bluuebunny/crossref_metadata_2025_split huggingface dataset:
Wnt5a Signaling in Atherosclerosis, its Effect on OxLDL Uptake and Foam Cell Differentiation
Authors: Ackers Ian , Szymanski Candice , Duckett K Jordan , McCall Kelly , Malgor Ramiro 
Published: June 2021

Atherosclerosis is a chronic inflammatory disease involving plaque buildup in the arteries and can lead to heart attack and stroke. A key step in the pathogenesis of atherosclerosis is oxLDL uptake by macrophages. Our lab has shown that oxLDL up regulated transcription of Wnt5a in macrophages. Wnt5a and Frizzled 5 (Fz5), a Wnt5a receptor, are highly expressed in foam cells in atherosclerosis. Our hypothesis is that Wnt5a stimulates oxLDL uptake and that macrophages show differential expression of Wnt5a receptors.In vitro studies: THP‐1 cell line was cultured and differentiated into macrophages. Cells were treated with oxLDL, nLDL, and rWnt5a in combinations to determine their effects on oxLDL uptake. Expression of receptors involved in oxLDL uptake such as CD68, CD36, and scavenger receptor (SR) were analyzed using immunofluorescence, western blot and PCR. To study Wnt5a signaling ROR2, Fz5, and Wnt5a were evaluated in THP‐1 cells as well as in human atherosclerotic tissue samplesIn THP‐1 cells oxLDL and Wnt5a increased CD68, CD36, SR and Fz5 at both protein and RNA level. IHC results show that Wnt5a is expressed by macrophages, foam cells, smooth muscle cells (SMC), and endothelial cells. But Fz5 is expressed only by some macrophages, and almost negative in SMC. We also found that ROR2 followed a similar expression pattern to Fz5 in macrophages while SMC near the lipid core were positive for ROR2 but negative for Fz5.In conclusion, our results suggest that oxLDL, through Wnt5a signaling, plays a role in foam cell formation and oxLDL uptake. In addition, expression of Wnt5a receptors is variable between cell types in atherosclerosis.
Correlates of Renal Atrophy in Murine 2 Kidney 1 Clip Hypertension
Authors: Kashyap Sonu , Ghossan Roba , Zaia Paula , Grande Joseph 
Published: June 2021

Renal artery stenosis (RAS) is a common disorder associated with both cardiovascular and renal morbidity and mortality. Recent studies have shown that stenting of the stenotic kidney fails to improve renal or cardiovascular outcomes, prompting a search for more effective therapies. Strain‐specific variability in renal outcomes is a well recognized problem associated with murine models of chronic injury. The objective of this study was to define strain specific differences in renal outcome and correlation with parameters of injury in a murine model of RAS.We used a 0.25 mm polytetrafluoroethylene cuff to establish RAS in mice (n=292 total). We examined the correlation between age, time following surgery, body weight, blood pressure, heart and kidney weights, ratio of STK/CLK weight, and plasma angiotensin level in 3 murine strains. Percent atrophy in the STK as a function of time following RAS surgery is summarized in the table below:

Strain
1 Week
2 Week
4 Week
6 Week

129
21%
31%
XX
45%

C57BLKS/J
25%
51%
63%
72%

C57BL/6J
XX
XX
39%
41%

The right kidney weight and the STK/CLK weight ratio correlated with %atrophy in all strains, with the strongest correlation observed in C57BL/6J mice. In C57BL/6J mice, blood pressure correlated with %atrophy. CLK weight correlated with heart weight, with the strongest correlation observed in 129Sv mice. In BLKS mice, the time following surgery and age correlated with CLK weight and %atrophy in the STK; blood pressure correlated with plasma angiotensin level.We conclude that RAS produces severe chronic renal injury in the STK of all strains studied, with the most severe injury observed in C57BLKS/J mice.The STK/CLK weight ratio correlated most strongly with %atrophy and can be used to predict development of chronic renal damage in all strains. Hypertrophy of the CLK, as measured by CLK weight, strongly correlated with heart weight.
Effects of GAT107, an Allosteric Agonist and Positive Modulator of α7 Nicotinic Acetylcholine Receptors, on Chronic Inflammatory and Neuropathic Pain in Mice
Authors: Bagdas Deniz , Wilkerson Jenny , Lichtman Aron , Thakur Ganesh , Damaj M Imad 
Published: June 2021

