bluuebunny/crossref_metadata_2025_split huggingface dataset:
Epigenetic Regulation of PTEN in Hepatic Cancer
Authors: Qadir Ximena Vanegas , Han Chang , Wu Tong 
Published: June 2021

The pathogenesis of hepatocellular carcinoma (HCC) remains misunderstood offering limited therapeutic options for patients with hepatic cancer.Treatment of HCC cell line (Huh‐7) with DNMT inhibitor, Decitabine, and HDAC inhibitor, Trichostatin A, increased the expression of PTEN protein. Methylation‐specific qPCR assay showed Decitabine/TSA treatment resulted in hypermethylation of CpG islands located on the PTEN gene promoter. Furthermore, miR‐185 targets DNMT1, thus upregulating the expression of PTEN in Huh‐7. Forced overexpression of miR‐185 abolished the expression of DNMT1 protein and enhanced the level of PTEN protein in Huh‐7 stable cells. MiR‐185 xenograft tumor volume from SCID mice was significantly decreased versus control. Likewise, Ki‐67 immunostaining in Huh‐7 control stable cell line showed a higher proliferative. Xenograft protein samples showed increased PTEN expression and decreased DNMT1 activity. DNMT‐1 3'UTR reporter plasmids were constructed/cloned showing the wild type‐3'UTR of DNMT1 was significantly depleted in Huh‐7 miR‐185 stable cell line versus control in luciferase assay experiments. Co‐transfection of Huh‐7 miR‐185 ORF versus control showed DNMT1 restoration via western blot, Invasion and Cell Proliferation Assay. In‐situ hybridization showed a substantial decrease of staining expression in human HCC tumor samples versus normal liver tissue probed for miR‐185.Our results demonstrate a novel epigenetic regulation of PTEN by miR‐185, which is critical in hepatic carcinogenesis.
p38MAPK inhibition: a new combined approach to reduce neuroblastoma resistance under etoposide treatment
Authors: Marengo Barbara , Ricciarelli Roberta , De Ciucis Chiara , Furfaro AnnaLisa , Colla Renata , Canepa Elisa , Traverso Nicola , Marinari Umberto Maria , Pronzato Maria Adelaide , Domenicotti Cinzia 
Published: June 2021

Neuroblastoma (NB), the second most common pediatric tumor, is characterized by biological and clinical heterogeneity. Considering the limited success of chemotherapy, the aim of our study is to identify and target key molecular pathways associated with NB chemoresistance. In this context, we analyzed the main survival and death pathways triggered by etoposide, a commonly used chemotherapeutic agent, in HTLA‐230, a MYCN‐amplified NB cell line isolated from a high‐risk patient. Etoposide induced a concentration‐dependent reduction of cell viability and, at very high doses, totally counteracted cell tumorigenicity and neurosphere formation. In addition, etoposide activated p38 Mitogen‐activated protein Kinase (MAPK), AKT and c‐Jun N‐terminal kinase. Therefore, the treatment with etoposide combined with SB203580, an inhibitor of p38MAPK activity, decreased cell viability and tumorigenicity, counteracted neurosphere generation and slowed down the cell migration and invasion. In this context, the expression of COX‐2, ICAM‐1 and CXCR4 was down regulated, the formation of capillary‐like structures was prevented, by generating a phenotype inadequate for tumor development.. Collectively, our results suggest that p38MAPK inhibitors, in combination with standard chemotherapy, could be a novel strategy to counteract NB resistance and relapse.Grants: PRIN 2009M8FKBB_002; Genoa University.
Both supraspinal and spinal mechanisms may be involved in the maintenance of gastric mucosal integrity in the rat.
Authors: Gyires Klara , Tóth Viktoria , Király Kornél , Barna István , Zádori Zoltán Sándor 
Published: June 2021

