bluuebunny/crossref_metadata_2025_split huggingface dataset:
HYPE‐mediated AMPylation as a novel therapeutic target for neurodegeneration
Authors: Camara Ali 
Published: April 2020

A major hallmark of Parkinson’s disease (PD) is the deposition of the intrinsically disordered protein α‐synuclein (αSyn) into intracellular inclusions termed Lewy bodies. HYPE—the sole human representative of a conserved family of adenylyltransferase enzymes—has been shown to covalently modify (AMPylate) αSyn in vitro. Remarkably, HYPE‐mediated AMPylation ameliorates many of the neurotoxic phenotypes of αSyn implicated in the progression of PD, such as αSyn fibrillation and membrane permeability. These potentially cytoprotective phenomena conferred by HYPE’s adenylyltransferase activity make it an attractive therapeutic target. Unfortunately, wild‐type HYPE is intrinsically inhibited, showing only basal AMPylation levels relative to a constitutively active mutant (E234G‐HYPE). To this end, we set out to screen both FDA‐approved and proprietary small‐molecule compound libraries towards the identification of novel manipulators of HYPE AMPylation. Employing fluorescence polarization (FP) of a labelled ATP analogue (ATP‐FAM) on a 384‐well microplate platform, we’ve developed a robust, high‐throughput assay suitable for monitoring changes in AMPylation. First‐pass selection of our combined ~10,000 compound libraries yielded promising hit percentages: 0.3 for activators of WT HYPE, and 1.2 for inhibitors of E234G‐HYPE. Challenging neuronal cell culture models of PD with these hits provides molecular insights into αSyn‐induced neurotoxicity, and paves the path for novel therapeutic strategies in combating PD.Support or Funding InformationThis project was made possible with support from Grant # 11000704 (S. Mattoo, PI), Grant # UL1TR002529 (A. Shekhar, PI), and Grant # TL1TR002531 (T. Hurley, PI), from the National Institutes of Health, National Center for Advancing Translational Sciences, Clinical and Translational Sciences Award.
Effects of ECG Summarization Methods on the Characterization of Drug‐Induced QT Prolongation – a Demonstration of Need for Standard Methods of ECG Beat Summarization6.3.6
Authors: Ether Nicholas , Bailie Marc , Leishman Derek , Lauver Adam 
Published: April 2020

A vital aspect of drug‐development is the pre‐clinical screening of drug‐induced cardiotoxicity. The effect a drug has on the ECG QT‐interval (QTi) is an especially vital screening due to the positive relationship between QT prolongation and arrhythmic events, such as Torsade de Pointes. Consequently, the accurate recording, measurement, and calculation of QTi and heart rate corrected QTi is necessary for cardiotoxicity studies. However, many aspects of these processes are subjective, and techniques vary greatly between researchers and across studies. Examples of subjective aspects of QT prolongation (QTp) characterization include, but are not limited to: the method of heart rate correction for QT (QTc), the number of data points considered, the number of ECG beats or amount of time averaged per data point, and the specific time points evaluated. While the effects of different QTc methods has previously been explored in the literature, the efficiency and accuracy of various ECG summarization methods still warrants further refinement. We evaluated the effects of different methods in order to begin identifying potential industry standards that can minimize the number of data points needed without compromising model prediction. In order to demonstrate the effect different summarization methods have on the identification of drug‐induced QTp, we used Ponemah software to analyze previously collected ECG telemetry data from non‐human primates. The subjects were administered vehicle, three doses of dofetilide, vehicle again, and three doses of quinidine, with a week between each dose. Data was summarized using various methods, including 1, 15, 30, 60 and 120 minute continuous averages as well as 1‐minute averages taken at 1‐hour intervals. We then quantified the effect size of the doses on QTi using restricted maximum likelihood (REML) estimation. Using these estimates, differences in results between the summarization methods were then compared. After finding dramatic differences, we performed power analyses on each summarization method to identify the most accurate and efficient methods. Results suggest that characterization of the dose response and power varied depending on the ECG summarization method used. Using continuous interval averages was more consistent and had greater power than selecting a 1‐minute average at various intervals. Further improvement is achieved by taking pharmacokinetics into account when selecting time intervals. These results confirm the need for careful consideration of ECG summarization methods during study design in order to avoid missing vital data or introducing researcher bias. It also provides evidence to support the recommendation for the introduction of a uniform industry standard for analyses of cardiovascular data. 6.3.6Support or Funding InformationThis work was funded in part by a research grant from Eli Lily and Co, Indianapolis, IN
CARING PROFESSIONAL HEALTH: A POWERFUL OF YOGA
Authors: Rossi Lia Mara , Galdeano Denival Soares , Marques L. S. , Barros Mirna Duarte 
Published: April 2020

