RESUMO: O objetivo deste estudo foi avaliar a composição e as atividades quimiopreventiva de câncer e antioxidante da semente, polpa e casca de uvas híbridas Maximo (IAC 138-22) colhidas em diferentes estágios de maturação. Os teores de compostos fenólicos totais, antocianinas e taninos foram determinados a partir dos extratos etanólicos. O conteúdo de fenólicos e taninos totais foi determinado pelo método de Folin-Ciocaulteau, e o teor de antocianinas totais foi determinado pelo método do pH diferencial. A atividade antioxidante foi avaliada por diferentes métodos químicos, e a viabilidade celular (macrófagos e melanomas), e atividade quimiopreventiva de câncer foi testada em culturas de células por ensaio de indução de quinona redutase, ensaio de atividade inibidora de NF-κB induzido por TNF-α e ensaio de atividade inibidora de aromatase. A uva híbrida apresentou alto teor de fenólicos nas sementes e na casca, teor que reduziu significativamente com o amadurecimento. Os resultados mostraram que o amadurecimento dos frutos afetou significativamente a composição e as atividades antioxidantes e quimiopreventivas de câncer. Com exceção do teor de antocianinas, os resultados mais significativos foram observados para as sementes verdes, com o estádio mais verde apresentando os melhores resultados. As sementes verdes apresentaram a maior atividade antioxidante, mesmo quando comparadas ao resveratrol livre. Na avaliação da atividade quimiopreventiva do câncer, as sementes verdes mostraram capacidade de inibir a atividade do NF-κB e da aromatase, fatores importantes para o estágio inicial do desenvolvimento do câncer. Além disso, nenhum extrato apresentou atividade citotóxica. Portanto, as sementes verdes desta uva apresentam resultados promissores dadas as fortes atividades antioxidantes e quimiopreventivas do câncer aqui confirmadas, sugerindo efeitos benéficos à saúde.

ABSTRACT: This study evaluated the composition and the cancer chemopreventive and antioxidant activities of the seed, skin, and pulp of Maximo hybrid grapes (IAC 138-22) harvested at different ripening stages. The content of total phenolic compounds and tannins were determined by the Folin-Ciocaulteau method, and total anthocyanins were determined using the differential pH method. Different chemical methods assessed the antioxidant activity. Cell viability (macrophage and melanoma skin cancer) and cancer chemopreventive activity were tested in cell lines by quinone reductase induction assay, TNF-α-induced NF-κB inhibitory activity assay, and aromatase inhibitory activity assay. The hybrid grape had high phenolic contents in both seeds and skin, significantly reduced upon ripening. The results showed that fruit ripening significantly affected both the composition and the antioxidant and cancer chemopreventive activities. Except for the anthocyanin content, the most significant results were observed for green seeds, with the greenest stage showing the best results. The green seeds showed the highest antioxidant activity, even when compared to free resveratrol. In the assessment of cancer chemopreventive activity, the green seeds showed the ability to inhibit N-FκB and aromatase activity, important factors for the early stage of cancer development. Besides, no extracts showed cytotoxicity. Therefore, this grape’s green seeds showed strong antioxidant and cancer chemopreventive activities confirmed herein, suggesting beneficial health effects.

Plants have been used for centuries as therapeutic resources for various pathological conditions and are occasionally considered as the only resource available to some communities. However, knowledge on the potential of traditional medicinal plants have been lost over the years. Strategies that can guarantee the prospection of popular knowledge regarding medicinal plants to stimulate the spread of information between generations are therefore of enormous value. Thus, the present study aimed to investigate the use and knowledge on medicinal plants of students from a public school in Vila Velha (Brazil) and their associated community, including parents. In this urban locality, data were obtained by the application of a semi-structured questionnaire to 206 people from the school community. Although homegrown cultivation seems to be an uncommon practice in urban contexts, the use of medicinal plants by the sampled population was significant (80%), as well as the interviewees’ confidence on its therapeutic action (90%). It is worth mentioning that more than 70% of the respondents presented their parents and/or grandparents as their main source of information, highlighting the importance of popular knowledge valorization.

