Caracterização e quantificação de marcadores químicos do extrato hidroetanólico das folhas de Kalanchoe brasiliensis Cambess

Kalanchoe brasiliensis Cambess (Crassulaceae), commonly known as saião , coirama branca , folha grossa , is originally from Brazil and commonly found in São Paulo to Bahia, mainly in the coastal zone. Regarding of biological activities, most preclinical studies were found in the literature, mainly about the anti-inflammatory activity of extracts obtained from leaves and / or aerial parts of K. brasiliensis. As regards the chemical constitution, it has been reported mainly the presence of flavonoids in the leaves of the species, but until this moment did not knows which are the active compounds. Although it is a species widely used in traditional medicine in Brazil, there is no monograph about the quality parameters of the plant drug. In this context, this study aims to characterize and quantify the chemical markers of hydroethanolic extract (HE) from the leaves of K. brasiliensis, which can be used in quality control of plant drug and derivatives obtained from this species. The methodology was divided into two parts: i. Phytochemical study: to fractionate, isolate and characterizate of the chemical (s) marker (s) of the HE from the leaves of K. brasiliensis; ii. To Developed validate of analytical method by High Performance Liquid Chromatography (HPLC)-diode array detector (DAD) to quantify the chemical (s) marker (s) of the EH. i. The EH 50% was prepared by turbo extraction method. It was then submitted to liquid-liquid partition, obtaining dichloromethane, n-butanol and ethyl acetate (AcOEt) fractions. The AcOEt fraction was selected to continue the fractionation process, because it has a chemical profile rich in flavonoids. The acOEt fraction was submitted to column chromatography using different systems for obtaining the compound Kb1. To identify this compound, it was submitted to UV analysis ii. For quantitative analysis, the EH was analyzed by HPLC, using different methods. After selecting the most appropriate method, which showed satisfactory resolution and symmetrical peaks, it was validated according to parameters in the RE 899/2003. As result, it was obtained from the AcOEt fraction the compound Kb1 (2.7 mg). Until this moment, the basic nucleus was characterized by UV analysis using shift reagents. The partial chemical structure of the compound Kb1 was identified as a flavonol, containing hydroxyls in 3 , 4 position (ring A), 5 and 7 free (ring B) and a replacement of the C3 hydroxyl by a sugar. As the analysis were performed in the HPLC coupled to a DAD, we observed that the UV spectrum of the major peaks of EH from K. brasiliensis shown similar UV spectrum. According to the literature, it has been reported the presence of patuletin glycosydes derivatives in the leaves of this species. Therefore, it is suggested that the compound Kb1 is glycosylated patuletin derivative. Probably the sugar (s) unit(s) are linked in the C3 in the C ring. . Regarding the development of HPLC analytical method, the system used consists of phase A: water: formic acid (99,7:0,3, v / v) and phase B: methanol: formic acid (99,7:0,3, v / v), elution gradient of 40% B - 58% B in 50 minutes, ccolumn (Hichrom ®) C18 (250x4, 0 mm, 5 μm), flow rate 0.8 mL / min, UV detection at 370 nm, temperature 25 ° C. In the analysis performed with the co-injection of thecompound Kb1 + HE of K. brasiliensis was observed that it is one of the major compounds with a retention time of 12.47 minutes and had a content of 15.3% in EH of leaves from K. brasiliensis. The method proved to be linear, precise, accurate and reproducible. According to these results, it was observed that compound Kb1 can be used as a chemical marker of EH from leaves of K. brasiliensis, to assist in quality control of drug plant and its derivatives

Avaliação in vitro da eficácia de produtos homeopáticos contendo momordica charantia através de bioensaios

