Características da base de conhecimentos de professores no ensino de biologia celular a partir de um curso de formação continuada

Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES)

Indução de morte celular por apoptose decorrente da irradiação com Raios-X: um ensaio com células mamárias humanas tumorais

Apoptosis is a form of programmed cell death selectively removes abnormal cells, and thus contributes to maintaining the balance of the dynamics of cell reproduction. Therefore the verification of the occurrence of apoptotic cell death after a pathological stimulus is crucial for the analysis of the maintenance of normal cell cycle of a given tissue or organ. In this experiment were used cells lines human mammary tumor MDAMB231, T47, MCF7, which were irradiated with X-rays at a dose of 5 Gy in a time interval of 15 seconds, and filtration of 1mm aluminum. Samples containing the cells were grown in a specific culture medium, containing fetal bovine serum and growth factor, and two samples were prepared with each of the cell lines, one to be irradiated, and another that has not been irradiated, which denoted by negative control of the irradiation. The primary goal of the experiment was to verify and compare the rates of apoptosis in each cell lines, in which were irradiated and that were not irradiated, using flow cytometry as a method for detecting apoptotic cell death in together with specific markers annexin V and propidium iodide. Data from the readings made by flow cytometry were analyzed and interpreted using the software WinMDI statistical graph. By comparing the indices relating to the readings of positive and negative for specific markers of apoptosis, based on differences in the statistical data presented lectures regarding the cellular irradiated and not irradiated, collude cells in question once... (Complete abstract click electronic access below)

Influência da atividade de forragem, números de adultos e de larvas, na diferenciação de castas de Mischocyttarus cerberus styx Richards, 1940 (Hymenoptera, Vespidae, Mischocyttarini)

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Diferenciação hemogasimétrica, hemoconcentradora e da glicemia em testes de exaustão muscular usando cinco, quatro ou três exercícios

Em trabalho anterior, observou-se que diferentes grupamentos musculares submetidos ao teste de exaustão (TE) com pesos respondem semelhantemente com hemoconcentração, acidose e aumento da glicemia, entretanto, com variações grupomuscular dependentes. Nesse estudo, é comparado o efeito do impacto provocado pelo TE envolvendo, conjuntamente, 5, 4 ou 3 exercícios por meio da análise desses marcadores bioquímicos. Foram estudados 21 indivíduos do sexo masculino (20-32 anos) voluntários, familiarizados com treinamento em musculação, estratificados em três grupos de acordo com o número de exercícios: grupo 5 ( n= 6) 5 exercícios (Extensão do cotovelo, desenvolvimento pela frente, supino reto, levantamento terra e agachamento), grupo 4 (n= 7) 4 exercícios (Supino reto, rosca direta com barra, agachamento hack, tríceps no pulley), grupo 3 (n= 8) 3 exercícios (Agachamento hack, supino reto, rosca direta ) . Todos foram submetidos à sobrecarga inicial de 80% de uma repetição máxima (1RM), até a exaustão, em seguida, com auxílio externo e sem interrupção do exercício, a carga foi reduzida em 20% do peso, até a impossibilidade de continuidade de realização do exercício com a carga mínima (20% de 1RM). O sangue coletado em veia cubital antes (At) e imediatamente após (Ap) o TE foi analisado para indicadores hemogasimétricos (pH, pO2, pCO2, HCO- 3), hemoconcentração (Ht) e glicemia. O efeito do TE sobre cada variável (At/Ap) foi avaliado comparando-se os grupos por meio de ANOVA-medidas repetidas para significância de p=0,05. Houve significância (p<0,05) na elevação do Ht, glicose e pO2 e redução do pH, PCO2 e HCO- 3, Entretanto houve diferença significativa entre os grupos apenas em relação ao HCO- 3, no qual este apresentou maior variação no G4 em relação ao demais. Assim, esses marcadores sistêmicos respondem a exaustão muscular de modo semelhante ao envolvimento de 3,4 ou 5 grupamentos musculares

Análise da expressão protéica de moléculas de adesão celular em melanomas cutâneos