INTRODUCTIONHealth professionals and medical science students have high levels of stress, anxiety and performance‐related musculoskeletal disorders. Long workloads and strenuous shifts, feelings of empathy for the suffering of patients and families, demands for cleanliness and hospital conduct, and the need for many hours of study cause mood swings and emotional instability, diminish immune system function and increase psychosomatic diseases in students and health professionals.The goal of this study was to evaluate the effects for health professionals and students that participated in YogaSanta Program, an opened and free program of Yoga.MATERIAL and METHODSThe World Health Organization Quality of Life assessment (WHOQOL‐BREF) was sent by e‐mail for health professional as well as for Medicine, Nursing e Speech Therapist students from Santa Casa de São Paulo School of Medical Sciences(São Paulo – Brazil). Who have shown WHOOQL‐BREF score under 50% were invited to participate of “YogaSanta Program”. YogaSanta aims “to take care of people who care for people” and offer free Yoga sessions for all. We selected 22 adults that practice one‐hour yoga session twice a week for 3 months consecutively. The sessions included Pranayama (breathing exercises), Asanas (postures), Yoganidra (relaxation) and Dhyana (meditation). All participants completed Visual Analogue Scale (VAS) for assessing stress‐level in two different moments: (VAS after = VAS‐A) immediately after Yoga class and (VAS before = VAS‐B) just before Yoga class. VAS varies from zero to 10 where “zero” corresponds to “no stress” and “10” corresponds to the “worst stress level”. VAS‐B and VAS‐A were compared through the t‐paired test and the results are shown in this study.RESULTSFrom 22 participants, 6 occupied administrative functions the hospital, 13 were students and two were faculty members. The mean ± standard deviation (SD) of participants age was 32.3 ± 12.2 yrs (Range: 22 – 60 yrs). The mean ± SD of VAS‐B and VAS‐A were compared (EVA‐B 3.34 ± 1.4 vs. VAS‐A 1.89 ± 1.4) and showed a low correlation = 0.574 with p<0.001.DISCUSSION and CONCLUSIONSome previous articles suggest that physiological benefits like muscle strength, flexibility, blood circulation and oxygen uptake, hormone functions and the relaxation induced by meditation helps to stabilize the autonomic nervous system and this help yoga practitioners become more resilient to stressful conditions and reduce a variety of important risk factors for different diseases, especially cardio‐respiratory disorders.Based on this and the results presented above, we believe that the YogaSanta program is an efficient option for producing improvements in psychological well‐being and decreasing mood disorders in health professionals and health students, thus contributing to improving performance and quality of life and help to stress reduction.
Amelioration of metaflammation induced in rats by exogenous hydrogen sulfide: Focus on mesenteric adipocyte oxidative stress
Authors: Zayachkivska Oksana , Revenko Oleg , Bula Nazar , Savytska Maryana , Yaschenko Antonina 
Published: April 2020

