bluuebunny/crossref_metadata_2025_split huggingface dataset:
A Chinese herbal health formula, “Gan‐Lu‐Yin”, attenuates balloon injury‐induced neointima formation and suppresses vascular smooth muscle cell migration via inhibition of matrix metalloproteinase‐2/9
Authors: Huang Shih‐Huan , Chien Yi‐Chung , Sheu Ming‐Jyh 
Published: June 2021

BACKGROUNDVascular smooth muscle cell (VSMC) migration contributes to neointimal hyperplasia and restenosis after vascular injury. Gan‐Lu‐Yin (GLY), which is a traditional Chinese herbal medicine frequently prescribed to those patients with cancer, allergy, inflammation and also used as an immunoregulatory agent. To determine whether GLY possesses antimigratory properties, we stimulated rat VSMCs with 10% fetal bovine serum, and determined their molecular mechanisms related with migration properties. Also, animal experiments were conducted to determine if GLY could attenuate the neointima formation after balloon injury.METHODSVSMC were treated with different concentrations of GLY, and then analyzed with Flow cytometric analysis, zymography, transwell, and western blotting. SD rats subjected to balloon injury were stained with H&E.RESULTSBased on the results, we found that thickness of neointima was significantly attenuated by GLY. Non‐cytotoxic doses of GLY inhibited VSMCs migration was through its negative regulatory effects on phosphorylated ERK1/2, PI3K/AKT, and FAK. The data showed GLY has sufficient effects to inhibit the migration of VSMCs cells. In addition, GLY might block injury‐induced vascular neointima formation via the inhibition of VSMCs migration, without apoptosis.CONCLUSIONSThis study provides a better understanding of how GLY affects VSMC migration and its role in balloon injury‐induced neointima formation.
Characterization of a novel, high‐affinity and selective fluorescent antagonist for the 5HT1A receptor
Authors: Fatokun Amos Akintayo , Middleton Richard J. , Thompson Kevin S.J. , Briddon Stephen J. , Hill Stephen J. 
Published: June 2021

The use of fluorescent ligands in G‐protein‐coupled receptor pharmacology to study ligand‐receptor interactions is now a viable alternative to the use of traditional radioligands, as it is non‐invasive, allows the study of interactions in real‐time and in single cells and is environmentally friendly. We have characterized a new, fluorescent antagonist acting selectively at the 5HT1A receptor. Using Chinese Hamster Ovary (CHO) cells stably expressing the 5‐HT1A receptor and a cyclic AMP response element‐coupled reporter gene (human secreted placental alkaline phosphatase (SPAP)), we calculated the log KB value of the ligand from the SPAP assay as −8.49 ± 0.14 (n=3). The ligand showed no significant affinity at closely‐related 5‐HT receptors. Confocal imaging on the ImageXpress Ultra confocal plate reader revealed clear membrane binding at concentrations consistent with the affinity value from the functional assay and a concentration‐dependent displacement of this binding by an unlabeled competitor, NAN‐190. These data show that this molecule is a high‐affinity, selective fluorescent antagonist at the 5HT1A receptor and thus can be used to probe the pharmacology and kinetics of ligand‐receptor interactions at this receptor.This research was supported by the award to AAF of a Nottingham Advanced Research Fellowship and a Marie Curie International Incoming Fellowship.
Role of voltage‐dependent calcium channels in stretch‐induced lymphatic vessel contractions
Authors: Lee Stewart S , von Der Weid Pierre‐Yves 
Published: June 2021

