Efeitos toxicogen?ticos do resveratrol em c?lulas de c?ncer de bexiga com diferentes status do gene TP53.

Programa de P?s-Gradua??o em Ci?ncias Farmac?uticas. CIPHARMA, Escola de Farm?cia, Universidade Federal de Ouro Preto.A atividade antitumoral do resveratrol, composto polifen?lico encontrado principalmente nas uvas, tem sido estudada em diversos tipos de c?ncer. No c?ncer de bexiga, seus efeitos antiproliferativos j? foram demostrados, tanto in vitro quanto in vivo, entretanto pouco se sabe sobre seu mecanismo de a??o. Nesse contexto, o presente estudo teve como objetivo avaliar o potencial antineopl?sico e os poss?veis mecanismos de a??o molecular do resveratrol em c?lulas tumorais de bexiga com diferentes status do gene TP53 (RT4 - TP53 selvagem; 5637 e T24 - TP53 mutado). Avaliou-se os efeitos do resveratrol em rela??o a citotoxicidade, altera??es morfol?gicas, sobreviv?ncia clonog?nica, genotoxicidade, mutagenicidade, produ??o de esp?cies reativas de oxig?nio, progress?o do ciclo celular, taxas de apoptose/necrose, migra??o celular, metila??o global, imunocitoqu?mica para p53 e PCNA e express?o dos genes AKT, CDH1, CTNNBIP1, DNMT1, FGFR3, HAT, HDAC1, HOXB3, mTOR, MYC, PLK1, RASSF1A, SMAD4 e SRC. Os resultados mostraram que o resveratrol possui efeito citot?xico em todas as linhagens estudadas, sendo a linhagem TP53 selvagem mais sens?vel ? a??o desse composto. Em rela??o aos efeitos a longo prazo, observou-se que o resveratrol causou redu??o do n?mero de col?nias em todas as linhagens, juntamente com redu??o da express?o do gene PLK1. Aumento significativo de danos prim?rios ao DNA foi detectado em todas as linhagens e provavelmente ocorreu devido aos efeitos pr?-oxidantes do resveratrol. Aumento significativo das taxas de apoptose foram encontradas na linhagem TP53 selvagem ao mesmo tempo em que ocorreu redu??o da express?o dos genes AKT, mTOR e SRC. A redu??o da migra??o celular foi acompanhada pela redu??o da express?o do gene SMAD4 e aumento da express?o de CDH1. Al?m disso, os efeitos antiproliferativos do composto na linhagem TP53 selvagem foram acompanhados de modula??o dos genes HAT, HOXB3 e DNMT1. Nas linhagens mutadas para o gene TP53 pode-se observar que o resveratol leva a parada do ciclo celular na fase S, juntamente com redu??o da express?o do gene PLK1. Tamb?m foi observada inibi??o da migra??o celular acompanhada do aumento da express?o dos genes CDH1 e CTNNBIP1. Adicionalmente, houve modula??o de vias relacionadas a HOXB3, RASSF1A e HAT na linhagem T24. Concluindo, o resveratrol possui atividade antiproliferativa independente do status de TP53 em c?lulas tumorais de bexiga, entretanto, os efeitos antineopl?sicos observados devem-se a diferentes mecanismos de a??o.The antitumor activity of resveratrol, a polyphenolic compound found mainly in grapes, was verified in several cancers. The antiproliferative effects of resveratrol were also demonstrated in bladder cancer, however it is necessary to better understand its mechanism of action. In this context, the present study aimed to evaluate the antineoplastic potential and resveratrol molecular mechanisms of action in bladder tumor cells with different TP53 gene status (wild type RT4 - TP53; 5637 and mutated T24 - TP53). The effects of resveratrol on cytotoxicity, morphological changes, clonogenic survival, genotoxicity, mutagenicity, reactive oxygen species production, cell cycle progression, apoptosis/necrosis rates, cell migration, immunocytochemistry for p53 and PCNA, global methylation and AKT, CDH1, CTNNBIP1, DNMT1, FGFR3, HAT, HDAC1, HOXB3, mTOR, MYC, PLK1, RASSF1A, SMAD4 and SRC gene expression were evaluated. Resveratrol was cytotoxic in all cells lines studied and the TP53 wild-type cells was more sensitive to the action of this compound. Regarding long-term effects, resveratrol caused a reduction in the number of colonies in all cell lines, along with a reduction in PLK1 gene expression. Significant increase in primary DNA damage was detected in all cell lines and probably occurred due to pro-oxidant effects of resveratrol. Significant increase in apoptosis rates was found in the wild type TP53 cells and this was accompanied by AKT, mTOR and SRC downregulation. Cell migration inhibition was accompanied by SMAD4 downregulation and CDH1 upregulation. In addition, modulation of HAT, HOXB3 and DNMT1 gene expression also contributed to antiproliferative effects of resveratrol in the wild type TP53 cells. In the TP53 mutated cells, cell cycle arrest at S phase with PLK1 downregulation was observed. Cell migration inhibition was accompanied by CDH1 and CTNNBIP1 upregulation. Additionally, there was modulation of the HOXB3, RASSF1A and HAT gene expression in T24 cells, as well as nuclear PCNA reduction. In conclusion, resveratrol had antiproliferative activity in bladder tumor cells; however, the mechanisms of action are dependent on TP53 status

