bluuebunny/crossref_metadata_2025_split huggingface dataset:
Nanoscale‐Organization of Key Molecular Complexes Differentially Alter Vascular Smooth Muscle Function in Male and Females
Authors: Matsumoto Collin , Tajada Sendoa , Santana Luis Fernando 
Published: April 2020

Arterial smooth muscle contraction is the result of both electro‐mechanical and pharmaco‐mechanical coupling. During electro‐mechanical coupling, membrane depolarization activates dihydropyridine‐sensitive CaV1.2 channels, raising intracellular calcium concentrations ([Ca2+]i) and results in contraction of arterial smooth muscle. Pharmaco‐mechanical coupling activation of G‐protein coupled receptors (GPRCs) by angiotensin II (AngII) produces inositol 1,4,5‐triphosphate (IP3) and protein kinase C (PKC) activated diacylglycerol (DAG). This leads to an increase in [Ca2+]i via sarcoplasmic reticulum IP3 receptors and PKC dependent Ca2+ channels in the sarcolemma. It is known that male and females respond differently to antihypertensive drugs. However, it is unknown whether the nano‐scale organization of key molecular complexes between sexes play a role in smooth muscle physiology. Our data show that Ca2+ influx through CaV1.2 channels is larger in female than in male arterial myocytes. Furthermore, male smooth muscle responds greater to GPCR activation by AngII than female. We observed sex‐specific nano‐organization of CaV1.2 and AngII related signaling proteins angiotensin II receptor type 1 (AngIIR1), AKAP150 and PKC in arterial smooth muscle. We propose that there are sex‐based differences in smooth muscle physiology that are a result of different nano‐organization of signaling complexes.Support or Funding InformationT32 HL086350
Role of Dopamine D2 Receptor in the Leptin Receptor‐expressing Neurons in Modulation of Compulsive‐like Eating Behavior
Authors: Go Bok Soon , Kim Wanyeon 
Published: April 2020

Leptin, a hormone secreted by adipose tissue, is known to modulate satiety, caloric intake and body weight. There are six alternative spliced isoforms of leptin receptors (LepRa‐f). LepRb is highly expressed in the hypothalamic arcuate nucleus (ARC) which is involved in the regulation of food intake and body weight. Leptin activates pro‐opiomelanocortin‐expressing (POMC) neurons and inhibits agouti‐related peptide (AgRP)‐expressing neurons in the ARC.A recent study shows that dopamine modulates neuronal activities of arcuate POMC neurons and AgRP neurons. Dopamine inhibits arcuate POMC neurons via dopamine D2 receptor but activates AgRP neurons through dopamine D1 receptor. Dopamine is a neurotransmitter that plays a key role in mood, motivation and satiety. Dysregulation of dopamine neurotransmission is involved in the drug addiction. Food addiction is a recently proposed concept and compulsive eating behavior is shown in food addiction as well as an obesity. Negative reinforcement in drug addiction is a concept that negative affect associated with drug withdrawal promotes drug taking to ‘self‐medicate’ or remove those negative states. In the present study, we generated mice lacking D2 receptor in cells expressing LepRb (D2R flox; LepRb‐Cre; D2R cKO). Mice were provided palatable food for more than two weeks to induce food‐addiction like phenotypes in mice. To examine the role of D2 receptor in the leptin receptor‐expressing neurons in modulation of compulsive‐like eating behavior, light‐dark conflict test (LD test) was performed during the palatable food withdrawal period. In addition, palatable food withdrawal‐mediated anxiety/depression‐like behavior were measured during marble burying test (MBT), elevated plus maze (EPM), open filed test (OFT), and forced swim test (FST). No significant changes were found in palatable food (PF) intake in the light box during LD test, suggesting that D2 receptor in cells expressing LepRb would not be involved in the regulation of compulsive eating behavior. Furthermore, there was no significant changes in anxiety/depression‐like behavior measured by MBT, EPM, OPT and FST during the PF withdrawal period.Support or Funding InformationThis research was supported by Basic Science Research Program through the National Research Foundation of Korea (NRF) funded by the Ministry of Education (2019R1I1A1A01060007).
Expansion of NK cells by co‐crosslinking of 4‐1BBL and IL15R
Authors: Kim Jongil 
Published: April 2020