Lymph drainage maintains tissue fluid homeostasis and facilitates immune response. It is promoted by phasic contractions of lymphatic vessels, which increase in rate with increase in luminal pressure. The contractions depend on activation of voltage‐dependent calcium channel (VDCC), which have not been characterized in lymphatic vessels. We used pressure‐ and wire‐myography, electrophysiology, PCR and immunofluorescence imaging to investigate the electrophysiological properties of L‐type and T‐type VDCCs and their role on stretch‐induced lymphatic contractions.Members of the VDCC family were expressed at the messenger RNA and protein level in rat mesenteric lymphatic vessels. The stretch‐induced increase in force was significantly attenuated in the presence of L‐type VDCC blockers nifedipine and diltiazem, while the T‐type VDCC blockers mibefradil and nickel significantly decreased the stretch‐induced increase in contraction frequency. Furthermore, nifedipine and diltiazem depolarized, while mibefradil and nickel hyperpolarized lymphatic muscle membrane potential.Our data suggest a delineating role of VDCCs in stretch‐induced lymphatic vessel contractions. We propose that activation of T‐type VDCC depolarizes membrane potential, regulating the frequency of lymphatic contractions via opening of L‐type VDCCs, which drive the strength of contractions.Supported by NIH
Homocysteine disrupts actin organization and vascular outgrowth by an iNOS‐dependent mechanism during angiogenesis
Authors: Mayo Jamie Nadine , Chen Cheng‐Hung , Bearden Shawn E. 
Published: June 2021

ObjectiveHyperhomocysteinemia (HHcy) impairs angiogenesis. The goal of this study was to test the hypothesis that HHcy impairs angiogenic growth through an iNOS dependent mechanism.MethodsWe used an ex‐vivo adult mouse choroid tissue model of VEGF‐stimulated angiogenesis. Mouse microvascular endothelial cells were studied in culture in parallel experiments.Results20μM homocysteine (Hcy) treatments significantly decreased the area of angiogenic outgrowth (8.9mm2 ± 0.31 vs 4.1mm2 ± 0.58; p<0.05). Hcy 1) canted the orientation of actin filaments (orientation in relation to explant center: 38.6ܰ±13.0ܰ vs 95.4ܰ± 6.5ܰ; p<0.05), 2) decreased the occurrence of cellular extensions beyond the leading edge of outgrowth (7 ± 0.89 vs 2.2 ±0.52; p<0.05), and 3) reduced the motility of dispersed cells (19.8μm/hr ±1.4 vs 13.1μm/hr ±1.7; p<0.05). Inhibition of iNOS with 1400W (10μM) rescued the effect of Hcy on area of outgrowth, occurrence of cellular extensions, and cell motility.ConclusionsHcy decreases angiogenic outgrowth by inhibiting motility and disrupting actin filament organization in part via an iNOS‐dependent mechanism. This work was supported by NIH grant #HL106548(SEB).
Dematin binds to F‐actin filaments in activated platelets.
Authors: Azim Anser C , Hill Angela 
Published: June 2021

BackgroundDematin is expressed in hematopoietic cells including human platelets. A spectrin based cytoskeleton uniformly supports the resting membrane platelets. Dematin is expressed as a 52 kDa and 49 kDa protein in human platelets but its role in the platelet activation, binding to polymerized actin in resting and activated platelets is poorly understood. We hypothesize the role of dematin in stabilizing human platelet cytoskeleton.MethodWe isolated human platelets from anti‐coagulated blood as described earlier (Azim AC et al Blood. 2000, 95(3):959–64.). Briefly, platelet‐rich plasma was obtained by centrifuging the blood at 200 g for 15 min. Platelets were separated from the plasma by gel filtration over a Sepharose‐2B column pre‐equilibrated with platelet buffer (145 mM NaCl, 10 mM Hepes, 10 mM glucose, 0.3% BSA, 0.5 mM NaH2PO4, and 5 mM KCl, pH 7.4). Platelets were used after leaving them at 37°C for 30 min after treatment with Thrombin or simply with glass activation.ResultsWe show by immune‐gold microscopy and western blot analysis the distribution of dematin in resting and activated platelets.DiscussionOur studies confirm that platelets express dematin and show that dematin is a structural protein of the resting platelet cytoskeleton and is bound to F‐actin filaments. This study also demonstrates distribution of actin and dematin after PAR‐1 receptor ligation of platelets by thrombin by immunochemical and electron microscopic techniques.
The triggers of lipopolysaccharide‐induced hypothermia and hypotension lie outside the brain
Authors: Steiner Alexandre A , Lewis Kevin , Al‐Saffar Hiba , Liu Elaine , Schober Alexandra , Corrigan Joshua J , Shibata Keita 
Published: June 2021