Cutaneous melanoma is a malignancy caused by the uncontrolled proliferation of melanocytes. Has been increasing in incidence and mortality over the years. This type of tumor may arise in the skin or any other place where there was migration of neural crest cells. Melanocytes are located in the basal layer of the epidermis forming the epidermal melanin unit, involving a melanocyte to 36 keratinocytes. The proliferation of melanocytes depends on this relationship with the keratinocytes, gap junctions, E-cadherin and desmoglein. The basal membrane is responsible for promoting the integrity of the skin, their adhesion structures shown to be altered in several skin diseases. Some molecules that play the role of adherence in this membrane are the laminins, type IV collagen and type XVII and perlecan. Analysis of protein expression of cell adhesion molecules in melanoma by immunohistochemistry on tissue microarray (TMA). We selected paraffin blocks of patients who were diagnosed with cutaneous melanoma in the period 1995 to 2010, reaching a sample of 124 cases. All these have gone through review of preparation for subsequent histological analysis of tissue microarray (TMA). The protein expression will be analyzed by immunohistochemistry, using markers anti-laminin, anti-collagen type IV and type XVII and anti-perlecan. There was a predominance of female patients, Caucasians and southeastern Brazil. There were frequent in type of extensive superficial ulcers with Breslow depth greater than 4 mm, Clark level IV and mitotic index greater than 6 mitoses per cga. The patients, most of them were female with mean age of 57.13 years and Caucasian. The most frequent histological type was extensive superficial followed by acral lentiginous. Predominated cases very deep, with Breslow thickness greater than 4 mm, Clark IV, ulcerated and on average by 9 cga mitosis, leading to worse prognosis in this sample. It...(Complete abstract click electronic access below)

Desenvolvimento de equipamento LED para aplicação em cultura celular

O estágio realizado em parceria entre o Departamento de Física e Biofísica do IBB-UNESP e o Hemocentro de Botucatu da FMB-UNESP teve como objetivo desenvolver um novo equipamento de luz LED capaz de interagir com culturas celulares. Desta forma foi desenvolvido um equipamento com luz azul e luz vermelha. A luz azul é responsável por inibir o crescimento celular, a luz vermelha é responsável por aumentar o crescimento celular. O equipamento consiste de uma fonte, e dois conjuntos com três LEDs cada. A cultura celular escolhida foi a de Miemola Murino (NS1), uma linhagem que possui alta taxa de crescimento e que foi mantida em estufa e meio de cultura, (simulando condições normais de sobrevivência), sendo irradiada três vezes ao dia. As amostras foram coletadas e contadas em câmara de Neubauer. Os resultados obtidos, quando comparados com o controle, não apresentaram significativa diferença de crescimento quando irradiados com luz LED vermelho. Porém quando comparados o grupo controle com o grupo irradiado com LED azul, notou-se uma significativa redução da quantidade de células a partir do sétimo dia de irradiação. Acredita-se que a ineficácia dos LEDs vermelhos ocorreram pois utilizamos células com metabolismo bastante elevado, dificultando um aumento ainda maior da taxa de crescimento. A redução da população de células azuis ocorre pois a radiação interage com a membrana celular, causando a quebra dessa membrana e conseqüente morte da célula. Outra via para a morte é a lesão causada no DNA das células, que ocorre quando a luz azul é absorvida pela molécula

Diferentes métodos de diagnóstico molecular na diferenciação das espécies de Sarcocystis spp

