bluuebunny/crossref_metadata_2025_split huggingface dataset:
Transformation of mouse embryonic fibroblasts by dual expression of EBV LMP‐1 and HPV‐16 E6
Authors: Kinjo Takao , Shimabuku Tetsuya , Tamanaha Ayumi , Kitamura Bunta , Tanabe Yasuka , Oshiro Yoshihide 
Published: June 2021

Background and ObjectiveFrom the high prevalence of Epstein‐Barr virus (EBV) and high‐risk human papillomavirus (HPV) infection among oral cancer patients in Okinawa, southwest islands of Japan, we hypothesize that dual EBV and HPV infection is related to oral carcinogenesis. The purpose of this study is to explore the mechanisms of transformation induced by dual EBV and HPV infection.MethodsWe constructed mouse embryonic fibroblasts expressing singly or dually EBV and HPV genes, and compared transformation properties.ResultsSingle expression of EBV latent membrane protein‐1 (LMP‐1) and dual LMP‐1+HPV E6 expression induced increased cell proliferation and resistance to apoptosis, whereas expression of a single viral gene did not exhibit these properties. Dual LMP‐1+E6 expression showed anchorage independence and tumor formation in nude mice, whereas expression of LMP‐1 alone did not. Furthermore, dual LMP‐1+E6‐expressing cells induced increased NF‐κB signaling, and the knockdown of LMP‐1 or E6 expression eliminated the increased cell proliferation, anchorage independence, and NF‐κB activation.ConclusionsThese data suggested that the singular expression of these viral genes is insufficient for inducing transformation and that dual LMP‐1+E6 expression, which is associated with increased NF‐κB activity, is required for transformation.
Morphologic evaluation of dental pulp vascularity during DMBA experimental administration
Authors: Marto Carlos Miguel , Costa Eduardo , Farinha Rodrigo , Cabrita António Silvério 
Published: June 2021

7,12 ‐ Dimethylbenz(a)anthracene (DMBA) is a well known carcinogen and immunosuppressor. Oral administrated DMBA circulates in the bloodstream and rich vascularized tissues like dental pulp can be particularity sensible to this compound effect. In this study we evaluate dental pulp morphologic changes during DMBA per os experimental administration.Control and test groups were constituted by ten male Wistar rats each, 8 weeks old. Control group was maintained without manipulation and test group was submitted to daily administration of 20mg/kg DMBA dissolved in olive oil and administrated by oral gavage, both during 4 weeks. All animals were sacrificed at the end of 4 weeks period and a complete necropsy was performed. Slides of dental pulp were obtained from the lower incisors dental root medium third and hematoxylin‐eosin coloration was performed. Slides were observed in light microscopy and morphometric analysis was performed, evaluating areas of dental pulp vascularity, calcification and presence of inflammatory cells. T‐student test was used in statistical analysis with a significance level of 0,05.Results show no statistic difference between control and test group and none inflammatory process was observed. We hypothesized that these results may be due the immunosuppressor activity of DMBA, administrated dose or even protective role of olive oil so more studies must be performed.
Dietary modulation of BALT on a rat mammary tumor model
Authors: Lopes Ana Isabel Calado , Pereira Paula , Cabrita António Silvério 
Published: June 2021

