bluuebunny/crossref_metadata_2025_split huggingface dataset:
1′‐Acetoxychavicol acetate accelerates human skin wound healing
Authors: Mori Atsuki , Sugawara Koji , Tsuruta Daisuke , Matsui‐Yuasa Isao , Kojima‐Yuasa Akiko 
Published: June 2021

IntroductionChronic wounds are recently increasing in number among diabetic or bedridden person. It is important to find effective treatments for them. 1′‐acetoxychavicol acetate (ACA) is the compound obtained from ginger family such as Alpinia galangal. ACA has been reported to have various physiological effects, anti‐cancer, anti‐obesity, and anti‐oxidant. Here, we focus on the effect of ACA on wound healing.MethodsWe performed scratch assay in the presence of mitomycin‐C to block HaCaT keratinocytes (KCs) cell proliferation. We performed an in vitro scratch assay with MEK/ERK inhibitor (U0126) or PI3K inhibitor (LY294002), to examine how the inhibition of MEK/ERK or PI3K affects HaCaT KCs migration. Furthermore, we performed human wounded skin organ culture with ACA. We measured the length of epithelial tongue.ResultsACA significantly promoted wound closure in the presence of mitomycin‐C. This result showed that ACA stimulates HaCaT KCs migration without modifying proliferation. MEK/ERK or PI3K‐inhibited HaCaT KCs in the presence of ACA showed a reduction of migration as compared to the ACA alone treated cells. These results suggest that ACA could promote wound closure via inducing ERK/MAPK and PI3K activity. Furthermore, ACA stimulated human skin re‐epithelialisation of organ cultured human wounded skin. This result fits with our results of scratch assay using HaCaT KCs, ACA significantly stimulated wound closure in situ. This suggested that ACA had wound healing‐promoting effect in human skin.ConclusionWe found that ACA had wound‐healing promoting effects by stimulating ERK/MAPK and PI3K activation. ACA might be a novel agent for treating chronic wounds.
Sensitive Imaging Techniques Demonstrate LCPUFA‐induced Improvements of Brain Structure in Mildly Obesogenic Mice
Authors: Schoemaker M. , Arnoldussen I. , Noordman H. , Zerbi V. , Wielinga P. , Kleemann R. , Gross G. , van Tol E. , Kooistra T. , Kiliaan A 
Published: June 2021

Obesity is associated with impaired cognition. Could early nutrition with Long‐Chain Polyunsaturated Fatty Acids reduce potential detrimental effects of an obesogenic diet on brain structure in a humanized mouse model for mild obesity?Four‐week old male ApoE*3Leiden mice were fed regular chow with or without arachidonic acid (ARA) and docosahexaenoic acid (DHA). From 14 until 26 wks of age, mice received a mildly obesogenic high‐fat/high‐carbohydrate (HFHC) diet. At 13 and 26 wks of age, mice were examined in an 11.7 Tesla Bruker Biospec magnetic resonance imaging (MRI)‐scanner. GLUT‐1 fluorescent immunohistochemistry was performed.Early ARA&DHA counteracted HFHC‐induced changes in cerebral blood flow in mice at 26 wks of life, indicating improved cerebrovascular function, supported by effects on GLUT‐1 positive blood vessels. Increased mean diffusivity in the motorcortex of HFHC‐fed mice, indicative for gray matter loss, was inhibited by early ARA&DHA (P蠄0.05). Resting state functional MRI analysis showed decreased functional connectivity between ventral hippocampus, motorcortex and somasensory cortex in HFHC‐fed mice vs. control mice. Mice early fed ARA&DHA were protected against these detrimental effects.Early life ARA&DHA protects vascular function, gray matter integrity and functional connectivity between brain regions in the context of mild obesity later in life. Sensitive imaging techniques detect subtle diet effects on brain structure.
Dilute Russell's Viper Venom Clotting Time (DRVVT) is Useful for the Routine Monitoring of Newer Non‐Vitamin K Anticoagulants
Authors: Brayne Mark , Kola Jake , Hoppensteadt Debra , Iqbal Omer , Fareed Jawed 
Published: June 2021