Adaptive transfer therapy using NK cell has been widely applied for cancer treatment. However, previous strategies on expansion and activation of NK cells is unsuitable for clinical application, because previous expansion strategies require feeder cells. Here, we established a method based on enhancement of NK cell function using removable magnetic protein G beads. We developed the fusion proteins such as 4‐1BB/igG1Fc and IL‐15Rα/IgG1Fc, then attached to magnetic protein G beads. The effect of immobilized 4‐1BB/IgG1Fc and IL‐15Rα/IgG1Fc on NK cell was assessed. We could confirm that fusion protein titration, proliferation, cytotoxicity, expression of activating receptors and IFN‐γ in NK cells were increased by immobilized fusion proteins on day 12. In summary, these results suggest that NK cells stimulated with bead platform can be efficiently without feeder cell, and appropriate for clinical safety. These bead platforms can be applied to a variety of diseases and cell types by changing the molecules to attach to the beadsSupport or Funding InformationProf. KyuBum KwackDepartment of Biomedical Science, College of Life Science, CHA University, Seongnam‐si, Gyeonggi‐do, Republic of Korea.
Glutathione peroxidase‐1 regulates adhesion and metastasis of triple‐negative breast cancer cells via FAK signaling
Authors: Lee Eun Kyung 
Published: April 2020

Triple‐negative breast cancer (TNBC) cells, which do not express genes for estrogen receptor (ER), progesterone receptor (PR), and Her2/neu, develop highly aggressive and metastatic tumors resistant to chemo‐ and hormonal therapies. We found that expression of glutathione peroxidase‐1 (Gpx1) is silenced in the non‐TNBC cells but resumed in the TNBC cell lines. Such restored Gpx1 expression plays a vital role in the metastasis of TNBC cells by regulating cell adhesion. Transcriptomic and signaling pathway analyses demonstrate that depletion of Gpx1 essentially impairs cell adhesion/spreading by down‐regulating FAK/c‐Src activation. Mechanistically, the substrate of Gpx1, H2O2, directly inhibits the FAK kinase activity. As a result, depletion of Gpx1 greatly suppresses lung metastasis of TNBC cells in vivo. Overall, our study identifies that Gpx1 is a redox safeguard of FAK kinase and allows for a therapeutic process to effectively control the metastasis of deadly malignant TNBC.Support or Funding InformationNational Research Foundation of Korea (2018R1A2B3006323 and 2017M3A9B6073098)National R&D Program for Cancer Control (1420280).
The Role of BCAT1 on Pediatric Dilated Cardiomyopathy
Authors: PIRES DA SILVA Julie , MIYANO Carissa , GARCIA Anastacia , SUCHAROV Carmen 
Published: April 2020

