Quantificação de macro e micro nutrientes em algumas etnovariedades de cubiu (Solanum sessiliflorum Dunal)

Considerando a ampla variabilidade genética de cubiu (Solanum sessiliflorum Dunal), quantificaram-se os macro e micro-nutrientes, objetivando a ampliação da tabela de composição química de alimentos típicos da região amazônica. Os frutos provenientes da Estação Experimental de Hortaliças Alejo von der Pahlen (EEH) do Instituto Nacional de Pesquisas da Amazônia (INPA), localizados no km 14 da Rodovia AM 010 em Manaus, AM, foram processados no Laboratório de Alimentos e Nutrição do INPA. Avaliaram-se oito etnovariedades de cubiu identificados como: 2 I, 3 I, 6, 7, 12, 14, 17, 29 I e III em estádio de maturação comercial. Os teores de elementos minerais foram quantificados pela técnica de Ativação por Nêutrons Instrumental e a fibra alimentar pelo método enzímico-gravimétrico. Os resultados demonstram ser o cubiu um fruto com baixo conteúdo energético (média de 33 kcal), com conteúdo de fibra alimentar total na ordem de 1,6%. Em relação aos macros elementos minerais, a etnovariedade 6, apresentou a maior concentração em potássio (513,5±3,1mg), cálcio (18,9±0,6mg) e a etnovariedade 2 I em Fe (564,4±58,1µg) e Cr (99,3±8,3µg). A menor concentração foi constatada na etnovariedade 12 para os elementos K (229,0±4,5mg), Na (53,7±5,5µg) e Zn (89,3±4,7µg). Apesar das variações em relação as diferentes etno variedades e conseqüentemente concentrações em elementos minerais, o cubiu, pode estar contribuindo para atingir as recomendações desses nutrientes.

Effects of eucalyptol on house fly (Diptera: Muscidae) and blow fly (Diptera: Calliphoridae)

The effects of eucalyptol were evaluated against the house fly, Musca domestica L., and blow fly, Chrysomya megacephala (F.). The bioassay of adults, using topical application, indicated that M. domestica males were more susceptible than females, with the LD50 being 118 and 177 µg/fly, respectively. A higher LD50 of C. megacephala was obtained; 197 µg/fly for males and 221 µg/fly for females. Living flies of both species yielded a shorter life span after being treated with eucalyptol. The bioassay of larvae, using the dipping method on the third instar, showed that M. domestica was more susceptible than C. megacephala, with their LC50 being 101 and 642 µg/µl, respectively. The emergence of adults, which had been treated with eucalyptol in larvae, decreased only in M. domestica. Having the volatile property, fumigation or impregnated paper test of eucalyptol or the efficacy of repellence or attractiveness merits further investigations to enhance bio-insecticidal efficacy.

Análise derivativa de dados hiperespectrais medidos em nível de campo e orbital para caracterizar a composição de águas opticamente complexas na Amazônia

A técnica de análise derivativa de dados espectrais foi usada para estudar a variação dos constituintes opticamente ativos (COAs) na água, por meio de dados de campo e de imagens do sensor orbital Hyperion/EO-1. A imagem Hyperion usada neste estudo foi adquirida no dia 23 de junho de 2005, no final do período de cheia. Uma campanha de campo foi realizada entre 23 e 29 de junho de 2005, para coletar dados espectrais e limnológicos in situ. A imagem foi pré-processada visando eliminar faixas de pixels anômalos e convertida de valores de radiância para reflectância de superfície, portanto, corrigidos dos efeitos de absorção e espalhamento atmosféricos. Uma análise da correlação foi realizada para examinar a associação da reflectância e de sua primeira derivada espectral com as concentrações dos COAs. Melhores resultados foram obtidos após a diferenciação dos espectros, o que ajudou a reduzir a influência de efeitos indesejáveis, provindos de diferentes fontes de radiância, sobre as medidas de reflectância da superfície da água realizadas em ambos os níveis de aquisição de dados. Por meio de ajustes de regressões empíricas, considerando o conjunto de dados Hyperion, a primeira derivada espectral em 711 nm explicou 86% da variação da concentração de sedimentos inorgânicos em suspensão (µg.l-1) e a primeira derivada espectral em 691 nm explicou 73% da variação na concentração da clorofila-alfa (µg.l-1). As relações de regressão foram não-lineares, pois, em geral, as águas que se misturam na planície de inundação Amazônica se tornam opticamente complexas. A técnica de análise derivativa hiperespectral demonstrou potenciais para mapear a composição dessas águas.

