Navegando por Palavras-chave "cell culture"

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    Avaliação da ação do methotrexate nas células em cultura de endométrio de mulheres com e sem endometriose
    (Universidade Federal de São Paulo (UNIFESP), 2015-04-30) Kleine, João Paulo Ferreira de Oliveira [UNIFESP]; Silva, Ismael Dale Cotrim Guerreiro da [UNIFESP]; Universidade Federal de São Paulo (UNIFESP)
    The Endometriosis is today a benign disease that affects about six million Brazilian. According to the Brazilian Association of Endometriosis, between 10% and 15% of women of reproductive age (13-45 years) can develop it and 30% are likely to become sterile. The disease is characterized by the presence of the endometrium, this tissue lining the uterus, outside the uterine cavity, or other organs of the pelvis as fallopian tubes, ovaries, intestines, and bladder. However, such growths may also occur in other parts of the body. The exact causes of endometriosis are unclear, but some possible causes for the problem are more speculated retrograde menstruation, growth of embryonic cells, poor immune system and many others. Although the setting is very well established, there are many features not defined yet endometriosis as its etiology, why cause infertility? Why some women complain of extreme pain and other not describe any pain? Why the disease affects so many different locations? And, along with other obscure features, which is the best treatment? This study aimed to define a possible treatment / action induced by chemotherapy by methotrexate in vitro cultures of endometrial cells, evaluating its role in proliferation, apoptosis and cell cycle of women endometrial cells with and without endometriosis after treatment. We selected 10 patients treated at Algia industry and Pelvic Endometriosis Department of Gynecology, Federal University of São Paulo, Paulista School of Medicine (UNIFESP-EPM) and Hospital São Paulo (HSP), 5 women with endometriosis grade IV (severe) indicating laparoscopic surgery and 5 women without any symptoms or suspected endometriosis to up the control study group. All after clinical examination or laparoscopy had a small portion of shaved endometrium / collected for in vitro culture of the same and subsequent treatment with methotrexate chemotherapy. The results show that the cultured cells showed a faster growth of patients with endometriosis compared to those of the control group independent of treatment. The initial evaluation with hormone therapy had no effect on the samples as much control as in endometriosis cells. Methotrexate has not decreased cell proliferation in both the control group and in the endometriosis group and the same has not demonstrated increased apoptosis in both the control group and in the endometriosis group.
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    Determination of angiotensin I-converting enzyme activity in cell culture using fluorescence resonance energy transfer peptides
    (Elsevier B.V., 2007-04-15) Sabatini, R. A.; Bersanetti, P. A.; Farias, S. L.; Juliano, L.; Juliano, M. A.; Casarini, D. E.; Carmona, A. K.; Paiva, A. C. M.; Pesquero, J. B.; Universidade Federal de São Paulo (UNIFESP)
    An assay using fluorescence resonance energy transfer peptides was developed to assess angiotensin I-converting enzyme (ACE) activity directly on the membrane of transfected Chinese hamster ovary cells (CHO) stably expressing the full-length somatic form of the enzyme. the advantage of the new method is the possibility of using selective substrates for the two active sites of the enzyme, namely Abz-FRK(Dnp)P-OH for somatic ACE, Abz-SDK(Dnp)P-OH for the N domain, and Abz-LFK(Dnp)-OH for the C domain. Hydrolysis of a peptide bond between the donor/acceptor pair (Abz/Dnp) generates detectable fluorescence, allowing quantitative measurement of the enzymatic activity. the kinetic parameter K-m for the hydrolysis of the three substrates by ACE in this system was also determined and the values are comparable to those obtained using the purified enzyme in solution. the specificity of the activity was demonstrated by the complete inhibition of the hydrolysis by the ACE inhibitor lisinopril. Therefore, the results presented in this work show for the first time that determination of ACE activity directly on the surface of intact CHO cells is feasible and that the method is reliable and sensitive. in conclusion. we describe a methodology that may represent a new tool for the assessment of ACE activity which will open the possibility to study protein interactions in cells in culture. (c) 2007 Elsevier Inc. All rights reserved.
