bluuebunny/crossref_metadata_2025_split huggingface dataset:
Brevetoxin Induced Dendritic Arborization and Functional Recovery in a Murine Post‐Stroke Model
Authors: Sequeira Erica , Pierce Marsha , Gomez Dina , Murray Thomas 
Published: November 2023

RATIONALEStroke is a leading cause of long term disability rendering most survivors incapable of walking without assistance. This condition triggers structural and physiological changes in neuronal circuits adjacent to the infarct region, termed peri‐infarct area, that plays a key role in recovery and represents a potential therapeutic target. Pyramidal neurons are abundant in the cerebral cortex and form excitatory glutamatergic synaptic contacts. These glutamatergic neuron dendritic domains play a role in excitability and neuronal plasticity. Synaptic plasticity is largely mediated through N‐methyl D‐aspartate receptors (NMDARs) and calcium [[Ca2+]] dependent signaling pathways. Intracellular sodium [[Na+]i] acts as a signaling molecule in as much as it exerts and regulates NMDAR channel activity. Brevetoxin‐2 (PbTx‐2) is a voltage‐gated sodium channel (VGSC) gating modifier that increases [Na+]i, upregulates NMDAR function and increases downstream Ca2+ dependent signaling pathways.OBJECTIVEPurpose of this study was to develop a sensitive and a reproducible behavioral test to evaluate motor movements post‐stroke in mice and to test the effect of PbTx‐2 on recovery of movement. In addition, this study aimed to examine the effect of PbTx‐2 on dendritic arbor complexity of cerebral cortical layer V pyramidal cells.METHODSA stroke was produced by administration of an intraperitoneal injection of Rose Bengal dye followed by a 12.5 min light illumination of the caudal forelimb area of the motor cortex region. Hydrogel surgery was then performed on Day 5 post stroke which involves drilling the skull to inject Hydrogel vehicle or Hydrogel + 100 pmol PbTx‐2 using a Hamilton syringe. A Pasta Matrix Reach Task was used to determine the number of pieces of pasta retrieved post‐stroke and post‐Hydrogel treatment from a 5×5 matrix of pasta. Brain slices were imaged using Leica TCS SP8 MP confocal microscope that produces 3D images which were then analyzed with Imaris 8.3 software to measure dendritic arbor complexity by Sholl analysis.RESULTSThe results demonstrated that the number of pastas retrieved was significantly impaired on Day 1 post‐stroke with a subsequent improvement following treatment with 100 pmol PbTx‐2 as compared to the control mice. In addition, PbTx‐2 treated layer V pyramidal neurons also had a greater dendritic arbor complexity in comparison to controls.CONCLUSIONPreclinical research in a murine stroke model provided a behavioral assessment of neural insult‐induced functional deficits and PbTx‐2 induced motor recovery. The Pasta Matrix Reach Task proved to be a sensitive behavioral measurement for the study of neural impairment and recovery. Increased dendritic arbor complexity in layer V pyramidal cells indicates that PbTx‐2 can potentially aid neuroplasticity at the peri‐infarct region.Support or Funding InformationNIH R01NS053398
Identifying a Phospholipase C Epsilon Region that Interacts with Gbeta Gamma
Authors: Madukwe Jerry , Malik Sundeep , Smrcka Alan 
Published: November 2023

Many hormones and neurotransmitters function by activating isozymes of Phospholipase C (PLC) family through G protein coupled receptors (GPCRs). PLCs hydrolyze membrane phospholipids to generate downstream signaling molecules essential for normal function of the cell. Chronic PLC activation has been implicated in diseases that affect the body. The PLCɛ isoform has been shown by our laboratory to be scaffolded at the nuclear envelope yet responds to plasma membrane (PM) endothelin 1 (ET‐1) receptor stimulus. ET‐1 stimulates phosphatidyl inositol 4‐phosphate (PI4P) hydrolysis in the perinuclear Golgi apparatus generating diacyglycerol and activating a downstream nuclear pool of PKD in neonatal cardiac ventricular myocytes (NRVM). This signaling process leads to reactivation of the fetal gene program and pathologic myocyte hypertrophy (Zhang et al. Cell, 2013).Small GTPases and Heterotrimeric G protein activate PLCɛ. We have shown that the PM endothelin‐1 receptor stimulation of Golgi PI4P depletion is dependent on Gβγ dimer (Malik et al. Mol. Biol. Cell, 2015). Other groups have shown that PLCɛ co‐expressed with Gβγ and a select group of constitutively active small GTPases produce marked increases in inositol phosphate accumulation in COS7 cells. Domains of PLCɛ that interacts with small GTPases for its activation have previously been defined. However, it is not known whether PLCɛ interacts with Gβγ directly for its activation, or which region is required. Our laboratory has previously shown that Gβγ directly interacts with PLCβ2 in vitro in a region within its catalytic domain. We hypothesize that a region within the catalytic domain of PLCɛ interacts with Gβγ directly and propose to identify this region.We reexamined PLCβ2 and PLCɛ activation by Gβγ, Gq and small GTPases (hRasG12V) in COS7 cells co‐transfection assay and observed marked increases in inositol phosphate accumulation. We then identified a region of PLCɛ that aligns with the catalytic domain of PLCβ2. We are currently making mutations within the catalytic domain and other regions of PLCɛ which will be tested for Gβγ activation using the COS7 cell co‐transfection assay. We will also determine if purified Gβγ can interact with and activate PLCɛ in vitro.
Quercitrin, a natural flavonoid glycoside, protects against cisplatin‐induced injury in rats without hindering cisplatin beneficial cytotoxic activity
Authors: Arafa Elshaimaa A. , Mohamed Wafaa , Shehata Basim , Hassan Hossam , Raslan Mai , Elsherbiny Gamal , Elgendy Abelnasser 
Published: November 2023

