A sensitive microextraction by packed sorbent-based methodology combined with ultra-high pressure liquid chromatography as a powerful technique for analysis of biologically active flavonols in wines

A new approach based on microextraction by packed sorbent (MEPS) and reversed-phase high-throughput ultra high pressure liquid chromatography (UHPLC) method that uses a gradient elution and diode array detection to quantitate three biologically active flavonols in wines, myricetin, quercetin, and kaempferol, is described. In addition to performing routine experiments to establish the validity of the assay to internationally accepted criteria (selectivity, linearity, sensitivity, precision, accuracy), experiments are included to assess the effect of the important experimental parameters such as the type of sorbent material (C2, C8, C18, SIL, and C8/SCX), number of extraction cycles (extract-discard), elution volume, sample volume, and ethanol content, on the MEPS performance. The optimal conditions of MEPS extraction were obtained using C8 sorbent and small sample volumes (250 μL) in five extraction cycle and in a short time period (about 5 min for the entire sample preparation step). Under optimized conditions, excellent linearity View the MathML source(Rvalues2>0.9963), limits of detection of 0.006 μg mL−1 (quercetin) to 0.013 μg mL−1 (myricetin) and precision within 0.5–3.1% were observed for the target flavonols. The average recoveries of myricetin, quercetin and kaempferol for real samples were 83.0–97.7% with relative standard deviation (RSD, %) lower than 1.6%. The results obtained showed that the most abundant flavonol in the analyzed samples was myricetin (5.8 ± 3.7 μg mL−1). Quercetin (0.97 ± 0.41 μg mL−1) and kaempferol (0.66 ± 0.24 μg mL−1) were found in a lower concentration. The optimized MEPSC8 method was compared with a reverse-phase solid-phase extraction (SPE) procedure using as sorbent a macroporous copolymer made from a balanced ratio of two monomers, the lipophilic divinylbenzene and the hydrophilic N-vinylpyrrolidone (Oasis HLB) were used as reference. MEPSC8 approach offers an attractive alternative for analysis of flavonols in wines, providing a number of advantages including highest extraction efficiency (from 85.9 ± 0.9% to 92.1 ± 0.5%) in the shortest extraction time with low solvent consumption, fast sample throughput, more environmentally friendly and easy to perform.

Comparison of prophylactic and therapeutic use of short-chain fatty acid enemas in diversion colitis: a study in Wistar rats

OBJECTIVES: To study the effect of short-chain fatty-acids on atrophy and inflammation of excluded colonic segments before and after the development of diversion colitis. INTRODUCTION: Diversion colitis is a chronic inflammatory process affecting the dysfunctional colon, possibly evolving with mucous and blood discharge. The most favored hypotheses to explain its development is short-chain fatty-acid deficiency in the colon lumen. METHODS: Wistar rats were submitted to colostomy with distal colon exclusion. Two control groups (A1 and B1) received rectally administered physiological saline, whereas two experimental groups (A2 and B2) received rectally administered short-chain fatty-acids. The A groups were prophylactically treated (5th to 40th days postoperatively), whereas the B groups were therapeutically treated (after post-operative day 40). The mucosal thickness of the excluded colon was measured histologically. The inflammatory reaction of the mucosal lamina propria and the lymphoid tissue response were quantified through established scores. RESULTS: There was a significant thickness recovery of the colonic mucosa in group B2 animals (p = 0.0001), which also exhibited a significant reduction in the number of eosinophilic polymorphonuclear cells in the lamina propria (p = 0.0126) and in the intestinal lumen (p = 0.0256). Group A2 showed no mucosal thickness recovery and significant increases in the numbers of lymphocytes (p = 0.0006) and eosinophilic polymorphonuclear cells in the lamina propria of the mucosa (p = 0.0022). CONCLUSION: Therapeutic use of short-chain fatty-acids significantly reduced eosinophilic polymorphonuclear cell numbers in the intestinal wall and in the colonic lumen; it also reversed the atrophy of the colonic mucosa. Prophylactic use did not impede the development of mucosal atrophy

Purificação, caracterização e avaliação da atividade antiproliferativa de um inibidor de quimotripsina tipo kunitz de semente de Eryhrina velutina