Homeopathic medicines have been used for over two hundred years without the examination of their effects on in vivo and in vitro assays, due to the peculiarity of homeopathic preparations, the high dilution, which creates a challenge for the use of usual analytical techniques of quality control of medicine.Although there is scarcity of literature and variety of experiments, recently there have been some studies with few in vitro assays which have shown positive responses when evaluating the mechanism of action of homeopathic medicines which are able to act on a specific system.The present study aims to evaluate the efficacy of homeopathic products containing Momordica charantia through bioassays.Homeopathic products were tested by the MTT to assess cytotoxicity in RAW 264.7 (macrophage-like cells) and in tumor cells HeLa (human cervical adenocarcinoma cells), CHO K1 (Chinese hamster ovary cells), PANC-1 (human pancreas cancer cells) and PC-3 (human prostate cancer cells), dosage of inflammatory mediators NO, TNF-α and IL-6 released by RAW 264.7 cells, analysis of the death process and cell cycle changes of PC-3 by flow cytometry. The data demonstrate that homeopathic products of Momordica charantia did not show cytotoxicity to RAW 264.7, increased the production of inflammatory mediators by RAW 264.7 synergistically with LPS, showed cytotoxicity to PC-3 with change in its cell cycle inhibiting its proliferation, being the 30CH the most potent sample. Correlation studies were conducted in order to evaluate the possible in vitro applicable models to the quality control of homeopathic products with Momordica charantia. The data showed that the best applicable models in assessing the quality are the MTT to assess cytotoxicity in RAW 264.7 and PC-3 in 24 hours for Momordica charantia fruit products and dosage of NO production by RAW 264.7 with and without LPS

Desenvolvimento e validação de método analítico para determinação de WE010, WE014 e WE017

The synthetic guanylhydrazones WE010 (3,5-di-tert-butil-4-hidroxibenzaldehyde-guanylhydrazone), WE014 (4-bifenilcarboxialdehydeguanylhydrazone) and WE017 (3,4-diclorobenzaldehydeguanylhydrazone) showed high cytotoxic activity in terms of percentage inhibition of cancer cells growth. However, further progress in the development of these drug candidates requires precise and convenient methods for their qualitative and quantitative analyses. The aim of this study was to develop and validate High Performance Liquid Chromatography with diode-array detection (HPLC-DAD) and Ultra Fast Liquid Chromatography with diode-array detection (UFLC-DAD) methods suitable for as simultaneous as isolated determination of studied guanylhydrazones, based on the optimization of chromatographic parameters and obtaining reduced detection times. The chromatographic analyses of analytes by HPLC were performed on C18 ACE analytical column (150 mm x 4.6 mm), with a particle size of 5.0 μm. Among all the conditions assayed, the best results of separation were obtained with a mixture of methanol:water (60:40, v/v) as the mobile phase at a flow rate 1.5mL/min and pH of 3.5 adjusted at acetic acid. The UFLC method was developed by experimetal desing techniques in order to find optimal chromatographic analytical conditions, which were achieved on XR-ODS analytical column (50 mm x 3.0 mm), with a particle size of 2,2 μm, maintained at 25 ºC. The mobile phase was consisted of methanol:water (65:35, v/v) with 0.1% triethylamine (TEA) and pH of 3.5 adjusted at acetic acid, at a flow rate 0.5 mL/min. The procedure were validated following evaluating parameters such as specificity, linearity, limits of detection (LD) and quantification (LQ), precision, accuracy and robustness, giving results within the acceptable range. Although the UFLC method shows better sensitivity (lower values of LD and LQ), robustness (lower rates of relative standard deviation) and minimize spending time and solvent, both developed methods were adequately applied to the analysis of guanylhydrazones molecules, may be used in routine of quality control laboratories. Keywords: guanylhydrazones, HPLC/DAD, UFLC/DAD, validation of analitical method

Testes bioquímicos para avaliação do vigor em sementes de Mimosa caesalpiniaefolia Benth

Mimosa caesalpiniaefolia Benth. is a forest species of the Mimosaceae family, recommended for recovery of degraded areas. The evaluation of vigor by biochemical tests have been an important tool in the control of seed quality programs, and the electrical conductivity and potassium leaching the most efficient in the verifying the physiological potential. The objective, therefore, to adjust the methodology of the electrical conductivity test for seeds of M. caesalpiniaefolia, for then compare the efficiency of this test with the potassium in the evaluation of seed vigor of different lots of seeds M. caesalpiniaefolia. To test the adequacy of the electrical conductivity were used different combinations of temperatures , 25 °C and 30 ºC, number of seeds , 25 and 50, periods of imbibition , 4 , 8 , 12 , 16 and 24 hours , and volumes deionized water, 50 mL and 75mL. For potassium leaching test, which was conducted from the results achieved by the methodology of the adequacy of the electrical conductivity test, to compare the efficiency of both tests , in the classification of seeds at different levels of vigor, and the period 4 hours also evaluated because the potassium leaching test can be more efficient in the shortest time . The best combination obtained in experiment of electrical conductivity is 25 seeds soaked in 50 mL deionized or distilled water for 8 hours at a temperature of 30 ° C. Data were subjected to analysis of variance, the means were compared with each other by F tests and Tukey at 5 % probability, and when necessary polynomial regression analysis was performed. The electrical conductivity test performed at period eight hour proved to be more efficient in the separation of seed lots M. caesalpiniaefolia at different levels of vigor compared to the potassium test

