Avaliação da indução de apoptose pela desintegrina recombinante DisBa-01 em macrófagos e células tumorais

O câncer é causado pela proliferação descontrolada de células, demonstrando uma capacidade coletiva de invasão e metástase. Durante a metástase, as células malignas precisam resistir a anoikis, uma apoptose celular gerada por falta de adesão. No câncer, as integrinas influenciam as células do hospedeiro associadas ao tumor bem como as próprias células tumorais, tendo o potencial de modular a progressão tumoral, a sobrevivência celular, a invasão e a metástase. A integrina αvβ3 é expressa em diversos tumores humanos, mas está em níveis muito reduzidos ou mesmo ausente nos tecidos normais, sendo considerada um alvo privilegiado na terapia anti-tumoral. Células derivadas da medula óssea, como o monócito/macrófago, também expressam esta integrina, ainda que em níveis reduzidos. A desintegrina recombinante DisBa-01 atua principalmente sobre as integrinas αvβ3 e αIIbβ3, e parece possuir a capacidade de inibir a adesão celular de linhagens possuindo a integrina αvβ3 à vitronectina. Sendo assim, observou-se a apoptose que pode ser gerada pela inibição da adesão celular ou pela própria presença da desintegrina, um antagonista de integrina que afeta a adesão celular, em linhagem neoplásica e em linhagem imortalizada de macrófagos, utilizando os métodos de marcação com anexina V e técnica de TUNEL, bem como a presença de células viáveis através da técnica de MTT. As linhagens celulares utilizadas nesse experimento foram expostas a desintegrina DisBa-01 por 24 horas para realização dos testes e as populações celulares aderentes e não aderentes foram analisadas separadamente. Verificou-se uma diminuição na viabilidade celular quando ocorre perda de adesão e na presença da proteína, mas foi observado um resultado não significativo para a ocorrência de apoptose. A desintegrina DisBa-01 ocasiona uma diminuição na viabilidade celular, entretanto parece não ser por apoptose gerando anoikis

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 atividade citotóxica de espécies vegetais em linhagens celulares humanas

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

Redução bacteriana via terapia fotodinâmica

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

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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Avaliação do Efeito Fotodinâmico sobre o Crescimento in vitro de Pythium insidiosum

Photodynamic Therapy (PDT) is a therapeutic method which employs a photosensitizer and light to cause cellular death. The chemical compounds have low or none toxicity for hosts cells. Under the incidence of light, in an appropriated wavelength, these chemical compounds produce reactive oxygen which affects the biomolecules of the target-cells. The specific illumination of the affected area increases the selectivity of the therapy, since the photodynamic process occurs only in the irradiated area. Pythiosis, for instance, is a life-threatening emerging disease caused by a fungus-like organism called Pythium insidiosum. The disease occurs in man and other animals, being mostly observed in horses. Human pythiosis may present as ophthalmic, cutaneous-subcutaneous and systemic forms of lesions. Due to the fact that P. insidiosum is not a true fungus, it is refractory to most antifungal drugs and the treatment of the disease is difficult. Extensive surgery procedures, such as limb amputation, are the treatment of choice, however relapses may occur frequently. Although not totally effective, the use of immunotherapy associated to surgery have shown some results. Considering that pythiosis is an emerging disease few explored in its etiological and therapeutic aspects, which are limited and few effective, it is of great importance to encourage the development of researches for new strategies of treatment. In this sense, it was evaluated the effect of PDT on in vitro growth of the pathogen employing two chemical compounds as photosensitizer, porphyrin and chlorine, at different concentrations in combination with several energetic dosages. Porphyrin showed inhibition of growth at 25mg/mL with 100J/cm2 of energetic dosage and chlorine showed similar results employing low concentrations (0,7, 1,0 and 1,3mg/mL) with 70J/cm2 of energetic dosage... (Complete abstract click electronic access below)

Terapia transfusional em pequenos animais

Blood transfusion is a very important therapeutic and profilatic procedure. However, the most part of the transfusions in animals in Brazil are done with total blood. The transfusion therapy made in an appropriate way, with hemotherapic products, can save lives and warranties a better health to patients. In all transfusion procedures, the cross match test must be done because there are lots of different blood group types, based on erythrocytes antigen. A slow infusion rate and intensive patient monitoration help to prevent transfusion reactions, mainly hemolytic reactions

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)

Confecção e caracterização de filmes finos de boro para a medida da taxa de reação 10B(N, a)7LI na terapia por captura de nêutrons pelo boro

The Boron Neutron Capture Therapy (BNCT), based on the 10B(n,α)7Li reaction, represents a promising modality for the treatment of cancers that are resistents to conventional treatments. So, it is necessary to find drugs (boron compounds) with high selectivity for each type of cancer, the neutrons source should be well characterized and the rate of 10B(n,α)7Li reaction should be measured with great accuracy as possible. This study aimed to develop a method for manufacturing thin films of boron, for measure the 10B(n,α)7Li reaction, and analyze the uniformity of the films. Five thin films of boron were manufactured with three different concentrations of boric acid, heated to transform the acid in boron, irradiated with thermic neutrons coupled to CR-39 detectors, in BNCT line at the reactor IEA-R1 IPEN/CNEN, São Paulo. After the irradiation, the detectors were chemically attacked with NaOH to reveal the tracks. The methodology presented is effective because it resulted in deposition of boron as thin film enabling the quantitative analysis of 10B(n,α)7Li reaction. The analysis of the uniformity of density of the induced tracks in CR-39 shows that, in most of the films, there is no uniformity in surface distribution of boron, but when the film is divided, we obtain some uniform sectors