Agonists and positive allosteric modulators (PAMs) of α7 nicotinic acetylcholine receptors (nAChRs) are currently being considered as novel therapeutic approaches for pain modulation. Recent discovery of compounds that can function as dual allosteric agonists and PAMs (ago‐PAMs) added a new pharmacological approach for the regulation of α7 nAChR function.GAT107 [(+)‐enantiomer of racemic 4‐(4‐bromophenyl)‐3a,4,5,9b‐tetrahydro‐3H‐cyclopenta[c]quinoline‐8‐sulfonamide] is such an agent which has been shown to be an α7‐selective ago‐PAM.The present study investigated whether GAT107 attenuates chronic inflammatory and neuropathic pain in mice. We found that systemic administration of GAT107 significantly reversed pain behaviors in formalin test, complete Freud adjuvant (CFA)‐ and lipopolysaccharide‐induced inflammatory pain, chronic constriction injury‐induced neuropathic pain, without motor impairment. GAT107 failed to show antinociceptive effects on acute thermal stimulus. The antinociceptive effects of GAT107 were mediated by of α7 nAChRs. Our sites studies suggest that spinal but not peripheral sites mediate in the effect of GAT107 in CFA model. Finally, GAT107 blocked pain‐induced aversion after acetic acid injection, using a conditioned place aversion test. The present results suggest that α7 nAChR modulation by ago‐PAMs may provide new targets in inflammatory and neuropathic pain.This work was supported by VCU Massey Cancer Center, TUBITAK (2219).
Ocimum basilicum essential oil complexed with β‐Cyclodextrin increases fos protein expression in CNS areas and produces analgesic profile
Authors: Quintans Lucindo , Nascimento Simone , Araujo Adriano , De Lucca Waldecy , Antoniolli Angelo , Quintans Jullyana , Bonjardim Leonardo 
Published: June 2021

Ocimum basilicum leaves essential oil (LEO) is rich in monoterpenes, such as linalool and geraniol, and it have shown analgesic effects, but short half‐life and water insolubility are restraint for LEO therapeutic approaches. β‐cyclodextrin (β‐CD) has been applied to improve pharmacological profiles of essential oils or related compounds. Thus, we assessed the antihiperalgesic effect of LEO, alone, or complexed in β‐CD (LEO/β‐CD), on chronic non‐inflammatory muscle pain (an animal model for fibromyalgia) and evaluate the c‐Fos expression in the mice CNS). Mice were treated every day with either LEO/β‐CD (25, 50 or 100 mg/kg, p.o.), LEO (25 mg /kg, p.o.), tramadol (TRM 4mg /kg, i.p.) or vehicle (saline), and 60 minutes after treatment parameters were assessed. Oral treatment with LEO or LEO‐βCD, at all doses tested, produced a significant reduction of mechanical hyperalgesia (p<0.001) and they are able to significantly (p<0.05) increase Fos protein expression in nucleus raphe magnus and periaqueductal gray. Our results provide evidence suggesting that LEO and LEO/β‐CD produces analgesic effects through an activation of CNS areas, such as periaqueductal grey.Financial Support: CNPq/Brazil, CAPES/Brazil, FAPITEC/SE/Brazil and FINEP/Brazil
B Cell‐Targeted Therapy with Anti‐CD20 Monoclonal Antibody Reduced Secondary Tissue Damage and Enhanced Behavioral Recovery Following Experimental Spinal Cord Injury in Mice
Authors: Cuzzocrea Salvatore , Impellizzeri Daniela , Bruschetta Giuseppe , Cordaro Marika , Crupi Rosalia , Esposito Emanuela 
Published: June 2021

