Análise da Expressão de APE1 após Estresse Oxidativo Induzido em Células Proficientes e Deficientes na Via de Reparo por Excisão de Nucleotídeos

Riboflavin is a vitamin very important in aerobic organisms, as a precursor of many coenzymes involved in the electron transporter chain. However, after photosensitization of riboflavin with UV or visible light, it generates reactive oxygen species (ROS), which can oxidize the DNA. The repair of oxidative lesions on DNA occurs through the base excision repair pathway (BER), where APE1 endonuclease plays a central role. On the other hand, the nucleotide excision repair pathway (NER) repairs helix-distorting lesions. Recently, it was described the participation of NERproteins in the repair of oxidative damage and in stimulation of repair function fromAPE1. The aim of this research was to evaluate the cytotoxic effects of photosensitized riboflavin (RF*) in cells proficient and deficient in NER, correlating with APE1 expression. For this propose, the cells were treated with RF* and it was performed the cell viability assay, extraction of whole proteins, cells fractionation, immunoblotting, indirect immunofluorescence and analysis of polymorphisms of BER gens. The results evidenced that cells deficient in XPA and CSB proteins were more sensitive to RF*. However, XPC-deficient cells presented similar resistance to MRC5- SV cells, which is proficient in NER. These results indicate that XPA and CSB proteins have an important role on repair of oxidative lesions induced by RF*. Additionally, it was evidenced that single nucleotide polymorphisms (SNPs) in BER enzymes may influence in sensitivity of NER-deficient cell lines. Concerning the APE1 expression, the results showed that expression of this protein after treatment with RF* only changed in XPC-deficient cells. Though, it was observed that APE1 is recruited and is bound to chromatin in MRC5-SV and XPA cells after treatment with RF*. The results also showed the induction of DNA damage after treatment with RF*, through the analysis of-H2AX, since the treatment promoted an increase of endogenous levels of this phosphorylated protein, which acts signaling double strand-break on DNA. On the other hand, in XPC-deficient cells, regardless of resistance of RF*, the endogenous levels of APE1 are extremely reduced when compared with other cell lines and APE1 is not bound to chromatin after treatment with RF*. These results conclude that RF* was able to induce cell death in NERdeficient cells, where XPA and CSB cells were more sensitive when compared with MRC5-SV and XPC-deficient cells. This last result is potentially very interesting, since XPC-deficient cell line presents low levels of APE1. Additionally, the results evidenced that APE1 protein can be involved in the repair of oxidative damage induced by RF*, because APE1 is recruited and bound strongly to chromatin after treatment.

Efeitos biológicos in vitro de agentes físicos utilizados em fisioterapia

The uses of radiobiocomplexes labeled with technetium-99m contributed to health science advances. Stannous chloride (SnCl2) has been used as a reducing agent for the labeling process. Cytotoxic and genotoxic effect of the SnCl2 have been described in several studies and with this experimental models alterations in molecular and cellular level can be evaluated. In the last years the physicals therapists acquired new devices which emits electromagnetic radiation such us Extremely Low Frequency Pulsated Electromagnetic Fields (E.L.F. P.E.M.F.), radiofrequency, Intense Pulsed Light (I.P.L.) and others which emits sonic waves such us Biorresonance. Scientific evidence of the effects and dosage is important to protect public health and to reach exposition levels that result in significant biological effects. The aim of this project is to verify the effects of these physical agents in plasmid DNA and E. coli AB1157 cultures in presence or absence of SnCl2 and the effects in blood constituents labeled with technetium-99m. Wistar rats blood was exposed to the cited sources and the labelling of blood constituents with 99mTc was carried through. Cultures of E. coli AB1157 and plasmidial samples DNA had been also exposed the physical agents. The results suggest that these agents are capable of altering neither the survival of E. coli cells or plasmid DNA electrophoresis mobility. The multidiscipline character was clearly in this study due the interaction between Nuclear Medicine department of the UERJ and the Laboratory of Physical Agents of the Maimonides University in Argentina until the union between the teacher (biomedical and physiotherapist) and student (physiotherapist), besides collaborators of the area of Physics and Biology, promoting new ideas and perspectives and also adding the knowledge of different areas and origins