A chymotrypsin inhibitor was purified from Erythrina velutina seeds by ammonium sulphate fractionation, affinities chromatographies on Trypsin-Sepharose, Quimotrypsin-Sepharose and reversed phase C-18 FPLC/AKTA system. The inhibitor, named EvCI, shown molecular mass of 17 kDa, as determined by SDSPAGE. 2D-PAGE showed four isoinhibitors with pI values of 4,42, 4,63, 4,83 and 5,06, with molecular mass of 17 kDa each. The aminoacid sequence of EvCI was determined by MALDI-TOF-MS and showed a high similarity with other Kunitz-type inhibitor of Erythrina variegata. EvCI competitively inhibited chymotrypsin, with Ki of 4 x10-8 M, but did not inhibited trypsin, pancreatic elastase, bromelain and papain. The inhibitory activity of EvCI was stable over wide pH and temperature ranges. In the presence of DTT 100 mM for 120 min, EvCI lost 50 % of activity. Cytotoxicity was studied in HeLa, MDA, HepG2, K562 and PC3 cells after 72-h incubation period. EvCl inhibited HeLa cells growth with an IC50 value of 50 μg/ml. Subsequent studies in HeLa cells analysis of cell death by annexin V/PI double-staining and cell cycle, using flow cytometry. The results provide evidence for a cytostatic activity of EvCl and support further studies on potential application of this inhibitors as an antiproliferative agent in combined therapy against cervical cancer

Influência da adição de cálcio nas propriedades de pastas geopoliméricas destinadas a cimentação de poços de petróleo

Nowadays, the search for new technologies that are able to follow the upcoming challenges in oil industry is a constant. Always trying properties improvements of the used materials, looking for the best performance and greater life time. Besides the search for technologies that show an improvement of performance, the search for materials environmentally correct along the whole production process. In Oil well cementing, this search for new technologies passes through the development of slurry systems that support these requests and that are also environmentally friendly. In this context, the use of geopolymer slurries is a great alternative route to cementing oil wells. Besides having good properties, comparable to Portland cement slurries, this alternative material releases much less CO2 gas in the production of their root materials when compared the production of Portland cement, which releases tons of CO2. In order to improve the properties of geopolymer slurries has been added Calcium Oxide, as observed in other studies that slurries where the Calcium is present the values of compressive strength is greater. The addition has been realized based in the CaO/SiO2 molar ratio of 0.05, 0.10 and 0.15. Have been performed compressive strength tests, thickening time, rheology and fliud loss control test of the slurries, following NBR 9831, as well as the physical chemical characterization of XRD, SEM and TG. Has been observed in most of the tests the slurries follow a tendency until the ratio of 0.10, which inverses in the ratio 0.15. This behavior can be explained by two phenomena that occur simultaneously, the first one is the break of the polymer chains and a consequent increase in molucules mobility, which prevails until the ratio of 0.1, and the second is possible approach of the chains due to the capacity of the calcium ions stabilize the charges of two different aluminum. There is only one linearity in the mechanical behavior that can be attributed to the appereance of the C-S-H phase. Based on this, it is concluded that the phenomenon of breaking the polymer chains predominates until the ratio of 0.1, causing an increase of the filtrate volume, lower rheological parameters and increasing thickening time. From the ratio of 0.15 the approach of the chains predominates, and the behavior is reversed

Neustonic distribution of decapod planktonic stages and competence of brachyuran megalopae in coastal waters

Settlement rate may not reflect larval supply to coastal waters in different marine invertebrates and demersal fishes. The importance of near-shore oceanography and behaviour of late larval stages may be underestimated. The present study conducted neustonic sampling over station grids and along full-length transects at two embayments in south-eastern Brazil to (1) compare diurnal and nocturnal occurrence of most frequent decapod stages to assess their vertical movements, (2) describe the formation of larval patches and (3) measure competence of crab megalopae according to their distance to recruitment grounds. Several shrimp species apparently undergo a diel vertical migration, swimming crab megalopae showed no vertical movements and megalopae of the intertidal crab Pachygrapsus transversus revealed a reversed vertical migration. During the day, crab megalopae aggregated in convergence zones just below surface slicks. These larvae consisted of advanced, pre-moult stages, at both mid-bay and near-shore patches. Competence, measured as the time to metamorphosis in captivity, was similar between larval patches within each taxon. Yet, subtidal portunids moulted faster to juveniles than intertidal grapsids, possibly because they were closer to settlement grounds. Megalopae of Pachygrapsus from benthic collectors moulted faster than those from bay areas. These results suggest that alternative vertical migration patterns of late megalopae favour onshore transport, and actual competence takes place very close to suitable substrates, where larvae may remain for days before settlement. Lack of correlation between larval supply and settlement for Pachygrapsus suggests that biological processes, besides onshore transport, may play an important role in determining settlement success of coastal crabs.