Traumatic injury to the spinal cord activates B cells, which culminates in the synthesis of autoantibodies. The functional significance of this immune response is unclear. Antibodies produced after SCI caused pathology, in part by activating intraspinal complement and cells bearing Fc receptors. These data indicate that B cells, through the production of antibodies, affect pathology in SCI. There is increasing appreciation of the important role of B cells in spinal cord trauma and consequently, increasing interest in treating these disorders through B cell‐depletion therapy. The purpose of this study was to investigate the effects of anti‐CD20 mAb B cell depletion therapy within the first 24 hours of SCI.In this study, the T6‐T7 vertebrae in three groups of mice were injured by a 50‐g clip‐induced compression method, and the anti‐murine CD20 IgG2a antibody (clone 18B12, 1 mg/kg) was intravenously administered starting 1 hour and 6 hours postinjury. Animals were sacrified at 24 hours. The anti‐CD20 antibody treatment caused significant attenuation of leukocyte infiltration, reactive oxygen species‐associated enzymes, and secondary tissue damage. Basso‐Beattie‐Bresnahan (BBB) scores were significantly higher in anti‐CD20‐treated mice than controls 10 days postinjury. Our data demonstrate an important role of B cells which could possibly lead to B cell‐based strategies for the treatment of SCI.
Small Molecule Inhibitors of Constitutively Active Gsa
Authors: Pals‐Rylaarsdam Robin , Carroll Julie , Ceja‐Ruiz Victor , Turcic Kyle , Jenkins Evan , Habbal Renee , Raub Matthew , Cruz Joseph , Walters D Eric 
Published: June 2021

Constitutive activation of Gsα is seen in the genetic disorder McCune‐Albright Syndrome, as well as some endocrine adenomas and in tissues affected by cholera toxin. Random mutagenesis of constitutively active Gα subunits in yeast identified three sites that, when mutated, suppressed the constitutive activity caused by R201H: F142, R231, and L266. Because R231 is on the surface of the protein, near the GTP‐binding site it was selected as a candidate site for drug targeting, with the goal of developing inhibitors of the constitutively active Gs alleles. Docking molecules in silico to the R231 binding pocket on the surface of Gs identified commercially available molecules with high potential to bind to the suppressor site. Eighteen molecules with high predicted binding affinity were tested for the ability to decrease the elevation in basal cAMP observed in HEK cells overexpressing the constitutively active Gs‐R201H allele. Two molecules, 6‐oxo‐N‐{[2‐(pyrrolidin‐1‐yl)pyridin‐4‐yl]methyl}‐1,6‐dihydropyridazine‐3‐carboxamide and N‐{[3‐(3‐methylphenyl)phenyl]methyl}‐6‐oxo‐1,6‐dihydropyridazine‐3‐carboxamide were able to significantly reduce cAMP levels by at least 10% from baseline. Neither of these molecules significantly reduced basal cAMP levels in cells transfected with wild‐type Gs constructs. These molecules will form the basis for further studies to develop antagonists of mutant Gs proteins associated with human disease.This work was Supported by NIH grant 1R15ED020190‐01
The Evaluation of Bac7 Labeled Nanocarriers to Deliver Antiretrovirals as a Pre‐Exposure Prophylaxis (PrEP) Therapy for HIV Prevention
Authors: Nelson Antoinette , Adler Derek , Myers Dan , Szekely Zoltan , Zhang Xiaoping , Sinko Patrick 
Published: June 2021