Little is known about the neuroimmune mechanisms responsible for the switch from fever to hypothermia observed in severe forms of systemic inflammation. We evaluated whether bacterial lipopolysaccharide (LPS) acting directly on the brain could promote a fever‐hypothermia switch as well as the hypotension that is often associated with hypothermia in models of systemic inflammation. At an ambient temperature of 22ºC, freely moving rats received intracerebroventricular (i.c.v.) injections of LPS at doses ranging from 0.5 to 25 μg. Despite the use of such high doses, the prevailing thermal response was fever. Nevertheless, an analysis of individual responses revealed that two out of eighteen rats displayed some decrease in body temperature. To investigate if a hypothermic response could be hidden within the prevailing febrile response, rats were pretreated with a cycloxygenase‐2 inhibitor (SC‐236, 3.5 mg/kg i.v.) known to block fever. This strategy also failed to reveal any consistent hypothermic response following i.c.v. LPS. At the doses tested, i.c.v. LPS was similarly ineffective at inducing hypotension. Additional doses of LPS did not need to be tested because the 25‐μg dose was already sufficient to induce both hypothermia and hypotension when administered peripherally (i.a.). These findings led to the conclusion that, unlike fever and other sickness symptoms, hypothermia and hypotension are triggered exclusively by LPS acting outside the brain.Support:AHA, FAPESP
Effects of acute stress on central and peripheral immune or inflammatory mediators
Authors: Hamdan Jameel N , Shelton Cecilia A , Barron Kristina I , Gosselink Kristin L 
Published: June 2021

Previous studies have shown that stress can induce inflammatory responses in the body, and contribute to deficits in learning and memory. Whether or not this is a causal relationship, however, is not well understood. This study investigated immune and inflammatory changes induced by stress; we hypothesized that cytokine secretion and white blood cell (WBC) profiles would be altered by stress exposure. Adult male rats were restrained for 30 min, and brain tissue and blood harvested 2h after the end of the stress. Flow cytometry was used to count T‐ and B‐cells from the WBC fraction of the blood. Blocks of brain tissue containing hippocampal and hypothalamic regions were dissected, homogenized in PBS, and a Milliplex MAP kit used to measure multiple cytokines in these samples and in the plasma. Acute stress did not significantly alter cytokine levels in any of our tissues. A significant reduction was seen, however, in the number of B‐cells in acutely stressed rats compared to controls (p<0.05). In conclusion, our data support our hypothesis in part, as changes in immune or inflammatory factors were seen after acute stress. Future studies will confirm and extend these data, determine the role of B‐cells during stress, and examine these responses in repeatedly stressed animals.Supported by: 8G12MD007592
SYMPATHOVAGAL IMBALANCE IN TAKOTSUBO CARDIOMYOPATHY
Authors: Norcliffe‐Kaufmann Lucy J , Kaufmann Horacio , Martinez Jose , Reynolds Harmony 
Published: June 2021

