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Tityus serrulatus venom and its toxins Ts1 and Ts5 increase cytosolic ca^(2+)concentration in isolated vascular smooth muscle cells

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Journal of Biophysical chemistry 2012年第4期3卷 287-294页

作者： Mario dos A.neto Flavio Vasconcelos Lusiane M.Bendhack Eliane c.Arantes Faculty of Inga-UNINGA MaringaBrazil Laboratory of Pharmacology Department of Physics and ChemistryFaculty of Pharmaceutical Sciences of Ribeirao PretoUniversity of Sao PauloRibeirao PretoBrazil Laboratory of Animal Toxins Department of Physics and ChemistryFaculty of Pharmaceutical Sciences of Ribeirao PretoUniversity of Sao PauloRibeirao PretoBrazil Laboratory of Toxicology Faculty of PharmacyInstitute of Health SciencesFederal University of ParaBelemBrazil

Voltage-gated Na+ channel (Nav channel) scorpion toxins are classified as α- and β-neurotoxins. Ts5 (α-neurotoxin) and Ts1 (β-neurotoxin) from Tityus serrulatus venom (TsV) interact with Nav channels, increasing Na+ influx and activating voltage-dependent ca2+ channels. This study aimed to investigate the effect of TsV, Ts1 and Ts5 on the cytosolic ca2+ concentration ([ca2+]c) in rat aortic smooth muscle cells. Toxins were isolated by ion exchange chromatography (Ts1) followed by RP-HPLc (Ts5). The rat aortic smooth muscle cells were isolated in Hanks buffer pH 7.4 and loaded with 5 μmol/L of Fura-2AM (45 minutes at 37℃), in order to measure [ca2+]c by fluorescence of Fura-2/AM (ratio 340/380 nm). The fluorescence was measured in one single cell (excitation: 340 and 380 nm;emission: 510 nm). TsV (100 and 500 mg/mL) and its toxins Ts1 and Ts5 (50 and 100 mg/mL each) led to a concentration-dependent increase in [ca2+]c. Tetrodotoxin (1 mmol/L), a Nav channel blocker, and verapamil (1 mmol/L), a voltage-operated ca2+ channel blocker, inhibited the increase in [ca2+]c induced by TsV (500 mg/mL). In conclusion, TsV and its toxins induce a concentration-dependent increase in [ca2+]c that probably occurs through interaction with Nav channels, thus inducing depolarization and consequent ca2+ influx. This assumption is based on the fact that this effect is inhibited by tetrodotoxin and verapamil, showing a direct action of TsV toxins on aorta smooth muscle cells.

关键词： Ts1 Ts5 Na^(+) channel ca^(2+) channel Tityus serrulatus

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The Exogenous Application of Abscisic Acid Induce Accumulation of Anthocyanins and Phenolic compounds of the ‘Rubi’ Grape

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American Journal of Plant Sciences 2017年第10期8卷 2422-2432页

作者： Francisco José Domingues neto Adilson Pimentel Junior cristine Vanz Borges Silvia Regina cunha Daniel callili Giuseppina Pace Pereira lima Sergio Ruffo Roberto Sarita Leonel Marco Antonio Tecchio School of Agriculture S&#227 o Paulo State University (UNESP) Botucatu Brazil Institute of Biosciences S&#227 o Paulo State University (UNESP) Botucatu Brazil Department of Agronomy Londrina State University (UEL) Londrina Brazil

The abscisic acid (ABA) is related to the formation of certain polyphenols, such as anthocyanin, improving the nutritional and commercial quality of the red grapes. In this study, we verified whether the exogenous application of?S-ABA favors the accumulation of anthocyanin, the total phenolic compounds and the antioxidant activity of the?‘Rubi’?grape. The experiment was performed during the cycle 2014/2015 with the?‘Rubi’?vine (Vitis vinifera?L.) grafted in the rootstock 420-A. The experimental design was composed of randomized blocks, with four treatments and six blocks, in a total of 24 experimental parcels, constituted of one plant each. We used the?S-ABA isomer in different concentrations and times of application, 400 mg·L-1?S-ABA in the beginning of the ripening (BR);400 mg·L-1?at BR + 200 mg·L-1?at 25 days after first application (DAFA);400 mg·L-1at BR + 400 mg·L-1?at 25 DAFA;besides the control (0 mg·L-1). After 40 days of the first application, the samples were harvested and we verified that the?S-ABA induced the accumulation of anthocyanins and total phenolic compounds in the?‘Rubi’?grape. From these results we can recommend two applications of?S-ABA, 400 mg·L-1?at BR + 200 mg·L-1?at 25 DAFA.