RESUMO Objetivo: Avaliar o efeito da pasteurização nas propriedades antioxidantes e oxidantes do leite humano. Métodos: Foram utilizadas 42 amostras de leite cru e após a pasteurização, para avaliação da atividade antioxidante pelos métodos da capacidade redutora do ferro e sequestro dos radicais derivados do ácido 2,2’-azino-bis (3-etilbenzotiazolina-6-sulfônico). A peroxidação lipídica (PL) foi estimada pela determinação das substâncias reativas ao ácido tiobarbitúrico e pela avaliação dos produtos proteicos de oxidação avançada. Resultados: Não se observou diferença significante no leite humano cru nem após a pasteurização em relação às propriedades antioxidantes determinadas pelo método da redução do ferro (50,0±3,4% e 48,8±3,0%, respectivamente) e pelo sequestro dos radicais derivados do ácido 2,2’-azino-bis (3-etilbenzotiazolina-6-sulfônico) (28,9±1,5% e 31,2±1,3%, respectivamente). Os resultados médios de malondialdeído [MDA] (62,6±4,1 e 64,3±3,6 µM/mg) e produtos de oxidação proteica (59,4±3,4 e 54,2±3,8 µM/L) entre os grupos leite fresco e leite pasteurizado, respectivamente, não evidenciaram diferença significante. Conclusões: Os dados demonstraram que o leite humano possui importante atividade antioxidante e que o processo de pasteurização não interfere nessa propriedade, evitando assim a peroxidação dos lipídios e a formação de produtos avançados de oxidação proteica.

ABSTRACT Objective: To evaluate the effect of pasteurization on antioxidant and oxidant properties of human milk. Methods: 42 samples of milk before and after pasteurisation were used to evaluate the antioxidant activity by the ferric reducing capacity and by scavenging the 2,2’-azino-bis 3-ethylbenzthiazoline-6-sulfonic acid radical. Lipid peroxidation was estimated by the concentration of malondialdehyde product using the thiobarbituric acid reactive substances assay and by the evaluation of advanced oxidation protein products. Results: No significant difference was observed in fresh human milk and after pasteurization in relation to antioxidant properties determined by the ferric reducing capacity (50.0±3.4% and 48.8±3.0%, respectively) and by scavenging the 2,2’-azino-bis 3-ethylbenzthiazoline-6-sulfonic acid radical (28.9±1.5% and 31.2±1.3%, respectively). The results of malondialdehyde (62.6±4.1 and 64.3±3.6 µM/mg) and protein oxidation products (59.4±3.4 and 54.2±3.8 µM/L) of fresh and pasteurized milk, respectively, did not exhibited any significant difference. Conclusions: This data showed that human milk has an important antioxidant activity and that the pasteurizing process does not influence the antioxidant capacity, avoiding the peroxidation of breast milk lipids and the formation of advanced protein oxidation products.

ABSTRACT The objective was to investigate the total saponin and protodioscin concentrations and the cytotoxicity in vitro, of five samples of the plant Tribulus terrestris, commercially available in the metropolitan region of Vitória - Espirito Santo, Brazil, and to compare them with the aqueous extract of the plant. The chromatographic profile and quantification of protodioscin in commercial samples and plant extract were evaluated by LC-MS/MS. The percentage of total saponins were determined by the colorimetric method. Extracts and protodioscin cytotoxicity were analyzed by the MTT assay in three cell lineages: fibroblasts (L929), ovarian cancer (Ovcar3) and murine hepatoma (Hepa1c1c7). All extracts displayed high levels of total saponins (207.2 to 780.3 mg g-1 of dry extract). The chromatographic profile revealed a wide diversity of compounds, and the saponin protodioscin was detected in only two extracts. One extract displayed high cytotoxicity, with IC50 values of 157.0, 38.2 and 7.4 µg mL-1 for the Ovcar3, Hepa1c1c7 and L929 cell lines, respectively. The other extracts displayed cytotoxic effects only at concentrations equal to or greater than 125.0 µg mL-1. Surprisingly, the most cytotoxic extract displayed the highest protodioscin concentration. Therefore, it is suggested that these products be marketed with caution, and followed-up by a certified healthcare professional.

ABSTRACT Seaweeds are related to anti-inflammatory, anti-bacterial and anti-noceptive effects. This work aimed to verify the potential of seaweed Padina gymnospora (Kützing) Sonder 1871 to improve wound healing in vitro. P. gymnospora was collected at a bethonic area in Espirito Santo. Methanolic extract of P. gymnospora was obtained by percolation. To determine cytotoxicity, colorimetric MTT tests were performed against normal fibroblasts (L929), macrophages (RAW 264.7) and human ovarian carcinoma (OVCAR-3) cell lines using concentration range of 12–110 µg ml-1. To evaluate in vitro wound healing, monolayer of fibroblasts L929 was seeded and artificial wounded. Cell proliferation was blocked by 5 µg ml-1 Mytomycin C. Nitric oxide inhibition was quantified with Raw 264.7 by Griess reaction. Minimal inhibitory concentration (MIC) against Staphylococcus aureus was determined. Eletrospray ionization with Fourier transform ion cyclotron resonance mass spectrometry (ESI-FT-ICR MS) was applied to detail composition of P. gymnospora methanolic extract. No cytotoxic effect in all cell lines was detected until the maximum concentration of 110 µg ml-1. P. gymnospora promoted significantly migration at the concentration of 25 µg ml-1 (p < 0.05). A prominent inhibition of nitric oxide formation was achieved in a concentration of 20 µg ml-1 of methanolic extract of P. gymnospora (62.06 ± 1.20%). Antibacterial activity against S. aureus could be demonstrated with MIC of 500 µg ml-1. ESI-FT-ICR MS analysis indicated eleven molecules between then, linolenic, oleic and linoleic acid. P. gymnospora favored wound repair in vitro what could be related to its fatty acid composition. In addition, its antimicrobial effect, and NO inhibition activity contribute for a new approach of P. gymnospora as a promise natural product for treatment of cutaneous wound.

