bluuebunny/crossref_metadata_2025_split huggingface dataset:
Collaborative Work Mediated by the Use of Google Drive as a Teaching Strategy Learning in Subjects of Research in Students of the Health Area
Authors: Arroyo Ana Gabriela Campos , Matías Flora María Cabrera , Escobar Omar David Orlanzzini 
Published: April 2020

Scientific research in nutrition is essential to improve the quality of life of populations, identify health problems, propose health strategies and prevention plans for the most prevalent diseases. So, it is necessary to encourage students to interest in subjects related to research using innovative teaching and learning strategies that favor individual and collaborative learning.ObjectiveIdentify how the use of Google Drive contributes as a teaching‐learning strategy in the subject of research and development of degree projects in students in the area of health.Materials and methodsThe approach is mixed, concurrent and exploratory. 117 students from the Faculty of Nutrition Sciences of the University of Morelia were considered. At the beginning of the course, all students were asked to create a Gmail account in order to have access to all Google Drive tools. Once this was done, they were explained in face‐to‐face classes how to access and what elements the Google Drive platform has, individual folders with last name and first name were created and during the whole semester the students worked personally in their folders, developing activities in a way individual or in teams according to the programmed topic. One of the main activities carried out was the development of its own research protocol. Once the semester was completed, a questionnaire was applied through Google Forms, which consisted of two sections. The first section with identification questions and the second section with 15 closed and open questions to identify how the use of Google Drive. Contributes as a teaching‐learning strategy in research subjects.Results97.9% of the students considered that Google Drive is a good teaching‐learning strategy, 95.7% indicated that it helped them learn and facilitated the realization of their research projects, and 97.9% indicated that it helped them to work on collaborative form.ConclusionThe use of Google Drive can be a teaching‐learning strategy that contributes to working in a collaborative, real‐time, comfortable, dynamic way and that provides a new way in which the teacher and the student can keep a personalized follow‐up in each of the research projects that are carried out.Support or Funding InformationResearch supported and endorsed by the Santander University and University of Morelia.
G Protein Gamma Subunits: Phosphorylation‐Mediated Governors of G protein Signaling in Humans?
Authors: Toosi Zahra Nassiri , Choudhury Ajanta , Su Xinya , Torres Matthew P. 
Published: April 2020

Intracellular heterotrimeric G proteins (consisting of Gα, Gβ, and Gγ subunits), interface with 7‐transmembrane G protein coupled receptors (GPCRs) and function as the primary transducers of extracellular signals such as hormones, neurotransmitters, as well as pharmaceutical compounds. Upon binding their cognate ligands, GPCRs promote the dissociation of Gα and Gβγ subunits, which are then free to interact with effector proteins that promote intracellular signaling responses to the stimulus. Within this context, Gγ subunits are thought of primarily as membrane anchors for Gβ subunits, which must reside at the plasma membrane to facilitate functional protein interactions. However, emerging evidence suggests that Gγ subunits serve as phosphorylation‐dependent governors of G protein signaling. We have recently demonstrated this in the budding yeast model system, in which negative feedback phosphorylation of the intrinsically disordered N‐terminal tail of Gγ/Ste18 disrupts Gβγ/effector interactions and inhibits the normal GPCR‐dependent activation of MAPKs (Choudhury S. et al, Cell Rep. 2018). More recently, we have discovered that the yeast Gγ subunit undergoes combinatorial phosphorylation in response to several different forms of extracellular stimuli, including osmotic stress, pH stress, glucose deprivation, and cell cycle progression, in addition to GPCR activation. In light of these discoveries, we have hypothesized that Gγ phosphorylation plays a similar role in mammalian GPCR pathways. Indeed, we show that, like Gγ/Ste18, all mammalian Gγ subunits have an intrinsically disordered N‐terminus and most also undergo multi‐site phosphorylation. Here we provide new evidence to evaluate the hypothesis that Gγ phosphorylation is a governor of G protein signaling in mammals by demonstrating that phosphorylation is readily detectable on many different human Gγ subunits (Gγ3, 4, 5, 7, 10, 12) and by systematically testing the requirement for phosphorylation in proper Gβγ‐specific signaling pathways including the activation of PKD1 kinase by phospholipase C β2/3.Support or Funding InformationNIH
Alteration of KCa Channel Function by Homocysteine in Human Internal Mammary Artery
Authors: Sun Wen-Tao , Xue Hong-Mei , Hou Hai-Tao , He Guo-Wei , Yang Qin 
Published: April 2020