The newer non‐vitamin K anticoagulants such as rivaroxaban (Bayer Healthcare) (R), apixaban (BMS/Pfizer) (A) and dabigatran (Boehringer Ingleheim) (D) have been approved to manage thrombotic and cardiovascular disorders. This study was designed to determine which of the available clotting assays could be used to monitor these new oral anticoagulants in plasma.MaterialsCitrated blood was drawn from ten donors and platelet poor plasma (PPP) was obtained. The PPP was supplemented with A, R and D in a concentration range of 0‐1.0 ug/ml. In addition, plasma samples, containing varying concentrations of either A, R or D were also blindly analyzed. The plasma samples were analyzed using PT/INR (Innovin, Seimens, Deerfield,IL), APTT reagent (Platelin, TCoag, Ireland) dilute Russell's viper venom time (DRVVT) and DRVVT confirm (HemosIL, Instrumentation Laboratories (Bedford, MA).ResultsIn the PT and APTT assays a concentration dependent modest increase in clotting time was observed. In contrast, both the DRVVT and DRVVT confirm assays the concentration‐response curves were straight lines with a steep slope. Therefore, it was possible to extrapolate the concentrations of the unknown samples using both of these assays. In addition , there was a good correlation between the calculated concentration and the actual concentration in the range of 50 to 1000 ng/ml (r2=0.82) with both assays.ConclusionsThese results suggest the dRVVT and DRVVT confirm assays are sensitive to A, D and R at concentrations > 50 ng/ml. Therefore, A,D and R can be monitored by these assays. Both of these assays are commercially available and can be readily adapted to any automated instrument.
Visualization and Objective Quantification of Autonomic Circuitry in Mouse Area Postrema Using a Genetic Method
Authors: Gokozan Hamza , Corcoran Sarah , Czeisler Catherine , Gygli Patrick , Catacutan Fay , Baig Faisal , Otero Jose 
Published: June 2021

The transcription factor PHOX2B is a master regulator of autonomic nervous system function as well as the specification of noradrenergic neurons. Incomplete maturity and/or absence of noradrenergic neuron networks are a proposed mechanism of infant breathing disorders including apnea of prematurity, sudden infant death syndrome, and congenital central hypoventilation syndrome (CCHS).PHOX2B mutations result in the “chemosensation” loss characteristic of CCHS, resulting in poor respiratory drive secondary to hypercarbia (high pCO2). The objective of our work is to identify PHOX2B derived structures in the developing brain and generate new animal models capable of tracing the neuronal connections between neuronal nuclei during embryonic and postnatal development.We utilized a PHOX2B‐CRE X ROSA‐GFP mouse to trace the appearance of PHOX2B‐derived cells in nuclei of the hindbrain. In order to trace the connections between these nuclei other hindbrain nuclei, we utilized a transgenic mouse in which targeted neurons display traceable processes and synapses. This transgenic mouse reagent (Jackson Labs stock number 010590) shows CRE‐mediated expression of tdTomato in cell bodies and GFP expression at synapses. Using these conditional gene targeting approaches we were able to conclude that neurons derived from PHOX2B progenitor domains innervate certain hindbrain structures.By using our transgenic approach, we were able to (1) identify PHOX2B derived progenitors generate both neurons and glia, (2) show PHOX2B derived neuronal synapses in Ventral Medulla and Area Postrema. We also present an unbiased stereology method which can be used to determine the complexity of PHOX2B neuronal network.
Role of SLMAP Genetic Variants in Susceptibility of Diabetes and Diabetic Retinopathy in Qatari Population
Authors: Upadhyay Rohit , Crystal Ronald , Ding Hong 
Published: June 2021