ObjectiveThe objective of this study is to engineer nanocarriers (NCs) capable of translocating through colorectal mucus and tissue and delivering drugs into cells of the distal colon. NCs will be delivered in quickbreak foams to provide sufficient colorectal coverage and be used as a colorectal PrEP therapy to prevent HIV transmission.MethodsFITC‐labeled poly(ethylene glycol) NCs were conjugated to the protease inhibitor amprenavir (APF) or cell‐penetrating peptide (CPP) Bac7 (BPF). APF and BPF were used to treat Caco‐2 cells in vitro and cell uptake was studied via flow cytometry. Mice were also treated with enemas containing APF or BPF and tissue retention was investigated. Foam formulations were studied for their ability to distribute NCs evenly throughout the distal colon. Histology and toxicity studies were also completed to assess the safety of the foam formulation.ResultsBPF uptake was shown to be 4‐fold higher than APF in vitro. Similar results were seen in vivo and BPF was retained within colorectal tissue for up to 5 days suggesting transport into the lamina propria. Ex vivo tissue samples show complete coverage of the distal colon by foams and NCs. Preliminary toxicity data of foam components display promising results based on histological observations.ConclusionBac7 CPP increased the translocation of NCs across the colorectal mucus and increased uptake into colonic cells both in vitro and in vivo. The current foam formulation is effective in providing uniform coverage of the distal colon and causes minimal tissue damage.AcknowledgementsFunding by NIH Grants T32GM8339, 1R01AI084137 and R37AI/DK‐051214
cAMP‐signaling via EPAC1 mediates vascular endothelial cell adaptation to fluid‐shear stress
Authors: Rampersad Sarah , Hubert Fabien , Umana Maria , Freitag Silja , Butler Nathalie , Brzezinska Paulina , Wudwud Alie , Maurice Donald 
Published: June 2021

Herein, we report a novel role for cAMP‐signaling in human aortic endothelial cell (HAEC) adaptation to fluid‐mediated shear stress (FSS). Our results showed that increases in cAMP enhanced elongation and alignment of HAECs cultured under LSS for 48hrs and that this effect was largely mediated by the cAMP‐effector molecule, exchange protein activated by cAMP (EPAC)‐1, but not Protein Kinase A (PKA). Additionally, major HAEC cAMP‐hydrolyzing enzyme, phosphodiesterase (PDE)4D, was also shown to play a crucial role in integrating FSS signals in these cells.Our results demonstrated that flow‐mediated increases in atheroprotective genes, Kruppel‐like factor, endothelial nitric oxide synthase and thrombomodulin mRNA expression were reduced in EPAC‐1 and PDE4D knockdown HAECs exposed to both LSS and HSS. Furthermore, EPAC‐1 knockdown increased leukocyte extravasation across HAEC monolayers in vitro and activation of EPAC significantly reduced LPS‐induced leukocyte rolling flux in mice.VECs have numerous mechanosensors which facilitate the integration of FSS signals from the flowing blood. One such sensor consists of platelet‐endothelial cell adhesion molecule (PECAM)‐1, vascular endothelial‐cadherin (VECAD) and vascular endothelial growth factor receptor (VEFGR)‐2. Our current data suggests that EPAC1 and PDE4D can additionally mediate flow‐sensing through this mechanosensory complex. All together, this data provides important mechanistic insight providing therapeutic targets in endothelial dysfunction and atherosclerosis.
Tideglusib, an allosteric inhibitor of Glycogen Synthase Kinase (GSK)‐3β increases Angiogenic activities of Endothelial Cells
Authors: Baruah Jugajyoti , Wary Kishore 
Published: June 2021

RationaleThe regulation of blood vessel growth is of paramount importance in ischemic cardiovascular and peripheral limb diseases. However, there is no effective pharmacotherapy to induce rapid revascularization of ischemic tissues.GoalTo address if newly discovered allosteric inhibitor of GSK‐3β enzyme, Tideglusib (TDG) regulates angiogenic phenotypes of endothelial cells.Methods and ResultsThe treatment of cultured ECs with Wnt3a or TDG stabilized β‐catenin polypeptide species in a time dependent manner. Quantitative RT‐PCR showed increased Cyclin‐D1, Nanog, c‐myc and Vascular Endothelial Growth Factor Receptor‐2(VEGFR‐2) transcripts. Accordingly, TDG increased expression of not only transcription factor Nanog, but also Cyclin‐D1 and VEGFR‐2 proteins. Microscopic analyses of ECs stained with anti‐VE‐cadherin antibody provided evidence of the disappearance of VE‐cadherin from the adherens junctions, while increased accumulation of β‐catenin in the nucleus. Pretreatment of ECs with TDG followed by Propidium iodide (PI) and Annexin‐V labeling showed increased survival and decreased cell death (p<0.05). Importantly, TDG treatment and the addition of 5‐bromo‐2′‐deoxyuridine (BrdU) followed by microscopy showed significant increase in the incorporation of BrdU in ECs. In addition, in a scratch assay and Boyden‐chamber cell migration experiments indicated the ability of TDG to induce migration of ECs. Furthermore, in an aortic ring assay TDG increased angiogenesis significantly.SummaryThus, for the first time, we show that TDG up‐regulates expression of Nanog, Cyclin‐D1 and VEGFR‐2 to augment neovessel formation, a critical biological process required for re‐establishing blood flow to the ischemic tissues.
Acid Inhibitors Exacerbate H.pylori‐Induced Gastritis Both In Vitro And In Vivo
Authors: Yanaka Akinori 
Published: June 2021