BackgroundPediatric dilated cardiomyopathy (DCM) is a devastating and poorly understood disease with most clinical treatment paradigms extrapolated from the adult population. Our studies showed that aspects of metabolism and mitochondria function are dysregulated in pediatric hearts. One potential factor which may be implicated in pediatric cardiac dysfunction is the Branched‐Chain Amino Acid Transaminase 1 (BCAT1), an enzyme highly expressed in pediatric non‐failing hearts and down‐regulated in response to DCM‐induced heart failure. While BCAT1’s role in cardiac dysfunction is unclear, in cancer cells and macrophages, BCAT1 is important for mitochondria function. The objective of this study is to investigate the role of BCAT1 in the heart, which may result in age‐specific therapies.MethodsWe measured metabolite composition of the pediatric DCM and non‐failing (NF) hearts. Activity and level of BCAT1 by enzymatic assay, RT‐qPCR and Western Blot in adult and pediatric and DCM and NF hearts was also measured. Pathologic changes in gene expression and mitochondrial function were assessed in Neonatal Rat Ventricular Myocytes (NRVMs) by RT‐qPCR and Seahorse assay in the presence or absence of BCAT1 inhibitor.ResultsAnalysis of metabolite content showed a dramatic accumulation of branched‐chain amino acids (BCAAs) in pediatric DCM hearts. BCAT1 mRNA and protein levels are increased in pediatric NF hearts compared to adult NF hearts, and decreased in pediatric DCM hearts. In addition, inhibition of BCAT1 in NRVMs results in pathologic gene expression and inhibition of pro‐inflammatory cytokines. Finally, Seahorse assay showed a decrease of mitochondrial respiration in NRVMs treated with BCAT1 inhibitor.ConclusionsDecreased levels of BCAT1 are detrimental to the pediatric failing heart. These findings suggest that BCAT1 could be a specific therapeutic target for the pediatric population.Support or Funding InformationR01 HL139968 and HL126928
The Personalized Precise Exercise Training Holistic Integrative Protocol (PPET‐HIP) for Patient with Myocardial Incompetence and Catecholamine‐Sensitive Ventricular Arrhythmia after her Sudden Cardiac Arrest
Authors: Song Ya , Sun Xingguo 
Published: April 2020

INTRODUCTIONSThe patient was a 23yrs female nurse in Fuwai‐hospital. After 2d overtime work, she had suddenly cardiac arrested followed by successful CPR and 5hr unconsciousness. Afterward she was diagnosed as catecholamine‐sensitive ventricular tachycardia, myocardial insufficiency and etc; and some physicians suggested installing of cardiac pacemaker with ICD. However, based on HIPM, we believe her main problem was the mismatching of supply‐demand and poor integration among systems, and Personalized Precise exercise Training Holistic Integrative Protocol (PPET‐HIP)can reverse those. We suggested give us ~100d PPET‐HIP and then decide for pacemaker‐ICD.METHODSAfter performing symptom limited maximum cardiopulmonary exercise testing (CPET) for objectively quantitative evaluation, we generated her PPET‐HIPΔ50% Power for intensity titration. According to results of continuous beat‐by‐beat blood pressure and continuous blood glucose monitoring during 30min Δ50% Power intensity titration we did select qid, 6d/w intensive management.RESULTSAfter 1m of intensive management, her continuation duration of PPET‐HIP was increased from 1–2min to 6–7min. The total exercise duration was increased from 17 to 24min, and resting was decreased from 8–10 to 3–4 times/30min. In 1st CPET, ventricular premature (PVC) began from loading intensity over 75w/min until peak loading127.5W/min , and total PVC was 119beat/130s.After three‐month PPET‐HIP,CPET showed PVC began from loading intensity over 100w/min until peak loading 162.5W/min , and total PVC was 68beat/170s. Comparing with the 1st one, three‐month CPET’s key parameters of peakVO2, AT, peak loading, peak CO and oxygen uptake efficiency plateau (OUEP) were increased by 20.1%, 15.2%, 21.5% and 22.5%, 21.7% and 5.7% respectively, only VE/VCO2 was decreased by 5.4%; all indicated improvement of functional status. The she has been believed that she couldn’t be nursing work again. However, after leaving Fuwai, she has become a clinical nurse again and working in cardiac rehabilitation, which is very meaningful for her.CONCLUSION

Under the guidance of HIPM, we could safely and effectively manage patients by PPET‐HIP.
Increased functional capacity parameters in CPET by 5–23%.
Increased threshold of PVC and decreased rate of PVC during CPET.
Decreased number of PVC and decreased rate of PVC in 30min training of PPET‐HIP.Support or Funding InformationNational Natural Science Foundation of China (81470204);Fuwai Hospital, National Cardiovascular Institute of Chinese Academy of Medical Sciences (2012‐YJR02);Research on Clinical Characteristics of the Capital (Z141107002514084);Research and Outcome Promotion of Clinical Characteristics in the Capital (Project No. Z161100000516127)
Gene regulation networks in muscle differentiation in lamin A associated muscular dystrophy
Authors: Dutta Subarna 
Published: April 2020