Atividade inseticida do óleo essencial de Tanaecium nocturnum (Barb. Rodr.) Bur. & K. Shum (Bignoneaceae) sobre Sitophilus zeamais Motsch. (Coleoptera: Curculionidae)

O óleo essencial extraído de folhas frescas de Tanaecium nocturnum (Barb. Rodr.) Bur.& K. Shum por destilação de arraste a vapor foi avaliado quanto à toxicidade a Sitophilus zeamais Motsch., principal praga do milho armazenado. Papel de filtro e grãos de milho foram impregnados pelo óleo para se avaliar o efeito por via de contato (papel-filtro) e fumigação, respectivamente. Para avaliação do efeito da aplicação tópica 0,5 µl das diferentes concentrações do óleo foram aplicadas em adultos do inseto. A partir de uma ampla faixa de concentrações, foram determinadas as mais promissoras para os bioensaios definitivos. Na determinação das dose/concentrações-letais (DL50 e CL50) foi utilizada a análise de Probit, realizando-se também, uma análise de regressão linear conjunta de todos os dados de mortalidade. O óleo de T. nocturnum foi considerado tóxico para S. zeamais baseado nos seguintes valores: CL50 de 14,1 ng.cm-2 e CL50 de 1.321,6 ng.g-1 de grãos para os efeitos de contacto (papel-filtro) e fumigação, respectivamente, e DL50 de 14,7 µg.mg-1 de inseto para efeito tópico. Porcentagens de mortalidade próximas a 100 % foram obtidas nas concentrações de: 2 e 5 % (m/v) (contato), 3 4, e 5 % (m/v) (fumigação) e 10 % (m/v) para o efeito de aplicação tópica. O presente estudo mostrou que o ácido cianídrico, liberado do óleo essencial de T. nocturnum por hidrólise, pode ter atividade inseticida para S. zeamais e que concentrações acima de 4 % (m/v) são promissoras no controle do inseto.

Cytotoxic evaluation of essential oil from Casearia sylvestris Sw on human cancer cells and erythrocytes

A Casearia sylvestris (Flacourtiaceae) é uma planta popularmente conhecida como "guaçatonga" e é usada por povos indígenas da América do sul (Brasil, Peru e Bolivia) no tratamento de muitas doenças, incluindo câncer. Estudos citotóxicos mostraram que esta planta apresenta um possível e interessante potencial antitumoral devido à presença de moléculas chamadas casearinas. Além disso, a composição do óleo essencial mostrou uma alta concentração de sesquiterpenos de alto potencial citotóxico. Neste trabalho, nós verificamos que o óleo essencial da C. sylvestris apresentou uma boa citotoxicidade seletiva contra as linhagens de células tumorais HeLa, A-549 and HT-29 (CD50 63,3, 60,7 e 90,6 µg.ml-1, respectivamente) quando comparada às células não-tumorais Vero (CD50 210,1 µg.ml-1) e macrófagos de camundongos (CD50 234,0 µg.ml-1). Além disso, o óleo causou hemólise em sete diferentes tipos de eritrócitos, indicando que a C. sylvestris precisa ser usada com cuidado. Também foram testados padrões de β-cariofileno e α-humuleno que mostraram citotoxicidade similar àquelas apresentadas pelo óleo, indicando que estes compostos podem ser os responsáveis pelos efeitos tóxicos que foram observados neste estudo.

Thiamin, selenium, and copper levels in patients with idiopathic dilated cardiomyopathy taking diuretics

OBJECTIVE: To analyze the association of thiamin, selenium, and copper serum levels with cardiac function in patients with idiopathic dilated cardiomyopathy using diuretics, and also to compare them with levels in control patients with no evidence of disease. METHODS: The study comprised 30 patients with heart disease and 30 healthy control individuals. Thiamin was analyzed by measuring the activity of erythrocytic transketolase and the effect of thiamin pyrophosphate. Selenium and copper serum levels were measured by hydride generation and flame atomic absorption spectrophotometry, respectively. RESULTS: Thiamin deficiency was observed in 10% of the control individuals and in 33% of the patients with heart disease (p=0.02). The mean selenium and copper serum levels in control individuals and patients with heart disease were, respectively, 73.2±9.9 µg/L (56.5 to 94.5 µg/L) and 72.3±14.3 µg/L (35.5 to 94 µg/L) (p=0.77); 1.1±0.4mg/L (0.6 to 1.8mg/L) and 1.2± 0.4mg/L (0.6 to 2.2mg/L) (p=0.27). No association between the levels of these nutrients and cardiac function was observed. CONCLUSION: Thiamin deficiency was significantly more frequent in patients with heart disease. No significant difference was observed between the mean selenium and copper serum levels in control individuals and in patients with heart disease. The results suggest possible benefits with thiamin replacement in patients taking diuretics.