Emprego dos ácidos graxos de cadeia curta na colite de derivação fecal : estudo em ratos Wistar

Diversion colitis is a chronic inflammatory process affecting the dysfunctional colon, after a colostomy. It is postulated that nutritional deficiency of the colonic epithelium by the absence of short-chain fatty acids (SCFA) is one of the factors responsible for the appearance of DC and that their employment could reverse the morphological changes of the mucosa. The treatment of choice for fecal diversion colitis (DC) is the reconstruction of the intestinal tract, although they suggested therapeutic options using enemas. This study evaluates the effect of SCFA in atrophy and inflammation in excluded colonic segments before and after the installation DC. Forty Wistar rats were divided into four groups (n = 10 for each group), submitted colostomy with distal colon exclusion. Two control groups (A1 and B1) received rectally administered physiological saline, whereas two experimental groups (A2 and B2) received rectally administered short-chain fatty-acids. The A groups were prophylactically treated (5th to 40th days postoperatively), whereas the B groups were therapeutically treated (after postoperative day 40), for 07 days. Histological sections stained with HE were used for histological analysis of the thickness of the colonic mucosa excluded (t- Student p ≤0.05). Inflammatory reaction of the lamina propria and mucosa were measured with scores previously established (Mann Whitney p ≤ 0.05). There was a significant thickness recovery of the colonic mucosa in group B2 animals (p = 0.0001), which also exhibited a significant reduction in the number of eosinophilic polymorphonuclear cells in the lamina propria (p = 0.0126) and in the intestinal lumen (p = 0.0256). Group A2 did not prevent the mucosal atrophy and significant increases in the numbers of lymphocytes (p=0.0006) and 50 eosinophilic polymorphonuclear cells in the lamina propria of the mucosa (p = 0.0022). Therapeutic use of short-chain fatty-acids significantly reduced eosinophilic polymorphonuclear cell numbers in the intestinal wall and in the colonic lumen; it also reversed the atrophy of the colonic mucosa. Prophylactic use did not impede the development of mucosal atrophy

Renal and cardiovascular effects of Bothrops marajoensis venom and phospholipase A(2)

Bothrops marajoensis is found in the savannah of Marajo Island in the State of Par S and regions of Amapa State, Brazil. The aim of the work was to study the renal and cardiovascular effects of the B. marajoensis venom and phospholipase A(2) (PLA(2)). The venom was fractionated by Protein Pack 5PW. N-terminal amino acid sequencing of sPLA(2) showed amino acid identity with other lysine K49sPLA(2)s of snake venom. B. marajoensis venom (30 mu g/mL) decreased the perfusion pressure, renal vascular resistance, urinary flow, glomerular filtration rate and sodium tubular transport. PLA(2) did not change the renal parameters. The perfusion pressure of the mesenteric bed did not change after infusion of venom. In isolated heart, the venom decreased the force of contraction and increased PP but did not change coronary flow. In the arterial pressure, the venom and PLA(2) decreased mean arterial pressure and cardiac frequency. The presence of atrial flutter and late hyperpolarisation reversed, indicating QRS complex arrhythmia and dysfunction in atrial conduction. In conclusion, B. marajoensis venom and PLA(2) induce hypotension and bradycardia while simultaneously blocking electrical conduction in the heart. Moreover, the decrease in glomerular filtration rate, urinary flow and electrolyte transport demonstrates physiological changes to the renal system. (C) 2009 Elsevier Ltd. All rights reserved.