Takotsubo cardiomyopathy is an acute reversible cardiac dysfunction syndrome associated with high circulating catecholamine levels. Our objective was to investigate whether abnormal cardiovascular control might play a role in the pathophysiology.We studied autonomic cardiovascular reflexes in 10 women who had takotsubo (33±7 months after being hospitalized) and 10 age/BMI matched healthy women.In the women with takotsubo, indices of vagal modulation of heart rate induced by respiration were uniformly reduced (expiratory:inspiratory ratio: p<0.01, pnn50%: p<0.02, rMSSD: p<0.03). Cognitive (stroop test: p<0.03) and emotional arousal (event recall: p<0.05) produced exaggerated pressor responses, without detectable ECG changes. Pressor responses to hemodynamic stimuli were also amplified (Valsalva SBP overshoot: p<0.05). Takotsubo women had increased BP variability in the short‐term (St. Dev. SBP: p<0.01). Ambulatory recordings captured an exaggerated morning surge in SBP after awakening from sleep (p<0.05). Cardiovagal baroreflex gain was significantly lower in the takotsubo women (sequence analysis: p<0.01, regression method: p<0.001, transfer function gain: p<0.001).Women with takotsubo have heightened sympathetic responsiveness, labile BP and reduced vagal modulation of the heart. This shift in sympathovagal balance could play a role in the pathophysiology.
COX‐2 induction is ROS dependent in rats during 3 days unilateral ureteral obstruction
Authors: Østergaard Martin , Nilsson Line , Frøkiær Jørgen , Nørregaard Rikke 
Published: June 2021

Oxidative stress may contribute to cellular dysfunction of the kidney with unilateral ureteral obstruction (UUO). Previously we have shown induction of cyclooxygenase type 2 (COX‐2) in renal medullary interstitial cells (RMIC) during UUO. This study investigated the role of oxidative stress or ROS in the induction of COX‐2 during UUO.Rats were subjected to 3dUUO and treated with inhibitors of the ROS production (NADPH‐oxidase inhibitor diphenylene iodonium (DPI) and the complex‐1 inhibitor rotenone (ROT)). Clearance experiments were performed. The kidneys were removed and prepared for immunoblotting or immunohistochemistry (IHC) analyses.Bodyweight (BW) was unchanged in 3dUUO rats subjected to DPI whereas ROT decreases BW. Hematoxylin and eosin (HE) staining of the kidney showed lower degree of dilatation due to DPI or ROT treatment in 3dUUO rats. COX‐2 and the oxidative stress marker heme‐oxygenase type 1 (HO‐1) was both increased in inner medulla in response to 3dUUO. This was attenuated by DPI or ROT administration. IHC confirmed these findings. COX‐1 was not changed in 3dUUO rats subjected to DPI or ROT. Plasma urea was not affected by DPI or ROT treatment in response to 3dUUO. HE staining of heart, liver, and brain did not reveal any obvious changes due to administration of DPI or ROT.These results suggest that oxidative stress contribute to the regulation of COX‐2 in inner medulla in rats subjected to 3 days UUO.
Sulfur Dioxide Derivative Prevents Left Ventricular Hypertrophy and Electrophysiological Alterations
Authors: YARAÞ Nazmi , Özdemir Semir , Dalaman Uđur , Öztürk Nihal , Olgar Yusuf 
Published: June 2021

SO2and derivatives generally accepted as toxic gasseous molecules and inhalation of itdrives biological systems severe conditions. On the other hand, they are used as safepreservative in foods. Recently, SO2was discovered to be produced in thecardiovascular system and to influence important biological processes.It was reported by Liang Y.et al that endogenous SO2progressively inhibit hypertrophycaused by isoproterenol application to rats via increasing antioxidation capacity. In this study we aimed to investigate underlying molecular and electro physiological mechanisms of this inhibition of ventricular hypertrophy. Wistar rats were administrated 20 mg/kg isoproterenolfor seven days subcutaneuosly to drive hypertropic cardiomyopathy and some animals was taken SO2 donor sodium metabisulfite together with isoproterenol through i.p. at a dose of 85 mg/kg. At the end of the experiment, animals were sacrified by heart exicition and followed by enzimatic dissociation of single left ventricular myocytes. Iso admistrated animal heart (6.69±0.41 mg/kg bodyweight) were hypertrofied compared to control animals hearts(3.94±0.12 mg/kg). On the other hand, metabisulfite inhibits hypertrothy of the rats hearts (4.27±0.32 mg/kg).Single cardiomyocytes were examined electrophysiologicaly by voltage clamp in order to examine effect of SO2on potassium currents (IK1, Ito and Iss), action potential (AP) and L‐type calcium current (ICaL). While metabisulfite had no effect on potassium currents, prolonged AP and reduced ICaL ameliorate to the control level. In conclusion, SO2derivative sodium metabisulfite prevent cardiac hypertrophy generated by adrenergicstimulation both electrophysiologicaly and histologicaly. This work supported by Akdeniz University Scientific Research Unit.
Blood‐brain barrier leak precedes brain histopathology and cognitive impairment in mild hyperhomocysteinemia
Authors: Rhodehouse Bryce C , Mayo Jamie N , Beard Richard S , Chen Cheng‐Hung , Bearden Shawn E 
Published: June 2021