Terapia do diabetes mellitus em cães

Diabetes Mellitus is caracterized by a disturbance on endocrine pâncreas with reduction in serum levels of insulin. The deficiency or absence of insulin promotes alterations in the metabolism of carbohydrates, lipidis and proteins. The most common clinical signs of disease are polyuria, polydipsia, polyphagia and loses of weight. The diagnosis is made based on clinical symptoms, with laboratory confirmation through persistent hyperglycemia on fasting and glycosuria. Treatment includes insulin therapy, diet, exercise and oral hypoglycemic agents in an attempt to reverse the catabolic effects associated with deficiency or antagonism of insulin and restore normal homeostasis of the metabolism of proteins, lipids and carbohydrates

Diagnósticos de Enfermagem Prevalentes de Pacientes em Pós-Operatório de Cirurgia Bariátrica em uma Unidade de Terapia Intensiva

The goal of this study is to characterize the clinical outline of patients in postoperative of bariatric surgery attended in an Intensive Care Unit and identify the nursing diagnosis presented by these patients in accordance to Taxonomy II of North American Nursing Diagnosis Association (NANDA). This is a transversal, descriptive and prospective study that was desenvolved at the Intensive Care Unit of the Clinical Hospital of Botucatu Medical School – UNESP. The population was composed by all the patients who were submitted to the bariatric surgery and were attended at the Intensive Care Unit in the period between June and August of 2010, totalizing 13 patients. The data were collected based on the Nursing History that is composed by the Anamnesis, Physical Examination and Complementary Exams. The results were worked out: a) among the 13 patients studied, 10 are women (76,9%) and 3 men (23,1%); the majority is between 30 and 40 years old; 11 people of 13 are morbid obese, which means that they have Body Mass Index higher than 40 kg/m2; b) It was identified 22 nursing diagnosis and 14 of these were found in 100% of the patients. They belong to the dominions of Activity/Rest, Comfort, Elimination and Changing, Security and Protection, Functions and Relationships. Therefore, in relation to the factors, it was conclude that nursing diagnosis help nurses to understand the unsettled human answers or the risks to disturbance in an individual way, which contributes to plan specific interventions to these patients. It was possible to verify that the stage of the nursing diagnosis that is part of the nursing process should be executed by the nurses systematically, because it has been realized by the nurses’ clinical idea, but not in a systematical way

Morte e morrer: opinião dos profissionais de saúde de uma unidade de terapia intensiva pediátrica

Presently, the dying process and death most often occur in hospitals and, particularly, in Intensive Care Units (ICU), where patients’ lives are prolonged thanks to advanced technological devices and highly efficient medicines. To learn about the opinion of health care professionals working at a Pediatric Intensive Care Unit in relation to the dying process and dying. This is a descriptive quantitative study. A questionnaire was applied to the unit’s staff members from June to August, 2011. Data were statistically analyzed. Twenty-five professionals answered the questionnaire, and 72% faced death as a natural life process. 60% felt compassion, but that feeling did not interfere with how they cared for patients. Concerning their professional training, 52% reported not to have received any concerning patients’ caregivers in the dying process or death; therefore, they experienced such situation when they were already working, and 76% reported to be interested in updating courses on that theme. Further discussion about this topic during academic education is necessary. It is also necessary to provide health care professionals with specialization courses, debates and experience exchange so that they can better understand and deal with their feelings and limitations in face of death and thus give better care to patients and relate to patients’ families during the dying process of a loved one

Intervenção Farmacêutica na Terapia Antirretroviral (TARV) em Pacientes Não cumpridores

No Brasil, 190.000 pacientes HIV+ utilizam a terapia antirretroviral. Embora a distribuição da medicação seja gratuita, o problema enfrentado é a não adesão do paciente à terapia, que deve ser igual ou superior a 95%. A não adesão causa o aumento da carga viral, queda dos linfócitos CD4+ e resistência viral à medicação. A complexidade da farmacoterapia, as reações adversas à medicação e a falta de conhecimento sobre HIV e os hábitos necessários para cumprir a terapia comprometem a adesão. Embora o acompanhamento farmacêutico seja eficaz na promoção do cumprimento da terapia, faltam estudos que avaliem esta influência na adesão a TARV. Propor um plano de Intervenções Farmacêuticas e Educativas para promover o cumprimento da TARV. Trata-se de um estudo prospectivo, qualitativo e semi-experimental, realizado com pacientes não cumpridores da terapia segundo o autorrelato. Foram realizados seis encontros, sendo o primeiro para anamnese, aplicação de questionários de avaliação da adesão e qualidade de vida e consulta a exames clínicos; os quatro seguintes para intervenções farmacêuticas e educativas, e o sexto para a avaliação dos mesmos critérios do primeiro encontro, e da satisfação quanto à intervenção. Os encontros contemplaram: intervenções educativas sobre a síndrome, a terapia e mudanças no estilo de vida, proposta de um instrumento para auxiliar na tomada da medicação e recomendação ao médico sobre a necessidade da avaliação laboratorial. Apenas dois pacientes participaram de todos os encontros. Estes pacientes apresentavam carga viral indetectável e contagem de linfócitos T CD4+ desejável, relataram não utilizar estratégias para lembrar-se de tomar a medicação e possuiam pouco conhecimento sobre a doença e a terapia. Durante os encontros, estabeleceram-se plano de metas individuais... (Resumo completo, clicar acesso eletrônico abaixo)