ObjectivesHyperhomocysteinemia is an independent risk factor for cardiovascular disease. We previously in porcine coronary arteries demonstrated that disruption of KCa channel activity is involved in homocysteine‐induced impairment of dilatory function. Whether homocysteine affects KCa channels in human arteries remains uninvestigated. In the present study, we aimed to investigate the effect of homocysteine on the expression and functionality of the members of KCa channel family, including large‐ (BKCa), intermediate‐ (IKCa) and small‐ (SKCa) conductance KCa channels in human internal mammary artery (IMA).MethodsResidual IMA segments collected from patients undergoing coronary artery bypass grafting (CABG) were used for myography study for KCa subtype‐mediated relaxation and contraction. Western blot and immunohistochemistry were employed to detect the expression and distribution of KCa subtypes in IMA.ResultsBKCa subtype in the KCa channel family significantly regulated the vasoreactivity of IMA. Blockade of BKCa channels inhibited acetylcholine‐induced relaxation and enhanced U46619‐induced contraction. In comparison, both IKCa and SKCa subtypes played a minor role in the control of relaxation and contraction of IMA. Exposure to homocysteine compromised the vasodilating activity of the BKCa subtype in IMA, evidenced by the suppressed relaxant response to the BKCa channel opener and the decreased impact of the BKCa channel blocker on acetylcholine‐induced relaxation and U46619‐evoked contraction. The functional loss of BKCa was associated with a downregulation of the protein expression of BKCa β1 subunits in the smooth muscle of IMA. Inhibition of endoplasmic reticulum (ER) stress showed protective effect against homocysteine on the function and expression of BKCa channels. In contrast, the role of IKCa and SKCa subtypes in mediating endothelium‐dependent relaxation was augmented by homocysteine though no alterations in the regulatory capacity on vasoconstriction were observed. Homocystine did not alter the protein expression level of IKCa and SKCa subtypes.ConclusionsHomocysteine compromises the vasodilating activity of BKCa channels in IMA, which may be attributed to the loss of BKCa β1 subunits mediated by ER stress. Unlike BKCa, IKCa and SKCa subtypes are unessential for vasoregulation in IMA, the loss of BKCa functionality in hyperhomocysteinemia enhances the role of IKCa and SKCa in mediating endothelial dilator function. Targeting BKCa may form a strategy to improve the postoperative graft performance in CABG patients with hyperhomocysteinemia who receives IMA grafting.Support or Funding InformationSupported by NSFC 81870227 & 81870288, Tianjin Science and Technology Commission (18PTZWHZ00060) & Binhai New Area Health Bureau (2018BWKZ005), and Non‐profit Central Research Institute Fund of Chinese Academy of Medical Sciences 2018TX31002 & 2019XK310001.
Role of Nrf2 in Regulation of Expression of CYP 2C11 and CYP 4X1 Epoxygenases and Production of Epoxyeicosatrienoic Acids (EETs) in the Rat Brain
Authors: Gebremedhin Debebe , Guerts Aron , Lombard Julian H. 
Published: April 2020

Nuclear factor (erythroid‐derived 2)‐like‐2 (Nrf2) is the master antioxidant and cell‐protective transcription factor native in the rat brain. The CYP epoxygenases 2C11 and 4X1 are expressed in the brain and metabolize arachidonic acid to epoxyeicosatrienoic acids (EETs) that are known to induce cerebral vasodilation and increase nutritive blood flow to areas of heightened neuronal activity and also provide antioxidant and anti‐inflammatory actions. The influence of Nrf2 gene deletion on CYP 2C11 and 4X1 epoxygenase expression and activity in the rat brain is not known. In this study we used isoform specific antibodies and Western blotting to determine the effect of Nrf2 gene knockout (KO) on the expression of proteins for CYP 2C11 and CYP 4X1 epoxygenases in whole brain and isolated cerebral arteries. We also measured the production of epoxyeicosatrienoic acids (EETs) in homogenates of whole brain and isolated cerebral arteries of Nrf2 wild type (WT) and Nrf2 KO rats. CYP epoxygenase expression and EETs production were reduced in whole brain and arteries of the NRF2 KO rats compared to WT controls. These findings indicate that knockout of the Nrf2 gene leads to a marked reduction in the expression of proteins for CYP 2C11 and CYP 4X1 epoxygenases and in the production of 8, 9‐, 11,12‐ and 14,15‐EETs compared to those measured in the Nrf2 WT rat brain issues and cerebral arteries. The results of the current study suggest that CYP 2C11 and 4X1 epoxygenases may be components of the Nrf2‐induced phase 2 genes that are known to limit oxidative stress and provide cellular defense in the brain. We postulate that Nrf2 may regulate functions of the cerebral circulation and provide protection against oxidative stress, in part, via modulation of the expression level of CYP 2C11 and 4X1 epoxygenase proteins and the production of EETs.Support or Funding InformationNIH R01 #HL‐128242 and NIH #R21‐OD018309
Estrogen Replacement Improves Mesenteric Arterial Relaxation in UC Davis Pre‐diabetic Rats: Possible Involvement of Estrogen Receptor Beta
Authors: Razan Md Rahatullah , Akther Farjana , Karimi Habibullah A. , Amissi Said , Graham James L. , Stanhope Kimber L. , Havel Peter J. , Rahimian Roshanak 
Published: April 2020