A type lamin has came into the limelight due to it’s involvement in the different plethora of diseases, collectively known as laminopathies. According to the previous reports, lamin A has involvement in disrupting differentiation of overexpressed R453W mutant of lamin A associated autosomal EDMD. In our immunofluorescence studies, we have ventured myoblast cells expressing a severe skeletal muscular dystrophy mutated lamin A, had a significant lower capacity to differentiate than wild type one. Anomalies in the reulations of protein expression levels of MyHC, MyoD were also checked by western blot analysis. These convey that, A type lamins play a crucial role by influencing expression of genes necessary for muscle cells differentiation .The abnormalities due to muscular dystrophy may lead to impairment of gene circuitry ,which are related to myoblast fusion necessary for the formation of mature myotubes. In our RNA sequencing study, we evaluated a thorogh gene expression profiling in the context of complex, highly coordinated process of muscle differentiation in stably transfection of wild type and mutant in C2C12 myoblast cells, which were differentiated for a specific time period. In the expression profiling of seven day time course, the genes which are showing highest scores in the disputes between wild type versus mutants ,were analysed further to get eloquent idea on cell signaling pathways were involved in. We identified dysfunctional pathways to compare wild type versus mutant of muscular dystrophy during differentiation of myoblast to myotube transition. These data provide a model in the muscular dystrophy pathogenesis to interlace the impacts of genetic modifiers in the differentiation potentiality.Support or Funding InformationThis work was funded by DAE, SERB‐DST.
Propofol Postconditioning Ameliorates Hypoxia/Reoxygenation induced H9c2 Cells Apoptosis and Autophagy via upregulating Forkhead Transcription Factors under Hyperglycemia
Authors: Han Ronghui , Han Hong , Xie Xiang , Chen Hao , Liu Danyong , Zhang Liangqing , Xia Zhengyuan 
Published: April 2020

Administration of propofol, an intravenous anesthetic with antioxidant property, immediately at the onset of post‐ischemic reperfusion (propofol postconditioning, P‐PostC) has been shown to confer cardioprotection against ischemia‐reperfusion injury , while the underlying mechanism remains incompletely understood. Also, whether or not P‐PostC can confer cardioprotection in diabetes or under hyperglycemic condition has yet to be explored. The FoxO transcription factors were reported to play critical roles in promoting cardiomyocyte survival during conditions of oxidative stress through induction of antioxidants and cell survival pathways (J Biol Chem. 2011;286(9):7468–78) and was also involved in hypoxic postconditioning mediated neuroprotection. However, the role of Foxo in postconditioning mediated protection in the heart and in particular in high glucose condition is unknown. We hypothesized that P‐PostC may attenuate hypoxia/reoxygenation (H/R) injury in cardiac origin H9c2 cells by attenuating apoptosis and autophagy through upregulating FoxO1 and FoxO3a under hyperglycemia. Rat heart‐derived H9c2 cells were exposed to high glucose (HG) for 48 hours, then subjected to H/R (composed of 8 hours of hypoxia followed by 12 hours of reoxygenation) in the absence or presence of postconditioning with various concentrations of propofol at the onset of reoxygenation. Our data showed that HG with or without H/R decreased cell viability, increased lactate dehydrogenase (LDH) leakage and the production of reactive oxygen species (ROS) in H9c2 cells, all of which were significantly reversed by propofol (P‐PostC), especially at the concentration of 25μM. Moreover, we found that propofol (P25) decreased H9c2 cells apoptosis and autophagy that were associated with increased FoxO1 and FoxO3a expression. The protective effects of propofol (P25) against H/R injury were reversed by silencing FoxO1 or FoxO3a. Taken together, these results indicated that propofol postconditioning attenuated H9c2 cardiac cells apoptosis and autophagy induced by H/R injury through upregulating FoxO1 and FoxO3a under hyperglycemia.Support or Funding InformationSupported by National Natural Science Foundation of China (NSFC) grant (81970247)
Increased Levels of Cortisol in Patients Undergoing Total Joint Arthroplasty may be Associated with Stress and Inflammatory Processes
Authors: Haddad Sandra , Khanna Rajan , Schmitt Daniel , Slovacek Hannah , Siddiqui Fakiha , Hoppensteadt Debra , Liles Jeffrey , Wanderling Christopher , Fareed Jawed , Hopkinson William 
Published: April 2020