Several neuropeptides given intracerebroventricularly (i.c.v.) or into motor nucleus of vagus induce gastric mucosal defense in the rat.AimTo study 1. if central modulation of the level of endogenous gastroprotective compounds may affect gastric mucosal defense, 2. if gastroprotection can be elicited also by microinjection of neuropeptides into lateral hypothalamus (LH) or intrathecallally (i.t.).MethodsGastric mucosal damage was induced by acidified ethanol. β‐endorphin level was determined by RIA.Results1. Gastroprotection was induced via inhibition of the degradation of anandamide and 2‐arachidonoyl‐glycerol by URB 597 (1.5–29.5 nmol i.c.v) and JZL 184; (0.3–1.3 nmol i.c.v.) resp., as well as by the inhibition of the degradation of endomorphins by diprotin A (1 μmol i.c.v.). 2. Neonatal monosodium glutamate treatment reduced significantly the immunoreactive β‐endorphin content in the nucleus of the solitary tract and the mucosal protective effect of opioid peptides. 3. Gastric mucosal protection was elicited both by i.t. administration of endomorphins and β‐endorphin (10 pmol) and by microinjection of different neuropeptides into LH.Conclusion1. Elevation of the level of endocannabinoids and endomorphins resulted in gastric mucosal defense, while reduction of the level of β‐endorphin counteracted the gastroprotective action of opioid peptides. 2. Gastroprotection could be elicited also by spinal mechanism.
RGS2 regulates urotensin II‐induced intracellular Ca2+ elevation and contraction in glomerular mesangial cells
Authors: Adebiyi Adebowale 
Published: June 2021

Urotensin II (UII), a kidney‐localized vasoactive peptide regulates renal functions, including renal blood flow, glomerular filtration rate, urine output, and electrolyte excretion. In the present study, the physiological functions of UII and regulation of UII receptor (UTR) in glomerular mesangial cells (GMCs) were investigated. RT‐PCR, Western immunoblotting, and immunofluorescence indicated the expression of UTR in murine GMCs. Mouse UII (mUII) elevated intracellular Ca2+ concentration ([Ca2+]i) and decreased murine GMC surface area by ~ 55 and 20%, respectively. mUII‐induced [Ca2+]i elevation and contraction in the cells were attenuated by SB 657510, a UTR antagonist, araguspongin B, an inositol 1,4,5‐trisphosphate receptor inhibitor, thapsigargin, a sarco/endoplasmic reticulum Ca2+‐ATPase inhibitor, and La3+, a store‐operated Ca2+ channel blocker. mUII‐induced [Ca2+]i elevation in murine GMCs was reduced by ~ 82% in Ca2+‐free medium. However, nimodipine, an L‐type Ca2+channel blocker did not alter mUII‐induced [Ca2+]i elevation and contraction in the cells. Co‐immunoprecipitation, co‐localization, and in situ ligation proximity assay demonstrated that UTR interacts with regulator of G‐protein signaling (RGS) 2 in murine GMCs. Biotinylation indicated that RGS2 is predominantly localized intracellularly in murine GMCs. However, mUII treatment elevated RGS2 surface expression in the cells by ~ 55%. siRNA‐mediated knockdown of RGS2 in murine GMCs increased mUII‐induced [Ca2+]i elevation and contraction by ~ 35 and 31%, respectively. These findings indicate that mUII induces store‐operated Ca2+ entry and contraction in murine GMCs. These data also suggest that mUII stimulates recruitment of RGS2 protein to GMC plasma membrane as a negative feedback mechanism to regulate UTR activation.Supported by the National Institute of Health
Green Tea and Cancer: A Qualified Health Claim Case Study
Authors: Berhaupt‐Glickstein Amanda , Hallman William K. , Nucci Mary , Hooker Neal 
Published: June 2021