Efeito da utilização da levedura desidratada de álcool em dietas para tilápia-do-nilo

Este trabalho teve como objetivo avaliar o efeito da levedura desidratada de álcool (Saccharomyces cerevisae) como substituto do suplemento vitamínico, em dietas de tilápia-do-nilo (Oreochromis niloticus). Trezentos alevinos machos, sexualmente revertidos, com peso médio de 1,5 g, foram distribuídos igualmente em 20 aquários de fibra de vidro, equipados com filtro biológico. Cinco dietas isoprotéicas com 32% de proteína bruta na primeira fase (40 dias) e com 28% de proteína bruta na segunda fase (74 dias) foram fornecidas ad libitum, duas vezes ao dia. A dieta-controle não conti-nha levedura, e as demais apresentavam 10% de levedura. O delineamento foi inteiramente casualizado, com cinco tratamentos e quatro repetições. Não houve influência significativa dos tratamentos sobre o ganho de peso e taxa de crescimento específico. Foi observada, nos peixes alimentados com dietas sem suplemento vitamínico, alta mortalidade. Os peixes que receberam levedura apresentaram menor conteúdo corporal de proteína, e maior, de lipídeos. A levedura desidratada de álcool pode ser usada como fonte de vitaminas hidrossolúveis em dietas para alevinos de tilápia-do-nilo.

Break: o grito corporal da periferia

In order to question a sociology as it considers appropriate the body dimensions, which contributes in a mean singular in relation to body, not only as a matter, but mainly as a producer of knowledge, our research shows a reflection on corporality as the production of knowledge and to understand different perspectives and social logic here in contemporaneity, whose rationality seems to be placed in question through a performance expressed, a lot of 'breaks', including the space, which the dancer break appropriates of a reversed form and not more than linear. Our problem is the possibility of a greater understanding of this young body with dance the break, is not only 'breaking' his body, but fluent in their own body fragments, producing probably a break in relation to a dominant power established, by creating a kind of 'social resistance' to provide another sense for his life, out of the senses by the dominant of a social field. This is the area of production of knowledge, of the 'body' that cries keep our eyes, the challenge of dive in the depth of gestures, beyond the physical structure, restoring in a strong political constitution and cultural in so far as this body faces. The body as spectacular becomes 'cause and effect' of communication, this body is not only individual, it is not expression only for itself; when the young dance outside their walls individuals, makes it relational, turning to relate with the other, with the space, time and the world. The methodological point of view, to study this corporality in element break, the body as language of senses, we considered withdrawals in the field, observations of movements of the universe of break creative dance, performance and analysis of "attitudes" (expression peculiar world hip hop ), as well as my experience with dancing in the street interventions through 'urban' and our own attitudes to the object of research which challenges us academically and, it all, existentially

Iniqüidade em saúde: uma extensão das desigualdades sociais no sistema público de saúde do Brasil (2003-2006)

This issue analises the unevenness in the brazilian system of public health care as an extension of socials inequities. It is a theoretical study based in a historical method, using empirical camp from academic, corporation and institution researchs, along the period 2002-2006. Equality and effectiveness in health systems are analitical basic cathegories grap in the root of the doctrine, principles and organization of the Unique Health System, in which sectorial actions are inserted. Discuss the estructural prodution and determined those inequalities through some social determiners of health system: income, land, food securitiy, nutritional situation, basic sanitation, epidemiological inequities and public management policy. Carry out a thematic review over health social production, it formlation and the goals of social policies, as well as the insertion of the equality principle in the assistance system, in the frame of the running public health regulations. It uses reflections that enlighted the correlation between the process of political-institutional actions and equity on health assistance. Analized the pertinency of sectorial reorganizational strategies on basic attendance, confronting the hipothesis that those strategies reinforce social inequities in health system, because it organize diferential assistance levels over not equal baselines. The results show up that social inequalities, even remaining, have had a small decrease; that the selectiviness of actual public policies and the duplication of the health system, increases the differences within and between the social classes and configures the assistance as inequal. The basic care system has great shortages that also appeares in middle and complex assistance levels. As conclusion, it remarks that the health assintance system, even with it integrality has limits; structural problems on material conditions of living and health system could not be reversed only with institutional legal arragements; by the contrary, in border conditions, these strategies produce policies that reinforce inequities, neglecting the equity principle of the system in which frame, they work. One patina of this tim

Indicadores zootécnicos e econômicos da tilapicultura em tanques-rede de diferentes dimensões

The present study aimed to compare the main zootechnical and economic indicators of farming tilapia in superintensive system developed in two different net cages dimensions: 6 and 18 m(3). Were used 12,960 juveniles male reversed Nile tilapia (Oreochromis niloticus), strain supreme, with an initial average weight of 32.0 g (+/- 0.85) for 6 m(3) net cages and 88.5 g (+/- 0.96) for 18 m(3) net cages. The fish were fed three times a day with commercial extruded ration, crude protein levels from 40 to 32%, particle size from 2 to 8 mm and daily feeding rate starting with 8% of live weight (LW), decreasing up to 1% LW. It was conclued that, although no statistical difference in most indicators zootechnical (P>0.05), the production of tilapias in bigger net cages resulted in lower average total cost and higher rentability per kg of fish produced, 34.4% higher, resulting in better profitability.