Obtenção de insumo farmacêutico a partir das folhas de Cissampelos sympodialis Eichl e seu efeito sobre mediadores inflamatórios relevantes para a asma

Coordenação de Aperfeiçoamento de Pessoal de Nível Superior

O direito social fundamental de acesso à energia elétrica e a atuação estatal

Electrical energy today is an essential element in the life of any human being. Through the access to electrical energy it is possible to enjoy dignified conditions of life, having in mind the possibility of making use of minimal material conditions of life. The lack of access to electricity is directly linked to poverty and degrading conditions of life, in which are some communities in Brazil, especially the more isolated from urban centers. Access to the electric service is a determining factor for the preservation of human dignity, constitutional principle inscribe in the art.1 of the Federal Constitution, and the promotion of development, being a right of everyone and a duty of the State to promote universal access. For that reason, focuses mainly on the analysis of their setting as a fundamental social right and its importance for national development. For this, the theoretical and descriptive method was used, with normative and literary analysis, in particular the Constitution of 1988. This study also discusses the form of action of the State in the energy sector, to give effect to the fundamental social right of access to electricity, the characteristics of public service and the principles that guide it, in addition to the role of public policies in universalization of access, in particular the analysis of the Program Luz para Todos, and the function of regulation in the implementation of these policies and the provision of adequate public services.

O município brasileiro e o direito fundamental do acesso à justiça: a prestação da assistência jurídica municipal ao necessitado

This scholarly work aims to investigate the feasibility and constitutionality of access to justice through the provision of full and free legal assistance by the Brazilian municipalities. Investigates the historical aspects of federalism in a global context, emphasizing the contributions left by American federalism. In the Brazilian context, emphasizing the importance of municipalities as federal entities and their outstanding characteristics, while addressing regional issues of federalism. Leanings to the more detailed analysis of the Brazilian municipalities, contextualizing its legal status, its independence and its constitutional powers. It is emphasized in the same way, the relevant transformations of Brazilian municipalities over the last twenty years of this Constitution of the Federative Republic of Brazil in 1988, especially the various constitutional amendments that affected the local autonomy and budgetary aspects, fiscal and skills, bringing significant changes to the municipalities. It is an approach to the concept of justice and deepening the studies on the fundamental right of access to justice in its various connotations. In this vein, it is a study on the legal advice provided in Brazil, especially the powers of the Public Defender of the States and Union, as well as the provision of such public service by Brazilian municipalities and its relevance to citizens in need. At this point, it deepens the relevance of the theme of this dissertation earning the implications of municipal performance in the provision of legal assistance provided to the needy, and the activity of the Municipal Attorney or legal counsel in conducting such a task and its implications for legal and procedural especially on the constitutionality or otherwise of the conduct of such public service, confronting the constitutional articles that are correlated with the subject. Within this context, evaluates the municipal legal assistance under the test of constitutionality, in particular the assistance given by the Executive, through the Municipal Attorney or specialized secretariats and that provided by the Legislature, although it only has the typical functions of legislating and control the municipal accounts, comes in a few municipalities in Brazil deploying sectors with the performance of legal services to the needy. At this point the thesis, one wonders if some important aspects of this activity such as political influence and patronage, very common in day-to-day municipal prosecutors and legal advisers, public employees or occupying commissioned positions within the municipal administrative structure in several municipalities throughout Brazil. Finally, there will be a conclusion as to the constitutionality of the service being done by presenting proposals and recommendations that may improve the municipal legal aid, allowing a constitutional backing to this important service is being provided in capital cities and municipalities throughout the length of Brazil