Murine bronchus associated lymphoid tissue (BALT) is a mirror of systemic immunity since it varies greatly with immune status of the animal and plays a pivotal role in local immune responses at the lung surface. BALT enhanced histopathology (EH) is a semiquantitative screening tool to determine whether there is a potential effect on the immune system of rats under controlled conditions.168 Sprague Dawley virgin female rats with 42 days were randomly distributed for seven groups of 24 animals each. Besides group I, at 55 days the groups were induced for mammary cancer with 20mg of 1,12‐dimetilbenzantracene (DMBA). Dietary protocols were applied for 150 days as following. Group I: standard diet (SD), 300Kcal/100g; Group II: DMBA +SD; Group III: SD + beans 40g/Kg of SD, 341Kcal/100g; Group IV: SD + olive oil (50ml/Kg of SD); Group V: fiber supplement 4× the SD, 250Kcal/100g; Group VI: fiber supplement 4× the SD + beans 19,95gr + olive oil 25ml on 1Kg of SD; Group VII: SD + mediterranean lyophilized diet (41Kcal). The animals were ethically euthanized and the lungs were routinely fixed and stained with hematoxylin‐eosin. BALT was evaluated by EH.The BALT morphology of tumoral rats with mediterranean diet and caloric restrain was similar to the BALT of control group. The other tumoral groups had decreased BALT in size and lymphocyte number compared to control group.These results suggest that diet can interfere on BALT morphology.
Macrophage Phenotypes in Murine Aneurysm
Authors: Morgan Stephanie , Wang Qiwei , Harberg Calvin , Liu Bo 
Published: June 2021

A prominent pathological feature of abdominal aortic aneurysm (AAA) is the infiltration of mononuclear phagocytes such as macrophages. Accumulating literature has highlighted the importance of macrophage phenotypes and their role in vascular disease. Inflammatory (M1) macrophages produce proteases and proinflammatory cytokines, while alternatively activated (M2) macrophages produce anti‐inflammatory cytokines and extracellular matrix (ECM) components. Using an elastase‐induced model of murine AAA, we tested the hypothesis that aneurysm induction shifts M1/M2 balance toward inflammation. Leukocytes were isolated from the aortic tissue and analyzed by flow cytometry. Three days after surgery, arteries treated with elastase (aneurysm) or inactivated elastase (control) contained similar levels of total macrophages (7aad‐CD45+CD11b+F4/80+). However, as disease progressed, aneurysmal aorta displayed significantly higher number of macrophages than controls ~2.15 and 1.28 fold higher 7 and 14 days after surgery, respectively. Interestingly, the M1:M2 ratio (CD86+:CD206+) was higher in aneurysm arteries, largely due to a selective suppression of M2 polarization. These data suggest an imbalance between M1 and M2 macrophages may contribute to a chronic inflammatory state during aneurysm progression.Funding provided by NRSA T32 HL 07936 from the NHLBI.
DISTINCT EFFECT OF H2O2 IN HEALTH AND UNHEALTHY HEMATOPOIETIC CELLS SURVIVAL
Authors: PEDRO AMANDA NOGUEIRA , MUNHOZ THALYTA AP. CESÁRIO , DIAS CAROLINA CARVALHO , PAREDES‐GAMERO EDGAR JULIAN , ORIGASSA CLARICE C , EÇA LILIAN PIÑERO MARCOLIN , FERREIRA ALICE TEIXEIRA 
Published: June 2021

In the last years, many data has suggested the participation of oxidative stress in the regulation of hematopoietic cell homeostasis. Hematopoietic cells are particularly vulnerable to reactive oxygen species (ROS) in a concentration‐dependent manner. It is known that constitutively ROS levels differ between normal and malignant cells. In this respect it is the ideal balance in the redox status for both cell types in order to selectively cause death of cancer but not normal cells. In this work it was induced a pro‐oxidant environment by stimulating murine and fresh human umbilical cord blood (health models) as well as HL‐60 leukemic cells (unhealthy model) with different concentrations (0, 1, 5 and 10 μM) of hydrogen peroxide (H2O2) in vitro. It was observed that H2O2 (5 μM) induced cancer but not normal cells death (28% versus 0,5%). However, HL‐60 hematopoietic stem cells has increased in percentage at the same time progenitors cells diminished. In addition clonogenic capacity was great affected on these cells, enabling to generate myeloid clones. Differentiation was not modulated by the H2O2 in any cell type. HL‐60 loss viability had the participation of phosphor‐AKT which expression has diminished significantly. In summary, we found that at low concentration, H2O2 is able to kill hematopoietic cancer cells without affect normal cells.Supporting: FAPESP
Inhibition of prostaglandin E2 or thromboxane A2 promotes human aortic smooth muscle cell differentiation
Authors: Adedoyin Oreoluwa Olufunmilayo , Loftin Charles D 
Published: June 2021