ObjectiveOverexpression of SLMAP gene has been associated with diabetes and endothelial dysfunction of macro‐ and micro‐blood vessels. In this study, our primary objective is to explore the role of SLMAP gene polymorphisms and genetic variants of SLMAP in the susceptibility of type 2 diabetes, with or without type 2 diabetes associated retinopathy (DR), in the Qatari population.Research design and methodsA total of 341 Qatari subjects (non‐diabetic controls and type‐2 diabetic patients with or without diabetic retinopathy) were genotyped for SLMAP gene polymorphisms (rs17058639 C>T; rs1043045 C>T and rs1057719 A>G) usingTaqman SNP genotyping assay.ResultsSLMAP rs17058639 C>T polymorphism was associated with the presence of diabetic retinopathy among Qataris with type 2 diabetes. One‐way ANOVA and multiple logistic regression analysis showed SLMAP SNP rs17058639 C>T as an independent risk factor for DR development. SLMAP rs17058639 C>T polymorphism also had predictive role for severity of DR. Haplotype Crs17058639Trs1043045Ars1057719 was associated with the increased risk for DR among Qataris with type 2 diabetes.ConclusionsThe data suggests the potential role of SLMAP SNPs as a risk factor for the susceptibility of DR among type 2 diabetes patients in Qatari population.
Expression of GDNF Throughout Rat Brain After Intranasal Delivery of hGDNF Plasmid Nanoparticles
Authors: Aly Amirah , Harmon Brendan , Shah Parin , Sesenoglu‐Laird Ozge , Padegimas Linas , Cooper Mark , Waszczak Barbara 
Published: June 2021

Glial cell line‐derived neurotrophic factor (GDNF) gene therapy is a promising approach for CNS disorders such as Parkinson's disease (PD). We are examining intranasal administration of PEG‐CK30 DNA nanoparticles (NPs) encoding hGDNF, developed by Copernicus Therapeutics, Inc. We have shown that intranasal NPs incorporating hGDNF DNA (pGDNF), or a hGDNF‐eGFP fusion plasmid (pUGG), transfect brain cells in vivo, and that intranasal pGDNF NPs protect substantia nigra (SN) dopamine neurons in a rat model of PD.Here, we examined transfection in rat brain 7 days after intranasal delivery of these NPs. ELISA showed increased GDNF levels throughout the brain of rats given pGDNF relative to saline controls. Immunohistochemistry (IHC) revealed that the number of eGFP positive cells was significantly higher in rats given intranasal pUGG NPs compared to background fluorescence in controls, with the highest numbers in midbrain. Double label IHC showed that most eGFP positive cells were immediately adjacent to capillary endothelial cells staining for RECA‐1, and they were often contiguous to GFAP positive astrocytic endfeet enwrapping capillaries. When eGFP cells were observed adjacent to dopamine neurons in SN, or NeuN labeled cells in any region, these neurons were also found close to a capillary. This pattern suggests that pericytes are transfected after intranasal administration of DNA NPs, consistent with distribution by perivascular transport. These results confirm delivery and transfection of cells in rat brain after intranasal administration of Copernicus' DNA NPs and support use of intranasal hGDNF NPs as a non‐invasive gene therapy for PD.
Antithrombin Perfluorocarbon Nanoparticle Deposition Prevents Stent Thrombosis
Authors: Palekar Rohun , Vemuri Chandu , Marsh Jon , Arif Batool , Wickline Samuel 
Published: June 2021

BackgroundDespite pharmacologic modification of vascular implants, stent thrombosis remains clinically significant. The purpose of this research was to demonstrate the utility of antithrombin nanoparticles to prevent stent thrombosis.Methods: Perfluorocarbon nanoparticles conjugated to multiple copies of the direct thrombin inhibitor D‐phenylalanyl‐L‐prolyl‐L‐arginyl‐chloromethyl ketone (PPACK‐NP) were tested in static and in flow‐loop in vitro models for their ability to prevent thrombosis on bare metal stents. Thrombin‐adsorbed stents were treated with saline, free PPACK, control NP or PPACK‐NP, followed by exposure to platelet poor plasma (PPP) or platelet rich plasma (PRP) for 60 minutes. In PRP‐exposed groups, subsets were pretreated with either clopidogrel or vorapaxar.Results: Treatment of thrombin‐adsorbed stents with PPACK‐NP abrogated clot formation following PPP (p<0.05) or PRP (p<0.0005) exposure. Scanning electron microscopy of PRP exposed stents treated with PPACK‐NP revealed accumulation of adsorbed nanoparticles (Fig. A, arrow) on the surface with little fibrin or platelet deposition compared to control NP‐treated stents (Fig. B). Furthermore, PPACK‐NP treatment in combination with platelet inhibitors limited clot formation over that of clopidogrel (p<0.05) and vorapaxar (p<0.005) alone. Application of PPACK‐NP in an in vitro flow model confirmed the ability of PPACK‐NP to prevent the formation of an occlusive thrombus on stents compared to controls (Fig. C&D).ConclusionIn summary, our results demonstrate the ability of PPACK‐NP to prevent stent thrombosis and suggest the potential for enhancing stents with nanoparticle coating to improve patency.Research Support: NIH HL112303
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In silico and in vitro evaluation of benzothiazole derivatives with inhibitory activity on 11‐beta‐hydroxysteroid dehydrogenase type 1
Authors: C. Cabrera Perez Laura , Padilla Martinez Itzia , Cruz Alejandro , Correa Basurto Jose , Rosales Hernandez Martha , Loera Pineda Juan 
Published: June 2021