关键词： Vitis vinifera S-ABA Table Grape Bioactives compounds

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comparison of clonidine and Metoprolol Effectiveness on the control of Post-Operative Blood Pressure in Otorhinolaryngology-Head and Neck Surgeries: A clinical Trial

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Open Journal of Anesthesiology 2021年第2期11卷 49-58页

作者： Sérgio Menezes Silvio caldas neto Ruy Lins Filho Jayme Santos neto Rossana Lins Universidade Federal de Pernambuco Recife Brazil

Objectives: Adequate control of hemodynamic parameters in patients undergoing otolaryngology-head and neck (OLHN) surgeries is one of the objectives of peri-operative anesthesia management. In this context, we seek to evaluate the effectiveness of metoprolol compared to clonidine in the post-operative control of systolic (SBP) and diastolic (DBP) blood pressures, and heart rate (HR), when these medications are used in the immediate pre-operative period. Method: this is a randomized double-blind clinical trial. Patients over 18 years old, according to the American Society of Anesthesiologists Physical Status Classification (ASA) I or II, who would undergo OLHN surgery, presenting with greater than 140 mmHg (SBP) or 90 mmHg (DBP), upon arrival at the surgical ward, were included in the study. 46 patients were randomized into two groups (C or M) who received clonidine (75 or 150 mcg) or metoprolol (5 or 10 mg), respectively. The averages of SBP, DBP and HR were compared between groups upon arrival at the post anesthesia care unit (PACU) and four hours post-surgery. Results: In group C, SBP (127.7 ± 18.8 vs 137.3 ± 14.1 mmHg, p = 0.03) and DBP (73.1 ± 15.46 vs 82.6 ± 7.9 mmHg, p 0.01) were lower, when compared with group M. No difference was observed in HR immediately upon PACU arrival. In addition, for four hours post-surgery, no change was noted in the SBP, DBP or HR. Conclusion: metoprolol was less effective than clonidine in reducing systolic and diastolic blood pressures in the immediate post-operative period of OLHN surgeries.

关键词： clonidine Metoprolol Hypertension Post-Operative

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Nicotinic cholinergic receptors in esophagus:Early alteration during carcinogenesis and prognostic value

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World Journal of Gastroenterology 2016年第31期22卷 7146-7156页

作者： Marina chianello Nicolau Luis Felipe Ribeiro Pinto Pedro Nicolau-neto Paulo Roberto Alves de Pinho Ana Rossini Tatiana de Almeida Simao Sheila coelho Soares lima Departamento de Bioquímica Instituto de Biologia Roberto Alcantara GomesUniversidade do Estado do Rio de Janeiro Programa de carcinogênese Molecular Instituto Nacional de CancerCoordenacao de Pesquisa Gastroenterologia Hospital Universitário Pedro ErnestoUniversidade do Estado do Rio de Janeiro

AIM: To compare expression of nicotinic cholinergic receptors(cHRNs) in healthy and squamous cell carcinoma-affected esophagus and determine the prognostic ***: We performed RT-q PcR to measure the expression of cHRNs in 44 esophageal samples from healthy individuals and in matched normal surrounding mucosa, and in tumors from 28 patientsdiagnosed with esophageal squamous cell carcinoma(EScc). Next, we performed correlation analysis for the detected expression of these receptors with the habits and clinico-pathological characteristics of all study participants. In order to investigate the possible correlations between the expression of the different cHRN subunits in both healthy esophagus and tissues from EScc patients, correlation matrices were generated. Subsequently, we evaluated whether the detected alterations in expression of the various cHRNs could precede histopathological modifications during the esophageal carcinogenic processes by using receiver operating characteristic curve analysis. Finally, we evaluated the impact of cHRNA5 and cHRNA7 expression on overall survival by using multivariate ***: cHRNA3, cHRNA5, cHRNA7 and cHRNB4, but not cHRNA1, cHRNA4, cHRNA9 or cHRNA10, were found to be expressed in normal(healthy) esophageal mucosa. In EScc, cHRNA5 and cHRNA7 were overexpressed as compared with patient-matched surrounding non-tumor mucosa(EScc-adjacent mucosa; P ctively). Positive correlations were observed between cHRNA3 and cHRNB4 expression in all samples analyzed. Additionally, cHRNB4 was found to be differentially expressed in the healthy esophagus and the normalappearing EScc-adjacent mucosa, allowing for distinguishment between these tissues with a sensitivity of 75.86% and a specificity of 78.95%(P = 0.0002). Finally, cHRNA5 expression was identified as an independent prognostic factor in EScc; patients with high cHRNA5 expression showed an increased overall survival, in comparison with those

