bluuebunny/crossref_metadata_2025_split huggingface dataset:
Cardioprotective Effects of Melatonin, Berberine and Curcumin by Inducing the JAK2/STAT3 Signaling Pathway in Myocardial Ischemia Repercussion
Authors: Khan Naushad , Pandey Rajeev , Murthy Guru 
Published: April 2020

Ischemic heart disease and stroke are the World’s major cause of deaths, accounting for a combined 15.2 million deaths in 2016 as per WHO. Following Cardiac Ischemia, we are often faced with a bigger problem of damage induced by myocardial ischemia/reperfusion injury. It has been observed that reperfused myocardium displays more severe injury than myocardium subjected to the same ischemic duration that had not been reperfused. This observation has led to deduce that reperfusion could result in Myocardial injury more severe than one that could have occurred by Ischemia alone. However certain mediators like melatonin, berberine and curcumin have shown to induce JAK2/STAT3 signaling pathway and have shown a protective effect against myocardial ischemia reperfusion. In the present work we will be mainly focusing on the correlation between Melatonin, Berberine & Curcumin and cardiac JAK2/STAT3 receptors in cardioprotection against myocardial ischemic reperfusion.Support or Funding InformationCommunity Health and Research Center of Spartan (CHARCOS), Spartan Health Sciences University, School of Medicine, St. Lucia
Colonic Crypt‐Derived 3D Gut Organoid from Hypertensive Rat Maintain In vivo Characteristics
Authors: li Jing , Su Qing , Karas Marianthi M. , Richards Elaine M. , Raizada Mohan K. 
Published: April 2020

ObjectiveSubstantial evidence links gut dysbiosis to hypertension in both animal models and in patients with high blood pressure. Increased blood pressure and hypertension‐associated pathologies in normotensive animals transplanted with fecal microbiota from hypertensive animals, support the concept of microbiota‐gut communication as an important early signal initiating and establishing hypertension. We hypothesized that colonic cryp‐3D organoids would retain the characteristics of the in vivo gut epithelium form which they were derived, and would provide a model system to investigate epithelial‐microbiota communication in hypertension.MethodsColonic crypts were isolated from adult Wistar‐Kyoto (WKY) rats and spontaneously hypertensive rats (SHR). 3D organoid cultures were established in organoid growth medium with Matrigel matrix and cultured for 3, 5 and 7 days without passage. The shapes and number of organoids were assessed by bright‐field microscopy using Trypan blue. Growth, differentiation and cell types in organoids were characterized with the use of antibodies (Ki67, Alpi, KRT20 and Muc2). Data were compared between WKY rats and SHR organoids.ResultsReproducible 3D organoid cultures were successfully established from the crypts of adult WKY and SHR. They proliferated and differentiated, and predominately contained enterocytes and goblet cells. We observed significantly more organoids per crypt from normotensive vs hypertensive rats (WKY: 0.20 vs. SHR: 0.11; p<0.01). In vivo, WKY rats had 18% longer colons than SHR (0.07 cm/g vs. 0.059 cm/g; p<0.0001).ConclusionsThese observations demonstrate that 3D organoid cultures maintain characteristics of the colonic epithelium and show significant differences between WKY rats and SHR. Thus, these organoids represent an ideal ex vivo system to investigate the gut epithelium in hypertension, and to examine the effects of gut microbiome‐derived metabolites in blood pressure control and initiation and establishment of hypertension.Support or Funding InformationHL132448
Mediator Kinase as a Transducer and Therapeutic Target in Oncogenic WNT/β‐ CATENIN Signaling
Authors: Barron Lindsey , Clark Allison , Burleson Marieke , Boyer Thomas 
Published: April 2020