11‐beta‐hydroxysteroid dehydrogenase 1 (11‐βHED1) plays an important role in obesity and metabolic syndrome, due to it regulates cortisol levels. High cortisol levels produce gain of adipose tissue, release of proinflammatory substances, and free radicals (FR)1,2. Then the inhibition of 11‐βHED1 by thiazole derivatives has been reported. However, nowadays there are not a drug to inhibit the enzyme. For instance, inhibitors developed by Amgen/Biovitrum have not yet been approved due to it is not concluded, because, their clinical trials was halted3. For this reason, in present work, eleven benzothiazole derivatives were studied by in silico methods. Studies by blind docking allowed to determine that compound 1 (C1) has better affinity for 11‐βHED1 (PDB:4BB6) than compound 2 (C2), because interacts by hydrogen bond with Y177 in 11‐βHED1 dimer, suggesting that C1 could be used in obesity treatment, besides that in previous studies it demonstrated antioxidant properties that can counteract the FR generated during obesity.Additionally, the results of the in silico studies of C1 and C2 will be supported in vitro by NADPH production, which is proportional to inhibitory effect on 11‐βHED1.1Jan H et al., 2014; 2Celso E, 2011; 3Rob M., et al 2012.
sGC Stimulation Results in Potent Anti‐hypertensive Effects as a Stand‐alone or Combination Therapy in the Rat
Authors: Tobin Jenny , Jefferson Robert , Shea Courtney , Solberg Erik , Long Kimberly , Nakai Takashi , Moore Joel , Sheppeck James , Germano Peter , Wakefield James , Sykes Kristie , Rivers Samuel , Masferrer Jaime 
Published: June 2021

Soluble guanylate cyclase (sGC) stimulators have the potential to treat a variety of cardiovascular diseases via the nitric oxide (NO)‐sGC‐cyclic guanosine monophosphate (cGMP) pathway. Ironwood Pharmaceuticals has synthesized a number of novel sGC stimulators with the objective of creating a once‐a‐day oral anti‐hypertensive.A screening paradigm that evaluated sGC enzyme stimulation, pharmacokinetics (PK), and pharmacodynamics (PD) identified three novel sGC stimulators that induce sustained cardiovascular effects after oral dosing. Potency against sGC was evaluated using an in vitro cell based assay that measured cGMP production. Compounds were subsequently evaluated in PK and blood pressure PD in vivo rat models. All compounds were potent stimulators of sGC in vitro (EC50 = 40‐220nM), had sustained compound exposure in vivo (Thalf 7‐12 hours) after oral dosing, and significantly decreased mean arterial blood pressure (MABP) in vivo (蠅 10mmHg) after oral dosing.The potential for sGC stimulators to work in combination with reference anti‐hypertensive therapies was assessed in our in vivo PD assay. Doses of Losartan, Atenolol, Amlodipine or each of our sGC stimulators that induced a decrease in MABP (蠅 10mmHg) when dosed alone were chosen. Combining sGC stimulators with anti‐hypertensives resulted in an additional 5‐50% decrease in MABP over either therapy given alone. These data indicate that IWP sGC stimulators possess potent anti‐hypertensive effects in the rat and that their efficacy increases when dosed in combination with standard anti‐hypertensive agents.This research was funded by Ironwood Pharmaceuticals.
A “Histodilutional Barrier” Formed in Gastric Mucosa by VEGF Pretreatment Protects Ethanol‐Induced Hemorrhagic Gastric Erosions in Rats
Authors: Deng Xiaoming , Xiong Ximing , Szabo Sandor 
Published: June 2021