Mathematical modeling to optimize pine lumber yield

In Brazil, only 4% of the 7.84 million hectares of planted forests is devoted to the production of lumber wood, being in Santa Catarina state most of the wood used for this purpose is of the Pinus genus. This work aims to estimate the maximum utilization of logs due to the production of wood boards in the city of Canoinhas, Santa Catarina. For that, a sawmill of the region were consulted and the dimensions of the pieces produced were verified. The dimensions and classes of logs commonly traded in the region were also raised. As a result, 82 models were created in Maxitora software in diagram format. With the cutting models the sawmill had its performance optimized with the use of techniques of operational research in a cutting problem. The whole linear programming technique was used for an estimated demand for the quantity of each of the pieces produced. The results showed that only five models are required to meet such demand, so the yield is 43.18%

Polymeric micelles containing resveratrol: development, characterization, cytotoxicity on tumor cells and antimicrobial activity

Antimicrobial and antitumor activities of resveratrol, a compound found mainly in grapes, have already been demonstrated. However, its low bioavailability is a limiting factor for therapeutic application. Polymeric micelles can be an approach to solve this problem since they can encapsulate hydrophobic substances. We developed and characterized micellar formulations containing resveratrol and evaluated their cytotoxic and antimicrobial effects. The formulations were prepared by the cold dispersion method with different concentrations of F127 (5 or 10% w/w) and resveratrol (500 or 5000 µM). The formulations were characterized according to size, polydispersity index, pH, encapsulation rate and in vitro release. Cytotoxic effect was evaluated on a bladder cancer cell line and antimicrobial effect was evaluated on E. coli, S. aureus and C. albicans. One of the formulations (10% w/w of F127 and 5000 µM of resveratrol) was a monodispersed solution with high encapsulation rate, thus it was chosen for the cytotoxicity and antimicrobial assays. MS10+RES-3 was able to preserve the antimicrobial and cytotoxic activity of resveratrol. This is the first study that evaluated antimicrobial potential and cytotoxicity of micelles containing resveratrol on bladder cancer cells and the results showed that micellar nanostructures could ensure the maintenance of the biological activity of resveratrol

Synthesis, in vitro and in vivo anti-Trypanosoma cruzi and toxicological activities of nitroaromatic Schiff bases.

Chagas disease is a major health problem not only in Latin America but also in Europe and North America due to the spread of this disease into nonendemic areas. In terms of global burden, this major tropical infection is considered to be one of the most neglected diseases, and there are currently only two available chemotherapies: benznidazole and nifurtimox. Unfortunately, although these chemotherapies are beneficial in the acute phase of the disease, benznidazole and nifurtimox lead to significant side effects, including hepatitis and neurotoxicity. Therefore, the search for and development of more effective, safe and inexpensive anti-Trypanosoma cruzi drugs are required. In this work, a series of 10 nitroaromatic Schiff bases bearing different (nitro) aromatic rings-was synthesized. Subsequently, the in vitro and in vivo anti-T. cruzi activities of the Schiff bases were investigated, as well as the in vivo toxicity and the biological effects. The basic structure of the most promising in vivo Schiff base, 10 would be useful in the synthesis of new compounds for Chagas disease treatment

Avalia??o da atividade antitumoral in vitro de solu??es micelares contendo alil isotiocianato.