Colorectal cancer (CRC) is the second and third most commonly diagnosed cancer in women and men, respectively. Few treatment options exist for advanced stage disease, thus, the development of targeted therapies is crucial for treatment. The canonical wingless related integration site (WNT)/β‐CATENIN pathway is a driver in >90% of CRCs. The mediator kinase module (mediator complex subunit 12 (MED12), mediator complex subunit 13 (MED13), CYCLIN C, and cyclin dependent kinase 8 (CDK8)) is a key node of oncogenic activation within the WNT/β‐CATENIN pathway, since β‐CATENIN activates transcription through its direct physical interaction with MED12. Additionally, CDK8 is an oncoprotein required for β‐CATENIN‐dependent gene activation and CRC growth.The objective of this study was to investigate the effects of genetic (disruption of MED12:CYCLIN C interface) and chemical inhibition of Mediator Kinase activity on CRC cell oncogenic properties and gene regulation. Oncogenic properties were assessed by colony formation and cell proliferation assays. Chromatin immunoprecipitation sequencing and RNA sequencing were used to evaluate gene regulation. Interestingly, genetic but not chemical inhibition led to a significant reduction in oncogenic properties. Superenhancer alterations and differential expression of superenhancer‐associated genes were observed with both genetic and chemical inhibition. However, little overlap was found for down‐regulated superenhancer‐associated genes.Based on the significant reduction in oncogenic properties, genetic disruption of the MED12:CYCLIN C interface appears to be an attractive target for the development of targeted molecular therapies for CRC. The difference in effect on oncogenic properties with chemical versus genetic inhibition is likely explained by the differences in down‐regulated superenhancer‐associated genes. Concordantly, genes involved in the WNT/β‐CATENIN pathway were down‐regulated with genetic but not chemical inhibition.Support or Funding InformationLindsey Barron was supported by a CPRIT postdoctoral training fellowship, the CPRIT Research Training Award (RP170345) and is currently supported by an IRACDA Grant (K12GM111726).
Optimization of LC‐ESI‐MS Quantification of Ventromedial Hypothalamic Nucleus Estradiol Content by Central Composite Design
Authors: Bheemanapally Khaggeswar , Ibrahim Mostafa M.H. , Ali Md. Haider , Napit Prabhat R. , Briski Karen P. 
Published: April 2020

Brain structures exhibit characteristic patterns of estrogen receptor expression and regulatory functions and local capacity to generate neuroestradiol, necessitating quantification of estradiol content of individual loci. Estradiol acts on ventromedial hypothalamic nucleus (VMN) substrates to control behavior and glucostasis. Current research utilized Analytical Quality of Design to optimize uHPLC‐electrospray ionization‐mass spectrometry (LC‐ESI‐MS) sensitivity for analysis of estradiol in VMN samples obtained by high‐resolution microdissection techniques. A Design of Experiments matrix was developed to assess effects of critical independent variables on estradiol chromatogram area. A second‐order polynomial equation was generated by statistical response surface methodology using central composite design to identify synergistic versus antagonistic factors and to predict singular/combinatory effects on response area. Estradiol, estrone, and estriol were used as optimizing compounds; ethynyl estradiol served as internal standard. Lysates of VMN tissue harvested by micropunch dissection were transferred to methanol‐water conditioned solid phase extraction syringe cartridges and treated with 2% formic acid. Estradiol content was extracted with methanol, then vacuum concentrated. Samples were treated with 2‐fluoro‐1‐methylpyridinium‐p‐toluenesulfonate (FMP‐TS) prior to application to a C18 column [0.2ml/min mobile phase (25:75, v/v) flow rate; 0.1% formic acid, acetonitrile], followed by spectrometric analysis of estradiol and internal standard responses at m/z 364.2 and 388.2, respectively. Optimized mass spectrometric vaporizer (320°C) and ion transfer tube (266 °C) temperatures and sheath (22 psig), auxiliary (5 psig), and sweep (0.7 psig) gas pressures resulted in maximum tissue estradiol response. Outcomes validate a novel combinatory methodology for estradiol quantification in precisely‐defined brain components.Support or Funding InformationNational Institutes of Health DK‐109382
Investigating the Effects of Histone H3.3 Point Mutations in Neural Crest Cells and Craniofacial Development
Authors: Nzirorera Nadine , Jabado Nada , Majewska Loydie 
Published: April 2020