In 2005, the Food and Drug Administration (FDA) released an enforcement letter for qualified health claims (QHC) about the ingestion of green tea (food and supplement forms) and the reduced risk of breast cancer or prostate cancer. The QHCs for green tea were enforced shortly after the FDA release of the Consumer Health Information for Better Nutrition Initiative (CHIBN), which expanded the enforcement of QHCs from dietary supplements to conventional foods. Green tea and breast/prostate cancer was identified as a case study to assess the progression of QHCs since the CHIBN expansion. An in‐depth review of court cases, FDA enforcement letters, and correspondence between FDA and green tea manufacturers was conducted. The investigation demonstrated that QHC statements about green tea and breast/prostate cancer have undergone multiple iterations and as a result, are a potential source of confusion for food and supplement designers and manufacturers. The progression of QHC statements for green tea and breast/prostate cancer reflects other enforced QHC diet‐disease categories. The evidence suggests that current FDA enforcement of QHCs is ineffective and in need of repair.Grant Funding Source: Robert Wood Johnson Foundation Investigator Awards in Health Policy Research
17β‐Estradiol (E2) induces sulfotransferase (SULT) 2A1 expression through estrogen receptor (ER) α
Authors: Li Wei , Ning Miaoran , Kim Heesue , Jeong Hyunyoung 
Published: June 2021

Our recent microarray study showed that E2 increases mRNA levels of SULT2A1 in human hepatocytes. SULT2A1 sulfonates dehydroepiandrosterone and oxysterol, playing an important role in maintenance of steroid and lipid homeostasis. The objective of this study is to elucidate regulatory mechanisms underlying SULT2A1 induction by E2. In HepG2‐ER cells (HepG2 stably expressing ERα) and human hepatocytes, E2 induced SULT2A1 expression at both mRNA and protein levels (1.8–2.7‐fold). Co‐treatment of ERα‐degrading antiestrogen ICI182,780 abrogated the induction, indicating that ERα is required for SULT2A1 induction by E2. Results from luciferase reporter assays in HepG2 cells showed that E2 increased SULT2A1 promoter activity by 13‐fold. Transient transfection of ERα mutants harboring point mutations in the DNA‐binding domain significantly decreased the induction, suggesting involvement of the classical mechanism of ERα action in SULT2A1 regulation. Results from deletion assays revealed that the cis‐element responsible for the ER action is located in −256/+140 of SULT2A1. Together, the results demonstrated that E2 induces transcription of SULT2A1 via ERα, potentially through direct binding of ERα to the promoter region. The induction of SULT2A1, which may alter the intracellular concentration of oxysterol, potentially provides insights into the known lipid‐lowing effects of estrogen.This work was supported by the National Institute of Child Health and Human Development (HD065532).
A novel method to estimate AF‐219 levels in bladder interstitium derived from drug excreted into the urine
Authors: Smith Steven A , Black Dan , Song Di , Ford Anthony P 
Published: June 2021

The P2X3 antagonist AF‐219, being studied in bladder disorders, is eliminated by renal clearance. Local renal delivery of AF‐219 to the bladder may confer a therapeutic advantage, supplementing & prolonging effect of systemic (p.o.) administration. We therefore sought to estimate the bladder venous concentration of a drug, assuming that when a drug is instilled into the bladder, the sole source of any systemic exposure must be via diffusion through to the veins that drain it. Thus by comparing the plasma levels following continuous intravenous infusion or intra‐vesicular instillation, we can estimate the concentration of drug in the veins, approximating the interstitial levels in the bladder wall itself. This approach was tested with AF‐219, methylnaltrexone, tiotropium, antipyrine, & AF‐353, co‐administered in cassette format to rats. The local concentration of each compound in the blood draining the bladder was calculated from plasma concentrations, infusion rate, & bladder blood flow. With this method, the local concentrations in the blood draining the bladder were determined to be approximately 1.1%, 1.7%, 2.0%, 2.4%, and 10% of the urine concentration of AF‐219, methylnaltrexone, tiotropium, antipyrine, and AF‐353, respectively.Conclusiondue to its high urinary concentration, AF‐219 can achieve a pharmacologically relevant exposure in bladder wall beyond that driven systemically.
Growth inhibition and radiosensitization of human papillomavirus (HPV) positive head and neck cancer (HNC) by epidermal growth factor receptor inhibition is mediated by apoptosis
Authors: Kimple Randall J , Smith Molly A , Blitzer Grace C , Huang Hao‐Shun , Torres Alexandra D 
Published: June 2021