关键词： Nicotinic cholinergic receptors Esophagus Esophageal squamous cell carcinoma Tobacco Alcohol Gene expression

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In vitro induced pluripotency from urine-derived cells in porcine

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World Journal of Stem cells 2022年第3期14卷 231-244页

作者： Kaiana Recchia Lucas Simões Machado Ramon cesar Botigelli Naira caroline Godoy Pieri Gabriela Barbosa Raquel Vasconcelos Guimarães de castro Mariana Groke Marques Laís Vicari de Figueiredo Pessôa Paulo Fantinato neto Flávio Vieira Meirelles Aline Fernanda de Souza Simone Maria Massami Kitamura Martins Fabiana Fernandes Bressan Department of Surgery Faculty of Veterinary Medicine and Animal SciencesUniversity of São PauloPirassununga 13635-000São PauloBrazil Department of Pharmacology and Biotechnology Institute of BioscienceSão Paulo State UniversityBotucatu 18618-689São PauloBrazil Department of Veterinary Medicine Faculty of Animal Sciences and Food EngineeringUniversity of São PauloPirassununga 13635-000São PauloBrazil Embrapa Suínos e Aves Empresa Brasileira de Pesquisa AgropecuáriaConcordia 89715-899Santa CatarinaBrazil Department of Animal Sciences Faculty of Animal Sciences and Food EngineeringUniversity of São PauloPirassununga 13635-000São PauloBrazil

BAcKGROUND The generation of induced pluripotent stem cells(iPSc)has been a game-changer in translational and regenerative medicine;however,their large-scale applicability is still hampered by the scarcity of accessible,safe,and reproducible *** porcine model is a large biomedical model that enables translational applications,including gene editing,long term in vivo and offspring analysis;therefore,suitable for both medicine and animal *** To reprogramme in vitro into pluripotency,and herein urine-derived cells(UDcs)were isolated from porcine *** The UDcs were reprogrammed in vitro using human or murine octamer-binding transcription factor 4(OcT4),SRY-box2(SOX2),Kruppel-like factor 4(KLF4),and c-MYc,and cultured with basic fibroblast growth factor(bFGF)*** characterize the putative porcine iPScs three clonal lineages were submitted to immunocytochemistry for alkaline phosphatase(AP),OcT4,SOX2,NANOG,TRA181 and SSEA 1 *** transcripts related to the pluripotency(OcT4,SOX2 and NANOG)were analyzed via reverse transcription quantitative realtime polymerase chain reaction in different time points during the culture,and all three lineages formed embryoid bodies(EBs)when cultured in suspension without bFGF *** The UDcs were isolated from swine urine samples and when at passage 2 submitted to in vitro *** of putative iPScs were obtained only from UDcs transduced with the murine factors(mOSKM),but not from human factors(hOSKM).Three clonal lineages were isolated and further cultured for at least 28 passages,all the lineages were positive for AP detection,the OcT4,SOX2,NANOG markers,albeit the immunocytochemical analysis also revealed heterogeneous phenotypic profiles among lineages and passages for NANOG and SSEA1,similar results were observed in the abundance of the endogenous transcripts related to pluripotent *** the clonal lineages when cultured in suspension without

关键词： Induced pluripotent stem cells Noninvasive Pluripotency Reprogramming Urine Porcine

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Role of Selective cyclo-Oxygenase-2 Inhibitor celecoxib in canine Osteosarcoma cell culture