The risk of cardiovascular diseases (CVD) is lower in premenopausal women compared to men, and this lower risk is related to the presumptive cardioprotective effects of female sex hormones, estrogen in particular. However, premenopausal women may lose this protection with the onset of menopause and/or when they become diabetic. Little is known about the vascular function in presence or absence of estrogen at the early stage of diabetes.The objective of this study was to investigate the effects of 17β‐estradiol (E2) replacement on mesenteric arterial (MA) function in ovariectomized (ovex) UC Davis type 2 diabetes mellitus (UCD‐T2DM) rats at the pre‐diabetic stage. MA were obtained from four groups of ovex rats implanted with a subcutaneous pellet for 45 days: (1) Control (Ctrl), ovex + placebo, (2) Ctrl, ovex + E2 (1.5mg/pellet), (3) pre‐diabetic, ovex + placebo, (4) pre‐diabetic, ovex + E2. Endothelium‐dependent vasodilation (EDV) to acetylcholine (ACh, 10−8 to 10−5 M) in MA were measured before and after indomethacin (Indo, 10μM), a cyclooxygenase (COX) inhibitor; followed by addition of L‐NAME (200μM), a nitric oxide synthase (NOS) inhibitor. The MA expression of estrogen receptors (ERs) and plasma E2 concentration were determined by Western blotting and ELISA assay, respectively.E2 treatment significantly increased plasma estradiol concentration in both Ctrl and Prediabetic groups. Analysis of ERs showed a higher expression of ER‐beta only in E2‐treated prediabetic group. ACh responses were similar in Ctrl groups, regardless of E2 replacement. ACh induced relaxation was, however, markedly reduced in MA of placebo treated pre‐diabetic group compared with other groups. Treatment of pre‐diabetic with E2 enhanced relaxation to the same level as those from Ctrl rats. Blocking of both COX and NOS completely abolished the relaxation responses to ACh in pre‐diabetic groups, regardless of E2 replacement, indicating no contribution of EDHF‐type relaxation in these groups. However, the added effect of L‐NAME, in presence of Indo, in blocking ACh‐mediated vasodilation was much more prominent in E2‐treated prediabetic group when compared with placebo treated group, indicating an enhanced role of NO. On the other hand, both NO and EDHF remained important for EDV in Ctrl groups. Addition of L‐NAME, however, caused a much smaller reduction of EDV in E2 treated than in placebo treated arteries, suggesting EDHF is the predominant mediator of mesenteric vasorelaxation in this groups.Here, we show that EDV is impaired at pre‐diabetic stage, may be due to loss of EDHF. E2 treatment restores EDV in MA of pre‐diabetic group, possibly via elevated contribution of NO‐mediated relaxation by increased E2 signaling (via ER beta) in this group. The role of ER alpha remains to be investigated.Support or Funding InformationSupported by NHLBI.
Prenatal chronic intermittent nicotine aerosol exposure programming a sex dependent hypertensive phenotype via vascular eNOS uncoupling
Authors: Liu Bailin , Liang Xia , Jie Jian , Li Yong , Dasgupta Chiranjib , Huang Xiaohui , Zhang Lubo , Shao Xuesi M. , Xiao DaLiao 
Published: April 2020