Evaluation by ELISA of Anisakis simplex Larval Antigen Purified by Affinity Chromatography

In order to improve the specificity and sensitivity of the techniques for the human anisakidosis diagnosis, a method of affinity chromatography for the purification of species-specific antigens from Anisakis simplex third-stage larvae (L3) has been developed. New Zealand rabbits were immunized with A. simplex or Ascaris suum antigens or inoculated with Toxocara canis embryonated eggs. The IgG specific antibodies were isolated by means of protein A-Sepharose CL-4B beads columns. IgG anti-A. simplex and -A. suum were coupled to CNBr-activated Sepharose 4B. For the purification of the larval A. simplex antigens, these were loaded into the anti-A. simplex column and bound antigens eluted. For the elimination of the epitopes responsible for the cross-reactions, the A. simplex specific proteins were loaded into the anti-A. suum column. To prove the specificity of the isolated proteins, immunochemical analyses by polyacrylamide gel electrophoresis were carried out. Further, we studied the different responses by ELISA to the different antigenic preparations of A. simplex used, observing their capability of discriminating among the different antisera raised in rabbits (anti-A. simplex, anti-A. suum, anti-T. canis). The discriminatory capability with the anti-T. canis antisera was good using the larval A. simplex crude extract (CE) antigen. When larval A. simplex CE antigen was loaded into a CNBr-activated Sepharose 4B coupled to IgG from rabbits immunized with A. simplex CE antigen, its capability for discriminate between A. simplex and A. suum was improved, increasing in the case of T. canis. The best results were obtained using larval A. simplex CE antigen loaded into a CNBr-activated Sepharose 4B coupled to IgG from rabbits immunized with adult A. suum CE antigen. When we compared the different serum dilution and antigenic concentration, we selected the working serum dilution of 1/400 and 1 µg/ml of antigenic concentration.

In Vitro and in Vivo Assays of 3,5-Disubstituted-Tetrahydro-2H-1,3,5-Thiadiazin-2-Thione Derivatives against Trypanosoma cruzi

Cytotoxicity assays of 24 new 3,5-disubstituted-tetrahydro-2H-1,3,5-thiadiazin-2-thione derivatives were performed. The 17 compounds with higher anti-epimastigote activity and lower cytotoxicity were, thereafter, screened against amastigote of Trypanosoma cruzi. Out of these 17 derivatives S-2d was selected to be assayed in vivo, because of its remarkable trypanocidal properties. To determine toxicity against J774 macrophages, a method based on quantification of cell damage, after 24 h, was used. Cell respiration, an indicator of cell viability, was assessed by the reduction of MTT [3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide] to formazan. Anti-amastigote activity was estimated after 48 h by microscopic counts of May Grünwald-Giemsa-stained monolayers. Nifurtimox and benznidazole were used as reference drugs. For the in vivo experiences, mice were infected with 10(4) blood trypomastigotes and then treated during 15 days with S-2d or nifurtimox by oral route. All of the compounds were highly toxic at 100 µg/ml for macrophages and a few of them maintained this cytotoxicity even at 10 µg/ml. Of the derivatives assayed against amastigotes 3k and S-2d showed an interesting activity, that was held even at 1µg/ml. It is demonstrated that the high anti-epimastigote activity previously reported is mainly due to the non-specific toxicity of these compounds. In vivo assays assessed a reduction of parasitemia after administration of S-2d to infected mice.

Activity of Brazilian and Bulgarian propolis against different species of Leishmania

Extracts of propolis samples collected in Brazil and Bulgaria were assayed against four Leishmania species - Leishmania amazonensis, L. braziliensis, L. chagasi from the New World, and L. major from the Old World - associated to different clinical forms of leishmaniasis. The composition of the extracts has been previously characterized by high temperature high resolution gas chromatography coupled to mass spectrometry. Considering the chemical differences among the extracts and the behavior of the parasites, it was observed significant differences in the leishmanicidal activities with IC50/1 day values in the range of 2.8 to 229.3 µg/ml . An overall analysis showed that for all the species evaluated, Bulgarian extracts were more active than the ethanol Brazilian extract. As the assayed propolis extracts have their chemical composition determined it merits further investigation the effect of individual components or their combinations on each Leishmania species.

Inferências sobre atividade rizosférica de espécies com potencial para fitorremediação do herbicida tebuthiuron