INTRODUCTIONInhibition of cyclooxygenase‐2 (COX‐2) helps maintain vascular SMC differentiation when treating abdominal aortic aneurysms in a disease model. However, mechanisms of COX‐2‐dependent vascular SMC phenotypic modulation have not been identified. We assessed the hypothesis that COX‐2 induces SMC de‐differentiation by the action of prostaglandin E2 (PGE2) or thromboxane A2 (TXA2).METHODSCultured SMCs were treated with enzyme inhibitors or prostanoid receptor agonists or antagonists. Expression of α‐actin as a differentiation marker was examined by real‐time PCR, western blot or immunocytochemistry; and EIA performed to measure inhibitor effects on PGE2 production.RESULTSCelecoxib increased α‐actin expression and significantly reduced PGE2 synthesis in stimulated cells (p < 0.0001; n=3; unpaired Students t‐test). Differentiated cells treated with 1μM PGE2 showed significant reduction in differentiation (p < 0.05; n=3; one‐way ANOVA), and treatment with 300nM SQ 29548 (TXA2 receptor antagonist) promoted differentiation. Also, treatment of de‐differentiated cells with 10 μM seratrodast (TXA2 receptor antagonist) increased α‐actin expression by 52%, while 5μM IBOP (TXA2 receptor agonist) reduced differentiation compared with untreated controls.CONCLUSIONCOX‐2 inhibition may increase hASMC differentiation by inhibiting production of PGE2 or TXA2.Funding: NIH HL083122
A Single, Ultra‐High Dose Aminoglycoside Therapy in a Rat Model of E. coli Induced Septic Shock
Authors: Pisipati Amarnath , Anand Kumar , Ariano Robert , Blakley Brian , Esfahani Ehsan K 
Published: June 2021

IntroductionSeptic shock is a life threatening condition often due to Gram negative bacterial infections and is responsible for around 200,000 deaths every year in United States of America alone. Our research focuses on determining the potential of a single, ultrahigh dose aminoglycoside therapy for accelerated bacterial clearance with minimal to manageable side effects in a rat model of E. coli induced septic shock.Methods: Study in uninfected animalsGentamicin was administered intravenously at 80 and 160 mg/kg doses to anesthetized and intubated Sprague Dawley rats. Mean arterial pressure, heart rate, respiratory rate and percentage oxygen saturation were determined. The occurrence and time course of neuromuscular paralysis/ventilator dependency, renal injury and auditory toxicity after ultra‐high dosing of gentamicin was determined. Renal injury was determined by Creatinine clearance and histology. Auditory brain stem response studies were performed to assess the auditory toxicity.Septic shock induced animalsSeptic shock was induced in rats by surgical, intra‐peritoneal implantation of a gelatin capsule containing E. coli bort (ATCC 700973) in a fibrinogen and alfa‐cellulose matrix. Gentamicin was intravenously administered at 5, 10, 20, 40, 80 and 160 mg/kg doses.ResultsUltra high gentamicin dosed animals were found to develop neuromuscular paralysis with ventilator dependency for 20 – 40 minutes and recovered thereafter. Rats did not exhibit any auditory and renal toxicity. Complete eradication of bacterial counts from blood occurred following ultra‐high doses.ConclusionRats were able to tolerate single, ultra‐high doses of gentamicin with complete eradication of bacterial counts from blood.Research is supported by University of Manitoba
MicroRNA‐21 increases the resistance to CDDP in Hela cervical carcinoma cells by targeting PDCD4 and MSH2
Authors: Cheng JiaoYing , Nishi Hirotaka , Yonekawa Chinatsu , Takara Kohji , Isaka Keiichi 
Published: June 2021