Hyperhomocysteinemia (HHcy) is associated with blood‐brain barrier (BBB) permeability and Alzheimer's disease. We explored the relation among BBB permeability, brain histopathology, amyloid‐beta (Abeta) transport and deposition and cognitive function in HHcy. We used C57Bl/6 mice with heterozygous deletion of cystathionine β‐synthase (Het) and wild‐type littermates (WT) [young: 5–9 mos; old: 12–29 mos]. BBB permeability was greater in Het compared to WT at all ages (p<0.05). Old, but not young, Het mice had greater cognitive impairment than any other group in Morris water maze tests (p<0.05). Old Het mice also showed greater inflammation (CD45+ and VCAM+) and leukoaraiosis in the fornix compared to WT (p<0.05) with concomitant evidence of disrupted autophagy in the hippocampus (increased caspase‐cleavage of beclin‐1; p<0.05). There were no differences for these measures among young or WT mice. There was no difference in levels of Abeta transporters in isolated cerebral microvessels (RAGE, LRP‐1, Pgp) or Abeta deposition in the brain with age or by Hcy level.Conclusionin mild HHcy, increased permeability of the BBB precedes the onset of brain histopathology and cognitive dysfunction, which is independent of Abeta deposition. Protecting the BBB from the effects of Hcy early in life may reduce the incidence or severity of cognitive decline later in life. Supported by NIH HL106548 (SEB).
EFFECT OF RESISTANCE TRAINING ON CIRCULATING Th2 CYTOKINES IN HIV+ MEN RECOVERING FROM SUBSTANCE ABUSE
Authors: Duplanty Anthony Adam , Vingren Jakob L , Curtis John H , Lee Elaine C , Budnar Ronald G , McFarlin Brian K , Hill David W 
Published: June 2021

Human immunodeficiency virus (HIV) and substance abuse (drug and/or alcohol) independently impair the immune system. IL‐4 and IL‐10 are cytokines involved in B‐cell mediated adaptive responses relevant to HIV. IL‐4 suppresses the production of Th1 cells. IL‐10 increases with disease progression in HIV infected individuals and down regulates the expression of Th1 cytokines. Thus, changes in Th2 cytokines could have important implications for HIV+ individuals.PurposeTo examine the effect of resistance training on resting concentrations of circulating Th2 cytokines.MethodsSixteen men (42 ± 11 years, 180 ± 9 cm, 89 ± 21 kg) infected with HIV and enrolled in a 60‐day in‐patient substance abuse treatment program. Participants were assigned to either supervised resistance exercise 3 times per week (RT) or no exercise training (Control) for six weeks. Before (Pre) and after (Post) the 6‐week period, resting and fasted blood samples were obtained and analyzed for serum IL‐4 and IL‐10.ResultsIL‐10 did not change following the 6‐week period for RT (Pre: 1.9 ± 0.3 pg·ml−1; Post: 1.8 ± 0.4 pg·ml−1) or Control (Pre: 1.6 ± 0.2 pg·ml−1; Post: 1.5 ± 0.2 pg·ml−1). IL‐4 was undetectable (<0.08 pg·ml−1) in 9 of 16 participants.ConclusionA six‐week resistance training program does not elicit changes in the circulating Th1 cytokines IL‐4 and IL‐10 in HIV+ men undergoing substance abuse treatment.
The Repressor Element‐1 Silencing Transcription Factor (REST) regulates the Hypoxia Inducible Factor (HIF) network through a novel negative feedback loop
Authors: Cavadas Miguel Santos , Taylor Cormac , Cheong Alex 
Published: June 2021