Craniofacial development is an intricate process that requires the meticulous and coordinated expression of many genes. Histone proteins which are fundamental to both the structural integrity and organization of DNA into nucleosomes and consequently, chromatin play an important role in coordinated gene expression. Furthermore, the modification of histone tails is now known to be an essential mechanism for coordinated expression of a large number of genes. Recently, a point mutation in the gene h3f3a which encodes the histone variant Histone H3.3, was identified in a forward genetic screen in zebrafish. H3.3A was found to be important for neural crest cell induction and generation of cranial neural crest cell derived head bones. The authors proposed that incorporation of histone H3.3 was required for expression of genes important for cranial neural crest cell specification, whereas remodelling of this histone tail may be important for later stages in neural crest cell differentiation. However, it remains to be confirmed if disrupted modification of histone tails is also important in neural crest cells. We postulate that modification of the H3.3 tail is required for coordinated expression of genes important for neural crest migration and/or differentiation. To test this hypothesis, we used CRISPR/Cas9 to generate a mouse line with a conditional missense mutation in H3f3a. The mutation generated was previously shown to result in an abnormal increase in methylation of K27 and a decrease in methylation of K36 of the histone H3.3 tail. In this poster, we will present preliminary data using the Wnt‐1 Cre2 mouse line to drive expression of this dominant negative mutation in neural crest cells. Alcian Blue Staining will be used to analyze cartilage in E14.5 embryos and alizarin red will be used to stain bones formed in heads of E17.5 embryos. Bones will be analyzed in control and mutant embryos. Our result will enable us to uncover if K27 or K36 methylation is important in neural crest cell formation, migration and/or differentiation. This work will begin to unravel the role of epigenetics in neural crest cell formation.Support or Funding Information
Changes in Motor Function in Response to Living in an Extreme Underwater Saturation Environment under High Stress and Increased Workload
Authors: Ari Csilla , D’Agostino Dominic Paul , Bharwani Sahil , Rehsi Anand , Moss Sara , Schmer-Galunder Sonja , Fiore Stephen 
Published: April 2020

Understanding changes in individual motor function and how it relates to workload, personality types, and shared knowledge is vital to the success of team‐based missions. Such changes may present a challenge in long duration spaceflight and other missions in extreme environments. Therefore, we studied grip strength, dexterity, stress level, task load and the personality types of participants living in an extreme environment during the NASA Extreme Environment Mission Operation (NEEMO) 23 in June 2019. The Aquarius Reef Base is home to NEEMO missions and is located at 20 m (66 ft) with 2.6 atm (air) pressure inside the habitat, where CO2 levels are often elevated similar to the International Space Station.Motor strength was assessed using a grip dynanometer and dexterity was measured using a pegboard. The tests were performed by 6 subjects (2 males, 4 females) 1 day pre‐mission, during the mission (grip test twice) and 1 day post‐mission. Dexterity test was also performed at the beginning and at end of Extravehicular Activities (EVAs). NASA TLX tests measuring mental, physical, temporal effort, performance, general effort and frustration, were assessed immediately after returning from EVAs. To monitor changes in stress level, Short Stress State Questionnaire (SSSQ) and Generalized Anxiety Disorder (GAD7) scores were collected. Measures of social cognition, including Reading the Mind in Eyes and Big 5 personality tests, were performed once before the mission.There was a trend of decreased grip strength at the end of mission and post‐mission, compared to baseline and beginning of mission. The dexterity scores at the beginning of EVAs were significantly higher, compared to before and during mission. This suggests that tasks with fine motor function can be affected by context such as in‐water performance. At the end of the EVAs, the dexterity score was significantly higher compared to before mission and during mission as well. There was a trend of worse performance on the dexterity test at the end of EVA, compared to the beginning of EVA. A trend of increasing GAD7 scores at half time of the mission and post‐mission could be observed. There were several differences in NASA TLX scores between different subjects.These results indicate that strength and dexterity was influenced during EVAs. This may have been due to both physical and psychological stress during the extreme underwater mission in a saturation environment. Further examination of these trends is recommended.Support or Funding InformationKetone Technologies LLC provided partial funding for the study. SIFT provided partial salary for CA.
The “Function vs. Disease Dilemma” in Contemporary Medicine: Physical Frailty & Sarcopenia as a Prototypic Condition of New‐Generation Geriatric Medicine
Authors: Tosato Matteo 
Published: April 2020