Ocorrência de Bacillus cereus em leite integral e capacidade enterotoxigênica das cepas isoladas

Pesquisaram-se a presença de Bacillus cereus e a produção de enterotoxinas produzidas por esses microrganismos em 120 amostras de diversos tipos de leite. Bacillus cereus foi isolado e identificado em 22 (73,3%), 15 (50,0%), 29 (96,7%) e quatro (13,3%) amostras de leite em pó, cru, pasteurizado e UAT (longa vida), respectivamente. Para a detecção de enterotoxinas pela técnica da alça ligada de coelho, foram positivos, respectivamente, três (13,6%), um (7,1%) e 10 (35,7%) microrganismos isolados das amostras de leite em pó, leite cru e leite pasteurizado. Pelo teste de aumento de permeabilidade vascular, dois (9,1%), um (7,1%), um (3,6%) e um (4,0%) microrganismos isolados de leite em pó, cru, pasteurizado e UAT apresentaram-se enterotoxigênicos, respectivamente. O uso da técnica de aglutinação passiva em látex demonstrou a produção da toxina diarréica por três (33,3%), sete (63,6%), quatro (30,8%) e oito (80,0%) microrganismos isolados, respectivamente, de leite em pó, cru, pasteurizado e UAT. Os resultados indicam um risco potencial, podendo colocar em risco a saúde dos consumidores desses produtos.

Aspectos morfológicos do saco vitelino em roedores da subordem Hystricomorpha: paca (Agouti paca) e cutia (Dasyprocta aguti)

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

Adubação foliar com nitrato de potássio em algodoeiro

Em diversas lavouras de algodão, no Brasil e no exterior, tem ocorrido deficiência tardia de potássio. Com o objetivo de verificar a influência da adubação foliar com nitrato de potássio (KNO3) na nutrição e produção de algodão (Gossypium hirsutum L.), em vista do nível de nutrição potássica, da época de aplicação e dose do produto, em locais com diferentes potenciais de produtividade, foram desenvolvidos experimentos em dois anos agrícolas (2000/2001 e 2001/2002). Foram empregadas doses de 16 e 32 kg ha-1 de KNO3, divididas em duas ou quatro aplicações, a partir da primeira semana de florescimento do algodoeiro. em outro experimento, foi aplicado 32 kg ha-1 de KNO3, em quatro aplicações realizadas da segunda à quinta semana de florescimento. A diminuição dos teores de potássio nas folhas do algodoeiro é um processo normal na fase de senescência da folha em fim de ciclo e não foi revertido com uso suplementar de K. A adubação foliar com KNO3 pode, em algumas condições, como por exemplo, em plantas deficientes em potássio, aumentar o teor do nutriente na folha. Entretanto, isso não afeta o conteúdo do nutriente nos frutos, a produtividade e a qualidade da fibra.

Ideações e tentativas de suicídio em adolescentes com práticas sexuais hetero e homoeróticas

Esta pesquisa, que teve como população-alvo adolescentes com idade entre 12 e 20 anos, residentes em três municípios do interior Paulista, buscou conhecer as associações entre orientação sexual e ideações e tentativas de suicídio. Corroborando com as pesquisas internacionais, evidenciou-se que os não heterossexuais têm mais chances de pensarem e tentarem suicídio, comparativamente aos heterossexuais. Todavia, encontrou-se que, dentre o grupo de adolescentes que se assumiram não heterossexuais, os que estão mais vulneráveis são aqueles que se autodefiniram bissexuais e outros, os quais constituem o grupo de pessoas menos assumidas, dentre os não heterossexuais. do mesmo modo, constatou-se que os respondentes apresentam diversas opiniões e valores homofóbicos, sexistas e heterocentrados, o que revela ser o espaço escolar, onde se encontram esses jovens não heterossexuais, bastante carregado de posicionamentos discursivos discriminatórios. Conclui-se que a questão do suicídio é uma problemática de saúde pública e que a população de jovens não heterossexuais necessita de abordagens específicas para a prevenção e de atenção relativas a essa conduta.