Metaflammation (metabolic inflammation) is low‐grade chronic inflammatory status linking to nutrient excess. Visceral adipocytes in mesenterium (AM) related to high fructose diet (HFD) are sources for metaflammation process, seriously threatening human health. Novel pathogenesis of AM is still unclear and there are no specific effective drugs for treatment of AM. Recently hydrogen sulfide (H2S) demonstrated potent vasodilatation actions which is promised for AM that have reduced vascularization Therefore, the aim of this study was designed to investigate the effects and underlying mechanisms of H2S on the prevention and treatment of experimental rats with AM during HFD.MethodsThe study was done on rats with standard diet (SD, control group) and 28 days HFD, and acute water immersion restraint stress (WRS) was induced for evaluation AM adaptive response and testing therapeutic potential of exogenous H2S donor NaHS (100 μmol/kg/day) and the hybrid H2S‐Aspirin (ATB 340, H2S‐ASA, 17,5 mg/kg/day) vs conventional aspirin (ASA, 10 mg/kg/day) on AM were evaluated by damage index by electronic microscopic study, and glucose level, activity of Cystathionine gamma‐lyase (CSE), Cystathionine beta‐synthase (CBS), Thiosulfate‐dithiol sulfurtransferase (TST) and Carbonyl group (CG) level in blood were examined biochemically (Institutional Ethical Committee Approval: 23/04/2018 № 4).ResultsIn HSD group with NaHS treatment protected the endothelial and sub‐endothelial structures in mesenterium destructions were absent, the nucleus of endothelial cells it was observed an uneven condensation of chromatin. Effect of hybrid H2S‐aspirin characterized as a protective on AM against HFD and WRS‐induced injury, exhibited a downward trend in the inflammatory infiltration, lowering the level CG, which may indicate a reduction in oxidative stress under the influence of hybrid H2S‐aspirin. ATB‐340 showed an increase CSE, CBS and TST activities vs ASA pretreatment (p<0.05). The use of aspirin and NaHS has shown an increase on 67% CSE and CBS activities in HSD rats vs saline group.ConclusionThese findings suggested that exogenous H2S prevented and ameliorated AM metflammation via reduction of oxidative damage, and provided a theoretical basis for effective tool for prevention of HFD‐related AM against long‐term postprandial hyperglycemia.AcknowledgmentAuthors would like to thank Prof. JL Wallace for ATB 340 providing and his expert suggestions.
Hippo Signaling Pathway has a critical role in Zika Virus Replication and in the Pathogenesis of Neuroinflammation
Authors: Garcia Gustavo , Paul Sayan , Beshara Sara , V Krishnan Ramanujan , Ramaiah Arunachalam , Nielsen-Saines Karin , Li Melody M. H. , French Samuel W. , Morizono Kouki W. , Kumar Ashok , Arumugaswami Vaithilingaraja 
Published: April 2020

Zika virus (ZIKV) is a reemerging human pathogen that causes congenital abnormalities including microcephaly and eye disease. The cell/molecular basis of ZIKV and host interactions inducing ocular and neuronal pathogenesis are unclear. Here we noted that the Hippo/SWH signaling pathway, which controls organ size through progenitor cell proliferation and differentiation, is dysregulated following ZIKV infection. In human fetal retinal pigment epithelial (RPE) cells, there is an early induction of transcriptional co‐activator, YAP, which is later degraded with a corresponding activation of the TBK1‐IRF3 Type I interferon pathway. YAP/TAZ silencing results in reduced ZIKV replication, indicating a direct role of Hippo pathway in regulating ZIKV infection. Using an in vivo Ifnr1−/− knockout mice model, ZIKV infection was found to reduce YAP/TAZ protein levels while increasing pYAP Ser127 in the retina and brain. Hippo pathway is activated in major components of the blood‐brain barrier (BBB), incuding endothelial cells and astrocytes. In addition, our data shows AMPK signaling pathway’s role in regulating YAP/TAZ in ZIKV‐infected cells. Our data demonstrates ZIKV infection initiates a crosstalk among AMPK‐Hippo‐TBK1 pathways, which regulate antiviral and energy stress responses during oculo‐neuronal inflammation.Support or Funding InformationThis work was funded by California Institute for Regenerative Medicine (CIRM) Quest – Discovery Stage Research Projects Grant (DISC2‐10188) to V.A. at the University of California, Los Angeles.
Optimization of Click Chemistry for Bioconjugation
Authors: Reilly Quinn , Mulllin Alissa C. , Oza Javin P. 
Published: April 2020

Application of the expanded genetic code has created specialized proteins that enable new methods to investigate biological systems. Biomaterials and antibody‐drug conjugations and often employ “click chemistry” utilizing copper(I)‐catalyzed azide‐alkyne cycloaddition. We have functionalized green fluorescent protein (GFP) by incorporating a non‐standard amino acid (nsAA), p‐Azido‐phenylalanine. However, the literature on the rate and yield of this reaction remains incomplete. To enhance bioconjugation, we have explored and optimized reaction conditions and stoichiometry. We employed two corroborating assays to test single‐ and double‐click reaction events: size exclusion chromatography and a fluorescence polarization assay. We evaluated the efficiency of a single click reaction between a small molecule fluorophore and our functionalized GFP. Completed click reactions were run through a purification column and fluorescence readings of eluted samples were used to evaluate yield. Analysis of this single click event was also conducted through a fluorescence polarization assay. This allows the determination of the reaction rate and yield in real time. Additionally, yield of double click reactions between two functionalized proteins and a polyethylene glycol (PEG) molecule with two alkyne handles has been evaluated using SDS‐PAGE. Through these efforts, we report reaction yields that surpass previously reported values. These advances support the improved employment of click chemistry for broad applications in biofunctionalization such as the development of biomaterials and antibody‐drug conjugatesSupport or Funding InformationFunding and support provided by the Cal Poly Chemistry and Biochemistry Department, the Center for Applications in Biotechnology, and the National Science Foundation.
Discovery of a Fibulotalocalcaneus (Peroneotalocalcaneus) Muscle – Rare Variant of Fibularis Quartus
Authors: Herrold Christina B. , Angilello Allison E. , Burkett Jesse T. , Zdilla Matthew J. , Lambert H. Wayne 
Published: April 2020

