CoordenaÃÃo de AperfeiÃoamento de Pessoal de NÃvel Superior / A Gentamicina (GM) à um aminoglicosÃdeo amplamente utilizado contra infecÃÃes por microrganismos gram-negativos. LesÃo renal aguda (LRA) à a principal limitaÃÃo para a sua eficÃcia terapÃutica, ocorrendo em 10-20% dos pacientes. O evento capital à produÃÃo de espÃcies reativa de oxigÃnio (EROs) e alteraÃÃo na dinÃmica intra-renal e glomerular. O gengibre à fonte de substÃncias bioativas que podem ter efeito satisfatÃrio na renoproteÃÃo por aÃÃo antioxidante. A fraÃÃo ZOEB4 à enriquecida com os compostos com [6]-, [8]- e [10]- gingerol, isolada a partir do extrato do Zingiber officinale Roscoe, parece ser promissora diante desta nefrotoxidade, podendo auxiliar o rim na resposta antioxidante endÃgena, diminuindo o dano oxidativo e atenuando a resposta prÃ-inflamatÃria. O presente estudo visou investigar os possÃveis efeitos protetores da fraÃÃo ZOEB4 em um quadro de LRA pela administraÃÃo de GM 100 mg/kg i.p. Foram utilizados ratos Wistar, adultos machos, divididos em 6 grupos. Os grupos controles foram induzidos com NaCl 0,9% por 7dias e tratados oralmente com tween-80 2%, ou com a fraÃÃo ZOEB4 25 mg/kg; grupos induzidos a nefrotoxicidade com GM 100mg/kg por 7dias acrescido do tratamento oral com a fraÃÃo ZOEB4(6,25, 12,5 ou 25mg/kg) ou com tween 80 2%. O tratamento oral ocorreu durante 5 dias a partir do 5 dia de induÃÃo. Ao termino de cada tratamento foram coletados plasma, urina e rins para as anÃlises. Foram avaliados os parÃmetros bioquÃmicos indicativos de funÃÃo renal, funÃÃo tubular, perfil oxidativo, atividade das enzimas antioxidantes, a transcriÃÃo gÃnica de mediadores prÃ-inflamatÃrios atravÃs da reaÃÃo da polimerase em cadeia em tempo real (qPCR), alÃm da anÃlise histopatolÃgica. A GM alterou consideravelmente quase que todos os parÃmetros investigados: Clcr (0,7Â0,1 mL/min), ureia (73,1Â7,5 mg/dL) proteÃna urinÃria (93,5Â9,2 mg/dL), FENa (3,3Â0,5 %) e FEK (78,76 11,7 %), MDA renal(1,7Â0,3 nM/mg de prot.), GSH (101,6 23,4Âg/mg prot.), SOD (52,47Â7,7 U/mg prot.) e TNF-α (3,2 Â0,7) estabelecendo assim, dano renal nos animais. O tratamento com a fraÃÃo ZOEB4 na dose de 25mg/kg desencadeou proteÃÃo diante desta nefrotoxicidade, revertendo a diminuiÃÃo do Clcr (1,3 0,2 mL/min), diminuindo os nÃveis sÃrico de ureia (41,7Â0,03 mg/dL), de proteÃna urinÃria (51,36Â3,2 mg/dL), da FENa (1,82  0,21 %) e FEK (48,8Â4,6 %), MDA (0,67Â0,2 nM/mg de prot.), alÃm de aumentar os nÃveis de GSH (250,2Â27,7 ug/mg prot.) e a atividade da SOD (112,2Â6,4 U/mg prot.) e, inibiÃÃo da transcriÃÃo de TNF-α (1,5Â0,2). Desta forma, proporcionando proteÃÃo a funÃÃo renal. Estes resultados, acrescentado aos relatos de estudos prÃvios realizados com os compostos desta planta, indicam um futuro promissor para a utilizaÃÃo dos gingerois como um coadjuvante no tratamento diante de LRAs induzidas por aminoglicosÃdeos.

[6]-Gingerol, [8]-gingerol, and [10]-gingerol are phytochemical extracts from ginger that are thought to contribute to its health-benefitting properties. The objectives of this investigation were to explore and compare the in vitro anti-proliferative and cytotoxic effects of [6]-, [8]-, and [10]-gingerol on human and mouse mammary carcinoma cells, as well as their ability to inhibit T cell proliferation. [8]-Gingerol and [10]-gingerol induced mammary carcinoma cell death that did not require ROS production or caspase activation. All three gingerols inhibited the proliferation of mammary carcinoma cells and T cells, and the production of IFN-? by T cells. The production of IL-2 and the expression of early T cell activation markers, CD25 and CD69, were significantly decreased by [8]- and [10]-gingerol. The results demonstrated that [10]-gingerol was the most potent, followed by [8]-gingerol, then [6]-gingerol. Consequently, [8]- and [10]-gingerol warrant further investigation for the treatment of breast cancer and the control of inflammation.