Since VEGF increases vascular permeability (VP), we tested the hypothesis that it might create a perivascular “histodilutional barrier”, like histamine, hence delay the absorption of damaging chemicals.MethodsFasted Sprague‐Dawley rats were given VEGF164 (0.01, 0.04, or 0.2 ug/100g) by gavage either alone or 30 min before administration of 75% ethanol (1 ml/kg). Evans blue was injected i.v. 15 min before autopsy. Rats were euthanized 30 min after VEGF alone or 1 hr after ethanol administration. Scrapings of gastric mucosa were harvested for VP measurement spectrophotometrically (560 nm) of Evans blueextravasation in the gastric mucosa. Hemorrhagic gastric mucosal lesions were measured by an Imaging Analysis System and calculated as % of glandular stomach, and also examined by histology.Results0.04ug/100g VEGF treatment significantly increased gastric VP compared to the controls (p < 0.05). Pretreatment with 0.01, 0.04, or 0.2 ug/100g VEGF reduced ethanol‐induced lesions from 10.6±1.7% (controls) to 8.9±1.8, 0.9±0.2 (p <0.01), or 2.8±0.7% (p <0.05), respectively. Histologic examination showed mild or moderate gastric edema in lamina propria or submucosa after VEGF alone, and no or only superficial gastric erosions in the rats treated with VEGF + ethanol.Conclusions1) Pretreatment with VEGF offered dose‐dependent gastroprotection in rats exposed to ethanol. 2) The likely mechanisms might be related to a histodilutional barrier induced by VEGF. 3) Further studies are needed to elucidate the molecular mechanisms of this new form of gastroprotection.(Supported by VA Merit Review Grants to S. Szabo).
N‐cadherin Upregulation and EMT Progression in As3+ and Cd2+‐Transformed Urothelial Cells
Authors: Sandquist Elizabeth , Somji Seema , Sens Donald , Garrett Scott 
Published: June 2021

Environmental agents are common causes of bladder cancer. Specifically, As3+ and Cd2+ are carcinogens implicated in bladder cancer development. Studies from our laboratory show that As3+ and Cd2+ cause malignant transformation of immortalized bladder urothelial cells, which form tumors in nude mice. Microarray analysis of repeated metal transformation in parallel revealed that N‐cadherin was the most upregulated gene in As3+ transformants, and a top induced gene in Cd2+ transformed cells. The E‐ to N‐cadherin switch is a well‐known indicator of the epithelial‐to‐mesenchymal transition (EMT) in cancer. N‐cadherin is correlated with tumor stage, increased recurrence, and decreased survival. While factors mediating E‐cadherin are well established, little is known of the regulation of N‐cadherin. The goal of the present study was to determine if N‐cadherin upregulation is maintained in animal models, the mechanism by which metal transformation leads to sustained N‐cadherin expression, and if this elevation is involved in EMT. Both mRNA and protein levels of N‐cadherin were increased in As3+ and Cd2+ transformed cell isolates, with focal expression present in intraperitoneal tumor heterotransplants. Twist and Snail were also elevated, indicating EMT. N‐cadherin may be epigenetically regulated, as MS‐275 and 5‐azacytidine altered N‐cadherin levels. Exposure of nontransformed urothelial cells to As3+ induced N‐cadherin, suggesting that heavy metal exposure promotes bladder cancer progression and EMT.Grant Funding Source: ND INBRE: 8P20GM103442‐14 from NIGMS and R25 ES016250 from NIEHS
Novel Nickel Superoxide Dismutase Treatment Delays FeCl3‐induced Arterial Thrombosis in Rats
Authors: Sung Yi‐Ting , Chien Chiang‐Ting 
Published: June 2021