PURPOSETo determine the mechanism by which HPV+ HNC cells and patient‐derived tumorgrafts are sensitive to growth inhibition and/or radiosensitization by cetuximab.METHODSFour cell lines and several direct‐from‐patient tumorgrafts validated for HPV expression were used in a subcutaneous flank approach using athymic Foxn1nu female mice. Apoptosis and cell cycle distribution were assessed by caspase activation, flow cytometry, and western blot.RESULTSCetuximab caused significant delay in cell proliferation (p<0.005), decrease in colony formation (p<0.026), and minimal radiosensitization. In both a cell line xenograft and a direct‐from‐patient tumorgraft model of HPV+ HNCs cetuximab resulted in significant tumor growth delay (median time to tumor quadrupling: 15 vs. 24 days, p=0.02; and 42 vs. 89 days, p=0.0001, respectively). When combined with radiation, cetuximab resulted in a significant increase in time to tumor quadrupling compared to either cetuximab or radiation alone (p<0.001). Cetuximab's effects correlated with enhanced apoptosis and a G1 cell cycle arrest.CONCLUSIONSCetuximab is active against HPV+ HNC cells and primary tumors and shows modest radiosensitization. Increased apoptosis was seen following cetuximab treatment. Mechanisms of resistance and predictors of response to EGFR inhibition in HPV+ HNC are being investigated.
The effect of CBG (BDS) botanical cannabinoid extract on MCF‐7 human breast carcinoma cells
Authors: Javid Farideh , Fshinjavid Saeed 
Published: June 2021

Recently the cannabinoid pharmacology has received attention in cancer research (Alexander et al., 2009). The present study investigated the potential anti‐tumour activity of CBG (BDS), a botanical cannabinoid extract on breast tumour cells.MCF‐7 cells (American Type Culture Collection) were grown and maintained in RPMI 1640 medium plus 10% fetal bovine serum at 37°C, 5% CO2. The cells were plated in 96‐well culture plates at a density of 1×104 cells/well and allowed to adhere at 37°C for 24 hours. The following day, various doses of extract or vehicle in the absence and presence of AM251, SR144528 and capsazepine, were added to the cells and further incubated for 4 days. Then the supernatant was removed and MTT (3‐(4,5‐dimethylthiazol‐2‐yl)‐2, 5‐diphenyltetrazolium bromide) was added for 4 hours. The ability of cells to form formazan crystals by active mitochondrial respiration was determined after dissolving the crystals in DMSO. Cytotoxicity was expressed as a relative percentage of the absorbance measured at 540 nm.CBG (BDS) extract induced dose‐dependent cytotoxic effect (IC50 of 0.096 mg/ml). Pre‐treatment with AM251, SR144528 and Capsazepine, CB1, CB2 and TRPV1 receptor antagonists, respectively, did not reverse the cytotoxicity afforded by CBG (BDS). Single application of antagonists alone or vehicle did not affect the survival rate of the MCF7 cells.The data suggest the unlikely involvement of CB1, CB2 and TRPV1 receptors in mediating CBG (BDS)‐induced cytotoxicity.We thank GW Pharmaceuticals for providing cannabinoid extracts.
Different Responses of RT‐PCR Amplifications after Tumor Necrosis alpha (TNFa) Induction at Nuclear Factor kappa B (NFkB) Gene Silenced LNCaP Cells
Authors: Gonen‐Korkmaz Ceren , Sevin Gulnur , Gokce Goksel , ARUN MEHMET Z. , Yetik‐Anacak Gunay , Yildirim Gokce , Reel Buket , Ogut Deniz , Kaymak Aysegul 
Published: June 2021