BackgroundGastric cancer (GC) is strongly associated with H.pylori infection. But, GC tends to develop in hypochlorhydric, but not in hyperchlorhydric patients. Since potent acid inhibitors, such as proton pump inhibitors (PPI) are widely used for the treatment of GERD, it seems reasonable to assume that potent acid inhibitors may excerbate H.pylori‐induced gastritis, promote gastric atrophy, and increase risk of GC. Based on these hypothesis, we examined (1) if the increased gastric luminal pH enhances toxicity of H.pylori‐derived ammonia in vitro, (2) if PPIs exacerbates gastritis in H.pylori‐infected mice in vivo.MethodsA:In vitro study: 1. Intact sheets of in vitro bullfrog gastric mucosa were incubated in Ussing chambers in vitro. Transmucosal electrical resistance (TER) was monitored to evaluate mucosal integrity. 2. GSM06 cells, derived from murine gastric mucosa, were cultured with or without 5 mM NH4Cl. Cell viablity and gastric mucosal apoptosis were evaluated by MTT, and by capase‐3 assay, respectively. B: In vivo study: Six‐wk‐old female mice were cultured for 8 wks after inoculated with H.pyloi, SS‐1. Degree of gastritis was evaluated using updated Sydney sysytem. Colonization of H.pylori was also examined.ResultsA‐1. NH4Cl decreased TER at high luminal pH (>7.0), but not at low luminal pH (5.0). A‐2. NH4Cl decreased cell viality and enhanced apoptosis more at higher ambient pH (7.8), than at low pH (7.2). B‐1. PPI Tx exacerbated inflammation in corpus, but not in antral mucosa in H.pylori‐infected mice. B‐2. PPI Tx increased colonization of H.pylori only in corpus.ConclusionsHigh ambient pH induced by potent acid inhibitors enhances toxicity of H.pylori‐dervied ammonia, thereby exacerbates H.pylori‐induced gastritis. Potent acid inhibition may increase risk of GC in patients with H.pylori infection.
Maximizing the Max Test: A Method for Determining Cardiorespiratory Fitness and Dysfunction in Mice.
Authors: Petrosino Jennifer , Mauyra Santosh , Kalyanasundaram Anuradha , Wilson Jacob , Periasamy Muthu , Ziouzenkova Ouliana 
Published: June 2021