Nickel complexes, [Ni(H2BA(R)TPP)](ClO4)2 (R = Ph for 1 or iPr for 2), supported by a pentadentate ligand H2BA(R)TPP were synthesized to mimic the nickel superoxide dismutase (NiSOD). Diabetes may exacerbate oxidative stress‐induced thrombosis. We explored the antioxidant activity and its biomedical effects of the synthetic NiSOD in the FeCl3‐induced carotid arterial thrombosis in the type I diabetic rats.We determined the scavenging reactive oxygen species responses in vitro by a chemiluminescent amplification method. Female Wistar rats in vivo were used and divided into four groups: control rats, streptozotocin (STZ, 60 mg/kg) treated rats, control rats with NiSOD (10 μg/ml), and STZ rats with NiSOD treatment. We investigated the underlying mechanisms of NiSOD on arterial thrombosis by immunohistochemistry and western blotting.Our data showed that NiSOD dose‐dependently decreased O2‐. and H2O2. The FeCl3‐induced time to occlusion of the carotid artery was 21±5 min in control rats, 21±2 min in STZ rats, 24±4 min in control rats with NiSOD treatment and 28±6 min in STZ rats with NiSOD (10 μg/ml), respectively. By western blot and immunohistochemistry, FeCl3 increased reactive oxygen species production, chemokine (C‐X3‐C motif) ligand 1 (CX3CL1), intercellular adhesion molecule 1 (ICAM‐1), vascular cell adhesion molecule‐1 (VCAM‐1), 4‐hydroxynonenal (4‐HNE) and 3‐nitrotyrosine (3‐NT) expression in the damaged arterial endothelium. NiSOD treatment significantly depressed the level of reactive oxygen species, CX3CLl, ICAM‐1, VCAM‐1, 4‐HNE and 3‐NT in the endothelial areas of control and STZ rats.These data suggest that a synthesized NiSOD can protect oxidative stress‐induced arterial thrombosis through the actions of antioxidant, anti‐adhesion, anti‐inflammation.
Pretreatment of Human Polymorphonuclear Leukocytes (PMN) with a New Carbon Monoxide (CO)‐Releasing Molecule (CORM401) Inhibits PMN Migration across Vascular Endothelial Cells
Authors: Inoue Ken , Woo Martin , Patterson Eric , Potter Richard F. , Capretta Alfredo , Fraser Douglas D. , Cepinskas Gediminas 
Published: June 2021

BackgroundWe have demonstrated that carbon monoxide (CO)‐releasing molecules (CORMs) reduce septic inflammation by interfering with PMN recruitment to the inflamed organs. In this study, we assessed the effects/mechanisms of a newly synthesized Mn2+‐based water‐soluble CORM (CORM401) on PMN adhesion(A)/transendothelial migration(TM) in an experimental model of endotoxemia in vitro.MethodsHuman umbilical vein endothelial cells (HUVEC) grown on parallel flow channels were stimulated with LPS (1μg/ml; 6 hrs) and interacted with freshly isolated PMN (pretreated for 30 min with CORM401 or its inactive compound, iCORM401; 100μM) for 5 min in the presence of 1.0 dyn/cm2 shear stress. Subsequently, HUVEC:PMN co‐cultures were additionally perfused for 20 min with medium containing CORM/iCORM401. Video and confocal microscopy were used to analyze PMN A/TM.ResultsStimulation of HUVEC with LPS significantly increased PMN A (0.6+/‐0.1 vs 30.9+/‐7.6 cells/mm2; p<0.05) and TM (0.0 vs 41.7+/‐6.8%; p<0.001). Pretreatment of PMN with CORM401 had no effect on PMN A, however significantly reduced PMN TM (41.7+/‐6.8% vs 15.2+/‐4.3; p<0.01). The above findings were confirmed employing confocal microscopy.ConclusionCORM401 interferes with PMN recruitment by specific suppression of PMN migratory potential. (HSFO 393, IRF‐25‐12).