ObjectivesMicroRNA‐21 (miR‐21) has been found to be over‐expressed in many human cancers, including cervical cancer. Our aim is to investigate the association of miR‐21 with the chemosensitivity of HeLa cervical carcinoma cells to cis‐Diaminedichloroplatinum (CDDP).MethodsWith cervical carcinoma cell lines, HeLa and CDDP‐resistant HeLa (HeLa/CDDP), the cell growth was examined by MTT assays with miR‐21 or miR‐21 inhibitor transfection. TaqMan RT‐PCR was performed to detect the miR‐21 and mRNA expression of Programmed Cell Death 4 (PDCD4) and mutS homolog 2, colon cancer, nonpolyposis type 1 (MSH2). The protein expression of PDCD4 and MSH2 was detected with western blot assay. Luciferase assay was performed to analyze the potential targets of miR‐21.ResultsCell growth results suggested miR‐21 inhibitor inhibit CDDP‐resistant HeLa growth significantly. Compared with HeLa, MiR‐21 and mRNA of PDCD4 and MSH2 expression in HeLa/CDDP were higher, but the protein expression of PDCD4 and MSH2 in HeLa/CDDP were downregulated in western blot results. Transfection of miR‐21 inhibitor for HeLa/CDDP with PDCD4 3′ untranslated region (3′ UTR) or MSH2 3′UTR vector showed significant increase in luciferase activity compared with control.ConclusionsOver‐expression of miR‐21 may play critical roles for the CDDP‐resistance in cervical cancer, at least in part via targeting PDCD4 and MSH2.
2K‐1C Hypertensive Rats Treated With Tempol Show Decreased MMP‐2 Activity Possibly Due To Oxidative Stress Blockage
Authors: Antonio Raquel Carros , Ceron Carla Speroni , Rizzi Elen , Santos José Eduardo Tanus , Gerlach Raquel Fernanda 
Published: June 2021

Hypertensive structural and functional modifications are associated with increased MMP‐2 activity and reactive oxygen species (ROS). ROS can induce post‐translational modifications in proteins, such as nitrosylation, that are known to affect anzyme activity. ROS can also directly activate MMPs. This study examined whether the treatment with Tempol prevents raised systolic blood pressure (SBP) and the following biochemical findings of hypertensive heart tissue: ROS activity, total gelatinolytic activity, MMP‐2 activity, and presence of nitrotyrosine residues.Study protocol:2kidney‐1clip (2K‐1C) rats and Sham‐operated rats (Sham) were treated with vehicle or Tempol (T) (18 mg/kg/day). Tested used were in situ oxidative stress; in situ zymography; immunofluorescence for MMP‐2 and nitrotyrosine. Tempol attenuated hypertension (2K‐1C: 193± 3mmHg; 2K‐1C+T; 159± 2mmHg; p<0,05), and led to decreases in ROS, total gelatinase activity, MMP‐2 activity and amount of stained nitrotyrosine in cardiac tissue (p<0,05 for all comparisons between the 2K‐1C group versus 2K‐1C+T). Thus, nitrosylation of MMP‐2 or of its substrates may change MMP‐2 activity in hypertension. Results show one important benefit of the treatment with the antioxidant Tempol that needs better understanding.This study was supported by Conselho Nacional de Pesquisa (CNPq) Brasil.
Analysis and Impact of Mainstream Tobacco Smoke from Regular and Reduced Exposure Products on Blood Brain Barrier: Are we clearing this smoke cloud yet?
Authors: Naik Pooja , Cucullo Luca 
Published: June 2021