The fibularis (peroneus) quartus muscle is a variant muscle located in the lateral leg compartment that, when present, resides posterior to the fibularis longus and brevis muscles. Only found in approximately 11.5% of legs, the fibularis quartus arises from the lower one‐third of the fibula and descends to wrap posteriorly around the lateral malleolus. Confusion in the nomenclature arises due to the variations seen in regards to the distal insertion point of this variant leg muscle. The fibularis quartus can insert into the foot at a variety of locations, leading to many variations in the naming of the subtypes of this muscle. If the fibularis quartus muscle inserts into the retrotrochlear eminence and fibular (peroneal) trochlea of the calcaneus, it is called the fibulocalcaneus externus, which are the most common insertion sites. More distally, the fibularis quartus can insert into the cuboid bone, which is called the fibulocuboideus, though this variant is seen less frequently. Even further into the foot, a variation of the fibularis quartus may insert into the fifth metatarsal of the foot, changing its name to the fibularis digiti minimi (quinti). In this study, we examined the lateral leg compartment to see if a fibularis quartus may be present. In three of 20 legs (15%), a fibularis quartus muscle was identified. The first two examples were the more common fibulocalcaneus externus variant, in that this muscle inserted into the calcaneus. The third example was unique in that its distal tendon split to insert onto the calcaneus as well as the talus bone. The rare example of a fibulotalocalcaneus (peroneotalocalcaneus) muscle has only been reported once previously. This rare variant of the fibularis quartus is only the second known example of this exceedingly rare fibularis quartus variant. This finding is important clinically due to the presence of a fibularis quartus muscle being related to chronic lateral ankle pain and ankle instability. It is also important for radiologists, surgeons, and clinicians to be aware of these lateral leg muscle variants when looking at radiological imaging or in advance of surgical procedures in the lateral leg or foot.Support or Funding InformationT. Walley Williams Summer Anatomy Research Fellowship; WV Research Challenge Fund [HEPC.dsr.17.06]; WVU Research Apprenticeship Program (RAP)
SLC26A3 Deficiency Alters the Colonic Lamina Propria Lymphocytes
Authors: Jayawardena Dulari , Priyamvada Shubha , Kageyama Takahiro , Majumder Apurba , Sano Teuyuki , Dudeja Pradeep K. 
Published: April 2020

SLC26A3 or Down Regulated in Adenoma (DRA) is the key anion transporter involved in electroneutral NaCl absorption in the mammalian intestine. DRA knockout (KO) mice exhibit diarrheal phenotype and greater susceptibility to experimental colitis. A recent genome wide association study (GWAS) identified SLC26A3 polymorphism leading to lower DRA expression as a risk factor for ulcerative colitis. Additionally, we have shown that genetic deficiency of DRA in mice results in compromised barrier integrity and increased gut permeability. These studies render DRA to have a more complex involvement in IBD, beyond its transporter function. Enhanced intestinal permeability is associated with activation of mucosal immune system and may contribute to alterations in lamina propria lymphocytes consisting of innate (innate lymphoid cells ILC1, ILC2, ILC3) and adaptive immune system (Th1, Th2, Tregs and Th17). Thus, we sought to determine the total levels of lamina propria lymphocytes both innate and adaptive, to decipher their involvement in intestinal pathology observed in DRA KO mice. Ileal and colonic lamina propria lymphocytes were isolated from Wild type (WT) and DRA KO mice (n=7). Fluorescence Assisted Cell Sorting (FACS) analysis was carried out in these mice to determine the various subtypes of CD127+ ILCs and TCRb+/CD4+ Th Cells. Isolated lymphocytes were also stimulated to secrete cytokines using phorbol 12‐myristate 13‐acetate/PMA (50 ng/mL) and ionomycin (500 ng/mL) and were further analyzed with FACS. Our results demonstrated that GATA3+ colonic Th2 cells were significantly increased along with the corresponding CD127+/GATA3+ ILC2 in DRAKO mice compared to WT mice. These cells partake in mediating intestinal fibrosis and collagen deposition which are highly upregulated in DRAKO mice colons. However, tuft cell hyperplasia which may contribute to increased ILC2 was not observed in DRA KO mice. Similarly, the anti‐inflammatory CD127+/RORgt+ ILC3, which are important in maintaining intestinal epithelial integrity and intestinal cell homeostasis were significantly down‐regulated showing lower secretion of the important cytokine IL‐22 in DRAKO mice colons. However, the FOXP3+ Treg levels increased in colon of KO mice. The differences in lymphoid cells were mainly detected in the colon (where DRA expression is higher) and were less evident in the ileum. Overall, our data indicate that loss of DRA transporter results in altered intestinal lamina propria lymphocytes in the colon which may in part contribute to the higher susceptibility of DRA KO mice to inflammatory and infectious insults.Support or Funding InformationNIH/NIDDK, Department of Veterans Affairs
Isolation and Characterization of Antibiotic Resistant Bacteria from the Lane College Soil
Authors: McCullough Mariah , Baker Aliayah , Payne Quindarius , Van Stry Melanie 
Published: April 2020