Processo de desidratação da alga Gracilaria birdiae beneficiada em laboratório e artesanalmente: análise da atividade antioxidante e composição centesimal

The main aim of this study was to compare the procedure for dehydration of Gracilaria birdiae prepared handmade and laboratory, collected in the northern coast of Rio Grande do Norte. The sample was collected in the Rio do Fogo beach in march 2009. The sample collected followed by two processing, the first the material prepared in laboratory was air-dried at 50°C for 24 hours in air-flow oven. The second the handmade sample was air-dried on the sun during three days. The extract was prepared in three different solvents: ethanol, hydroethanol and water, resulting in ethanol, hidroethanol and aqueous extracts from handmade and laboratory sample. In according with results only the ethanol extract was fractionated yielding the fractions hexane, dichloromethane and ethyl acetate fractions. The different process to obtain Gracilaria birdiae resulted in the samples with different shades. The soluble solids content was higher in the laboratory sample. The chemical composition the both samples were characterized by presenting a considerable amounts of carbohydrates, with amior percentage protein and ash, respectively, in the handmade and laboratory sample. In two samples showed a low content of lipids and the lipid profile showed a higher proportion of monounsaturated fatty acids, with the absence polyunsaturated handmade sample. The phytochemical screening by chemical reactions showed the presence of flavonoids, tannins, alkaloids and saponins the laboratory sample, presenting a greater diversity of bioactive compounds. Through of the analysis by thin layer chromatography was possible to identify the phytosterols β-sitosterol and stigmasterol the both samples, also suggest the presence of β-carotene and chlorophyll α the laboratory sample. The levels of total phenolics and flavonoids were more significant in the ethanol extract of the laboratory sample. The in vitro lethality showed that extracts of the laboratory sample and handmade from 125 to 500 μg/ mL, respectively, were highly lethal. In the evaluation of antioxidant capacity by the system β-carotene/ácido linoleic method and by DPPH radical scavernging assay, the ethanol extract from the laboratory process showed significantly greater activity than the other extracts, being and the first and second methods, respectively, lower and equivalent to the synthetic antioxidant BHT. The handmade ethanol extract has not demonstrated skill in deactivating free radicals, but showed activity in inhibiting lipid peroxidation, although the values were significantly lower than the laboratory sample. We conclude that the dehydration process in the laboratory is the most efficient technique to maintenance of the chemical composition present in the seaweed, providing beneficial properties such as antioxidant capacity. We emphasize that this property can be explored with the objective of adding commercial value to the final product, which will promote the expansion of production of this seaweed in the community of Rio do Fogo

Testosterona e gonadotrofina coriônica humana estimulam a esteroidogênese em células da granulosa de folículo pré-ovulatório de égua?

Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)

Quantificação de células CD34+ do sangue do cordão umbilical de cães

Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)

A revenda de combustível e os limites constitucionais para a sua regulação: uma análise aplicada aos aspectos concorrênciais e ao controle de preços