ObjectiveMaternal cigarette smoking/nicotine use during pregnancy is associated with an increased risk of cardiovascular diseases and development of hypertensive phenotype in later life. However, the involved molecular pathway is remain unclear. The present study tested the hypothesis that cigarette smoking‐relevant nicotine inhalation during pregnancy causes a gender‐dependent change the balance of resistance vasculatures eNOS and prooxidant functions in adult offspring, leading to development of hypertensive phenotype in adult male offspring.MethodsThe time‐dated pregnant rats were exposed to chronic intermittent nicotine aerosol (CINA) via a novel lung alveolar region‐targeted aerosol method during pregnancy (E4 to E20). Experiments were performed in adult (~6 month‐old) offspring.ResultsCINA exposure enhanced vascular H2O2 production which was inhibited by eNOS inhibitor, L‐NAME in male offspring. CINA exposure selectively enhanced vascular NOX2 expression. CINA exposure enhanced angiotensin II (Ang II)‐induced BP response in male offspring, which was not affected by NOX2 inhibitor (gp91 ds‐tat) but significantly reversed by L‐NAME or BH4 treatment in vivo. In pressure myography experiments. CINA exposure had no effect on pressure‐induced myogenic tone but decreased the Acetylcholine (Ach)‐induced relaxation in adult male but not female offspring. In the presence of L‐NAME or H2O2 scavenger (PEG‐catalase), there were no differences of Ach‐induced relaxation between CINA and saline control offspring. Furthermore, pretreatment of eNOS cofactor BH4 enhanced Ach‐induced relaxation in CINA‐exposed male offspring, while no effect in saline control group. In addition, exogenous NO donor (SNP)‐induced mesenteric artery relaxations were similar between CINA exposed and saline control male offspring.ConclusionsThese findings provide novel evidence that maternal inhaled nicotine exposure could impair fetal vascular development and induce a gender‐dependent development of endothelial dysfunction phenotype in adult male offspring via enhancing vascular eNOS uncoupling. The uncoupled eNOS‐mediated ROS plays a predominant role in CINA‐induced hypertensive response in male offspringSupport or Funding InformationSupported in part by the NIH grants: R01HL135623, R03DA041492, and R01HD088039; TRDRP grants: 29IR0437 and T30FT0936
Combination of dapagliflozin and statins attenuate renal lipotoxicity in high‐fat high‐fructose diet‐induced insulin resistant rats
Authors: Thongnak Laongdao , Pongchaidecha Anchalee , Chatsudthipong Varanuj , Lungkaphin Anusorn 
Published: April 2020

High‐fat high‐fructose diet (HFF) can induce dyslipidemia and lipid accumulation in the kidney which are related to insulin resistance and metabolic syndrome. Dyslipidemia and lipid accumulation in obesity can lead to lipotoxicity and cellular damage particularly kidney injury by activating oxidative stress, fibrotic and apoptotic pathways. This study investigated whether dapagliflozin alone or combined with atorvastatin could improve metabolic parameters, lipid metabolism and lipid accumulation induced kidney injury in HFF‐induced insulin resistant rats. Male Wistar rats were fed with high‐fat diet with 10% fructose in drinking water for 16 weeks. After that, these rats were received drug treatments for 4 weeks; vehicle (HFF), SGLT2 inhibitor dapagliflozin (HFFD), atorvastatin (HFFA) and SGLT2 inhibitor combined with atorvastatin (HFFDA) groups. HFF rats demonstrated insulin resistance, dyslipidemia, and renal dysfunction associated with impaired renal lipid metabolism and lipid accumulation. Lipid accumulation in the kidney could promote renal dysfunction in HFF rats via the activation of oxidative stress, fibrosis and apoptosis. Dapagliflozin treatment could improve insulin resistance, lipid synthesis and lipotoxicity‐related renal oxidative stress, fibrosis and apoptosis leading to reversal of kidney dysfunction in HFF model. The combined treatment (HFFDA group) could also improve metabolic parameters which was not different from single drug treatment (HFFD or HFFA group). Notably, the reduced lipid accumulation in kidney which were linked to the improvement in lipid oxidation was better in the combined treatment group when compared to a single drug treatment. In summary, dapagliflozin combined with atorvastatin treatment exert the beneficial effects on metabolic parameters, lipid metabolism, and lipid accumulation induced kidney injury via the attenuation of oxidative stress, fibrosis and apoptosis in high‐fat high‐fructose diet‐induced insulin resistant model.Support or Funding InformationThailand Research Fund (RSA6080015 AL); the Royal Golden Jubilee PhD program (PhD/0063/2560 LT and AL); the Faculty of Medicine Research Fund, Chiang Mai University (AL and AP); the Graduate Research Scholarship Chiang Mai University (LT); and the Functional Food Research Center for Well‐being, Chiang Mai University (AL).
Structural and Molecular Characteristics of Axons in the Long Head of the Biceps Tendon
Authors: Streicher Johannes , Boesmueller Sandra , Gesslbauer Bernhard , Hirtler Lena , Bormann Daniel , Pastor Angel M. , Mittermayr Rainer , Blumer Roland 
Published: April 2020