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Open Journal of Pathology 2013年第4期3卷 144-149页

作者： Paulo R.O.Bersano Maria T.S.Alves Maria F.M.R.Gartner Adriana camargo Ferrasi Joao F.lima-neto Fernanda c.Landim Stephane c.Vexenat carlos Eduardo Fonseca Alves Glenda Nicioli da Silva Noeme S.Rocha Laboratory of Investigative and comparative Pathology School of Veterinary Medicine and Animal ScienceUniversidade Estadual PaulistaBotucatuBrazil Department of Pathology Paulista School of MedicineFederal University of Sao PauloSao PauloBrazil Institute of Molecular Pathology and Immunology of University of Porto PortoPortugal Hemocentro Paulista School of Medicine Federal University of Sao PauloSao PauloBrazil Laboratory of Reproduction and cellular Therapy Department of Animal Reproduction and Veterinary RadiologyUniversidade Estadual PaulistaBotucatuBrazil

Background: Experimental studies have shown that cyclo-oxygenase-2 (cox2) is related to the development and progression of tumors, since this enzyme is induced and expressed by cells such as macrophages, osteoblasts, “activated” endothelial cells, and tumor cells. The activity in tumors includes proliferation, cell transformation, tumor growth, invasion and metastasis and may play an important role in carcinogenesis of the canine osteosarcoma, since it has high expression in tissue fragments. The combination of selective cox2 inhibitors and other treatment modalities is the basis for a new anti-cancer therapy strategy. This in vitro study exposed primary cells of five different canine osteosarcoma cultures to selective cox2 inhibitor at increasing concentrations and times. Results: For cox2 negative cultures, despite the absence of differences, greater sensitivity of cells to treatment was observed. For cox2 positive cultures, a higher number of necrotic cells were observed (P ≤ 0.05), when compared with negative cultures. For exposure times with celecoxib doses, no difference (P > 0.05) was found between the three times analyzed for living, apoptotic and apoptotic/necrotic cells. There are similarities in the values of 24 h and 48 h, with slight reduction of living cells, increasing those undergoing apoptosis and apoptosis/necrosis. There was significance for necrosis (P ≤ 0.05). In 72 hours, a significant difference was observed between the other two previous values (P ≤ 0.05). It was found for the group of 100 μML-1, that there was a numerically greater signaling for apoptosis and lower (P = 0.08) for necrosis, and this point was the onset of the pharmacodynamic phenomenon, with drop in the values for living cells and increased number of necrotic cells, with a tendency (P = 0.08) for reducing the percentage of necrotic cells for the group of 100 μML-1 when compared to that of 10 μML-1. conclusions: For cox2 positive and negative cultures, there was difference for

关键词： canine Osteosarcoma cyclo-Oxygenase-2 celecoxib

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Use of Silicon in Mitigating Ammonium Toxicity in Maize Plants

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American Journal of Plant Sciences 2015年第11期6卷 1780-1784页

作者： cid Naudi Silva campos Renato de Mello Prado cassiano Garcia Roque Antonio Joao de lima neto Luiz Junior Pereira Marques Aridênia Peixoto chaves claudia Amaral cruz Department of Soils and Fertilizers Universidade Estadual Paulista/FCAV Jaboticabal Brazil Department of Soils and Fertilizers Universidade Estadual Paulista/FCAV Jaboticabal Brazil Universidade Federal de Mato Grosso do Sul Chapad&atildeo do Sul Brazil Universidade Federal de Vi&ccedilosa Vi&ccedilosa Brazil Department of Applied Biology and Agriculture Universidade Estadual Paulista/FCAV Jaboticabal Brazil Department of Plant Science Universidade Federal Rural do Semi-árido Mossoró Brazil M.Sc. in Agronomy Natal Brazil

Silicon is a beneficial element that can mitigate abiotic stresses, such as ammonium toxicity. The objective herein was to evaluate the effects of silicon (Si) on mitigating toxicity caused by excess ammonium in maize plants grown in nutrient solution. An experiment was conducted with maize plants (cultivar DKB 390 VT Pro II) grown in a greenhouse in pots (8 L) in a hydroponic system. The treatments were arranged in a 2 × 2 factorial, consisting of two ammonium concentrations (30 and 60 mmol·L-1) in the absence and presence of Si (10 mmol·L-1), arranged in a completely randomized design with six repetitions. At 28 days after applying the treatments the dry mass of shoots and roots was evaluated, along with accumulation of silicon and nitrogen in the shoots. The use of silicon resulted in increases in the studied variables, regardless of ammonium concentrations. Silicon reduced the effect of toxicity caused by ammonium excess in maize plants, resulting in greater growth and dry matter accumulation.