The gas retail represents the end of a section of the oil and natural gas derivative chain, for it is at this stage where the commercialization of those merchandises takes place towards the costumers. This process involves an enormous amount of economic agents, which reflects on an activity of great influence on the citizen's everyday. By the time of the gas retail price liberalization, in 2002, there were great expectations towards that measure, for the insertion of that segment in a competitive market was likely to create a decrease in prices. As there was not a drastic drop off in cost, the question was no longer the price itself, but, predominantly, the conduct taken by the economic agents that operate the market. Not in vain, the segment introduces a greater number of different procedures combined with the organs that compose the Brazilian System of Competition Protection. What is understood, however, is that many of these complaints are made in a lightly way, without a proper analysis of the market and its practices, that being why, in this paper, evidences the causes of these complaints and explained what, in fact, occurs in this market. Also, the organs that protect the free initiative in the sector use different methods to assess anticompetitive practices, which are counterproductive on the combat of anticompetitive practice, that being why the present paper analyzes the used methods on a critic perspective, choosing one which is believed to be the most adequate. The present work also tries to present the gas retail prices on a constitutional, free competition, free initiative and consumers defense perspective, analyzing the competition s aspects on the gas market; the shaping of the gas prices; the market boundaries; the anti-competitive practices under the gas market; and analyze the possibility, according to the defined economic standards in the constitutional text of existing a greater control or gas price indexing and/or regulation which limits the distributors and resellers profit on gas. Still, in consequence of this analysis, a study on Natal s market behavior will be developed in its competitive feature. That being said, moreover being a theoretical-descriptive study, data and statistics gathered is used, which will lead, willing to grasp an experiential study on a few aspects of the Potiguar gas retail market

Responsabilidade civil como função social na distribuição e revenda de combustível face ao direito do consumidor e ao meio ambiente

The civil liability of the distribution and retail fuel stems from the fact business activity developed to be high risk and can be said as risk inherent or latent danger that has predictability and normality characterized by placing the consumer in a position of vulnerability, such as the environment, both public policies defined and constitutional protection. Consumer protection as a fundamental right and the environment as the primacy of social order aims the welfare and social justice, as inserted right to the third dimension, are guarded by the State when it creates operational standards through public policies and indirectly intervenes in the economic order. This intervention is due to consumer protection and the environment are economic order principles, founded on free initiative and free competition, ensuring everyone a dignified existence which underlies the irradiation of fundamental rights in private initiative, before the commercial evaluation, mass consumption, the emergence of new technologies that link consumers to the environment before the protection of life, health and safety and ensuring a better quality of life for present and future generations. To repair this damage and the right to information are provided as fundamental rights that put the person at the epicenter of the relations and collective interest stands out against individual interests that to be done need public-private partnership. In such a way that the used methodology was an analysis of documents correlating them with bibliographic sources whose goals are to recognize the civil responsibility as limit to subjective right, having to develop a social function where guilt and risk grow distant and the damage is configured as a consequence of inherent risk, requiring the State interventional postures in fulfilling its public policy; prevailing in these risky activities the solidarity of those involved in the chain of production and socialization of damage forward those are provided directly of products of hazardous nature that put in a position of vulnerability the environment and the consumer

Efeitos da tricostatina A sobre a acetilação de histonas, proliferação celular e diferenciação de células tronco embrionárias murinas