Active and passive tobacco smoking (TS) are associated with dysfunction of vascular endothelial physiology (including the blood‐brain barrier) in a causative and dose dependent way largely related to the relative content of reactive oxygen and nitrogen species (ROS/RNS). ROS and RNS can mediate progressive endothelial damage causing alteration of extracellular matrix and apoptosis of endothelial cells. Accordingly, the tobacco industry is marketing several lines of “reduced‐exposure” (RE) tobacco products claimed to be less harmful. For this purpose we measured the ROS and nitrogen content of mainstream TS from different brands (including tobacco free and ultralight cigarettes) and assessed their effect on BBB endothelial cells. Our results showed that RE products have a higher content of hydrogen peroxide and RNS which correlated with higher release of MMP‐2 and IL‐6 from the BBB endothelium. These results were further supported by gene array analysis which also showed alterations in the gene expression of CYP enzymes, vascular adhesion molecules and signal transduction proteins (SHC4). Alteration in gene expression of hydroxysteroid (11‐beta) dehydrogenase 1 and 2, insulin induced gene 1, also suggests that TS could play a role in the pathogenesis of type 2 diabetes. Our results suggest that ‘light’ and ‘tobacco free’ cigarettes are equally or potentially more toxic than the regular products.This work was supported by NIH/NIDA R01‐DA029121–01A1 and A.R.D.F. to Dr. Luca Cucullo.
Voluntary Aerobic Exercise Destiffens Arteries of Old Mice Via Inhibition of NADPH Oxidase‐ and Superoxide‐Dependent Oxidative Stress
Authors: Eng Jason Samuel , Pham Bryant Thanh , Johnson Lawrence Cody , Giosca‐Ryan Rachel A , Seals Douglas R , Fleenor Bradley S 
Published: June 2021

We tested the hypothesis that voluntary aerobic exercise (wheel running) reverses age‐related large elastic artery stiffening by inhibiting NADPH oxidase‐ and superoxide‐dependent oxidative stress. Intrinsic mechanical properties were examined in thoracic (TA) and abdominal (AA) aortic segments from young (Y, 8 mo, n=8) and old (O, 26 mo, n=7) cage‐control male C57BL/6 mice, and old animals with access to voluntary running wheels (OVR, 26 mo, n=6) for 10 weeks. Compared with Y, O had greater elastic modulus (EM), an indicator of stiffness, in the TA (5899 ± 731 vs. 3307 ± 324 kPa, p<0.05) and AA (7796 ± 794 vs. 5268 ± 371 kPa, p<0.01), and increased aortic nitrotyrosine abundance (NT, 2.78 ± 0.61 vs. 1.00 ± 0.23 AU, p<0.05), a marker of oxidative stress. Voluntary running decreased EM in the TA (to 3712 ± 878 kPa, p<0.05) and AA (to 5988 ± 507 kPa, p<0.05), and reduced NT abundance (to 0.91 ± 0.25 AU, p<0.05). Additional TA segments were cultured for 72 hr with TEMPOL (100μm), a superoxide scavenger; apocynin (Apo, 1μmol), a NADPH oxidase inhibitor; or vehicle control (C). Treatment with either TEMPOL or Apo normalized EM in O mice to that observed in Y and OVR animals, without affecting EM in the latter groups. Our results demonstrate that voluntary aerobic exercise normalizes age‐associated stiffening of the TA and AA in mice via reversal of NADPH oxidase‐ and superoxide‐dependent oxidative stress.NIH AG013038, AG000279, HL007822
The role of differential oxygen distribution between the brain and peripheral tissues on tolerance to induced central hypovolemia
Authors: Lee Kay Victoria , George Mebin , Soller Babs R. , Ryan Kathy L. , Hinojosa‐Laborde Carmen , Convertino Victor A. , Rickards Caroline A. 
Published: June 2021