The Sarcocystis genus includes obligatory two-host life cycle protozoan parasites. It is the most numerous of the six genera of the Sarcocystidae family. The infection caused by parasites of this genus is a zoonotic and cosmopolitan disease known as sarcosistosis or sarcosporidiosis. The sarcositosis though frequently asymptomatic in its definitive hosts can be fatal in its intermediate hosts. The usual diagnoses of sarcosistosis takes place through a histological demonstration of schizonts in blood vessels and organs, and the presence of cysts in muscle tissue by necropsy or biopsy, this second method still more common and based on morphological features of the sarcocyst. However, these methods can be inadequate to a precise identification of the infector species once that, besides the genus being of numerous species, these often present similar morphological features. Another factor that makes the diagnostic more difficult is the non specificity of some Sacocystis species to their hosts. Consequently, molecular diagnostic methods have been used in order to identify the infector species and the parasite specific biological cycles, demonstrating also new species and coevolutive aspects between parasite and host. Among the most employed molecular techniques the Polimerase Chain Reaction (PCR), the nested-PCR and the Restriction Fragment Length Polymorphism (RFLP) stands out

Cultivo celular in vitro: importância para a pesquisa biomédica e dimensão da problemática de autenticação de linhagens celulares

Experimental models composed by human and animal cell lines are simplified and informative, allowing them to be widely used for biomedical research. Most laboratories that use in vitro cultivated cells maintain a variation of cell lines stored and cultivated. Therefore, misidentification and cross-contamination events can happen during cell lines handling. This problem can generate a repertoire of dubious results and papers, which may prejudice biomedical research. Recently it was created the International Cell Line Authentication Committee (ICLAC), which aims to spread knowledge about cross-contamination and misidentification of in vitro cell lines. Despite of the efforts spent trying to aware scientific community about the importance of the correct identification of cells, the number of papers based on misidentified cell lines it´s still worrying, compromising the reliability of out coming results and conclusions regarding them. The present study aims to analyze and discuss the main advantages and limitations of eukaryote in vitro cell lines use, characterizing the cell lines authentication problems. Therefore, compilation and critical analyses of literature data was realized, aiming to improve the understanding about this subject. Based on information about 445 cell lines with issues published by ICLAC it´s clear that contamination in human cell lines represented 89,2 % of mentioned problems. HeLa cell line was the responsible for most contamination, especially in 92 normal tissue cell lines, representing 44,6% of the contamination. These results reinforce the importance of periodic maintenance of cell lines cultures by labs and implementation of authentication methods as polymorphic STRs, besides obtaining cell lines from reliable sources and cell banks

Influência da luz LED na cultura celular de Mieloma múltiplo murino–NS1 in vitro

A monoclonal antibody (mAb) is an important tool in medical biotechnology and the production of biopharmaceuticals, especially for disease diagnosis and treatment of infections, because the antibodies have a significant advantage over chemical agents used in conventional therapies . The last thirty years the technology of production of monoclonal antibodies developed mainly the technique of obtaining in vitro, but also of their production is laborious, the cost is high. A major element of the high cost of production is the fact that the long-term culture consumes a large amount of imported inputs with high added value. A major contribution of this work is to promote cell growth more quickly and efficiently. Currently, a great race to discover new technologies and techniques to synthesize new antibodies and significantly increase the production of murine mAbs. New technologies such as laser and LED are innovations and widespread in modern life, so much so that its use has proliferated worldwide, primarily in the medical field. Recent studies show a series of results from the influence of the LED light in biological tissues such as: increasing the rate of cell proliferation, increased production rate of fibroblasts, increasing the rate of synthesis of RNA and DNA synthesis of ATP, etc. To assess the contribution of the LED in the culture of Myeloma NS1murino compared to the standard procedure. - NS1 cells were provided and followed the criteria of culture medium of the Laboratory of Cellular Engineering Center of Botucatu (POPs). The same amount of cells was grown in bottles of 25 cm2 polystyrene Tissue Culture Treated, specifically marked and kept in special medium RPMI 1640 Gibco BRL  supplemented with fetal bovine serum 10%, essential amino acids and non-essential, glucose, insulin and antibiotics. It was used in LEDs Cromatek wavelength of 630nm, 475nm and 530nm. The groups were... (Complete abstract click electronic access below)

Avaliação da proliferação celular na próstata de gerbilos vasectomizados, após indução química pela injeção intraperitoneal do carcinógeno N-metil-N-nitrosuréia (MNU)