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    Dimethylaminoethanol affects the viability of human cultured fibroblasts
    (Springer, 2007-12-01) Gragnani, Alfredo [UNIFESP]; Giannoccaro, Fabiana Bocci; Sobral, Christiane Steponavicius [UNIFESP]; Franca, Jeronimo Pereira de [UNIFESP]; Ferreira, Lydia Masako [UNIFESP]; Universidade Federal de São Paulo (UNIFESP)
    Background: in clinical practice, dimethylaminoethanol (DMAE) has been used in the fight against wrinkles and flaccidity in the cervicofacial region. the firming action of DMAE is explained by the fact that its molecule, considered to be a precursor of acetylcholine, alters muscle contraction. However, no experimental studies have confirmed this theory. Because the actual mechanism of DMAE action was not defined and there were no references in the literature regarding its direct action on fibroblasts, this study was performed to evaluate the direct action of DMAE on cultured human fibroblasts.Methods: Human fibroblasts obtained from discarded fragments of total skin from patients undergoing plastic or reconstructive surgical procedures performed within the Plastic Surgery Division at the Federal University of S (a) over tildeo Paulo were used for this study. the explant technique was used. the culture medium was supplemented with different concentrations of DMAE on the fourth cell passage, and the cell proliferation rate, cytosolic calcium levels, and cell cycle were evaluated. Statistical analysis was performed using analysis of variance (ANOVA) followed by a New-man-Keuls test for multiple comparisons.Results: A decrease in fibroblast proliferation was associated with an increase in DMAE concentration. A longer treatment time with trypsin was required for the groups treated with DMAE in a dose-dependent manner. in the presence of DMAE, cytosolic calcium increased in a dose-dependent manner. Apoptosis also increased in groups treated with DMAE.Conclusion: Dimethylaminoethanol reduced the proliferation of fibroblasts, increased cytosolic calcium, and changed the cell cycle, causing an increase in apoptosis in cultured human fibroblasts.
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    Evaluation of photodynamic therapy in adhesion protein expression
    (Spandidos Publ Ltd, 2014-08-01) Pacheco-Soares, Cristina; Maftou-Costa, Maira [UNIFESP]; Da Cunha Menezes Costa, Carolina Genuncio; De Siqueira Silva, Andreza Cristina; Moraes, Karen C. M.; Univ Vale do Paraiba; Universidade Federal de São Paulo (UNIFESP); Universidade de São Paulo (USP)
    Photodynamic therapy (PDT) is a treatment modality that has clinical applications in both non-neoplastic and neoplastic diseases. PDT involves a light-sensitive compound (photosensitizer), light and molecular oxygen. This procedure may lead to several different cellular responses, including cell death. Alterations in the attachment of cancer cells to the substratum and to each other are important consequences of photodynamic treatment. PDT may lead to changes in the expression of cellular adhesion structure and cytoskeleton integrity, which are key factors in decreasing tumor metastatic potential. HEp-2 cells were photosensitized with aluminum phthalocyanine tetrasulfonate and zinc phthalocyanine, and the proteins beta 1-integrin and focal adhesion kinase (FAK) were assayed using fluorescence microscopy. the verification of expression changes in the genes for FAK and beta 1 integrin were performed by reverse transcription-polymerase chain reaction (RT-PCR). the results revealed that HEp-2 cells do not express beta-integrin or FAK 12 h following PDT. It was concluded that the PDT reduces the adhesive ability of HEp-2 cells, inhibiting their metastatic potential. the present study aimed to analyze the changes in the expression and organization of cellular adhesion elements and the subsequent metastatic potential of HEp-2 cells following PDT treatment.
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    Nicotinic receptor/ionic channel complex (AChR) in androgen-dependent skeletal muscle cultures
    (Springer, 2002-02-01) Bielavsky, M.; Souccar, C.; Lapa, A. J.; Lima-Landman, MTR; Universidade Federal de São Paulo (UNIFESP)
    The kinetic properties of the nicotinic receptor/ionic channel complex (AChR) were compared in cell cultures obtained from androgen-dependent skeletal muscles of the perineal complex (P) and from muscles less dependent upon sex hormones (the thigh musculature, T). Because the development of P is delayed compared to other skeletal muscles in the rat, cultures were performed taking into account the age of the donor (4- or 6-day-old rats), and the time interval the cells remained in culture (7 days and 15 days). the ionic channel conductance (gamma) and the mean channel open time (2) were determined with the patch-clamp technique in the cell-attached configuration at room temperature. Cultures from P and T muscles were morphologically identical in size and shape, independent of the animals' age at plating or on the plating time. in all of them, the AChR was spread over the cell membrane. More than one AChR ionic channel conductance was observed in P and T cultures, and the prevalent value of gamma in either culture ranged from 30 pS to 35 pS. in P fibers from 4-day-old rats cultured for 7 days (P 4/7), the distribution of channel open times fitted a double exponential, while in T 4/7 they were fitted with a single exponential. in cultures from P and T muscles obtained from older rats (6 days old) and in those cells remaining in culture for a prolonged time (15 days), the channel open times also fitted a double exponential. Because P and T cultures lack trophic neuronal influences, the difference observed between the tau of P 4/7 and T 4/7 was thought to be the hormone requirement of P muscles to grow and differentiate. Likewise, the difference observed between T 4/7 and T 4/15 may indicate the need for neurotrophic influences to maintain higher T values in older cultures. Since this requirement is not found in cultured fibers, tau would tend to assume slower values approaching those of P without hormone activation.