关键词： Zea mays L. Beneficial Element Nitrogen Nutritional Disorder Abiotic Stress

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Zebrafish as an in Vivo Screen for Early Black cranberry Proanthocyanidin Biomolecular Activity

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American Journal of Molecular Biology 2014年第2期4卷 37-48页

作者： Nicholas J. Macedo catherine c. neto Anne M. Liberty Tracie L. Ferreira Department of Bioengineering The University of Massachusetts Dartmouth North Dartmouth USA Department of chemistry/Biochemistry The University of Massachusetts Dartmouth North Dartmouth USA

Antioxidants have been widely studied in various naturally occurring substances as a bioavailable cancer prevention treatment. Proanthocyanidins (PAcs), which are abundant polyphenols in Early Black (EB) cranberry (Vaccinium macrocarpon), are readily available and we have shown their anticancer activity in several cancer cell lines. This work focused on the activity of these compounds when incorporated into the zebrafish (Danio rerio) system. We began investigating the in vivo effect of these phytochemicals, the protective role of several other cranberry compounds, and the metabolic activity of the vertebrate model organism. Proanthocyanidin fractions were separated from fresh EB cranberry fruit by chromatography on Sephadex LH-20 in order to acquire a workable stock solution in DMSO. Various concentrations of proanthocyanidins in solution were tested against fish ranging in age from 1-cell stage to adult level of growth. Acridine orange apoptosis indicator dye was incorporated into the treatment protocol, and it was observed that irregular epithelial cell death was occurring in treated embryos but not in the control group. Further apoptosis assays were carried out utilizing Dihydroethidium (DHE) superoxide sensitive dye in the treatment protocol. Fluorescing red nuclei were visible along the outer surface of the epithelium cell layer;an indication of superoxide release within cells leading to the nicking of DNA within the nucleus. It was also possible to screen for superoxide release in PAcs treated ccD-cO18 and HT-29 cells using confocal microscopy and cell apoptosis was investigated by trypan blue cytotoxicity assay;cell apoptosis results were statistically significant as confirmed by ANOVA analysis. Results indicate that the phytochemicals may induce apoptosis in rapidly dividing cells.

关键词： Zebrafish In Vivo Proanthocyanidins

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Strategy Analysis of the Biggest World Supermarkets With Open capital and Which Act in the Brazilian Market

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chinese Business Review 2014年第5期13卷 333-346页

作者： Thyago de Melo Duarte Borges Joao Agra neto Luciano Queiroz de Araujo Junior Jamerson Viegas Queiroz Fernanda cristina Barbosa Queiroz Faimara do Rocio Strauh Nilton cesar lima christian Luiz da Silva Universidade Federal do Rio Grande do Norte (UFRN) Natal Brazil Universidade Tecnologica Federal do Parana (UTFPR) Curitiba Brazil

In view of the growth of retail, as well as the performance of large multinational supermarket branch in the Brazilian market, the purpose of this article is an analysis of the strategies of casino, carrefour, Walmart and open in the country capital, analyzing their operational, financial indicators socio-environmental initiatives. In order to achieve the main objective of this work, the construction of a method that validates this search was necessary. The main strategies and indicators of the three groups in four areas： corporate strategy, operational areo, financial areo, social and environmental issue, were analyzed. Study of qualitative, descriptive and exploratory character is realised through the importance of these three companies worldwide. As conclusions, it has been the expansion strategies that geared toward developing countries, adherence to new store formats, specializing in electronic commerce, as well as the superiority of Walmart （net sales） and casino （Profitability）.

关键词： Strategy Supermarkets Wal-Mart carrefour casino

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Risk factors associated with inflammatory bowel disease: A multicenter case-control study in Brazil

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World Journal of Gastroenterology 2020年第25期26卷 3611-3624页

作者： Valéria cristina Loureiro Salgado Ronir Raggio Luiz Neio Lucio Fernandes Boéchat Isabella Sued Leao Bianca do carmo Schorr JoséMiguel Luz Parente Daniela calado lima Eduardo Santos Silveira Júnior Genoile Oliveira Santana Silva Neogélia Pereira Almeida Andrea Vieira Maria Luiza Queiroz de Bueno Júlio Maria chebli érika Ruback Bertges Luísa Martins da costa Brugnara columbano Junqueira neto Stefania Burjack Gabriel campbell Luana Letiza Discacciati Joao Paulo Silva cézar Tiago Nunes Gilaad G Kaplan cyrla Zaltman Division of Gastroenterology Department of Internal MedicineClementino Fraga Filho HospitalFederal University of Rio de JaneiroRio de Janeiro 21940-230Brazil Institute for Studies in Public Health Federal University of Rio de JaneiroRio de Janeiro 21940-230Brazil Multidisciplinary Research Laboratory Clementino Fraga Filho HospitalInstitute of Thoracic DiseasesFaculty of MedicineFederal University of Rio de JaneiroRio de Janeiro 21940-230Brazil Division of Gastroenterology Department of Internal MedicineHospital UniversityFaculty of MedicineFederal University of PiauíPiauí64049-550Brazil Almeida Division of GastroenterologyInflammatory Bowel Disease Outpatient ClinicRoberto Santos General Hospital(HGRS)of the Bahia State Department of HealthBahia 40110-060Brazil Division of Gastroenterology Inflammatory Bowel Disease Outpatient ClinicIrmandade Santa Casa da Misericórdia of São PauloSão Paulo 01221020Brazil Division of Gastroenterology Department of Internal MedicineHospital UniversityFaculty of MedicineFederal University of Juiz de ForaMinas Gerais 36036-247Brazil Division of Gastroenterology Inflammatory Bowel Disease Outpatient ClinicFederal District Base HospitalBrasília 70330-150Brazil Gastrointestinal Physiology Institute of Nutritional ScienceNestle Research CenterLausanne 1000Switzerland Departments of Medicine and community Health Sciences University of CalgaryCalgary T2N4Z6Canada

BAcKGROUND The etiology of inflammatory bowel disease(IBD)is unknown,but it is believed to be *** hygiene hypothesis proposes that better hygiene conditions would lead to less infectious disease during childhood and favor the development of immune-mediated *** To test the hygiene hypothesis in IBD by assessing the environmental risk factors associated with IBD development in different regions of Brazil with diverse socioeconomic development *** A multicenter case-control study was carried out with 548 crohn’s disease(cD)and 492 ulcerative colitis(Uc)outpatients and 416 healthy controls,from six IBD centers within different Brazilian states at diverse socioeconomic development stages.A semi-structured questionnaire with 87 socioeconomic and environmental questions was *** regression model was created to assess the odds ratio(OR)with P value and 95%confidence intervals(cI).RESULTS Predictive variables for both diseases(cD and Uc)were women[odd ratios(OR)=1.31;OR=1.69],low monthly family income(OR=1.78;OR=1.57),lower number of cohabitants(OR=1.70;OR=1.60),absence of vaccination(OR=3.11;OR=2.51),previous history of bowel infections(OR=1.78;OR=1.49),and family history of IBD(OR=5.26;OR=3.33).Associated risk factors for cD were age(18-39 years)(OR=1.73),higher educational level(OR=2.22),absence of infectious childhood diseases(OR=1.99).The Uc predictive variables were living in an urban area(OR=1.62),inadequate living conditions(OR=1.48)and former smokers(OR=3.36).Appendectomy was a risk factor for cD(OR=1.58)with inverse association with Uc(OR=4.79).consumption of treated and untreated water was associated with risk of cD(OR=1.38)and Uc(OR=1.53),*** This is the first examining environmental exposures as risk factors for inflammatory bowel disease in *** of the variables associated with disease risk support the role of the hygiene hypothesis in IBD development.

关键词： crohn’s disease Ulcerative colitis Risk factors Environmental factors Hygiene hypothesis

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