Prostate cancer is the second leading cause of cancer death in the country. Due to this factor, the interest of showing what relationship exists between vasectomy surgery and the incidence of prostate carcinoma has started. Some epidemiological studies showed an increased risk of prostate cancer in vasectomized men (Emard et al., 2001). On the other hand other authors have argued that there is no correlation (Patel et al., 2005) and there are those who said that vasectomy is linked to reduced risk of prostate cancer (Ross, 1983). Faced with an analysis of published works, such discussion remains today. The vasectomy, or deferentectomia, is a contraceptive male method, which is the section of the vas deferens of man by preventing the sperm from being expelled along with the seminal fluid during ejaculation, and is one of the most simple, economical, uncomplicated post- operative. This study aims to evaluate the process of cell proliferation and to evaluate immuno-histochemically the expression of specific markers of PCNA (Proliferating Cell Nuclear Antigen) and Ki-67, in the prostate of the gerbil after chemical induction by intraperitoneal injection of the carcinogen N-methyl -N-nitrosourea (MNU), because the regulation of the functional balance between cell proliferation and apoptosis is associated with hyperplasia and prostatic carcinoma. The experimental procedure was performed at the Laboratory of the Department of Anatomy, along with collecting the data for later analysis

Interferência eletromagnética em equipamentos de diatermia por microondas por aparelhos telefonia móvel celular GSM

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Diferenciação miogênica “in vitro” de células tronco mesenquimais murinas de diferentes origens

Muscular dystrophy refers to a group of more than 30 genetical disorders characterized by progressive weakness and degeneration of the skeletal muscle. No effective therapy is available at present. Recent studies have reported that the transplantation of stem cells can offer an important potential therapy for genetic diseases. Adult bone marrow mesenchymal stem cells have been identified as a nonhematopoietic stem cell population capable of self-renewal with the ability to differentiate into many cell lineages, including bone, fat, cartilage and connective tissue. Because of their similarity with muscle progenitor cells, when they are injected in affected individuals, they are able to migrate into areas of skeletal muscle degeneration and participate in the regeneration process. The adipose tissue represents an alternative source of MSCs that, as the MSCs derived from bone marrow, are capable of in vitro differentiation into osteogenic, adipogenic, myogenic and chondrogenic lineages. The objective of this project is to investigate the “in vitro” myogenic potential of mesenchymal stem cells derived from murine bone marrow and adipose tissue. Four experimental groups were analyzed: mice from lineages Lama2dy-2J/J and C57black and, C2C12 lineage cells and transformed C2C12 expressing the eGFP protein. MSCs cultures were obtained by flushing the bone marrow femurs and tibials with α-MEM or by the subcutaneous and inguinal fat from the mice. Their characterization was done by flow cytometry and in vitro differentiation. Muscle differentiation was studied through the analysis of the expression of transcriptional factors involved in muscle differentiation and/or the presence and amount of specific proteins from muscle differentiated cell. The pluripotency from bone marrow MSCs of the two lineages was evidenced and, in the muscular differentiation... (Complete abstract click electronic access below)

Proliferação e Morte Celular (Apoptose) avaliadas em ratos Lewis após tratamento com mistura de praguicidas em baixas doses

The use of pesticides in Brazil has intensified over the years. And since 2009 it was ranked as the largest market for pesticides worldwide. The consequential diffuse contamination of the environment is therefore due to the increasing number of chemicals of different classes, origins and modes of action. Little is known about the action of pesticides on human health in situations of co-exposure. Possible toxic effects are not restricted to agricultural and industrial workers, but also the general population that may be exposed continuously to its residues in food and water. Although these pesticides are mostly present in the environment at low doses, it must be considered that possible cumulative or synergistic effects may occur when there are concurrent or sustained exposure for two or more of these agents, which can lead to late manifestation of subclinical damages, sometimes irreversible. Thus, the specific objective of this study was to assess the effect of carcinogenesis promotion of a mixture of pesticides at low doses and analyze the phenomena of cell proliferation and apoptosis in rat liver. A total of 50 male Lewis rats was separated into 5 groups for 8 weeks in a medium term hepatocarcinogenesis model. The three different classes of pesticides (dieldrin, dicofol, endosulfan, dichlorvos and permethrin), whose residues were detected by ANVISA during the period from 2001 to 2005 in tomatoes cultures, were added to the feed of rats initiated to hepatocarcinogenesis with diethylnitrosamine (DEN- 200mg/kg ip). We used two different mixtures, one with no toxic effects at doses (MEX1) referring to the NOEL (no-observed-effect level) and another at doses LOEL / LEL / LOAEL (Lowest-observed-effect level / Lowest-effect level / Lowest -observed-adverse-effect level), to the installation of adverse effects (MEX2), derived from chronic studies. All animals ...(Complete abstract click electronic access below)