IntroductionOsteoarthritis is a condition in which joint cartilage and its adjacent bone degenerate progressively over time due to inflammatory processes involving activation of matrix degrading enzymes. Total joint arthroplasty (TJA) is a common surgical treatment for osteoarthritis of the hip or knee. Cortisol is a steroid hormone in the class of glucocorticoids which is produced in the adrenal gland. Cortisol is generated in response to inflammation and physical or emotional stress. This study aims to investigate the cortisol levels in patients undergoing TJA.Materials and MethodsPlasma samples were prospectively collected from 71 TJA patients at baseline (pre‐surgery), 24 hours post‐operation, and 5 days post‐operation. Normal controls represented plasma samples from drug free health individuals (n=50). Cortisol and TNFa levels were measured in each sample using a commercially available ELISA kit. All results were compiled as group means ± SD.ResultsThe plasma cortisol level in normal individuals were 67 ± 11 ng/mL. In contrast, the pre‐surgical and 24‐hour post‐surgical samples showed a marked increase in cortisol levels (> 200ng/mL). The blood samples drawn at the 5th day after surgery showed a downward trend reaching near normal levels (74 ± 12 ng/mL). There was no significant difference between cortisol levels pre‐operatively and within 24 hours post‐TJA, however there was a significant difference in comparison to the levels on the 5th day and normal. The TNFa levels were elevated in TJA patients in the pre‐surgical (18.6 ±3.2 pg/ml), at 24 hours post‐surgical (24.2 ± 7.6 pg/ml) and on the 5th day (11.3 ± 3.1 pg/ml) in comparison to normals (5.1 ± 2.1 pg/ml).Discussion and ConclusionAt 5 days post‐operation, cortisol levels were significantly lower than at baseline or 24 hours post‐operation. This finding differs from previous findings, which reported increased levels post‐operatively relative to at baseline. The TNFa levels also showed elevated levels in the pre‐ and post‐surgical samples which followed a downward trend on the 5th day. These results are consistent with the previous reports that such biomarkers as IL‐6 and TNFa are upregulated in TJA patients prior to surgery suggesting a role of inflammatory process. Furthermore the increased level of cortisol may be related to not only the inflammation but pre‐surgical emotional stress in TJA patients. The near normalization of the cortisol levels on the 5th post‐surgical day is suggestive of differential down‐regulation of the inflammatory process and stress levels in TJA patients.
Ozanimod Attenuates Ischemia‐Induced Inflammation and Dysfunction in Human Brain Microvascular Endothelial Cells
Authors: Wendt Trevor S. , Li Yu Jing , Liu Qiang , Shi Fu-Dong , Gonzales Rayna J. 
Published: April 2020