<p>The aim of this study was to investigate the presence of acephate, carbaryl, carbendazim, chlorpyrifos, dichlorvos, fenpropathrin, methamidophos and permethrin in conventionally and organically grown tomatoes from Espírito Santo, Brazil, between May 2012 and April 2013. QuEChERS sample preparation was used and analysis was performed by LC-MS/MS and GC-MS. The method validation showed good linearity (R > 0.99), precision (RSD < 13%) and accuracy (89 - 123%), and the limits of quantification were below 0.01 mg kg^-1 for all pesticides. The pesticides acephate, carbaryl (0.23 mg kg^-1; exceeding maximum residue levels), carbendazim, chlorpyriphos, dichlorvos, fenpropathrin (0.41 mg kg^-1; exceeding maximum residue levels), methamidophos and permethrin (0.51 mg kg^-1, exceeding maximum residue levels) were found in conventional tomatoes. In organic tomatoes, only one sample exhibited the presence of permethrin (0.21 mg kg^-1). Considering that the pesticides found are toxic and carcinogenic, and given the high incidence of irregularities, it is important to implement government actions to ensure consumer safety.</p>

Avaliaram-se os efeitos do valdecoxibe, um inibidor seletivo da cicloxigenase-2 (COX-2), sobre os parâmetros sangüíneos da coagulação em ratos Wistar utilizando-se paralelamente a aspirina. Os animais foram divididos em grupos e submetidos à administração oral diária de 10 mg/kg animal para o valdecoxib, 100 mg/kg animal para a aspirina e veículo para o grupo controle, durante quatro semanas. A coleta do sangue foi efetuada após uma, duas, três e quatro semanas, e as concentrações plasmáticas do valdecoxibe determinadas por cromatografia líquida, obtendo-se valores médios de 101,1 - 113,5 - 164 e 184,6 ng/ml, respectivamente. As amostras de plasma foram também usadas para as análises hematológicas e os resultados comparados aos dos controles. O valdecoxibe apresentou redução significativa no TTPA (18%) após duas semanas, e redução do TP (12,2%) após três semanas (P<0.05). Os efeitos observados na contagem de plaquetas e nos níveis plasmáticos de fibrinogênio não foram significativos. As atividades anti-fator Xa e anti-fator IIa apresentaram redução, porém os resultados foram significativos somente para o anti-fator Xa na terceira semana (6,7%). Os resultados experimentais com as concentrações plasmáticas alcançadas demonstram que o valdecoxibe pode induzir alterações nos parâmetros hematológicos dos ratos. Esses dados poderão contribuir para a melhor compreensão dos efeitos colaterais e uso do fármaco com segurança.

The effects of the cyclooxygenase (COX)-2 selective inhibitor, valdecoxib, on blood coagulation parameters were evaluated, along with aspirin in male Wistar rats. Groups of animals were administered a daily oral dose of 10 mg/kg rat of valdecoxib, 100 mg/kg rat of aspirin and the vehicle alone during 4 weeks. Blood samples were collected at the end of 1, 2, 3 and 4 weeks of administration period and the plasma concentrations of valdecoxib were determined by RP-HPLC giving mean values of 101.1, 113.5, 164.0 and 184.6 ng/mL, respectively. The same plasma samples were used for the analysis of hematological parameters and the results compared to the controls. Valdecoxib induced significant activated partial thromboplastin time reduction (18%) after 2 weeks and prothrombin time reduction (12.2%) after 3 weeks (P<0.05). There were no significant changes in the platelet count and fibrinogen levels. The anti-factor Xa and anti-factor IIa activities showed slight reductions, but only significant for the anti-factor Xa on the 3rd week (6.7%). The results showed that valdecoxib at the dose tested with the plasmatic concentrations induced some changes in the hemostatic function of rats, which can be helpful to understand the side effects and the safe use of the drug.