Prostate cancer is the second leading reason of morbidity and mortality in men in the Western World. Androgen receptor positive and androgen responsive lymph‐node metastasis of prostate cancer cell line LNCaP was used in all further experiments. Western with NFkB specific antibody was done after TNFa induction at LNCaP cells and a significant increase was taken.LNCaP cells transfected with siNFkB construct for 1,2 and 4 days and induced with TNFa for 24 hours. RNAisolation, cDNA conversion and RT‐PCR amplifications were done with apoptosis related gene primers such as p53, Bcl‐2, STAMP family primers. Chromatine immunoprecipitation (ChIP) assay was done with NFkB specific antibody before and after TNF induction at LNCaP and normal prostate cell line WPMY1. Since both cell lines express STAMP1 and 2, promoter region located NFkB response‐elements were identified and shown with specific primers. However, no change was shown at STAMP2 RT‐PCR amplification at −/+ TNFa native/siNFkB; STAMP1 showed a significant decrease with TNFa induction, while a significant increase with siNFkB.Survival gene NFkB silencing caused anti‐apoptotic STAMP1 increase that repress p53, together with MDM2.Gene silencing differed a panel of cancer regulatory genes.
Informed target discovery for gene and stem cell therapy in acute lung injury
Authors: Zhou Dun Yuan , Amatullah Hajera , Shan Yue Xin , Gali Patricia , Hu Ping Zhao , dos Santos Claudia C 
Published: June 2021

IntroductionAcute lung injury (ALI)/sepsis‐induced acute respiratory distress syndrome (ARDS) accounts for 9% ICU deaths. There is an urgent need for specific treatments and mesenchymal stem cells (MSC) have reparative potential in sepsis and ALI.MethodsIn the murine model of CLP‐induced ARDS, microarray detected changes in global gene expression following sepsis and MSC treatment. LIMMA analysis identified mRNA and miRNA with significant transcriptional changes between sham, CLP‐operated, and CLP+MSC treated mice.ResultsWe confirmed our in silico data in our in vitro model. Adhesion molecules, occludin and claudin‐2, were putative targets of one miRNA of interest. In silico, occludin was reduced after CLP compared with sham, confirmed by western blots (in vivo and in vitro), and there was a 50% mRNA reduction. Similarly, claudin‐2 expression from the in silico data matched the protein and mRNA analyses (in vivo and in vitro). Subsequently, following transfection of the inhibitor of the miRNA of interest, occludin and claudin‐2′s mRNA levels were observed to be significantly up‐regulated.ConclusionmRNA and miRNA microarrays are informative in identifying candidate therapeutic tools for gene regulation in ARDS treatment. MSC administration after CLP in the murine model acts as a strategy that offers great insight in the detection of therapeutically relevant genes to treat ARDS/ALI.
Validation of an Early Life Candidate Biomarker for Childhood Asthma
Authors: Xu Haili , Radabaugh Timothy , Lu Zhenqiang , Billheimer Dean , Vercelli Donata , Halonen Marilyn , Lau Serrine S 
Published: June 2021

