homeostasis, although the exact mechanism is still unclear. Here we have studied a novel glucose sensing and endoplasmic reticulum (ER) stress‐responsive transcription regulator, TCF19, which regulates transcription of key metabolic and stress‐responsive genes through recognition of histone PTMs. Through biochemical and biophysical techniques, we have found that TCF19 Plant Homeodomain (PHD) finger has a unique preference for the lysine 4 trimethylation of histone H3, which is a transcription activation modification. Using siRNA mediated TCF19 depletion in human hepatoma cell line HepG2 and primary human hepatocyte‐like cell HepaRG, we observed a global effect on metabolic pathways and interestingly the gluconeogenic genes were significantly upregulated. Through endogenous immunoprecipitation, we found that TCF19 interacts with components of a histone deacetylase complex NuRD (Nucleosome Remodelling and Deacetylase) and Chromatin immunoprecipitation studies indicate their co‐recruitment onto promoters of gluconeogenic genes under high glucose conditions, suggesting that the observed repression is possibly mediated in concert with NuRD complex. In‐depth analysis revealed that TCF19 also represses gluconeogenesis in response to insulin stimulation in both HepG2 and HepaRG cells. Deletion mutants of TCF19 lacking the PHD finger were unable to repress gluconeogenesis in response to insulin, underscoring the importance of histone modification mediated regulation of insulin action. We have also observed that TCF19 associates with several other transcription regulators of metabolic genes which have an important role in ER stress and cell survival and has been widely linked to type 2 diabetes progression. Overall, our study presents a novel transcription regulator that integrates metabolic (glucose), as well as hormonal (insulin) signal. These findings seek to expand our understanding of the role of epigenetic changes in metabolic regulation, and in the long run, provide an opportunity for therapeutic intervention to treat metabolic disorders like diabetes.Support or Funding InformationThe study was supported by research grant entitled Biomolecular Assembly, Recognition and Dynamics (BARD) (Grant 12‐R&D‐SIN‐5.04‐0103) by the Department of Atomic Energy (DAE), Govt. of IndiaThis abstract is from the Experimental Biology 2018 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
Pathogenesis Related Protein 5 Expression is Induced by 1,4‐dimethylnaphthalene treatment of Potato Meristems
Authors: Pirritano Maria , Campbell Michael 
Published: June 2021

The agricultural compound 1,4‐dimethyl naphthalene (DMN) can be used to inhibit sprouting and prolong the storage of Solanum tuberosum. Potatoes treated with DMN exhibit characteristics that enable greater crop yields, including pathogen response, water retention, and greater shelf life. However, the molecular mode of action as to how DMN brings about these changes within the potato is not well understood. Two years of RNA‐seq data demonstrate extensive changes in gene expression within potato samples treated with DMN at different times throughout the year. Significant changes have been highlighted among a particular group of genes located on chromosome one. This group of genes codes for a pathogenesis‐related‐protein, PR5. As an extracellular protein, PR5 has an extensive signal transduction that responds to pathogens and abiotic stress. Among the PR5 family is a group of cytosine‐rich proteins called osmotins. Osmotin is a ubiquitous protein among several plant species and serves as a useful molecular marker for the systemic acquired resistance response, which is analogous to the innate immune system in plants. Gene expression measurements were taken over a six month period, during which dormancy was naturally breached. RNA‐seq data reveals 2‐3 fold increases between measurements taken before and as dormancy was breaking. After dormancy was breached, PR5 gene expression continued to rise significantly in comparison to previous measurements. These trends in gene regulation imply that the PR5 expression proceeds concurrently with sprout growth in DMN treated potatoes, and are related to the systemic changes seen from the outside of the potato. Phylogenetic analysis reveals the PR5 genes with the greatest changes in expression (7865 and 7870) demonstrate the greatest homology among their transcripts. Current analysis is being done using qPCR primers directed towards genes 7865 and 7870. Expression levels from qPCR should mirror the RNA‐seq data, in addition to demonstrate more immediate expression following DMN treatment. It has been hypothesized that gene expression spikes in prominent PR5 genes (7865 and 7870) will signify a broad molecular mode of action DMN plays inside S.tuberosum.Support or Funding InformationSupported in part by a grant from the 1,4‐Group, Meridian, ID.This abstract is from the Experimental Biology 2018 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
Defining Molecular Calipers: Affinity and Exchange of Auxin Response Activators and Repressors
Authors: Carroll Margaret C. , Ellis Jamie P. 
Published: June 2021