ObjectivesThe present study aimed to evaluate the possible nephroprotective and hepatoprotective effects of quercitrin (a natural flavonoid glycoside) against cisplatin‐induced toxicity in rats. The study extended to evaluate the effect of quercitrin on the beneficial cytotoxic activity of cisplatin.BackgroundCisplatin is a widely used cytotoxic agent whose benefits are limited by its nephrotoxic and hepatotoxic potentials. Quercitrin has a wide variety of pharmacological actions including anti ‐ oxidant, anti‐inflammatory and immunomodulatory activities.MethodsCisplatin toxicity was induced by a single 7.5 mg/kg i.p injection. Quercitrin (10 and 20 mg/kg) was orally administrated for 1 week starting one day before cisplatin. Assessment of cisplatin injury was performed by measuring serum creatinine (Cr) and blood urea nitrogen (BUN) as renal injury markers, while serum alanine transaminase (ALT) and aspartate transaminase (AST) were assessed as hepatic injury markers. In addition, serum total cholesterol (TC) and triglycerides (TG) were determined as dyslipidemic markers. Renal and hepatic tissue malondialdehyde (MDA), reduced glutathione (GSH) and total nitrate/nitrite (NOx) levels were measured as oxidative and inflammatory markers., Western blot analysis of renal and hepatic phospho‐Akt (p‐Akt) and nuclear factor‐kappa B (NF‐κB) expression were determined as tissue inflammatory and immunological markers, coupled with histopathological examination of renal and hepatic tissues.Animal handling protocol was approved by Ethics Committee of the Faculty of Pharmacy, Beni‐Suef University, following guidelines of the National Institute of Health (NIH) Guide for Care and Use of Laboratory Animals.ResultsQuercitrin significantly ameliorated cisplatin‐induced renal and hepatic injury evidenced by corrections of serum Cr, BUN, ALT, AST, TC and TG levels as well as renal and hepatic MDA, GSH, NOx contents and p‐Akt and NF‐κB expression levels, in addition to correction of histopathological alterations. Cytotoxicity assay on breast adenocarcinoma (MCF‐7) cells using the thiazolyl blue tetrazolium bromide (MTT) revealed that quercitrin did not inhibit the cytotoxic activity of cisplatin in vitro.ConclusionsOur data showed the ability of quercitrin to protect against cisplatin‐induced nephrotoxicity, hepatotoxicity and dyslipidemia without interfering with Cisplatin's beneficial anticancer activity, suggesting the effective use of quercitrin during cisplatin course treatment permitting dose scaling required in some resistant cancer types.
Fructose diet decreases APAP‐induced hepatotoxicity in mice
Authors: Tripathi Ashutosh , Cho Sung‐Joon , Chlipala George E. , Green Stefan , Jeong Hyunyoung 
Published: November 2023