Submitted by Regina Correa (rehecorrea@gmail.com) on 2016-10-03T20:53:43Z No. of bitstreams: 1 TeseACBMM.pdf: 12490231 bytes, checksum: 097b90957f143d8d696733602a92d242 (MD5) / Approved for entry into archive by Ronildo Prado (ronisp@ufscar.br) on 2016-10-05T13:41:11Z (GMT) No. of bitstreams: 1 TeseACBMM.pdf: 12490231 bytes, checksum: 097b90957f143d8d696733602a92d242 (MD5) / Approved for entry into archive by Ronildo Prado (ronisp@ufscar.br) on 2016-10-05T13:41:20Z (GMT) No. of bitstreams: 1 TeseACBMM.pdf: 12490231 bytes, checksum: 097b90957f143d8d696733602a92d242 (MD5) / Made available in DSpace on 2016-10-05T13:48:36Z (GMT). No. of bitstreams: 1 TeseACBMM.pdf: 12490231 bytes, checksum: 097b90957f143d8d696733602a92d242 (MD5) Previous issue date: 2015-03-20 / Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq) / Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES) / Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP) / Breast cancer is the leading cause of death in Brazil and worldwide, between the types of cancer, triple negative TNBC (Triple Negative Breast Cancer), which do not express hormone receptors, represents 20% of the cases and presents relapses within 3 years. However, the major cause of death in cancer is due the formation of metastasis. The TNBC has a greater propensity to form lung and brain metastasis. Furthermore, currently few treatments are available. Search for new target treatments, with fewer side effects is very important to treat this disease. Natural products are rich sources of molecules with antitumoral activity; among them are the gingerols, from ginger and resveratrol, from grapes. Nevertheless, not much is known about antitumor activity of [10]-gingerol (10G). Therefore, the aim of this work was to investigate the potential use of 10G as an antimetastatic molecule in in vitro and in vivo experiments. In this work, 10G had antiproliferative activity on several breast cancer cell lines (4T1BM2, 4T1BM2 shRNAβ3, 4T1Br4, MDA-MB-231, MDA-MB-231 BrM) and on a normal cell line, bEnd.3. The natural compounds affected tumor cell morphology and RSVT was able to inhibit cell migration and adhesion through vitronectin. 10G induced apoptosis via the extrinsic pathway through the increase of caspase-9, -3 and -7 expression and decrease of Bcl-2 expression, induced cell cycle arrest in G1 and subG0 phase. In in vivo models, 10G inhibited primary tumor growth and lung metastasis, as well as bone and brain metastasis. Therefore, this study was able to provide new data as 10G can be acting in tumor cells and its possible clinical application. / O câncer de mama é o que mais afeta as mulheres no Brasil e no mundo, entre os tipos de câncer de mama, o triplo negativo TNBC (Triple Negative Breast Cancer), que não expressa nenhum receptor hormonal, corresponde a 20% dos casos e apresenta recidivas em três anos. Entretanto, a maior causa de morte no câncer é devido a formação de metástases. O TNBC possui uma maior propensão a metástases pulmonares e cerebrais. Além disso, atualmente poucos tratamentos estão disponíveis. Buscas para novos medicamentos alvo para o TNBC e com menos efeitos colaterais são de grande importância para o tratamento dessa doença. Produtos naturais são fontes ricas em moléculas com atividades antitumorais, dentre elas os gingeróis, presentes no gengibre e o resveratrol (RSVT), presente nas uvas. Entretanto, pouco se sabe da atividade antitumoral do [10]-gingerol (10G). Este trabalho teve como objetivo investigar o potencial uso do 10G como molécula antimetastática em experimentos in vitro e in vivo. Neste trabalho verificou-se a atividade antiproliferativa do 10G e do RSVT em diversas linhagens de câncer de mama (4T1BM2, 4T1BM2 shRNAβ3, 4T1Br4, MDA-MB- 231, MDA-MB-231 BrM) e um controle de célula normal, a bEnd.3. Os produtos naturais (PNs) usados afetam a morfologia celular nas células tumorais e o RSVT é capaz de inibir a adesão e migração via vitronectina. O 10G induziu apoptose da linhagem 4T1Br4 através da via extrínseca, com o aumento da expressão de casspase-9, -3 e -7; induziu a parada do ciclo celular nas fases G1 e sub G0. Em modelos de metástase in vivo o 10G inibiu o crescimento do tumor primário e de metástases pulmonares, além de inibir metástases ósseas e cerebrais. Portanto, este trabalho foi capaz de fornecer dados de como o 10G atua nas células tumorais e sua possível aplicação clínica.