Auxin response factors (ARFs) regulate transcription of developmental genes in the auxin hormone pathway which affects a broad range of valuable plants including food crops and biofuel targets. Both transcription activators and repressors within the ARF family consist of three domains including a protein interaction domain, intrinsically disordered middle region and a DNA binding domain containing plant‐specific B3 domain and the ability to dimerize. The many layers of interactions, as well as a multiplicity of conserved factors within a single plant, suggest a more complex transcriptional regulation system than a simple on/off activation switch. Arabidopsis thaliana is a well‐understood model from which we chose an example transcription repressor (ARF1) and activator (ARF5). Both are present concurrently during seed embryogenesis, bind the same high‐affinity Auxin response element (AuxRE) sequence, and form homodimers in response to DNA constructs with AuxRE everted repeats (ER). We combined genomic analysis with biophysical studies to explore ARF binding and exchange on biologically relevant constructs. Through genomic analysis, we analyzed frequency and spacing of canonical and other high‐affinity AuxRE sites including both everted and direct repeats. The results reveal a much wider range of spacings between the repeats in the genome than tested in the literature with clear indications of periodicity. Utilizing a combination of fluorescence spectroscopy and gel filtration, we directly tested the association and exchange of AFR1 and ARF5 on DNA of varying repeat spacing and AuxRE sites. Control experiments indicate that fluorophore attachment does not affect dimerization or DNA binding of the proteins. FRET studies showed that dimerization clearly increases in the presence of AuxRE DNA. Varying DNA constructs indicates both binding site affinity and the spacing between sites affects the amount of dimer formed, but that a repeated sequence is not necessary to cooperatively form dimers. While ARF1 and ARF5 have different spacing preference, the high‐affinity binding site can overcoming spacing preference. The differences in dimer formation indicate that the spacing between everted repeats effects the protein's ability to bind DNA. The variation in spacing discovered in the Arabidopsis genome paired with the difference in activity with different spacings indicates that spacing motifs regulate ARF factors.This abstract is from the Experimental Biology 2018 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
Inactivation of Gene α‐1,3‐Galactosyltransferase in Bovine Aortic Smooth Muscle Cells Using CRISPR‐Cas9
Authors: Wang Xiaodan , Apple Jason , Huang Yan 
Published: June 2021

People who get bites from Amblyomma americanum usually develop a severe allergic reaction to red meat. The main reason caused this allergy is because of the presence of α‐1,3‐galactosyl epitopes on the membranes of most mammalian cells. Since human beings do not create galactose‐alpha‐1,3‐galactose (alpha‐gal), the patients' immune system is free to start attacking alpha‐gal when they eat red meat. The epitope transferase coded geneα‐1,3‐galactosyltransferase (GGTA1) was already identified, and it can be knocked out to solve the allergy of eating red meat. GGTA1 gene has successfully knocked out in pigs. However, there is no evidence to show the GGTA1 gene has been knocked out in beef. In this study, we utilized bovine aortic smooth muscle cells (BAOSMC) as the module for the research of GGTA1 gene knockout.The clustered regularly interspaced short palindromic repeats (CRISPR) systems are widely used in performing targeted genome editing in cultured cells. The use of CRISPR‐associated (Cas) systems as an RNA‐programmable DNA targeting and editing platform is followed by a synthetic single‐guide RNA (sgRNA), which can simply edit genome sequences like other tools such as transcription activator‐like effector nucleases (TALENs) and zinc‐finger nucleases (ZFNs).We used the gRNA design tool and selected 5′‐GGCCTGACGGTTTTCGCCGT‐3′ as the target gRNA sequence from the coding DNA sequence of Bos taurus alpha‐galactosyltransferase 1 (glycoprotein). The gRNA was constructed in the pSpCas9 BB‐2A‐GFP (PX458) vector provided by GenScript USA Inc. Vectors were amplified and transfected into BAOSMC by GenePORTER2 transfection reagent when the cells were 80% confluency. Green fluorescent can be viewed after 24 hours transfection (Fig. 1). The transfection efficiency can reach about 70% to 80%.Cells were collected in PBS at pH7.4 after 24 hours transfection. Total protein was extracted then the enzyme‐linked immunosorbent assay was used to examine the GGTA1 production. By normalized with the total protein concentration, the GGTA1 protein level in the transfected cells was 17.9 ± 7.25% lower (P< 0.05) than in the control cells, showing a significant inhibition of GGTA1 gene expression in the cells by CRISPR‐Cas9 gene edition method (Fig. 2).Our data show that the gRNA sequence that we chose was suitable for the GGTA1 gene knockout in BAOSMC. Moreover, the CRISPR‐Cas9 system was proved can be applied in the genome editing of bovine cells.Support or Funding InformationThis study is funded by Arkansas Beef Council.This abstract is from the Experimental Biology 2018 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.