Acetaminophen (APAP) is a commonly used over‐the‐counter analgesic and antipyretic. APAP overdose can cause hepatotoxicity, and is responsible for ~50% of acute liver failure in US. However, risk factors for severe liver injury after APAP dose remain unclear. Previous studies have shown conflicting effects of fructose diet on the susceptibility to APAP‐induced hepatotoxicity (1, 2). The aims of the present study were to evaluate the role of fructose diet in modulating the extent of APAP‐induced liver injury and explore potential roles of differential gut microbiota in any effects. Male C57BL/6J mice (8 weeks old) were given 30% (w/v) fructose in water (or regular water) for 8 weeks, followed by oral administration of APAP (600 mg/kg) or vehicle control. The animals were sacrificed 24 h after APAP dosing, and tissues were collected. APAP‐induced liver injury (determined by serum levels of liver enzymes) was decreased by two‐fold in mice pretreated with fructose. Fructose‐treated group exhibited (~1.5 fold) higher basal glutathione levels and (~2 fold) lower basal (mRNA and activity) levels of CYP1A2 and CYP2E1, suggesting decreased bioactivation of APAP and increased detoxification of toxic metabolite in fructose‐treated mice. Also, mRNA expression levels of GCLC (glutamate‐cysteine ligase; a rate‐limiting enzyme in synthesis of glutathione) exhibited a trend of increase in fructose group. mRNA expression of HSP70 (heat shock protein 70; a chaperone protein previously known to have a protective effect against APAP hepatotoxicity) was also found increased in fructose group. High‐throughput, next‐generation sequence (NGS) analysis of bacterial 16S rRNA gene amplicons from the cecal samples of vehicle group showed that fructose diet altered the composition of gut microbiota, leading to increased α‐diversity (Mann‐Whitney, p=0.043) and a significant difference in overall bacterial community (Bray‐Curtis, ANOSIM: R = 0.581, p = 0.001). However, no bacteria taxa were found to be significantly correlated with hepatic mRNA level of CYP2E1 or CYP1A2, or glutathione levels. These findings suggest that fructose diet decreases APAP‐induced liver injury in mice, in part by reducing metabolic activation of APAP and inducing detoxification to toxic metabolites. Fructose diet also alters gut microbiota composition in mice although its role in modulation of APAP toxicity remains unclear.Support or Funding InformationThis study was supported by Chicago Biomedical Consortium.
Capsaicin suppresses the metastasis of human NSCLC in vivo
Authors: Nolan Nicholas A , Akers Austin T , Colclough Kate W , Hurley John D , Williams Cathryn D , Rollyson William D , Brown Kathleen C , Denning Krista L , Brown Linda G , Egleton Richard D , Dasgupta Piyali 
Published: November 2023

Non‐small cell lung cancer (NSCLC) is characterized by aggressive clinical course, rapid doubling time and a propensity for extrapulmonary metastasis. A substantial proportion of NSCLC presents with metastatic disease at the time of their diagnosis. Metastasis is a complex multistep process. One of the earliest events of the metastatic process is the invasion of malignant cells through the surrounding stroma into the blood and lymph. The long‐term goal of our laboratory is to identify nutritional compounds (as single agents or in combination with standard chemotherapy), which will suppress metastasis of NSCLCs. Capsaicin is the pungent ingredient of chili peppers. Several convergent studies have shown that capsaicin inhibits the invasion and metastasis of several types of human cancers including melanoma, prostate cancer, and cholangiosarcoma. The present study examines the anti‐invasive properties of capsaicin and two capsaicin‐like compounds, capsiate and capsiconiate, found in a select variety of chili peppers on NSCLC. We measured the anti‐invasive activity of these compounds by the Boyden chamber assay and spherical invasion assays. We found that capsaicin and capsiate displayed equivalent anti‐invasive activity in NSCLC cells. In contrast, capsiconiate did not suppress the invasion of NSCLC cells. Furthermore, we tested the anti‐metastatic activity of capsaicin in a syngenic mouse model of metastasis. We observed that the dietary administration of capsaicin potently decreased the area of lung metastatic foci relative to control untreated mice. Our data may form the basis of novel nutrition‐based therapeutic regimens in NSCLC metastasis.Support or Funding InformationFunding for our study was supported by the an NIH R15‐AREA Grant (2R15CA161491‐02), WVU‐MU Partnership Grant, and a NASA Undergraduate Fellowship to NAN
PDE1C regulates the dynamics of actin‐based structures in migrating human arterial smooth muscle cells
Authors: Brzezinska Paulina , Payne Darrin , Rampersad Sarah , Mackeil Jodi , Burke‐Kleinman Jonah , Maurice Donald 
Published: November 2023