Programa de P?s-Gradua??o em Ci?ncias Farmac?uticas. CIPHARMA, Escola de Farm?cia, Universidade Federal de Ouro Preto.O alil isotiocianato (AITC), composto majorit?rio da semente de mostarda, vem sendo bastante estudado quanto a sua atividade antineopl?sica. Neste trabalho foram desenvolvidas solu??es micelares de Pluronic? F127 (F127) contendo AITC e o potencial antitumoral destas formula??es foi avaliado in vitro em linhagens celulares de carcinoma de bexiga com diferentes status do gene TP53 (RT4 ? TP53 selvagem e T24 ? TP53 mutado). As formula??es foram preparadas pelo m?todo de dispers?o a frio (DF) e pelo m?todo de reidrata??o de filme polim?rico (RFP). A quantidade de AITC incorporada as formula??es foi determinada por cromatografia l?quida de alta efic?cia acoplada a detector UV. Os resultados demonstraram que pelo m?todo DF, aproximadamente 96,5% do AITC adicionado durante a prepara??o foi incorporado na formula??o final. Entretanto, pelo m?todo RFP ocorreu perda de mais de 75% da quantidade de AITC adicionada, mostrando que a prepara??o de micelas por esse m?todo n?o foi vi?vel. A avalia??o da citotoxicidade e da prolifera??o celular foi realizada 24 e 48 horas ap?s tratar as c?lulas das duas linhagens de tumor de bexiga por 3 horas com as formula??es de AITC produzida pelo m?todo DF. C?lulas tratadas com AITC solubilizado em Tween (AITC livre) e c?lulas tratadas com os excipientes da formula??o foram usadas como controles. Ap?s 24 horas do tratamento, as formula??es contendo AITC nas concentra??es 0,0925; 0,125; 0,25 e 0,5?M causaram citotoxicidade na linhagem RT4; entretanto, na linhagem T24, somente a concentra??o mais alta (0,5 ?M) promoveu diminui??o no n?mero de c?lulas. Nenhuma citotoxicidade foi observada para o AITC livre. Ap?s 48 horas, todas as concentra??es das formula??es contendo AITC levaram a redu??es significativas da viabilidade celular em ambas as linhagens celulares. Al?m disso, as formula??es foram mais citot?xicas para as c?lulas RT4 do que para as c?lulas T24, contribuindo com a hip?tese de que c?lulas carregando o gene TP53 mutado s?o mais resistentes a quimioter?picos. Posteriormente realizou-se o ensaio de sobreviv?ncia clonog?nica. Tanto as solu??es micelares contendo AITC quanto o AITC livre foi capaz de reduzir o n?mero de col?nias formadas. Assim, podemos afirmar que solu??es micelares aceleram a a??o de AITC nas c?lulas tumorais avaliadas. Desta forma, as formula??es desenvolvidas podem ser uma abordagem interessante para a futura administra??o oral dessa subst?ncia no tratamento do tumor de bexiga.The allyl isothiocyanate (AITC), major compound of mustard seed, has been widely studied for its antineoplastic activity. This study aimed to develop micellar solutions of Pluronic? F127 (F127) containing AITC and the cytotoxic potential of those formulations were evaluated in vitro in two bladder transitional carcinoma cell lines with different TP53 gene status (RT4, with wild-type TP53; and T24, mutated TP53). The formulations were prepared by the cold dispersion method (DF) and the polymeric film rehydration method (RFP). The amount of AITC incorporated in the formulations was determined by high performance liquid chromatography coupled to a UV detector. The results showed that nearly 96.5% of the AITC initially added was found in the final formulation prepared by the DF method. However, when the formulation was prepared by the RFP method, more than 75% of the initial amount of AITC was lost, limiting the preparation of AITC micelles by this method. The cytotoxicity and cell proliferation assessment was done 24 and 48 hours after treating both bladder carcinoma cell lines (T24 and RT4) for 3 hours with the formulations produced by the DF method. Cells treated with AITC solubilized in Tween (free AITC) and with the formulation excipients were used as controls. After 24 hours of treatment, formulations containing the AITC concentrations of 0.0925; 0.125; 0.25 and 0.5?M showed cytotoxicity to RT4 cell line, while in the T24 cell line, only the highest concentration (0.5 ?M) reduced the number of cells. No cytotoxicity was observed for free AITC. After 48 hours, all concentrations of the AITC formulations induced significant reductions in cell viability in both cells line. These results indicate that micellar solutions containing AITC potentiated the cytotoxic effects of this drug. Furthermore, the formulations were more cytotoxic for RT4 cells than for T24 cells contributing to the hypothesis that cells carrying the mutated TP53 are more resistant to chemotherapy, because of the role that this gene plays in the control of apoptosis. Subsequently we carried out the clonogenic survival assay. Both micellar solutions containing AITC and free AITC were able to reduce the number of colonies formed. We can say that micellar solutions accelerate the AITC action in the tumor cells evaluated. Thus, the formulations developed can become an interesting approach for further studies and future administration of this substance in the treatment of bladder tumor