Realizou-se a identificação do fator estimulador da colônia de granulócitos humanos recombinante em produtos farmacêuticos por eletroforese em gel de poliacrilamida não redutora e imunodetecção com anticorpos específicos, que apresentaram banda única na região de, aproximadamente, 19 kDa. A avaliação de potência baseada na contagem do número de neutrófilos em camundongos com neutropenia forneceu valores entre 88,4 - 122,4 % em relação à potência declarada. A precisão expressa pela ponderação, calculada nos ensaios independentes, forneceu valores entre 141 e 432. As amostras lote-a-lote cumpriram os requisitos dos testes de toxicidade e endotoxinas bacterianas. Os resultados dos ensaios biológicos e imunológicos demonstram a qualidade dos produtos farmacêuticos em uso clínico, e as especificações sugeridas contribuem para assegurar a inocuidade e eficácia terapêutica dos produtos biológicos.

The identification of rhG-CSF was carried out in pharmaceutical preparations by non-reducing polyacrylamide gel electrophoresis and western blotting with specific antibodies, showing a single band in the 19 kDa region. The potency was assessed by the neutropenia mouse bioassay giving values between 88.4 and 122.4% of the stated potency. The precision index expressed by weight was between 141 and 432 for the independent assays. Batch-to-batch, the samples met the requirements for the safety test and bacterial endotoxins test. The biological and immunological results showed the quality of the products in clinical use and the specifications established contribute to assuring the safety and efficacy of biological medicines.

Realizou-se a avaliação biológica de potência de heparinas convencionais contra o 5º Padrão Internacional de heparina suína pelos ensaios preconizados da inibição da coagulação do plasma ovino (ICPO), tempo de tromboplastina parcial ativada (TTPA) e anti-fator Xa. Correlacionaram-se os resultados, demonstrando que o ICPO fornece potências em média 10,72%, significativamente superiores aos outros procedimentos. Analisou-se a influência de diferentes lotes de plasma sobre o resultado do ensaio do ICPO observando-se variação de até 7,32%. Como alternativa, padronizou-se o ensaio do anti-fator IIa e efetuou-se a determinação de potência das amostras obtendo-se valores reprodutíveis, comparáveis ao método do anti-fator Xa, que demonstram a viabilidade da inclusão como ensaio harmonizado para o controle da qualidade. Os resultados de potência, fornecidos lote a lote, em geral, cumprem as especificações farmacopéicas e demonstram a qualidade que garante a segurança e eficácia clínica dos medicamentos.

The biological evaluation of unfractionated heparins was carried out against the 5th International Standard of porcine heparin using the pharmacopoeial-recommended assays: the sheep plasma coagulation inhibition assay (SPCIA), activated partial thromboplastin time (APTT) and anti-factor Xa activity. The SPCIA method gave mean potencies which were 10.72% significantly higher than the other procedures. The effect of differences in plasma batches revealed potency variations of up to 7.32%. The anti-factor IIa assay was standardized as an alternative method for the potency assessment of heparin samples. The values obtained were in good agreement with the anti-factor Xa assay results, demonstrating the feasibility of the method as a harmonized assay for the quality control of medicines. Analysis of the potency estimates showed that generally batch-to-batch variations complied with pharmacopoeial specifications, demonstrating the overall quality of the products and assuring their safety and clinical efficacy.

Realizou-se a identificação de eritropoietina humana recombinante em produtos farmacêuticos comerciais por eletroforese em gel de poliacrilamida e detecção com anticorpos específicos, demonstrando-se típica banda larga, semelhante ao padrão de rhEPO da Farmacopéia Européia. Igualmente por focalização isoelétrica e imunodetecção, observaram-se 5-6 isoformas características de acordo com o laboratório produtor. A avaliação de potência efetuada através de ensaio biológico em camundongos normocitêmicos forneceu valores entre 67,6% e 119,4% em relação à declarada. A precisão dos ensaios combinados, calculada pela ponderação, variou de 200 a 389. Os testes de endotoxinas bacterianas, toxicidade e pH apresentaram valores variáveis conforme o lote. Concluiu-se destacando a importância dos testes e ensaios de controle para assegurar a qualidade lote a lote e garantir a eficácia terapêutica dos produtos farmacêuticos.

The identification of rhEPO in pharmaceutical products was carried out by polyacrylamide gel electrophoresis and detection with specific antibodies that revealed a typical single broad band similar to that obtained with the European Pharmacopoeia Biological Reference Preparation for erythropoietin. The isoelectric focusing with immunodetection revealed extensive heterogeneity with 5-6 isoforms, characteristic according to the manufacturer. The potency was assessed by the normocythaemic mouse assay with values within 67.6% and 119.4% calculated against the stated potency. The precision evaluated by the weight was within 200 and 389, for the combined assays. The toxicity, bacterial endotoxins, and pH gave variable results according to the batch. In conclusion, we emphasize the importance of the batch-to-batch tests and assays that would guarantee the quality and therapeutic efficacy of the pharmaceutical products.