Avaliação da atividade antiproliferativa dos extratos de Croton urucurana Baillon

O termo câncer refere-se a um conjunto de doenças que atingem e matam milhões de pessoas em todo o mundo. Esta doença é caracterizada pela proliferação descontrolada de células que não respondem mais aos estímulos externos e internos que controlam a proliferação, diferenciação e morte celular. O aumento da expectativa de vida da população é um dos fatores que contribuem para a crescente incidência de câncer no mundo já que o desenvolvimento desta doença está relacionado com o envelhecimento do organismo. O câncer é a segunda maior causa de morte na população mundial e diante de tal índice, fica clara a necessidade de desenvolvimento de novos quimioterápicos. Cerca de 60% das drogas anticâncer são derivadas de produtos naturais, demonstrando assim o grande potencial destes produtos no desenvolvimento de novas drogas anticâncer. Este projeto teve como objetivo avaliar a atividade anticâncer dos extratos brutos e frações obtidas da espécie vegetal Croton urucurana Bailon. Os extratos brutos diclorometânico (EBD) e etanólico (EBE) foram avaliados em ensaio de citotoxicidade in vitro em cultura de células tumorais humanas. Os dois extratos apresentaram atividade in vitro, porém somente o EBD foi submetido ao processo cromatográfico de fracionamento, sendo as frações obtidas deste processo submetidas ao teste antiproliferativo in vitro. O EBD também foi utilizado para avaliar sua atividade em modelo de tumor sólido de Ehrlich. Neste teste, esse extrato apresentou inibição do crescimento tumoral apenas com a menor dose. Como o EBE também apresentou atividade antiproliferativa in vitro, é importante dar continuidade aos estudos com este extrato, com o objetivo de avaliar sua atividade em modelos experimentais in vivo

Efeito de reguladores vegetais no desenvolvimento de plantas jovens de Tabebuia avellanedae Lorentz ex Griseb

Tabebuia avellanedae Lorentz ex Griseb (ipê-roxo), Bignoniaceae, ocorre na Floresta Tropical Atlântica e é de grande importância, uma vez que, devido às suas propriedades medicinais e lenho de grande valor comercial, já esteve sob ameaça de extinção. A mistura de reguladores vegetais é conhecida como bioestimulante que pode incrementar o desenvolvimento vegetal, estimulando a divisão, a diferenciação e o alongamento celular. Assim, neste trabalho avaliou o efeito da aplicação da mistura de reguladores vegetais do grupo das auxinas, giberelinas e citocininas no desenvolvimento de plantas jovens de ipê-roxo. Os tratamentos foram: testemunha, GA4+7 + BAP 100 mg L-1, GA4+7 + BAP 200 mg L-1, GA3 + IBA + cinetina 0,5% e GA3 + IBA + cinetina 1,0% os quais foram aplicados 4 vezes em intervalos de 30 dias. O efeito dos tratamentos foi avaliado pelas medidas de altura das plantas, diâmetro do caule, número de folhas, massa seca das folhas, caule e raízes e avaliação de trocas gasosas. O tratamento com GA3 + BAP 200 mg L-1 foi aquele que apresentou maior efeito no crescimento das plantas. Já para as avaliações fotossintéticas o tratamento com GA3 + IBA + cinetina foi aquele que apresentou a maior taxa de assimilação de CO2