Defensoria pública e sua legitimidade à tutela jurídica dos direitos humanos

The realization of human rights is a prerequisite to the development of peoples, this requires legal mechanisms and techniques to its consistent and effective promotion, protection and fulfillment. So, agree that there is an institution or public agency created for the purpose of protecting those who suffer most in the face of human rights violations: the needy. In Brazil, among other institutions and public agencies, the responsibility of the Public Defender to promote the protection of human rights. The constitutional system recognizes the institution in its essence the role of the state court, whose duty is to provide guidance and legal defense of the needy. The legal system as a whole sufraga the relevance of the Ombudsman as a mainspring of human rights. In the prison system, with the ultimate regulatory changes, such as Law 12.313 of 2010 which introduced changes to the Law 7.210 of 1984, the institution must ensure the correct and humane enforcement of sentences and the security measures pertaining to the needy. With the Complementary Law 132 of 2009, to systematize other duties of the Public Defender, highlighting their contribution to the movement of access to justice. Within the OAS, to adopt Resolution 2656, 2011, characterizing, with ruler and compass, the relevance of the Ombudsman access to justice and protection of human rights. In this step, the present study concerns the role of Defender in the legal protection of human rights, through monographic and deductive methods, as there remains a technical and theoretical connection between these two points themed legal phenomenon, since the rights humans, especially after the second half of the twentieth century, form the basis of the legal system of the major Western nations in the world. This led, therefore, the emergence of technical and legal institutions aimed at realizing human rights. This applies to the Defender. Access to justice and public service provision of legal assistance are human rights, therefore, essential to humans and necessary for social inclusion. Countries such as Brazil, marked by social inequality, depend on the structuring of institutions like the Defender, designed to promote citizenship to the Brazilian people

A Servidão Administrativa e o Estado Regulador: Compatibilidade com o Princípio Constitucional da Legalidade e a Limitação ao Direito Real de Propriedade

The economic changes occurred in the 90s, with the restructuring and privatization of various sectors of the economy have led to a redefinition of the State role, assuming a position of regulator and supervisor of public services in place to direct its role as straight intervenor. It is through the regulatory agencies, autarchies with special legal personality under public law, that the Regulator State will act. In this context, the first objective of this research is to analyze the legality of easements imposed by entities of the Direct Administration and Regulatory Agencies, whose execution is delegated to legal persons of private law, being those public service companies or mixed-economy societies. This examination in question the limits of servitude as a restrictive institute of property rights, observing the principles of function, supremacy of the public interests over the private ones, legality and the separation of powers. Defend the property rights like a fundamental right and your insurance as determining factor of economic development and social justice. Use the procedure in use will be the historiccomparative procedure, in order to demonstrate the legality of the public act as a maximum attempt to preserve the balance between the expansion of public services in various sectors of the economy, and the preservation of property rights, through regulation

Testes de vigor para avaliação de sementes de pepino

Dentro de um programa de controle de qualidade, a avaliação do vigor de sementes é fundamental e necessária para o sucesso da produção. O trabalho teve por objetivo comparar diferentes métodos para avaliação do potencial fisiológico de sementes de pepino. Para tanto, cinco lotes de sementes do híbrido Safira foram submetidos aos seguintes testes: germinação, primeira contagem de germinação, condutividade elétrica (4x50 sementes; 25ºC; 50 e 75mL; 12, 18 e 24h), germinação a baixa temperatura (4x50 sementes; 18ºC; contagens aos quatro e oito dias após a semeadura); envelhecimento acelerado tradicional (41ºC; 48, 72 e 96h) e com solução saturada de NaCl (41ºC; 72 e 96h). Pelos resultados obtidos, pode-se concluir que os testes de condutividade elétrica (50 e 75mL; 12, 18 e 24h) e de envelhecimento acelerado com o uso de solução salina (41ºC, por 72h), possibilitam a melhor identificação de lotes com diferentes níveis de qualidade fisiológica.