Hypoxia induces a number of metabolic changes with profound consequences for cell physiology and pathophysiology. Our understanding of the complex response to hypoxia has been focused on the Hypoxia‐Inducible Factor (HIF) and its regulation by prolyl‐hydroxylases (PHD). In solid tumours, both intra‐tumoral hypoxia and mutations in tumour suppressor genes can lead to the overexpression of HIF, a feature that has been associated with poor prognosis.The Repressor Element‐1 Silencing Transcription Factor (REST), is best characterized as a transcriptional repressor of neuronal genes in non‐neuronal tissues. More recently, REST was found to be affected by ischemia and to be down regulated in breast and colon cancer. This lead us to investigate if REST was an hypoxic sensitive transcription factor and if it interacts with the HIF pathway.Hypoxia induced an increase in REST translocation to the nucleus. siRNA knockdown of REST increased HIF, PHD2 and PHD3 protein expression. REST was found to be recruited to the HIF promoter, and HIF was recruited to the REST promoter in a hypoxia dependent way, indicating a novel negative feedback loop involving REST and HIF. In conclusion, REST was identified as a novel hypoxia‐sensitive transcriptional repressor that negatively regulates HIF, PHD2/3, acting as a novel negative feedback loop in the hypoxia response.
Klotho Protects Lung Epithelial Cells against Oxidative DNA Damage
Authors: Ravikumar Priya , Smith Blaine , Hsia Connie C.W. , Moe Orson W. 
Published: June 2021

Klotho is an anti‐aging protein through pleiotropic actions. We postulate that Klotho can protect the lung against O2 and Pi toxicity and tested the direct effects of Klotho in vitro using A549 lung epithelial cells exposed to hyperoxia (95–5% O2‐CO2) and increasing Pi (1 vs. 5 mM). Soluble or transmembrane full length Klotho was over‐expressed by transfection or recombinant soluble Klotho was added to the medium. DNA damage was assessed by ELISA assay of 8‐hydroxydeoxyguanosine (8‐OHdG) (OxiSelect™ Cell BioLabs Inc). Triplicate experiments were performed. The amount of 8‐OHdG (ng/ml, mean±SD) under each condition was compared by two‐way ANOVA. P ≤ 0.05 was considered significant.

O2
21%
95%

Klotho
−
+
−
+

Transmembrane Klotho

Pi 1mM
0.093±0.003
0.086±0.009
0.172±0.005*
0.076±0.006 †

Pi 5mM
0.326±0.009
0.300±0.010†
0.563±0.013*
0.323±0.013 †

Soluble Klotho

Pi 1mM
0.093±0.003
0.082±0.002†
0.166±0.005*
0.072±0.001 * †

Pi 5mM
0.322±0.011
0.292±0.005†
0.546±0.012*
0.324±0.011 * †

Conditioned Medium

Pi 1mM
0.089±0.002
0.079±0.002†
0.164±0.004*
0.070±0.004 * †

Pi 5mM
0.317±0.007
0.299±0.005 †
0.531±0.012*
0.303±0.014 * †

95% vs. 21% O2;
Klotho vs. control at the same Pi and O2 concentrations.
All differences between Pi conditions were significant.

Increasing either O2 or Pi concentration increased 8‐OHdG production, indicating DNA damage; the effects are synergistic. In pulmonary epithelial cells, either Klotho expression or addition ameliorates hyperoxia‐induced DNA damage but has only mild effects on Pi‐induced DNA damage. These findings suggest that increasing Klotho concentration in the lung may be beneficial in the prevention of oxidant DNA damage.SupportF32 HL103043, RO1 DK091392, DK092461, UO1 HL111146