Background: Embryonic stem cells are cells derived from early-stage embryos that are characterized by pluripotency and self-renewal capacity. The in vitro cultured murine embryonic stem cells can indefinitely propagate in an undifferentiated state in the presence of leukemia inhibitory factor (LIF). However, when stimulated, these cells can differentiate into cell lines derived from all three embryonic germ layers. The trichostatin A (TSA) is an epigenetic modifier agent and several studies have used the TSA to stimulate cellular differentiation. However, most of these studies only assessed one TSA concentration. Therefore, this study aimed to evaluate the effects of different TSA concentrations on histone hyperacetylation during in vitro cell differentiation of murine pluripotent embryonic stem cells, cultured with or without LIF, in the quest of to standardize their application on early cultures of embryonic stem cells.Materials, Methods & Results: Undifferentiated murine embryonic stem cells were plated in the presence of different TSA concentrations (0 nM, 15 nm, 50 nM and 100 nM) in the presence or absence of LIF. Thus, the treatments were evaluated in undifferentiated embryonic stem cells cultured in the presence of LIF (Control group: 0 nM LIF(+); Group 15 nM LIF+; Group 50 nM LIF+ and Group 100 nM LIF+), and in embryonic stem cells cultured in the absence of LIF (Control group: 0 nM LIF; Group 15 nM LIF(-); Group 50 nM LIF(-) and Group 100 nM LIF-). Treatment with TSA was performed for 24 h. After that the medium was replaced with fresh medium without TSA. Samples were collected at 0, 12, 24, 36 and 48 h after the beginning of the experiment. Three replicates were performed in each experimental group. The relative amount of Histone H3 lysine 9 acetylation was analyzed in all groups, as well as the cell proliferation in the embryonic stem cells cultured in the presence of LIF. In the control group (0 nM), the absence of LIF resulted in higher levels (P < 0.05) of H3lys9ac compared to the cultures supplemented with LIF. In the embryonic stem cells cultured in the presence of LIF, the 50 nM and 100 nM treatments resulted in higher levels (P < 0.05) of H3lys9ac when compared with 0 nM and 15 nM treatments. Evaluating the Hoechst area in the 0 nM group, it was observed that the number of cells increased (P < 0.05) according to the time of culture. Treatment with 15 nM also reflected a similar distribution, but the Hoechst area in 15 nM group was lower (P < 0.05) at 24 and 48h when compared to the observed in the control group. In the 100 nM treatment, was observed that the area of Hoechst was lower (P < 0.05) to that obtained in the control group at 12, 24 and 48h. In addition, it was observed that treatment with TSA induces greater cellular differentiation when compared to control groups in stem cells cultured in the presence of LIF as well as in the absence of LIF.Discussion: In the present study it was observed that TSA treatment increased the levels of histone acetylation in murine embryonic stem cells at a 50 nM concentration, making it possible to reduce the concentration recommended in the literature (100 nM). In addtion, it was concluded that the lower TSA concentrations utilized (15 nm and 50 nM) was less harmful to cellular proliferation than the 100 nM TSA concentration.

Implante de tubo de silicone com e sem colágeno na regeneração de nervos em eqüinos

Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)

Expressão da E-caderina em carcinoma de células escamosas e no tumor de células basais de cães

Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)

Entre orquídeas e girassóis :o laboratório de Enfermagem na visão de estudantes

To study the teaching/learning process about the Nursing procedures carried out in the laboratory, and learn both the sapiens and the demens dimensions of such process, is the main purpose of this study. The objectives are to: identify the major laboratory contributions to the teaching/learning process from the point of view of undergraduate students and the feelings they express; describe the difficulties they have identified; and analyze the relevance of the laboratory to this process. As part of the inquiry procedure, four core group meetings were held with 26 undergraduate students who had completed the course on Semiology and Semiotics in Nursing, which is the course where the Nursing laboratory is most needed as a learning space. The analysis, based on a qualitative approach, had as fundamental theoretical support studies made by Friedlander and Hayashida, who deal with learning/teaching in the Nursing laboratory, and by authors who favor humanization in teaching such as, among others, Freire, Maturana, Morin, Assmann. Results point toward the relevance of the Nursing laboratory as a facilitator for the learning/teaching process. In their speech the students repeatedly state that the development of procedures in simulated situations enable them to become more self-assured and technically prepared for caring. In addition, they emphasize that feelings such as fear, lack of confidence, anxiety, anguish and panic become diminished at the time of their clinic experience when they have had previous learning in the laboratory. They have also acknowledged that some difficulties of structural nature have become obstacles to a high-quality learning development. In summary, in spite of the difficulties that have been pointed out by the students concerning the use of the Nursing laboratory in the learning/teaching process, they also recognize that this is the locus par excellence where they can develop their skills and appease their anxieties

Um estudo da aplicação do conceito de risco na conscientização e conhecimento de estudantes de ensino técnico sobre riscos de segurança e saúde ocupacional em laboratório de soldagem

This dissertation presents a discussion on the level of risk awareness of students who have welding practice in the laboratories at CEFET/PI, both in Teresina and Floriano. Its main goal is to investigate risk factors involving students, teachers and employees that work in these laboratories. It s an empirical survey, held among a certain amount of students from the course in Mechanic Technology. For data analysis, the concept of risk was compared to the students direct measure of perception. The main results suggest the students must be better informed, more risk aware and more competent, in order to avoid accidents. They also point to a strong need for a more formal and effective performance to assure full consciousness about the risks involving the welding practice.