Temperatura e período de exposição de sementes de milho no teste de envelhecimento acelerado

O teste de envelhecimento acelerado (EA) apresenta possibilidades de uso nos programas de controle de qualidade de sementes de milho. Porém, para uso em rotina, há necessidade de padronização das condições de temperatura e período de exposição das sementes na realização do teste. Esta pesquisa teve por objetivo avaliar o efeito da combinação de duas temperaturas (42 e 45ºC) e dois períodos de envelhecimento (72 e 96h) nos resultados do teste de envelhecimento acelerado em sementes de milho. Para isso, sementes de seis genótipos de milho, tratadas e sem tratamento fungicida, com altos valores de germinação inicial, semelhantes entre si, dentro de cada genótipo, foram submetidas ao EA, de acordo com condições de temperatura e período de exposição pré-estabelecidos. Utilizou-se o delineamento experimental inteiramente casualizado com os tratamentos (lotes x germinação, antes e após o EA) dispostos em esquema fatorial, empregando-se o teste de Tukey a 5% de probabilidade na comparação das médias. Verificou-se que sementes de milho apresentam diferenças de sensibilidade ao envelhecimento acelerado, condicionadas pelo genótipo e tratamento fungicida. Contudo, a combinação 45ºC por 72h possibilita separar lotes de qualidade fisiológica superior e inferior, independentemente do genótipo. O período de exposição das sementes ao envelhecimento acelerado determina o teor de água atingido pelas mesmas.

Metodologia alternativa para condução do teste de envelhecimento acelerado em sementes de milho

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

Envelhecimento acelerado como teste de vigor para sementes de milho e soja

A avaliação do vigor de sementes tem sido ferramenta fundamental dentro do programa de controle de qualidade, sendo o teste de envelhecimento acelerado (EA) parte importante do processo. O trabalho foi desenvolvido com o objetivo de confirmar metodologia específica para avaliação do vigor de sementes, em laboratório, tendo como base o EA. Utilizou sementes de milho e soja, seis lotes para cada espécie, com padrões de qualidade (germinação) comercialmente aceitas. As condições de envelhecimento préestabelecidas foram 42ºC/96h e 45ºC/72h para milho e 41ºC/48 e 72h e 42ºC/48 e 72h para soja. Para cada combinação de temperatura e período de envelhecimento, as sementes foram colocadas em caixas plásticas, sobre tela, com 40mL de água destilada, distribuídas em camada única e com base no peso das mesmas. O envelhecimento foi feito usando-se dois tipos de câmara, BOD e jaquetada de água (CJ). Foi determinado o teor de água (TA) e a germinação das sementes antes e após o EA. A absorção de água pelas sementes considerando-se todas as combinações de período de exposição e temperatura, as duas câmaras e as duas formas de determinação do TA, foi maior quando as sementes foram acondicionadas em camada única em relação aquelas pesadas (massa constante), usando-se a câmara jaquetada. Para o milho, a combinação de 45ºC/72h com o acondicionamento das sementes em camada única foi o método mais eficiente para a separação dos lotes quanto ao vigor. Para soja, a combinação 42ºC/48h foi mais eficiente na separação dos lotes, não havendo interferência do método de acondicionamento das sementes, na tela. Conclui-se que a camada única e a câmara tipo BOD devem ser preferencialmente utilizadas no teste EA face aos menores custos de aquisição e manutenção da BOD, visando à separação de lotes quanto aos níveis de vigor, para milho e soja.

Avaliação do potencial fisiológico de lotes de sementes de soja

Vários procedimentos são utilizados para avaliar o potencial fisiológico de sementes, sendo fundamental a utilização daqueles que reflitam melhor o desempenho em campo. A pesquisa teve como objetivo avaliar o potencial fisiológico de sementes de soja por meio de testes de avaliação rápida, de vigor, de emergência de plântulas em campo e análise computadorizada de imagens (SVIS®), procurando identificar o procedimento mais eficiente na separação de lotes. Cinco lotes das cultivares BRS Valiosa RR e CD 208 foram avaliados em duas épocas (antes e após armazenamento) pelos testes de germinação, de avaliação rápida (embebição em água e pH do exsudato), de vigor (envelhecimento acelerado e condutividade elétrica), de emergência de plântulas em campo e análise computadorizada de imagens (SVIS®). Os testes de germinação em areia, de condutividade elétrica e de embebição em água foram eficientes na separação dos lotes, mas apenas este último possibilitou identificar diferenças consistentes entre os lotes nos dois períodos avaliados. Assim, o teste de embebição em água pode ser considerado promissor na composição de programas de controle de qualidade.

Alternative procedure for the cold test for soybean seeds

Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq)