bluuebunny/crossref_metadata_2025_split huggingface dataset:
Fic‐mediated deAMPylation is not dependent on homo‐dimerization and rescues toxic AMPylation in flies
Authors: Casey Amanda K. , Moehlman Andrew T. , Zhang Junmei K. , Servage Kelly A. , Krämer Helmut K. , Orth Kim 
Published: June 2021

Protein chaperones play a critical role in proteostasis. The activity of the major endoplasmic reticulum chaperone BiP (GRP78) is regulated by Fic‐mediated AMPylation during resting states. By contrast, during times of stress, BiP is deAMPylated. Here, we show that excessive AMPylation by a constitutively‐active FicE247G mutant is lethal in Drosophila. This lethality is cell autonomous as directed expression of the mutant FicE247G to the fly eye does not kill the fly but rather results in a rough and reduced eye. Lethality and eye phenotypes are rescued by the deAMPylation activity of wild‐type Fic. Consistent with Fic acting as a deAMPylation enzyme, its activity was both time and concentration dependent. Furthermore, Fic deAMPylation activity was sufficient to suppress the AMPylation activity mediated by the constitutively‐active FicE247G mutant in Drosophila S2 lysates. Further, we show that the dual enzymatic activity of Fic is, in part, regulated by Fic dimerization as loss of this dimerization increases AMPylation and reduces deAMPylation of BiP. We believe this mechanism leads to a molecular rheostat in the cell for induction of the UPR.Support or Funding InformationThis work was funded by National Institutes of Health (NIH) grant R01 GM115188, the Welch Foundation grant I‐1561, and Once Upon a Time…Foundation to K.O., NIH grants (EY010199 and EY021922) to H.K. and NSF Graduate Research Fellowship (1000176311) to A.T.M. K.O. is a Burroughs Welcome Investigator in Pathogenesis of Infectious Disease, a Beckman Young Investigator, and a W. W. Caruth, Jr., Biomedical Scholar and has an Earl A. Forsythe Chair in Biomedical Science.This abstract is from the Experimental Biology 2018 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
The loop diuretics bumetanide and ethacrynic acid inhibit voltage‐gated calcium and sodium channels in cultured cortical neurons
Authors: Katnik Christopher , Cuevas Javier 
Published: June 2021

One of the consequences of ischemic stroke is disruption of intracellular ionic homeostasis. Intracellular overload of both Na+ and Ca2+ have been linked to neuronal death in this pathophysiological state. The etiology of ionic imbalances resulting from stroke‐induced ischemia and acidosis includes the dysregulation of a number of plasma membrane transport proteins, such as increased activity of the sodium‐potassium‐chloride cotransporter 1 (NKCC1). Experiments using the NKCC1 inhibitors, bumetanide (BMN) and ethacrynic acid (EA), were carried out to determine if inhibition of this cotransporter affects the Na+ and Ca2+ overload observed following in vitro ischemia‐acidosis. Fluorometric Ca2+ and Na+ measurements were performed on cultured cortical neurons from embryonic (E18) rats. Both BMN and EA depressed ischemia‐acidosis induced [Ca2+]i overload, but did not appreciably alter [Na+]i increases observed under these conditions. Patch‐clamp electrophysiology was used to measure whole‐cell membrane currents to identify target(s) of BMN and EA, other than the electroneutral NKCC‐1, responsible for the effects of these drugs on [Ca2+]i. Voltage‐gated Ca2+ channels (VGCC) were inhibited by both BMN and EA with half‐maximal inhibitory concentration (IC50) values of 4 μM and 36 μM respectively. However, maximal concentrations of BMN failed to block ~40% of VGCC mediated currents, while EA completely abolished these responses. Similarly, voltage‐gated Na+ channels (VGSC) were blocked by BMN and EA with IC50 values of 13 μM and 30 μM, respectively. In contrast to the VGCC responses, both loop diuretics inhibited VGSC currents by ~80%. Experiments using the VGSC inhibitor, tetrodotoxin, showed that BMN was preferentially inhibiting TTX‐sensitive currents in these cells. Neither BMN nor EA affected currents mediated by acid‐sensing ion channels or ionotropic glutamatergic receptors. Taken together, our data suggest that these two loop diuretics are likely to decrease injury following ischemic stroke by effectively mitigating [Ca2+]i overload in neurons via the inhibition of both voltage‐gated Ca2+ and Na+ channels. Moreover, the depression of ischemia‐acidosis evoked [Ca2+]i overload by these compounds does not appear to be dependent on inhibition of NKCC1. Given that the inhibitory concentrations observed here for VGCC and VGSC are similar to those reported for block of NKCC1 and NKCC2 by these compounds, loop diuretics are likely to be inhibiting these channels at therapeutic doses used in humans. Therefore, blockade of these channels by loop diuretics may also contribute to other clinical effects of the compounds observed in animal studies and in humans.Support or Funding InformationAmerican Heart Association Grant‐In‐Aid, 17GRNT33661273 (JC)This abstract is from the Experimental Biology 2018 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
Inhibition of soluble epoxide hydrolase attenuates the kidney injury caused by ischemia/reperfusion in a murine model of acute kidney injury involved in GSK‐3β phosphorylation
Authors: Liu Jun‐Yan , Deng Bing‐Qing , Luo Ying , Kang Xin , Li Chang‐Bin , Lee Kin Sing Stephen , Hammock Bruce D. 
Published: June 2021

Acute kidney injury (AKI) causes severe morbidity and mortality for which new therapeutic strategies are needed. Inhibition of soluble epoxide hydrolase (sEH) has been reported previously to attenuate the kidney injury in cisplatin‐caused AKI. However, the effect of sEH inhibition on ischemia/reperfusion (I/R)‐caused‐AKI remains unknown. In addition, the mechanism underlying the benefit of sEH inhibition to AKI has not been well understood. Here we report that epoxyeicosatrienoic acids (EETs) are the major mediators responsible for the beneficial effect of sEH inhibition in I/R‐caused AKI. Inhibition of sEH resulted in a significant increase in plasma level of 14(15)‐EET. Administration of both 14(15)‐EET alone and TPPU (a potent sEH inhibitor) significantly prolonged the survival of AKI mice. Co‐administration of 14(15)‐EET with TPPU enhanced such effect in survival. Administration of 14(15)‐EET with or without TPPU to AKI mice significantly attenuated the kidney injury, which was supported by the histological analysis of renal tissue, plasma levels of creatinine and urea nitrogen, and renal levels of NGAL. Furthermore, 14(15)‐EET significantly reversed the I/R‐caused reduction in glycogen synthase kinase 3β (GSK3β) phosphorylation in murine kidney, dose‐dependently inhibited the hypoxia/reoxygenation (H/R)‐caused apoptosis of murine renal tubular epithelial cells (mRTECs), and reversed the H/R‐caused reduction in GSK3β phosphorylation in mRTECs. Therefore, inhibition of sEH attenuates kidney injury via, at least partially, 14(15)‐EET modulating the apoptosis of RTECs and GSK3β phosphorylation in mRTECs. This study provides AKI patients with promising therapeutic strategies.Support or Funding InformationThis study was supported in part by NSFC grants 81470588 and 81100090, as well as NIEHS Grant (R01 ES02710), NIEHS Superfund Grant (P42 ES04699), NIH/NHLBI grant (R01 HL59699‐06A1), and a Translational Technology Grant from the UC Davis Medical Center. K.S.S.L. is supported by NIEHS Grant R00 ES024806.This abstract is from the Experimental Biology 2018 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
MUSCADINE GRAPE SKIN EXTRACT INHIBITS ANDROGEN‐INDEPENDENT PROSTATE CANCER CELL GROWTH, INDUCING CELL‐CYCLE ARREST, AND DECREASING MIGRATION BY TARGETING HEAT SHOCK PROTEIN 40
Authors: Brown Collis 
Published: June 2021

Previously we demonstrated that muscadine grape skin extract (MSKE), a natural product, significantly inhibited prostate cancer cell growth by inducing apoptosis through the targeting of phosphatidylinositol 3‐kinase‐Akt and mitogen‐activated protein kinase survival pathways in androgen‐responsive transformed human prostate cancer epithelial cell lines. However, the preventive effect of MSKE on more aggressive androgen‐independent prostate cancer remains unknown. This study examined the effects of MSKE treatment using complementary PC‐3 prostate cancer cell culture and xenograft models. MSKE significantly inhibited PC‐3 human prostate cancer cell tumor growth in vitro and in vivo. The growth‐inhibitory effect of MSKE appeared to be through the induction of cell‐cycle arrest. This induction was accompanied by a reduction in the protein expression of Hsp40 protein and cell‐cycle regulation proteins, cyclin D1 and NF‐kBp65. In addition, MSKE induced p21 expression independent of wild‐type p53 induced protein expression. Moreover, we demonstrated that MSKE significantly inhibited cell migration in PC‐3 prostate cancer cells which was complimented by siRNA‐HSP40 knock‐down that also inhibited cell migration and growth. Overall, these results demonstrate that MSKE inhibits prostate tumor growth and migration, and induces cell‐cycle arrest by targeting Hsp40 and proteins involved in cell‐cycle regulation and proliferation. This suggests that MSKE may also be explored as a novel therapeutic in castration resistant prostate cancer.This abstract is from the Experimental Biology 2018 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
The α7‐nAChRs/heme oxygenase/carbon monoxide pathway arbitrates nicotine counteraction of the inflammatory and renal vasoconstrictor hyporeactivity in endotoxic rats
Authors: Wedn Abdalla M. , El‐Gowilly Sahar M. , El‐Mas Mahmoud M. 
Published: June 2021

The nicotinic/cholinergic antiinflammatory pathway plays integral roles in physiological and pathological regulation of inflammation. In this study, we tested the hypotheses that (i) nicotine guards against hemodynamic and renal vasoconstrictor dysfunction induced by endotoxemia in rats, and (ii) the α7‐nAChRs/heme oxygnase (HO) pathway constitutes a go‐between for the LPS‐nicotine interaction. Hemodynamic or renovascular functions were assessed 6 hr after i.p. administration of lipopolysaccharide (LPS, 5 mg/kg) with or without nicotine. LPS caused significant falls in systolic blood pressure (SBP) and reductions in cumulative renal vasoconstrictions induced by bolus injections of phenylephrine (PE, 0.002 – 4.42 nmol) or vasopressin (VP, 0.00092 – 0.27 nmol) into isolated perfused kidneys. LPS hypotension was demonstrated in both male and female rats, whereas the attenuation of renal vasoconstrictions was seen in male rats only. The reduced PE or VP responsiveness was reversed in preparations pretreated with nicotine (0.5, 1 and 2 mg/kg) in a dose‐dependent manner. This favorable action of nicotine disappeared after blockade of α7‐nAChRs (methyllycaconitine citrate, 2 mg/kg) or inhibition of HO‐1 (zinc protoporphyrin, 10 mg/kg). Moreover, the LPS attenuation of renal vasoconstrictions was improved upon concurrent treatment with pentoxfylline (TNFα inhibitor, 3 mg/kg), hemin (HO‐1 inducer, 10 mg/kg), tricarbonyldichlororuthenium (II) dimer (carbon monoxide‐releasing molecule, 10 mg/kg), in contrast to no effect for bilirubin (5 mg/kg). These data establish the first evidence that implicate the α7‐nAChRs/HO‐1/CO in the nicotine counteraction of renal vasoconstrictor hyporeactivity in endotoxemia.Support or Funding InformationSupported by the Science and Technology Development Fund, Egypt (STDF Grant No. 14895)This abstract is from the Experimental Biology 2018 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
The Aortic Function of Female UC Davis Type 2 Diabetes Mellitus (UCD‐T2DM) Rats
Authors: Akther Farjana , Razan Md Rahatullah , Tang Tia , Graham James L. , Stanhope Kimber L. , Havel Peter J. , Rahimian Roshanak 
Published: June 2021

UC Davis Type 2 Diabetes Mellitus (UCD‐T2DM) rat is a novel validated model of type 2 diabetes mellitus. UCD‐T2DM is characterized by polygenic, adult‐onset obesity and spontaneous β‐cell failure and, as a result, more closely models the pathophysiology of type 2 diabetes in humans than other rodent models. The objectives of this study were to determine the aortic function of female UCD‐T2DM rats and to investigate the underlying mechanism(s) of altered vascular function (if any) in this model. Endothelium‐dependent vasodilation (EDV) to acetylcholine (ACh, 10−8 to 10−5M) was measured in intact aortic rings pre‐contracted with phenylephrine (PE, 2μM). Endothelium‐independent vasodilation to sodium nitroprusside (SNP, 10−9 to 10−5 M) was assessed in endothelium‐denuded rings pre‐contracted with PE (2μM). Furthermore, constrictor response curves to PE (10−8 to 10−5 M) were generated before and after incubation with L‐NAME (200μM), a nitric oxide synthase (NOS) inhibitor. Expression of molecules associated with vascular reactivity and insulin signaling were also evaluated using Western blotting in aortic tissues.We demonstrated that the maximal relaxation to ACh was significantly reduced in aortic rings from female UCD‐T2DM rats compared to controls. The smooth muscle sensitivity to NO as measured by SNP‐induced relaxation was also reduced in this group. The responsiveness to PE was significantly enhanced in aorta of UCD‐T2DM group. Accordingly, the basal nitric oxide (NO), as indicated by the potentiation of the response to PE after L‐NAME, was reduced in diabetes group. Moreover, expression of eNOS, peNOS, pAkt, IRS1 and IRS2 were significantly reduced in the aorta of this disease model.These data, for the first time, show that the vascular function is altered in aortic rings of female UCD‐T2DM rats. Specifically, our data suggests that the decreased sensitivity of vascular smooth muscle to NO along with the enhanced contractile responsiveness to PE may in part contribute to the attenuated ACh responses in aorta of UCD‐T2DM female rats.Support or Funding InformationSupported by NIH/NIHLBIThis abstract is from the Experimental Biology 2018 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
Comparison of anesthetic effects of isoflurane used alone or combined with xylazine on induced cerebral ischemia
Authors: Kim Joo‐Heon , Won Jinyoung , Hong Yonggeun 
Published: June 2021