Actividad antitumoral de soluciones micelares que contienen isotiocianato de alilo: un estudio in vitro

Introducción: Varios productos naturales exhiben actividad antineoplásica prometedora contra las células cancerosas de vejiga, incluido el isotiocianato de alilo (AITC). Sin embargo, el AITC irrita las membranas mucosas e induce reacciones cutáneas vesiculares o eccematosas. Por tanto, las formulaciones farmacéuticas son necesarias para superar estos problemas. El objetivo era desarrollar soluciones micelares que contengan AITC e investigar su actividad antitumoral en líneas celulares de carcinoma de vejiga. Método: Las soluciones micelares se prepararon mediante el método de dispersión en frío. Posteriormente, evaluamos la citotoxicidad, la proliferación celular, la cinética del ciclo celular y los efectos a largo plazo de las micelas en las células del cáncer de vejiga. Resultados: Los ensayos de citotoxicidad y proliferación celular mostraron que hubo un aumento en la actividad de AITC cuando se encapsuló en micelas. También observamos la detención del ciclo celular en la fase S después del tratamiento con micelas AITC. Además, la formulación pudo mantener los efectos a largo plazo del AITC libre Conclusiones: Las soluciones micelares desarrolladas pueden convertirse en un enfoque interesante para la administración de AITC en el tratamiento del cáncer de vejiga.Introduction: Several natural products exhibit promising antineoplastic activity against bladder cancer cells, including allyl isothiocyanate (AITC). However, the AITC irritates the mucous membranes and induces eczematous or vesicular skin reactions. Thus, pharmaceutical formulations are necessary to overcome these problems. The aim was to develop micellar solutions containing AITC and investigate their antitumoral activity in bladder carcinoma cell lines. Method: The micellar solutions were prepared by cold dispersion method. Subsequently, we evaluated cytotoxicity, cell proliferation, cell cycle kinetics and long-term effects of micelles in bladder cancer cells. Results: Cytotoxicity and cell proliferation assays showed there was an increase in AITC activity when it was encapsulated in micelles. We also observed cell cycle arrest in the S phase after treatment with AITC-micelles. Furthermore, the formulation was able to maintain the long-term effects of free AITC. Conclusions: The micellar solutions developed can become an interesting approach for administration of AITC in the treatment of bladder cancer.FAPEMIG (Fundação de Amparo à Pesquisa do Estado de Minas Gerais) (Grant numbers PPM-00282-11 and CBB-APQ-01497-14)

Mathematical modeling to optimize pine lumber yield

In Brazil, only 4% of the 7.84 million hectares of planted forests is devoted to the production of lumber wood, being in Santa Catarina state most of the wood used for this purpose is of the Pinus genus. This work aims to estimate the maximum utilization of logs due to the production of wood boards in the city of Canoinhas, Santa Catarina. For that, a sawmill of the region were consulted and the dimensions of the pieces produced were verified. The dimensions and classes of logs commonly traded in the region were also raised. As a result, 82 models were created in Maxitora software in diagram format. With the cutting models the sawmill had its performance optimized with the use of techniques of operational research in a cutting problem. The whole linear programming technique was used for an estimated demand for the quantity of each of the pieces produced. The results showed that only five models are required to meet such demand, so the yield is 43.18%

Behavior of two Leishmania infantum strains?evaluation of susceptibility to antimonials and expression of microRNAs in experimentally infected J774 macrophages and in BALB/c mice.