Phenotypic remodeling and an associated increased migration by human arterial smooth muscle cells (HASMCs) are features of neo‐intimal hyperplasia that promote occlusive vascular diseases, including post‐angioplasty restenosis. Although it is established that cyclic adenosine 3′, 5′‐monophosphate (cAMP)‐mediated signaling inhibits overall HASMC migration, how this ubiquitous signaling system impacts the individual “steps” required for directed cell migration, including the dynamics of lamellipodial projections at their leading edges, is currently poorly defined. Using a modification of the trans‐well assay, our study highlights a critical role for a specific cyclic nucleotide phosphodiesterase, Phosphodiesterase 1C (PDE1C), in modulating lamellipodial dynamics in migrating HASMCs. Formation of HASMC lamellipodial projections in response to exposure of these cells to a chemotactic gradient (0.5% FBS) was assessed by quantifying the accumulation of actin‐rich structures on the underside of filters with 3 μm pores in 4hrs. When compared to vehicle‐treated controls, selective pharmacological inhibition of PDE1C (compound 33 (C‐33), Dart Neuroscience LLC) increased accumulation of lamellipodial structures significantly. Similarly, RNAi‐mediated knockdown of PDE1C using several distinct siRNA markedly increased accumulation of lamellipodial structures when compared to those formed in control siRNA transfected cells. In contrast, agents that increase intracellular cAMP by activating HASMC adenylyl cyclases (ACs) (i.e. forskolin) reduced accumulation of these actin‐rich structures in HASMCs exposed to a gradient of FBS. Most interestingly, lamellipodial projection accumulation in response to PDE1C inhibition, or knockdown, was also markedly antagonized by addition of forskolin. These results are consistent with the novel idea that PDE1C is acting locally at the leading edge of HASMCs to promote accumulation of lamellipodial structures and that this hyper‐localized effect is lost when HASMC cAMP levels are increased globally, using forskolin. Consistent with this, we show that PDE1C can associate with an A‐kinase anchoring protein AKAP79 and the cAMP effector Protein Kinase A (PKA). Overall, since each PDE1C and AKAP79 is found at the leading edge of migrating HASMCs, we suggest that PDE1C acts locally, via a PKA/AKAP79‐based signalosome, to regulate lamellipodial projection formation in migrating HASMCs.Support or Funding InformationCanadian Institutes for Health Research (CIHR)
Differential βarrestin1‐Mediated Signaling in Spontaneously Hypertensive versus Normotensive Rat‐Derived Astrocytes
Authors: Negussie Shmuel , Lymperopoulos Anastasios , Clark Michelle A. 
Published: November 2023

ObjectiveTo investigate the role of βarrestin1 in angiotensin II (AngII) type 1 receptor (AT1R)‐mediated signaling in astrocytes isolated from the brainstem of spontaneously hypertensive rats (SHRs) or normotensive rats.BackgroundThe βarrestins (βarrestin1 and‐2) terminate G protein signaling by G protein‐coupled receptors (GPCRs) and, at the same time, they initiate their own “second wave” of signaling from GPCRs, independently of G proteins. The AT1R is one such GPCR that signals via both G proteins and βarrestins in various cell types. It is unknown whether the AT1R signals via βarrestins in the brain. We sought to examine the involvement of βarrestins in the AT1R‐mediated Extracellular signal‐regulated kinase (ERK1/2) mitogen‐activated protein kinase (MAPK) activation and angiotensinogen (AGT) expression in Spontaneously Hypertensive Rat (SHR)‐versus normotensive rat‐derived primary astrocytes.MethodsPrimary cultures of brainstem astrocytes were isolated from the brains of 2–3 days old pups. 100 nM AngII and 10 μM SII (an AngII peptide analog which is a βarrestin‐biased agonist) were used to study AT1R‐mediated ERK1/2 activation and AGT expression. Protein analysis was done using Western blotting and mRNA measurement was done using real‐time PCR. The results in SHR astrocytes were compared to normotensive Wistar rat astrocytes. Protein analysis was done using Western blotting and mRNA measurement using real time PCR.ResultsOur results indicate that βarrestin1 is the major βarrestin isoform in astrocytes from both SHR and normotensive rat brainstem astrocytes. AngII activates ERK1/2 in SHR astrocytes more robustly than in normotensive rat astrocytes. On the other hand, SII strongly activates ERK1/2 in normotensive rat astrocytes and only minimally in SHR astrocytes. In addition, SII, similarly to AngII, upregulates cytoplasmic AGT protein synthesis in both SHR and normotensive rat astrocytes. However, SII has no significant effects on AGT mRNA levels in brainstem astrocytes from either rat model.ConclusionIn this study, we show that AngII‐AT1R‐induced ERK1/2 activation in SHR astrocytes appears to be almost exclusively G protein‐dependent, contrary to normotensive rat astrocytes. Moreover, the AT1R‐mediated βarrestin1‐dependent signaling pathway plays a role in the regulation of AGT protein synthesis in rat astrocytes derived from the brain stem region in both models.Support or Funding Information* This research was funded by Nova Southeastern University President's Faculty Research & Development Grant and Health Profession Division Grant.
Exploration of chemical components correlated with pharmacology of EHYT based on a LC‐HRMS metabolomics approach
Authors: Ha In Jin , Kim Eunok , Kim Kyuseok 
Published: November 2023