BackgroundMaintenance of cerebral perfusion and oxygenation is essential for survival from hemorrhage. As individuals vary in tolerance to reduced blood volume (e.g., central hypovolemia), we hypothesized that subjects with high tolerance (HT) will maintain cerebral oxygenation compared to low tolerant (LT) subjects, which will be associated with greater peripheral vasoconstriction.Methods13 human subjects were instrumented for cerebral (ScO2) and muscle (SmO2) oxygenation (near‐infrared spectroscopy, NIRS), then completed a step‐wise presyncopal‐limited lower body negative pressure (LBNP) protocol. Total hemoglobin (HbT) was calculated as an index of local peripheral resistance (inverse relationship) at the muscle.ResultsProgressive decreases in ScO2 were observed in LT subjects (n=5) from −45 mmHg LBNP (P≤0.01 compared to baseline), while ScO2 did not change throughout LBNP for the HT subjects (n=8; P≥0.44). SmO2 decreased at −30 and −60 mmHg LBNP (P≤0.01) in the HT group with concomitant increases in resistance (decreases in HbT; P≤0.03). In contrast, both SmO2 (P=0.01) and HbT (P=0.06) only decreased at −60 mmHg LBNP in the LT group.ConclusionsIn support of our hypothesis, increased tolerance to progressive central hypovolemia was associated with maintained cerebral oxygenation, due, in part, to earlier onset of peripheral vasoconstriction.Source of Funding: US Army MRMC
Effect of Cardiotonic Steroids on Monolayer Permeability and Junction Proteins in Lymphatic Endothelial Cells
Authors: Horvat Darijana , Cromer Walter , Kuehl Thomas , Zawieja David , Uddin Mohammad Nasir 
Published: June 2021

ObjectivePreeclampsia (preE) is a hypertensive disorder unique to pregnancy. Cardiotonic steroids (CS) such as marinobufagenin (MBG), cinobufotalin (CINO), and ouabain (OUB) are Na+/K+ ATPase inhibitors. MBG is elevated in a rat model and patients with preE. MBG causes a vascular leak syndrome in vivo and increases endothelial cell monolayer permeability. We evaluated the effect of these CS on monolayer permeability of lymphatic endothelial cells (LECs).MethodsLECs were isolated from a rat mesenteric collecting lymphatic vessel. The cells were treated with DMSO (basal), MBG, CINO, or OUB. Monolayer permeability of CS‐induced LECs was measured. The expression of β‐catenin and VE‐cadherin was measured by immunofluorescence. Western blot was used to measure β‐catenin and VE‐cadherin protein levels.ResultsMBG (≥ 1 nM, p<0.05) and CINO (≥ 10 nM, p<0.05) significantly increased the monolayer permeability of LECs compared to basal (DMSO) while OUB had no effect. MBG (p<0.05) and CINO (p<0.05) treatment significantly downregulated β‐catenin protein expression. However, CS did not cause any disruption of the LEC tight junctions. CINO downregulated the VE‐cadherin protein expression, but MBG did not.ConclusionsWe have demonstrated that bufadienolides MBG and CINO caused an increase in the monolayer permeability and downregulated the β‐catenin protein expression with no effect on tight junctions.
E. coli a‐haemolysin leads to ATP release and P2‐dependent calcium influx in human erythrocytes
Authors: Skals Marianne Gerberg , Leipziger Jens , Praetorius Helle 
Published: June 2021

The E. coli toxin á‐haemolysin (HlyA) lyses erythrocytes by inserting as pores in the membrane. Recently, we demonstrated that HlyA‐induced haemolysis is a sequential process with an initial volume reduction that precedes swelling and lysis. The initial volume reduction is triggered by an abrupt increase in [Ca2+]i.Moreover, we have shown that HlyA‐induced haemolysis requires extracellular ATP and P2 receptor activation. The present study investigates the mechanism for ATP release and [Ca2+]i increase that precede the volume changes in erythrocytes. By the luciferin/luciferase assay, we show that HlyA leads to ATP release within 5 minutes after toxin is added. ATP release was independent of pannexin‐ and volume‐dependent anion channels; two suggested ATP exit pathways. Thus, ATP may simply be released via the toxin pore. Through a combination of flow cytometry and live cell imaging, we show that the [Ca2+]i response consists of an abrupt increase followed by at slower, sustained increment. P2 antagonists do not interfere with the initial acute [Ca2+]i increase, but significantly reduces the slow phase of the HlyA‐induced influx of [Ca2+]i and cause a marked delay in the swelling of the cells. We suggest that ATP may be released via the toxin pore, which activates P2 receptors in the erythrocyte membrane. The influx of cations through P2 receptors contributes to net ion influx and thereby cell swelling and lysis.