Intraluminal suture method of middle cerebral artery occlusion (MCAo) has been extensively used in ischemic stroke study. Although the MCAo model has several advantages in comparing different stroke model, it has time‐consuming demands and requires high level of surgical skill. Low mortality and reproducible infarct size in MCAo model are affected by surgical technique, environmental variation and prolonged duration of surgery. The purpose of this study was to determine the effect of anesthetics during prolonged experimental surgery on reproducible stroke model. Animals were subjected to MCAo using intraluminal suture method and provide with isoflurane anesthesia or combined xylazine sedation (20 mg/kg). Animals were randomly divided into three groups: no anesthesia groups (control), inhaled anesthetics groups (isoflurane), inhaled anesthetics combined sedative group (isoflurane + xylazine). Inhaled anesthetics group was provided isoflurane during MCAo surgery (20 min), post‐occlusion period (60 min) and reperfusion (5–10 min), thus total duration of inhaled anesthesia was about 90 min. Inhaled anesthetics combined sedative groups was provided isoflurane during MCAo surgery (20 min) and reperfusion (5–10) min. Xylazine (20 mg/kg) was administered 5 min prior to MCAo and 30 min after MCAo by intramuscular injection. That total duration of inhaled anesthesia was about 30 min. Inhaled anesthetics group that anesthetized during all the post‐occlusion period demonstrated a poor prognosis and high mortality rate compared with control and inhaled anesthetics combined xylazine group. Although the stable respiration signal was shown without abnormal thorax movement, brain infarct size and neurologic deficits revealed no difference between groups. Our results indicated that a modified method of inhalation anesthesia combined xylazine sedation could improve the reproducibility of stroke model and reduce the risk of mortality.Support or Funding InformationFunding: NRF‐2013R1A2A2A01067169, NRF‐2017R1A2A2A01067169, KRIBB‐KGM4611714This abstract is from the Experimental Biology 2018 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
Inducible Adipose Tissue VEGF‐D Drives Lymphatic Expansion and Improves Systemic Insulin Sensitivity in Obesity
Authors: Chakraborty Adri , Lammoglia Gabriela M. , Rutkowski Joseph M. 
Published: June 2021

The metabolic syndrome etiology finds its roots in inflammatory adipose tissue expansion. Rapid adipose expansion, adipocyte insulin resistance, and tissue hypoxia result in dysregulated lipolysis and immune cell infiltration. Increased circulating cytokines and elevated lipid levels lead to ectopic lipid deposition and systemic insulin resistance. Lymphatic vessels regulate inflammation by clearing fluid, cytokines, antigens, and immune cells from the site of inflammation. We hypothesized that increased lymphatic vessel density in adipose tissue may ameliorate local inflammation and thus protect against the metabolic system in obesity. We generated transgenic mice capable of inducible, tissue‐specific expression of murine VEGF‐D under a tightly‐controlled tetracycline response element (TRE) promoter to stimulate lymphangiogenesis. Crossed with adipocyte‐specific adiponectin‐rtTA mice (Adipo‐VD), VEGF‐D overexpression by adipocytes induced de novo lymphangiogenesis in murine white and brown adipose tissues. Adipo‐VD mice gained equivalent weight on high fat diet feeding to their littermates, but demonstrated improved insulin sensitivity and elevated adipose metabolism. b‐adrenergic stimulation of Adipo‐VD mice white adipose lymphatic network promoted enhanced lipolysis and postulated a new route for increased glycerol flux into the circulation: lymphatic vessels. Beyond the increased lymphatic vasculature, adipose tissue architecture and adipocyte size were the same. While chemokine levels in the tissues were unchanged, we quantified fewer macrophage crown‐like structures and reduced macrophages and Tcells in the tissue. Quantitation of infused CD45.1+ leukocytes population in the CD45.2+ Adipo‐VD subcutaneous adipose tissue also indicated increased trafficking to the inguinal lymph node. Increased adipose tissue lymphatic density thus increases insulin sensitivity and enhances immune cell flux from expanded adipose tissue making lymphatics a novel target in the metabolic syndrome.Support or Funding InformationLipedema FoundationThis abstract is from the Experimental Biology 2018 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
The prevention of cardiometabolic risk factors is associated with the improvement in aerobic capacity
Authors: Vecchiatto Bruno , Martucci Luiz Felipe , Américo Anna Laura Viacava , Muller Cynthia Rodrigues , Evangelista Fabiana Sant'anna 
Published: June 2021