Since the late 1940’s antibiotics have been an asset to the treatment of bacterial diseases. Considering they are naturally occurring or synthetically made, they still slow down the growth of bacteria. β‐lactam antibiotics, like penicillin, kill sensitive bacteria by inhibiting the enzyme transpeptidase required for cell wall synthesis. The β‐lactamase producing bacteria are important contributors to resistant infections that have increasing concern in the battle against infectious diseases. Our goal for this project is to isolate and characterize antibiotic bacteria from the soil on the Lane College campus. In addition, we are characterizing beta‐lactamases present in these bacteria. We hypothesize that bacteria in the sample will be resistant to the beta‐lactam antibiotics. We collected a soil sample from Lane College campus and screened the presence of bacteria resistant to ampicillin. We selected three colonies for further analysis. We then used the β‐lactam antibiotics penicillin, amoxicillin, and ampicillin in the disk assay experiment to test for multidrug resistance. We also further characterized the bacteria with Gram staining. We concluded that our sample did contain multidrug antibiotic resistant bacteria. We will screen for the presence of the beta‐lactamases TEM‐1 and Bla‐1 with PCR followed by gel electrophoresis.Support or Funding InformationThis work is supported by NSF HRD 1623340.
PKA RIα Holoenzyme Structure Reveals ATP‐Dependent Dynamics and Allosteric Regulation in Isoform‐Specific Manner
Authors: Lu Tsan-Wen , Wu Jian , Aoto Phillip , Weng Jui-Hung , Ahuja Lalima G. , Sun Nicholas , Taylor Susan 
Published: April 2020

Protein Kinase A (PKA) is the master switch for cAMP‐mediated signaling. PKA holoenzyme (R2C2) comprised a cAMP‐binding regulatory (R)‐subunit dimer and 2 catalytic (C)‐subunits, while cAMP binding to the R‐subunits unleashes the activity of the C‐subunit. Of the 4 functional non‐redundant R‐subunits (RIα, RIβ, RIIα, RIIβ), RIα and RIβ contain PKA pseudo‐substrate sites, in contrast to RIIα and RIIβ have substrate sites in their inhibitor sequences.Understanding how protein kinases serve as molecular switches is one of the holy grails in kinase biology. We provide new molecular features of PKA RIα allostery by identifying two conformational states of the holoenzyme (R2C2), one with and the other without ATP.RIα, in contrast to RIIβ, contains PKA pseudo‐substrate sequence, and our structures, with and without ATP, show how ATP functions as an allosteric inhibitor of RIα activation. The conformational changes of RIα holoenzyme are driven by ATP. ATP also facilitates RIα holoenzyme locks into inactive state and become more resistant to cAMP.In contrast to the RIIβ holoenzyme, ATP has no effect on RIIβ holoenzyme conformations; however, it serves as a substrate and can facilitate the phosphorylation of the inhibitor sequence in RIIβ. When the RIIβ holoenzyme is phosphorylated, it is easier to be activated by cAMP and unleash kinase activity.The isoform‐specific quaternary structures reveal a novel interface that helps to define domain crosstalk and a previously unappreciated allosteric pathway for cAMP activation. The results allow us to further appreciate the diversity of PKA isoforms and to understand the mechanism for allosteric activation.