Strains of the same Leishmania parasite species, isolated from different host organisms, may exhibit unique infection profiles and induce a change in the expression of microRNAs among host macrophages and in model host mice. MicroRNAs (MiR) are endogenous molecules of about 22 nucleotides that are involved in many regulatory processes, including the vertebrate host immune response. In this respect, the infectivity and susceptibility to antimonials of two L. infantum strains, BH46, isolated from human, and OP46, isolated from symptomatic dog, were characterized in J774 macrophages and BALB/c mice. Parasite burden was assessed in the liver, spleen, and bone marrow using the serial limiting dilution technique. A higher parasite burden was observed in the spleen and bone marrow of animals infected with OP46 compared to BH46 strain. Our results also showed that OP46 was less susceptible to the antimonials. In addition, miR-122 and miR-155 expression was evaluated in the liver and J774 macrophages, and in spleens from infected animals, respectively. An increase was observed in the expression of miR-155 in J774 macrophages infected with both strains compared to uninfected cells, with a higher expression in cells infected with OP46. However, no difference in the expression of miR-122 and miR-155 was observed in the infected animals. Thus, this study shows that OP46 was more infective for mice, it caused a higher increase in miR-155 expression in infected macrophages and was less susceptible to the antimonials evaluated. These data suggest that alteration in miR-155 level likely plays a role in regulating the response to L. infantum

Antiproliferative and toxicogenomic effects of resveratrol in bladder cancer cells with different TP53 status.

The antitumor activity of resveratrol, a polyphenolic compound found mainly in grapes, has been studied in several types of cancer. In bladder cancer, its antiproliferative effects have already been demonstrated; however, its mechanism of action is not completely understood. The aim of this study was to evaluate resveratrol antitumor activity (12.5, 25, 50, 100, 150, 200, and 250 ?M) and its possible mechanisms of action in bladder tumor cells with different TP53 gene status (RT4, grade 1, TP53 wild type; 5637-grade 2 and T24-grade 3, TP53 mutated). Cell proliferation, clonogenic survival, morphological changes, cell cycle progression, apoptosis rates, genotoxicity, global methylation, immunocytochemistry for p53 and PCNA and relative expression profiles of the AKT, mTOR, RASSF1A, HOXB3, SRC, PLK1, and DNMT1 were evaluated. Resveratrol decreased cell proliferation and induced DNA damage in all cell lines. Regarding the long-term effects, resveratrol reduced the number of colonies in all cell lines; however, TP53 wild type cells were more resistant. Increased rates of apoptosis were found in the TP53 wild type cells and this was accompanied by AKT, mTOR, and SRC downregulation. In addition, the resveratrol antiproliferative effects in wild type TP53 cells were accompanied by modulation of the DNMT1 gene. In the TP53 mutated cells, cell cycle arrest at S phase with PLK1 downregulation was observed. Additionally, there was modulation of the HOXB3/RASSF1A pathway and nuclear PCNA reduction in the highest-grade cells. In conclusion, resveratrol has antiproliferative activity in bladder tumor cells; however, the mechanisms of action are dependent on TP53 status

Antiproliferative and toxicogenomic effects of resveratrol in bladder cancer cells with different TP53

The antitumor activity of resveratrol, a polyphenolic compound found mainly in grapes, has been studied in several types of cancer. In bladder cancer, its antiproliferative effects have already been demonstrated; however, its mechanism of action is not completely understood. The aim of this study was to evaluate resveratrol antitumor activity (12.5, 25, 50, 100, 150, 200, and 250 ?M) and its possible mechanisms of action in bladder tumor cells with different TP53 gene status (RT4, grade 1, TP53 wild type; 5637-grade 2 and T24-grade 3, TP53 mutated). Cell proliferation, clonogenic survival, morphological changes, cell cycle progression, apoptosis rates, genotoxicity, global methylation, immunocytochemistry for p53 and PCNA and relative expression profiles of the AKT, mTOR, RASSF1A, HOXB3, SRC, PLK1, and DNMT1 were evaluated. Resveratrol decreased cell proliferation and induced DNA damage in all cell lines. Regarding the long-term effects, resveratrol reduced the number of colonies in all cell lines; however, TP53 wild type cells were more resistant. Increased rates of apoptosis were found in the TP53 wild type cells and this was accompanied by AKT, mTOR, and SRC downregulation. In addition, the resveratrol antiproliferative effects in wild type TP53 cells were accompanied by modulation of the DNMT1 gene. In the TP53 mutated cells, cell cycle arrest at S phase with PLK1 downregulation was observed. Additionally, there was modulation of the HOXB3/RASSF1A pathway and nuclear PCNA reduction in the highest-grade cells. In conclusion, resveratrol has antiproliferative activity in bladder tumor cells; however, the mechanisms of action are dependent on TP53 status