Early diagnostic of cardiovascular morbidity plays a critical role preventing mortality. The Bruce protocol is an exercise diagnostic test that can determine cardiorespiratory fitness. This is a standardized protocol to evaluate cardiac function in humans, yet no such standardize test has been developed for widespread use with animal models.PURPOSEThe purpose of the study is to develop a similar method of testing V02max in mice. METHODS: Male WT mice (C57BL06, n=7) and mice with impaired cardiovascular function (Casq2‐null and obese WT, n=15) were used to optimize a progressive speed run test vs. our graded maximal run test and identify critical parameters indicating cardiovascular dysfunction.RESULTSOur graded test compared to the progressive lead to increased VO2max in reduced time in obesity model. With WT and Casq2‐null similar V02max was achieved in less time. Our graded test allowed for determination of a metabolic crossover parameter occurring sooner in dysfunctional animals compared to controls, as well as differential points of ventilatory threshold (VAT), time to VO2max, and %VO2max at VAT (P<.05). We determined that the crossover parameter is the most accurate predictor of cardiac dysfunction.CONCLUSIONSThe graded exercise protocol provided valuable metabolic and cardiovascular information for fast and noninvasive evaluation of cardiovascular dysfunction in animal models.
Effects of arginine vasopressin deficiency on innate immunity in the rat
Authors: Quintanar‐Stephano Andrés , Viñuela‐Berni Verónica , Organista‐Esparza Alejandro , Tinajero‐Ruelas Manuel , Huerta‐Carreón Erika , Kovacs Kalman , Berczi Istvan 
Published: June 2021

Immune and vascular endothelial cells possess arginine vasopressin (AVP) receptors. AVP is an important stimulating/regulatory hormone in acquired immunity, whereas its role on innate immunity (INIM) is not well known. Neurointermediate pituitary lobectomy (NIL) in the rat causes permanent low AVP serum levels. Here, we investigate the AVP and INIM relationship in NIL rats (3 weeks after surgeries) subject to INIM tests: Phagocytic index (PI) (peritoneal macrophages (Mθ) erytrophagocytosis), and Skin Evans blue extravasation‐histamine doses‐response test. In the Study 1, NIL group was compared against Intact control (IC), sham operated (SHAM) and anterior pituitary lobectomysed (AL) groups. In the study 2, NIL group was compared against IC, NIL+desmopressin (a synthetic analog of AVP) and IC+conivaptan (antagonist of V1a‐V2 AVP receptors) groups. Results: Study 1 showed that as compared with the IC and AL groups, PI was significant decreased in NIL animals. In the Evans blue extravasation‐histamine test study, a significant and similar increases of skin edema histamine doses‐dependent occurred in NIL and IC+conivaptan groups, whereas no significant differences in IC and NIL+desmopressin groups were appreciated.ConclusionAVP plays an important role in phagocytic activity of the peritoneal Mθ and for the stabilization of the vascular endothelial cells permeability during histamine inflammation.Supported by UAA‐PIFF 14‐1. Universidad Autónoma de Aguascalientes, México and the Jarislowsky and Lloyd Carr‐Harris Foundations.
The Effects of Bilirubin on Angiogenesis in Mice with Hindlimb Ischemia
Authors: Ikeda Yasumasa , Kanai Yusuke , Horinouchi Yuya , Izawa‐Ishizawa Yuki , Kihira Yoshitaka , Miyamoto Licht , Ishizawa Keisuke , Tsuchiya Koichiro , Tamaki Toshiaki 
Published: June 2021

BackgroundLow serum bilirubin levels are associated with peripheral artery disease. Bilirubin is known to exert anti‐oxidant effect and the enhancement of flow‐mediated vasodilation, however, bilirubin action on angiogenesis remains unclear. In this study, we examine the mechanism of bilirubin on angiogenic effect.Methods and ResultsWe employed C57BL/6J mice with unilateral hindlimb ischemia surgery and divided the mice into two groups (vehicle‐treated group and bilirubin treated group). The analysis of laser speckle blood flow demonstrated that the recovery of blood flow in response to ischemic hindlimb was promoted in mice with bilirubin treatment. The capillary density was significantly higher in ischemic lower limb of bilirubin‐treated mice. The phosphorylated levels of endothelial nitric oxide synthesis (eNOS) and Akt were augmented in ischemic tissues of mice with bilirubin treatment compared vehicle treatment. In in vitro experiments by using human aortic endothelial cells, bilirubin increased eNOS and Akt phosphorylation. Bilirubin also augmented cell proliferation, cell migration and tube formation. These bilirubin action on endothelial cell activation were abolished by LY294002, a phosphatidylinositol 3‐kinase inhibitor.ConclusionBilirubin promotes angiogenesis through endothelial cells activation via Akt‐eNOS‐dependent axis.