Physical inactivity reduces aerobic capacity and is directly associated with an increase in cardiometabolic risk factors. Here we investigated whether there is an association between increased aerobic capacity and the prevention of obesity and insulin resistance through aerobic exercise training (AET). Adult male C57BL6/J mice were assigned into chow‐fed controls (C, n=10), cafeteria diet (CAF, n=8), chow‐fed trained (T, n=9), and cafeteria diet plus trained (CAFT, n=8) groups. AET was performed simultaneously with diet and consisted of 8‐wk running session of 60 min at 60% of maximal speed, 5 days/wk. Experimental procedures were approved by Ethics Committee from School of Arts, Science and Humanities, University of Sao Paulo, Brazil (#001/2016). Both trained groups showed lower body weight at the end of protocol compared to C and CAF groups. However, only CAFT group had reduced body weight gain compared to the CAF group. Cafeteria diet induced glucose intolerance and insulin resistance in the CAF group, which was counteracted by AET in CAFT group. Indirect calorimetry measurement at rest revealed no difference in the variables oxygen uptake (VO2), carbon dioxide expiration (VCO2), respiratory exchange ratio (RER), energy expenditure (EE), carbohydrate and fat oxidation among groups. During maximal exercise tests, T and CAFT groups showed an increase in VO2máx (13% and 9.2%, p=0.07) and VO2peak (11.3% and 8.6%, p=0.07) compared to C and CAF, respectively. However no differences among groups were found for RER and rate of substrate oxidation. AET increased the time to exhaustion and maximal velocity in the T compared to C and CAF groups, and in the CAFT compared to CAF. The exercise intensity corresponding to the VO2máx (iVO2) was higher in the T group (2.30 ± 0.42 km⁄h) compared to C (1.86 ± 0.41 km/h) and CAF (1.73 ± 0.17 km/h) groups, and in the CAFT (2.23 ± 0.11 km/h) compared to CAF. Both trained groups showed late anaerobic threshold, improved the relative cost of running, and higher activity of citrate synthase and β‐HAD enzymes in the soleus muscle compared to C and CAF groups. In conclusion, these results revealed that the prevention of obesity and insulin resistance is associated with the improvement in aerobic capacity induced by AET.Support or Funding InformationFinancial Support: FAPESP # 2015/04948‐4; 2016/20659‐5.This abstract is from the Experimental Biology 2018 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
Relationship between Injury and Spirituality in Pilgrims on the Camino de Santiago
Authors: Rennyson Katelyn Elizabeth , Harris M. Brennan 
Published: June 2021

The purpose of this research was to determine whether there is a relationship between spirituality and the likelihood of injury and the impact of that injury on pilgrims walking along the Camino de Santiago; an ancient, 791 km pilgrimage route along the north of Spain from the French Pyrenees to Santiago de Compostela in Galicia, Spain.400 participants aged 18–80 were recruited along the Camino and through social media prior to and during the pilgrimage while on their journey towards Santiago. Participants completed a ten‐minute survey with questions pertaining to general demographics, injury and illness experiences on the walk, physical well‐being, and fitness. Spirituality was assessed using the FACIT‐Sp questionnaire. Data was analyzed using Excel and Qualtrics software. Research was approved by the Protection of Human Subjects Committee at William & Mary and complies with the principles and protections endorsed by Experimental Biology regarding the use of human subjects and informed consent.Results show that participants who scored a higher spirituality score were less likely to report that any injuries they experienced on the pilgrimage impacted their experience along the trail. The data also demonstrated that participants who were more spiritual were less likely to experience an injury while walking the pilgrimage.These data suggest that pilgrims who walk the Camino de Santiago for spiritual or religious reasons may experience less injuries overall and will feel less impacted by those injuries.This abstract is from the Experimental Biology 2018 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.