ObjectiveThe objective of our study is to validate a plasma proteomic signature, discovered by an antibody array approach, that accompanies asthma risk in early life.MethodsA previous antibody array study identified 4 asthma‐associated proteins (erythropoietin [EPO], galectin‐3, soluble GP130, and eotaxin‐3) in 11 plasma samples obtained at age 3 from children with asthma and 12 without (assessed at age 9) from a birth cohort enrolled nonselectively. We sought to validate EPO by assaying for its concentration using R&D Quantikine ELISA in the original group of asthmatics and non‐asthmatics, and also tested matched samples at age 5, an enlarged group of samples from the same cohort at age 3 and available matched samples collected at birth.ResultsMean Log EPO in age 3 plasma was decreased in asthma group (0.757 vs 0.969 IU/L; asthma vs no asthma; p=0.016). Matched samples at age 5 did not differ by group. Upon enlarging the age 3 sample size (asthma n=18; no asthma n=54) EPO concentration remained decreased in asthma group (0.795 vs 0.898 IU/L; p=0.028 [one‐tailed]); subjects from the enlarged group who also had samples at birth (asthma n=17; no asthma n=41) did not show a similar proteomic alteration (1.41 vs 1.45 IU/L; p=0.7).ConclusionOur study validates by a second assay that reduced plasma EPO at age 3 can serve as a candidate biomarker differentiating subjects with asthma from those without. (RC1ES018328)
Role of biased signaling by beta‐blockers in the development of asthma phenotype in a murine model
Authors: Thanawala Vaidehi Jatin , Forkuo Gloria , Al‐Sawalha Nour , Knoll Brian , Leff Paul , Bond Richard 
Published: June 2021

There is increasing evidence that the use of certain ß‐blockers may be beneficial in asthma therapy. ß‐blockers can act as inverse agonists (IA), antagonists (ANT), or agonists (AG) at ß adrenergic receptors (ßAR) based on the pathway studied. Using murine models, we have previously shown that at the Gs pathway, chronic ß2AR IA treatment but not ANT is beneficial. We now show that epinephrine is required for development of asthma phenotype in an ovalbumin sensitization and challenge (Ova S/C) model, using phenylethanolamine N‐transferase null mice (PNMT−/−), lacking the enzyme for epinephrine synthesis. We also show that in an enhanced murine asthma model by adding a protease activated receptor‐2 ligand (PAR2l) SLIGRL‐NH2 in addition to Ova S/C; the PNMT−/− mice do develop a partial asthma phenotype that is significantly less than Ova S/C wild type mice. In this PAR2l model, chronic administration of carvedilol, an IA at the ß2AR‐Gs pathway, but an AG at the ß2AR‐ßarrestin pathway, or alprenolol, an AG at both pathways, results in worsening of the asthma phenotype. The results indicate that IA at the canonical ß2AR‐Gs pathway is not sufficient to prevent the asthma phenotype, and suggest ligand bias for different pathways may play an important role in development of the asthma phenotype.RO1A179236 awarded to RAB
Fetal programming of metabolic health through a combined dietary and pharmacological pre‐pregnancy intervention in rats
Authors: Dennison Carol , Eslinger Amanda , Reimer Raylene 
Published: June 2021

Maternal obesity is associated with low birth weight, type 2 diabetes and obesity risk in offspring. Diets high in oligofructose (OFS) improve glucose control through glucagon‐like‐peptide‐1 (GLP‐1). Sitagliptin inhibits the degradation of GLP‐1in vivo. The aim of this study was to determine if a combined OFS and sitagliptin pre‐pregnancy intervention in obese female Sprague‐Dawley rats can improve the metabolic health of female offspring. Obese female Sprague‐Dawley rats were randomized to 1 of 6 groups: 1) AIN‐93; 2) 10% OFS; 3) Sitagliptin; 4) AIN‐93+OFS+Sitagliptin; 5) High fat & sucrose (HFS); 6) Caloric restriction (CR). A lean reference group was also included. Pups consumed AIN‐93 until 11wk when they were given HFS diet for 6 wk. Body weight and blood glucose were assessed. At birth the offspring of combination‐treated dams were significantly heavier than offspring in all other groups (p<0.04). Following the HFS diet challenge the blood glucose area under the curve for offspring of the combination group was less than the HFS and CR groups (p<0.05). Despite higher birth weight, the combined treatment significantly improved offspring glycemic control in adulthood. Funded by CIHR and ACHRI.Grant Funding Source: CIHR and ACHRI