Conversor CC/CA de alta freqüência baseado em inversores ressonantes com comutação seqüencial para excitação de uma tocha indutiva a plasma térmico

This work describes the study, the analysis, the project methodology and the constructive details of a high frequency DC/AC resonant series converter using sequential commutation techniques for the excitation of an inductive coupled thermal plasma torch. The aim of this thesis is to show the new modulation technique potentialities and to present a technological option for the high-frequency electronic power converters development. The resonant converter operates at 50 kW output power under a 400 kHz frequency and it is constituted by inverter cells using ultra-fast IGBT devices. In order to minimize the turn-off losses, the inverter cells operates in a ZVS mode referred by a modified PLL loop that maintains this condition stable, despite the load variations. The sequential pulse gating command strategy used it allows to operate the IGBT devices on its maximum power limits using the derating and destressing current scheme, as well as it propitiates a frequency multiplication of the inverters set. The output converter is connected to a series resonant circuit constituted by the applicator ICTP torch, a compensation capacitor and an impedance matching RF transformer. At the final, are presented the experimental results and the many tests achieved in laboratory as form to validate the proposed new technique

Projeto e controle de um sistema de aquecimento no Laboratório de Avaliação de Medição em Petróleo

The Oil Measurement Evaluation Laboratory (LAMP), located in the Federal University of Rio Grande do Norte (UFRN), has as main goal to evaluate flow and BS&W meters, where the simulation of a bigger number of operation variable in field, guarantees a less uncertain evaluation. The objective of this work is to purpose a heating system design and implementation, which will control the temperature safely and efficiently in order to evaluate and measure it. Temperature is one of the variables which influence the flow and BS&W accurate measurement, directly affecting the fluid viscosity and density in the experiment. To project the heating system it is of great importance to take the laboratory requirements, conditions and current restrictions into consideration. Three alternatives were evaluated: heat exchanger, internal resistance and external resistance. After the analyses are made in order to choose the best alternative for the heating system in the laboratory, control strategies were determined for it, PID control methods in combination with fuzzy logic were used. Results showed a better performance with fuzzy logic than with classic PID

Sistemas de automação e supervisão do laboratório de avaliação dos processos de medição de vazão e BSW

During a petroleum well production process, It is common the slmultaneous oil and water production, in proportion that can vary from 0% up to values close to 100% of water. Moreover, the production flows can vary a lot, depending on the charaeteristies of eaeh reservoir. Thus being, the meters used in field for the flow and BSW (water in the oil) measurement must work well in wide bands of operation. For the evaluation of the operation of these meters, in the different operation conditions, a Laboratory will be built in UFRN, that has for objective to evaluate in an automatic way the processes of flow and BSW petroleum measurement, considering different operation conditions. The good acting of these meters is fundamental for the accuracy of the measures of the volumes of production liquid and rude of petroleum. For the measurement of this production, the petroleum companies use meters that should indicate the values with tha largast possible accuracy and to respect a series of conditions and minimum requirements, estabelished by the united Entrance ANP/INMETRO 19106/2000. The laboratory of Evafuation of the Processes of Measurement of Flow and BSW to be built will possess an oil tank basically, a tank of water, besides a mixer, a tank auditor, a tank for separation and a tank of residues for discard of fluids, fundamental for the evaluation of the flow metars and BSW. The whole process will be automated through the use of a Programmable Logicat Controller (CLP) and of a supervisory system.This laboratory besides allowing the evaluation of flow meters and BSW used by petroleum companies, it will make possible the development of researches related to the automation. Besides, it will be a collaborating element to the development of the Computer Engineering and Automation Department, that it will propitiate the evolution of the faculty and discente, qualifying them for a job market in continuous growth. The present work describes the project of automation of the laboratory that will be built at of UFRN. The system will be automated using a Programmable Logical Controller and a supervisory system. The programming of PLC and the screens of the supervisory system were developed in this work