HYT (Hyeonggaeyeongyo‐tang) is a thirteen‐herb Traditional Chinese Medicine (TCM) and Korean medicine formula. Its modified prescription (Eunhwayeongyo‐tang; EHYT) is an herb medicine formula composed of the HYT and Lonicerae Flos (LF). Those have been traditionally used for acne, sinusitis, and otitis media otitis, tonsillitis, and a variety of otolaryngology symptoms. Nowadays, those prescriptions are used to treat diseases alone or combined with western medicine. However, it lacks in‐depth research of the constituents of EHYT and HYT. Establishing botanical medicine formula as globally‐accepted polychemical medicines required comprehensive chemical and biological profiling correlated with pharmacology. In this study, the production of pro‐inflammatory cytokines (IL‐1a, IL‐1b, IL‐6 and IL‐8) via Toll‐like receptor 2 (TLR2) and TLR4 in HaCat cells was demonstrated by RT‐PCR analysis. RT‐PCR analysis reveals EHYT inhibit LPS induced expression of chemokine IL‐8 in HaCaT cells. HYT and LF were shown the lower inhibition effects although those were treated on the induced cells in higher concentration than EHYT. Also, ultra high performance liquid chromatography coupled with quadrupole‐time‐of‐flight mass spectrometry and metabolomics were established to characterize the chemical profiling and explore significantly potent metabolites correlated with bioactivity. In total, potent 243 candidates were identified based on MS and MS/MS, and 36 compounds were characterized among three formula. A dicaffeoylquinic acid group was characterized as one significantly differential component among them. It demonstrated that UHPLC‐QTOF and chemometrics processing approach could be successfully applied for screening and comprehensive analysis of chemical components in complex constituents of herbal medicine. It reveals that EHYT consisting of HYT and LF has shown synergistic effects on anti‐inflammatory. LF may potentiate the anti‐inflammatory activity of HYT.Support or Funding InformationThis study was supported by a grant from the Traditional Korean Medicine R&D Project, Ministry of Health & Welfare, Republic of Korea. (HI16C0801)
Study of Systemic and Microvascular Responses to Progressive Hemolytic Anemia in Rats
Authors: Nugent William H , Carr Danuel A , Ormond Aimee , Selwood Simon P , Song Bjorn K 
Published: November 2023

Hemolytic Anemia (HA) is characterized by an increase in the ratio of red blood cell destruction to production, which results in reduced oxygen carrying capacity, tissue hypoxia, inflammation, and corresponding clinical manifestations. The microvasculature is the primary site of vascular resistance and gas exchange, and is well suited to studying the subclinical progression of HA. HA was induced in male Sprague Dawley rats by intraperitoneal administration of phenylhydrazine (PHZ) per the following dosing schedule: Group 1 (100 mg/kg on Day 1) and Group 2 (100 mg/kg on Day 1, and 50 mg/kg on Day 2). Group 3 (baseline; BL) was an untreated time control. Rats were surgically prepared for the following systemic and microvascular measurements: total hemoglobin (tHb), mean arterial pressure (MAP), heart rate, pulse pressure (PP), arteriolar contractile response to three doses of phenylephrine (PE; 0.01, 0.1, 1 mg/kg; I.V), and tissue oxygenation (PISFO2). PHZ reduced tHb by 47 and 75% for Groups 1 and 2 respectively. The 47% decrease in tHb for Group 1 was associated with the following changes from BL (Group 3): MAP decreased by 12%, PP increased by 121%, and PISFO2 decreased 23%. A 75% decrease in tHb (Group 2) resulted in the following changes from BL: MAP decreased by 34%, PP was no different, and PISFO2 decreased 40%. Heart rate remained unchanged between the three groups. The arteriolar contractile response profile to increasing concentrations of PE was right shifted, or less responsive, as tHb dropped between groups. The data indicate a robust cardiovascular compensatory response to severe hemolytic anemia, which serves to prevent peripheral tissue hypoxia. At a 75% reduction in tHb, though, this system begins to show signs of compromise as arteriolar diameters lose responsiveness to contractile stimuli and compensatory elevations in PP begin to decay. Yet, PISFO2 remains normoxic suggesting that as tHb is reduced, tissue hypoxia occurs precipitously not progressively.Support or Funding InformationSupport: Song Biotechnologies