Due to the worldwide demographic transition, healthcare systems are facing new demands. Health services—with their approach of mostly single acute conditions—are indeed confronted with an expanding older population characterized by specific medical needs related to multimorbidity and functional impairment. The European research project “Sarcopenia and Physical fRailty IN older people: multi‐componenT Treatment strategies” (SPRINTT) project is specifically designed to overcome existing barriers for efficient public health interventions against frailty, and promote the implementation of successful aging strategies across Europe. The just‐completed SPRINTT randomized clinical trial (RCT) compared the efficacy of a multicomponent intervention (based on long‐term structured physical activity, nutritional counselling, and an information and communication technology intervention) versus a Healthy Aging Lifestyle Education program for preventing incident mobility disability in community‐dwelling older persons with physical frailty and sarcopenia. For the RCT, we enrolled 1,517 community‐dwellers, aged 70 years and older (750 per treatment arm), non‐disabled older persons exposed to increased vulnerability to stressors. The identification of such population relied on three key elements: low muscle mass measured by DXA (identified using FNIH cutpoints); clinical signs of physical frailty (i.e., weakness, slow walking speed, and poor balance) assessed using SPPB; absence of major mobility disability (able to walk 400 meters within 15 minutes at baseline. The primary outcome was the incidence of mobility disability (i.e., incident inability to walk 400 meters). Secondary outcomes included, among others, changes in physical performance and function; ability of selected biomarkers to predict the rate of change in muscle mass; incidence of falls. The SPRINTT RCT was conducted in 15 study sites located in nine European countries. The mean age of our sample was 79 years (SD ± 5.8), 72% were women, BMI was 28.6 kg/m2 (± 5.9), with 37% of the population showing BMI >30 kg/m2, Handgrip strength was 20.1 kg (SD 8.2), SPPB score was 6.71 (SD 1.4), aLM in men was 21.0 kg (SD 4.06) and 15.0 kg in women (SD 1.98), the aLM/BMI was 0.71 (SD 0.06) in men and 0.51 (SD 0.07) in women. Data analysis is ongoing and final results may be available at the date of the EB meeting.Support or Funding InformationInnovative Medicines Initiative – Joint Undertaking (IMI‐JU 115621)
Mutation of Class‐Distinctive Residues in Gα13 to Identify Determinants of RH‐RhoGEF Binding Selectivity
Authors: O’Shea Corin R. , Tagliatela Alicia C. , Meigs Thomas E. 
Published: April 2020

Gα13 belongs to the G12/13 subfamily of heterotrimeric guanine nucleotide‐binding proteins that play a signaling role in cell growth and tumorigenic pathways, cytoskeletal rearrangements, and metastatic invasion. A structural bioinformatics analysis [Temple et al., 2010, PLOS Comput. Biol., 6(10): e1000962] identified a set of “class‐distinctive” residues in Gα13 which correspond to a different residue conserved in the other G protein subfamilies: Gs, Gi, and Gq. Using this information, we created a panel of Gα13 point mutants that replace each class‐distinctive residue with its putative ancestral form. In order to distinguish our mutant constructs from native Gα13 in cultured human embryonic kidney cells, we introduced a myc epitope tag to all Gα13 mutants, positioned within the αB–αC loop of the helical domain. Installation of this epitope tag was non‐disruptive to SRF signaling by Gα13, and allowed us to differentiate its recombinant and native forms in protein‐protein interaction experiments. While characterization of these Gα13 mutants is ongoing, we have identified a class‐distinctive Phe at position 234 for which ancestral substitution appears to cause selective uncoupling of protein binding within the Gα13‐responsive, RGS‐homology (RH) RhoGEFs. These findings shed light on the mechanism of Gα13 interaction with the individual RH‐RhoGEFs‐p115RhoGEF, PDZ‐RhoGEF, and leukemia‐associated RhoGEF‐ and further suggest that class‐distinctive Gα13 mutants may reveal binding determinants for additional effector proteins.Support or Funding InformationWe acknowledge support from the UNC Lineberger Comprehensive Cancer Center (University Cancer Research Fund), and the North Carolina GlaxoSmithKline Foundation.
Autophagic Burden on Calcium Signaling in Sympathetic Activation
Authors: Hayden Matthew R. , Wauson Eric , Tran Quang-Kim 
Published: April 2020