CoordenaÃÃo de AperfeiÃoamento de Pessoal de NÃvel Superior / Neste trabalho investigou-se o potencial biocatalÃtico do fungo Rhizopus stolonifer um fungo fitopatogÃnico em biotransformaÃÃes de metabÃlitos secundÃrios naturais Dois desses metabolitos, o diterpeno 3,12-dioxo-15,16-epoxi-4-hidroxicleroda-13(16),14-dieno e o derivado fenÃlico 6-gingerol foram biotransformados e seus respectivos produtos isolados e quantificados por CLAE A elucidaÃÃo estrutural dos produtos de biotransformaÃÃo pelo fungo Rhizopus stolonifer foram realizadas por anÃlise dos espectros de RMN 1H RMN 13C RMN 13C-DEPT 135 IV e EM Identificou-se que os produtos obtidos eram provenientes de uma biorreduÃÃo revelando o potencial deste fungo na reduÃÃo quimio e regiosseletivas de grupos carbonilas nÃo conjugados Realizou-se o estudo cinÃtico dos produtos de biorreduÃÃo por CLAE e ensaios citotoxicidade frente Ãs linhagens tumorais humanas OVCAR-8 (ovÃrio) SF-295 (glioblastoma) e HCT-8 (cÃlon) O produto biorreduzido 6-gingerdiol apresentou os melhores percentuais de inibiÃÃo do crescimento celular: 91,83; 70,90 e 78,56 respectivamente / In this work the biocatalytic potential of the fungus Rhizopus stolonifer a phytopathogenic fungus in biotransformations of natural secondary metabolites was investigated Two of these metabolites the diterpene 3,12-dioxo-15,16-epoxy-4-hydroxycleroda-13(16),14-diene and the phenolic derivative 6-gingerol was biotransformed and their products was isolated and quantified by HPLC The structural determination of biotransformation products was obtained by analysis of 1H NMR 13C NMR 13C-NMR DEPT 135 IV and MS spectrum The products were obtained by a biorreduction process revealing the potential of this fungus in selective reduction of unconjugated carbonyl groups The products from bioreduction were submitted to antitumor assay against human tumor cell lines OVCAR-8 (ovarian) SF-295 (glioblastoma) and HCT-8 (colon) The biotransformation product 6-gingerdiol showed the highest percentage of inhibition of cell growth: 91,83; 70,90 e 78,56, respectively

There is significant interest in the field of neuroscience to gain a better understanding of how neurons die in neurodegenerative diseases such as Alzheimer's and Parkinson's diseases. We have used the neuronal cell line NG115-401L with unique calcium signaling characteristics to test the hypothesis that improving calcium loading into the endoplasmic reticulum (ER) to increase ER calcium levels acts as a possible neuroprotective response. We approached this problem using both pharmacological and genetic approaches targeting the central mediator of calcium uptake in the ER localized sarco/endoplasmic reticulum Ca 2+ ATPase (SERCA) enzyme. The pharmacological studies involved use of the ginger root compound 6-gingerol, which to date is the best documented agent for activating SERCA enzymes in heart and skeletal muscle. However, in our experiments, gingerol did not appear to activate NG115-401L SERCA pumps; indeed, the compound produced a response more like that of a SERCA inhibitor inducing a rapid ER calcium depletion. In addition, gingerol stimulated robust calcium influx responses, an unexpected result given the NG115-401L neural cell line is uniquely deficient in calcium influx pathways. Our genetic approach involved expressing the stromal interaction molecule 1 (STIM1) protein in the NG115-401L cell, which is also an ER localized protein that serves as a pivotal calcium influx channel regulator. NG115-40lL neurons present a native deficiency of STIM1 expression in a background phenotype with well characterized perturbations in ER calcium regulation and control of calcium influx pathways. Thus, STIM1 may be predicted to increase ER calcium levels, conferring protection against neuron cell death due to ER calcium store defects. STIM1 expression reconstituted the corrupted calcium influx pathway in NG115-401L neurons, which conferred neuroprotective responses to ER calcium perturbation, mitochondrial oxidative stress and subsequent cell death. Our results argue for unique and undiscovered regulatory effects of gingerol on the ER calcium circulation system, and suggest that the expression of STIM1 in these neurons protects against ER stress and oxidative stress via reconstruction of cellular calcium homeostasis.