Sphingosine‐1‐phosphate receptor (S1PR) modulators elicit neuroprotection during stroke, however the mechanism(s) associated with beneficial outcome remains unknown. Endothelial dysfunction is a key pathophysiological contributor in acute ischemic stroke. Here, we researched the vascular specific role of the S1PR1 ligand, ozanimod, on endothelial function and health following hypoxia plus glucose deprivation (HGD), an in vitro ischemic‐like model. Specifically, we investigated whether ozanimod (0.5 nM) alters human male brain microvascular endothelial cell (HBMEC) proliferation, morphology, and integrity. HBMECs were exposed to HGD (1% O2) or normoxia (21% O2) for 3h at passage 7 using a humidified chamber (BioSpherix®) housed within a 5% CO2 incubator. For the proliferation studies, cells were treated with bromodeoxyuridine and incorporation of the synthetic nucleoside analog during normoxic or HGD exposure was measured via flow cytometric analysis. Morphology was assessed post normoxic or HGD exposure in HBMECs stained with crystal violet and visualized with brightfield microscopy. Trans endothelial electrical resistance was measured following HGD or normoxia exposure using polyethylene terephthalate 8 well plates and electric cell‐substrate impedance sensing (ECIS® Z Theta). Additionally, we investigated the impact of HGD on S1PR1 and cyclooxygenase‐2 levels using standard immunoblotting techniques. In some experiments, selective blockade with the antagonist, W146, verified S1PR1 activation involvement. We observed that S1PR1 was basally expressed in HBMECs and HGD increased expression suggesting S1PR1 as a potential target during acute ischemic injury. Ozanimod attenuated HGD‐mediated decreases in HBMEC proliferation and W146 reversed this response. S1PR1 blockade had no effect on cell proliferation following normoxia. Ozanimod also preserved HGD‐induced morphological changes. During electric cell‐substrate impedance assessment, ozanimod attenuated HGD mediated reduction in endothelial cell resistance and W146 abolished this effect suggesting S1PR1 dependency. Concomitantly, we observed that HGD reduced expression levels of zonula occludens (ZO)‐1 and claudin‐5. Ozanimod also decreased HGD‐mediated increases in proinflammatory mediator expression of cyclooxygenase‐2. In conclusion, ozanimod elicited protection against HGD in HBMECs and may play an important beneficial role against acute ischemic damage by improving brain endothelial integrity and viability via selective S1PR1 activation.Support or Funding InformationUniversity of Arizona Valley Research Partnership Grant VRP37 P2 (RJG, FS), American Heart Association 19AIREA34480018 (RJG).
DNA Repair in Colon Cancer Cell Lines.
Authors: O’Donoghue Tristan , Podlutsky Andrej 
Published: April 2020

Colorectal cancer remains the third most common cancer world‐wide, despite recent advances in early diagnosis and following treatment strategies. Genome stability plays important role in cancer’s origin and progression. The fidelity of DNA repair is of most importance for the colorectal cancer, which characterized by accumulation of mutations in oncogenes and tumor suppressor genes, and dozens of so called “passenger” mutations in genes that are not driving cancer progression. Studying of cell cultures established from colon‐cancer patients are important tool for understanding molecular mechanisms behind cancerous transformation and drug development. Two colon‐cancer cell lines are widely used in such research are HT29 and HCT116. We analyzed the nucleotide excision repair (NER) in these cells by monitoring DNA repair following UV‐C exposure. The NER pathway is one of several fundamental repair mechanisms that is guarding genome from accumulation of DNA damage. Following the single gel electrophoresis technique, we determine that HT29 cells are slower at repairing UV‐C induced damage compared to HCT116 cells or normal human fibroblasts, HDFa cells. This is an important funding, because it shows that colon‐cancer cell lines are different in stress response capacity. Such difference might be important for the drug development and our understanding of basic mechanisms of genome stability in cancer cell lines.Support or Funding Information1) Research reported in this publication was supported by the National Institute Of General Medical Sciences of the National Institutes of Health under Award Numbers UL1GM118991, TL4GM118992, or RL5GM118990. The content is solely the responsibility of the authors and does not necessarily represent the official views of the National Institutes of Health. UA is an AA/EO employer and educational institution and prohibits illegal discrimination against any individual:
www.alaska.edu/titleIXcompliance/nondiscrimination. 2) NASA EPSCoR, NNH15ZHA003C