Signaling via calcium plays essential roles in all aspects of cell functions. Macroautophagy is a catabolic process that regulates important aspects of cell metabolism. While many studies have indicated that calcium is involved at various steps in the regulation of autophagic flux, the impact of autophagic flux on calcium signaling is unknown. We conceptually hypothesized that an “autophagic burden” exists in which enhanced requirement of the cell to process autophagic cargos would exert negative impact on calcium signaling. To test this hypothesis, we utilized an “non‐stimulation” approach to increase the “autophagic burden” by heterologously expressing in cells the wildtype microtubule‐associated proteins 1A/1B light chain 3B (LC3) or a mutant version in which the C‐terminal glycine responsible for ATG4‐mediated cleavage for LC3 lipidation has been removed (LC3DG) and examined the impact on adrenergic receptor‐mediated calcium signaling. HEK293 cells produced oscillatory intracellular calcium signals in response to agonists of b adrenoceptors (isoproterenol), α1A adrenoceptor (A61603), or a combination thereof (10 mM norepinephrine, (NE)). Cells expressing a fusion between the RFP mKate2 and wildtype LC3 (mKate2‐LC3) responded to 10 mM NE with substantial reductions in the number of intracellular calcium oscillations in the physiological ranges of 300 – 600 nM, 600 – 900 nM, or 900 – 1200 nM, as compared to the non‐expressing cells examined in the same microscopic fields, recognized by absence of mKate2 fluorescence. In contrast, cells expressing the mKate2‐LC3DG produced the same number of oscillatory calcium signals compared to non‐expressing cells in the same microscopic fields across the same ranges of cytoplasmic calcium stimulated by NE. The data suggest that an “autophagic burden” exists that may dampen the effects of acute sympathetic stimulation significantly via reduction in adrenoceptor‐mediated calcium signals.Support or Funding InformationIOER Grant 091707 to Q‐KT
Lysine 33 polyubiquitin chain formation by the HECT Ubiquitin E3 Ligase AREL1
Authors: Ladda Emily Rose , Spratt Donald 
Published: April 2020

Apoptosis Resistant E3 Ligase 1 (AREL1) is a member of the Homologous to E6AP Carboxyl Terminus (HECT) E3 ubiquitin ligase family. All HECT E3 ubiquitin ligases contain a conserved HECT domain that facilitate the covalent attachment of ubiquitin onto their intracellular substrates. Ubiquitylation is involved in numerous cell signaling pathways that can result in proteasomal degradation, DNA damage repair, and cell trafficking, among others. The biological function of AREL1 is to inhibit intrinsic apoptosis by ubiquitylation of pro‐apoptotic inhibitor of apoptosis (IAP) antagonist proteins, SMAC, ARTS, and HtrA2, for proteasomal degradation. Interestingly, previous studies have found that AREL1 is the only HECT E3 ubiquitin ligase identified to date that is capable of building polyubiquitin chains via lysine 33 linkages, and the biological role of this linkage remains unclear. Using circular dichroism (CD), in vitro ubiquitin activity assays, disulfide complex formation, and multidimensional heteronuclear NMR spectroscopy, we are currently deciphering the unique molecular mechanism used by AREL1 to build the lysine 33 poly‐ubiquitin chains.Support or Funding InformationNIH NIGMS Maurine H. Milburn Fund Clark University Carlson School of Biochemistry and Chemistry
Structural and Mechanistic Characterization of HERC2 E3 Ubiquitin Ligase C‐lobe with Implications in Cancer, Prader‐Willi Syndrome, and Eye Color
Authors: Rich Kayla , Schwaegerle Noah , Spratt Donald 
Published: April 2020

HERC2 (HECT domain containing and RCC1‐like domain containing E3 ubiquitin ligase 2) is a HECT E3 ubiquitin protein ligase that is implicated in breast cancer, Prader‐Willi syndrome, and eye color. HECT E3 ubiquitin ligases covalently attach ubiquitin onto substrates to target proteins for proteasomal degradation, initiate DNA damage repair, control cell cycle and intracellular trafficking, and regulate transcription. HERC2 has proposed implications with DNA damage repair and proteasomal degradation. The HERC2 gene has been linked to the neurodevelopmental disorder Prader‐Willi syndrome and is part of the cis‐regulatory module for OCA2 (oculocutaneous albinism 2). To date, only a few substrates of HERC2 have been identified including BRCA1 (breast cancer 1). Using biochemical and biophysical techniques, such as circular dichroism and multidimensional heteronuclear NMR spectroscopy, the HERC2 C‐lobe structure was characterized. Our studies show that HERC2 is mainly α‐helical with a flexible acidic C‐terminal tail extension. We demonstrate that the C‐terminal tail extension is important for polyubiquitin chain formation through intramolecular interactions with ubiquitin. The truncation of the HERC2 C‐terminal tail decreases protein stability and function. These structural and mechanistic studies on the HERC2 C‐lobe are important for elucidating the cellular role of HERC2 along with its implications with disease.Support or Funding InformationNIH‐NIGMS FundingAbbVie summer research grantClark University