ObjectivesAlthough the long head of the biceps tendon (LHBT) is a major source of shoulder pain, the innervation is not sufficiently documented. Thus, the present study aimed to characterize structural and molecular features of nervous elements in the LHBT.MethodsThe proximal part of eleven LHBTs was harvested intraoperatively. There were 8 female and 3 male specimens. Age ranged from 66 to 86 years. For structural analyses, nervous elements were viewed in the transmission electron microscope. For molecular characterization, we used a series of neuronal markers: This included general neuronal markers [antibodies against neurofilament and protein gene product 9.5 (PGP9.5)] as well as specific neuronal markers[antibodies against myelin basic protein (MBP), calcitonin gene related product (CGRP), substance P (SP), tyrosine hydroxylase (TH), and growth associated protein 43 (GAP43)]. Anti‐neurofilament and anti PGP9.5 visualized the overall innervation. Anti‐MBP visualized myelination, anti‐CGRP and anti‐SP nociceptive fibers, anti‐TH sympathetic nerve fibers, and anti‐GAP43 nerve fibers during development and regeneration. Immunolabelled sections were analyzed in the confocal laser scanning microscope.All procedures performed in this study were approved by the local medical ethics committee (Austrian workers compensation boards’ research ethics committee and institutional review board of the Medical University of Vienna, No. 06/2017) and in accordance with the 1964 Helsinki declaration and its later amendments or comparable ethical standards. Informed consent was obtained from all persons included in this study.ResultsWe show that the LHBT contains unmyelinated as well as myelinated nerve fibers which group in nerve fascicles and follow blood vessels. Many myelinated and unmyelinated axons exhibit molecular features of nociceptive nerve fibers. Another subpopulation of unmyelinated axons exhibits molecular characteristics of sympathetic nerve fibers. Unmyelinated sympathetic fibers and unmyelinated nociceptive fibers express proteins that are found during development and regeneration.ConclusionPresent findings show that the LHBT of aged persons is richly innervated. The occurrence of growth protein GAP43 in nociceptive nerve fibers supports the hypothesis that in‐growth of nociceptive fibers is the source of chronic tendon pain.
The Cholesterol Synthesis Enzyme Lanosterol 14α‐Demethylase is Post‐Translationally Regulated by the E3 Ubiquitin Ligase MARCH6
Authors: Scott Nicola A. , Sharpe Laura J. , Capell-Hattam Isabelle M. , Gullo Samuel J. , Luu Winnie , Brown Andrew J. 
Published: April 2020

Cholesterol synthesis is a complex process with over twenty enzymes involved. Each enzyme within the pathway undergoes distinct regulation to maintain cholesterol homeostasis. Typically, enzymes within the pathway are degraded in response to a high sterol status, but some are highly stable. In this study, we demonstrate that an enzyme at the branch point in the pathway, lanosterol 14α‐demethylase (LDM), undergoes unique regulation. We show that the preceding enzyme, lanosterol synthase (LSS) is stable, whilst LDM is rapidly degraded. Surprisingly, this degradation is not triggered by sterols but by nitric oxide. Here we identify that the important E3 ubiquitin ligase, MARCH6, is responsible for facilitating the ubiquitination and proteasomal degradation of LDM and also the terminal cholesterol synthesis enzyme, 24‐dehydrocholesterol reductase (DHCR24). Our work highlights MARCH6 as the first example of an E3 ubiquitin ligase which targets multiple steps in a single biochemical pathway and indicates new facets in the control of cholesterol synthesis.Support or Funding InformationThis work was supported by the Australian Research Council Grant DP170101178. The PhD students were funded by the Research Training Program from the Australian Government Department of Education.