A ação da microbiota rizosférica, acelerando a degradação de compostos no solo, é conhecida como fitoestimulação e constitui-se em um dos principais mecanismos de fitorremediação de herbicidas no solo. Plantas tolerantes ao tebuthiuron, que sejam capazes de estimular sua microbiota rizosférica, podem ser de grande interesse na fitorremediação desse herbicida. O objetivo deste trabalho foi avaliar a atividade rizosférica de quatro espécies vegetais com potencial para fitorremediação de tebuthiuron e inferir a contribuição radicular no processo de descontaminação desse herbicida. Foi analisado o solo rizosférico de feijão-de-porco (Canavalia ensiformis), milheto (Pennisetum glaucum), mucuna-anã (Estizolobium deeringianum) e mucuna-preta (Estizolobium aterrimum) e de uma testemunha (sem planta), com e sem a presença do tebuthiuron, na dose de 0,73 µg g-1. A taxa de evolução de CO2 foi quantificada aos 1, 2, 3 e 10 dias da aplicação dos tratamentos (DAT). Os tratamentos com herbicida foram submetidos à contaminação com 40 µg g-1 de tebuthiuron. Após a aplicação, mediu-se a taxa de evolução de CO2 aos 1 e 50 DAT, empregando-se respirômetro de fluxo contínuo. A espécie feijão-de-porco apresentou sempre a maior taxa de evolução de CO2 em relação às demais espécies e à testemunha, seguido de mucuna-preta na dose comercial. Sob concentrações maiores que a dose comercial, os valores médios de taxa de evolução de CO2 foram maiores no solo rizosférico de feijão-de-porco, seguido por mucuna-preta e mucuna-anã. Feijão-de-porco foi a espécie com melhor desempenho nas avaliações realizadas; excetuando-se esta espécie, a contribuição rizosférica na fitorremediação de tebuthiuron em níveis acima da dose comercial não foi relevante.

Oral administration of a low dose of midazolam (75 microg) as an in vivo probe for CYP3A activity.

OBJECTIVE: We investigated whether the oral administration of a low dose (75 micro g) of midazolam, a CYP3A probe, can be used to measure the in vivo CYP3A activity. METHODS: Plasma concentrations of midazolam, 1'OH-midazolam and 4'OH-midazolam were measured after the oral administration of 7.5 mg and 75 micro g midazolam in 13 healthy subjects without medication, in four subjects pretreated for 2 days with ketoconazole (200 mg b.i.d.), a CYP3A inhibitor, and in four subjects pretreated for 4 days with rifampicin (450 mg q.d.), a CYP3A inducer. RESULTS: After oral administration of 75 micro g midazolam, the 30-min total (unconjugated + conjugated) 1'OH-midazolam/midazolam ratios measured in the groups without co-medication, with ketoconazole and with rifampicin were (mean+/-SD): 6.23+/-2.61, 0.79+/-0.39 and 56.1+/-12.4, respectively. No side effects were reported by the subjects taking this low dose of midazolam. Good correlations were observed between the 30-min total 1'OH-midazolam/midazolam ratio and midazolam clearance in the group without co-medication (r(2)=0.64, P<0.001) and in the three groups taken together (r(2)=0.91, P<0.0001). Good correlations were also observed between midazolam plasma levels and midazolam clearance, measured between 1.5 h and 4 h. CONCLUSION: A low oral dose of midazolam can be used to phenotype CYP3A, either by the determination of total 1'OH-midazolam/midazolam ratios at 30 min or by the determination of midazolam plasma levels between 1.5 h and 4 h after its administration.

Pharmacokinetics of midazolam in CYP3A4- and CYP3A5-genotyped subjects

OBJECTIVE: We investigated whether differences in pharmacokinetics of midazolam, a CYP3A probe, could be demonstrated between subjects with different CYP3A4 and CYP3A5 genotypes. METHODS: Plasma concentrations of midazolam, and of total (conjugated + unconjugated) 1'OH-midazolam, and 4'OH-midazolam were measured after the oral administration of 7.5 mg or of 75 micro g of midazolam in 21 healthy subjects. RESULTS: CYP3A5*7, CYP3A4*1E, CYP3A4*2, CYP3A4*4, CYP3A4*5, CYP3A4*6, CYP3A4*8, CYP3A4*11, CYP3A4*12, CYP3A4*13, CYP3A4*17 and CYP3A4*18 alleles were not identified in the 21 subjects. CYP3A5*3, CYP3A5*6, CYP3A4*1B and CYP3A4*1F alleles were identified in 20, 1, 4 and 2 subjects, respectively. No statistically significant differences were observed for the AUC(inf) values between the different genotypes after the 75- micro g or the 7.5-mg dose. CONCLUSION: Presently, CYP3A4 and CYP3A5 genotyping methods do not sufficiently reflect the inter-individual variability of CYP3A activity.

Exclusion of pulmonary embolism using C-reactive protein and D-dimer. A prospective comparison.

Our goal was to evaluate the diagnostic utility of C-reactive protein (CRP) alone or combined with clinical probability assessment in patients with suspected pulmonary embolism (PE), and to compare its performance to a D-dimer assay. We conducted a prospective study in which we performed a common immuno-turbidimetric CRP test and a rapid enzyme-linked immunosorbent assay (ELISA) D-dimer test in 259 consecutive outpatients with suspected PE at the emergency department of a teaching hospital. We assessed clinical probability of PE by a validated prediction rule overridden by clinical judgment. Patients with D-dimer levels &gt; or = 500 microg/l underwent a work-up consisting of lower-limb venous ultrasound, spiral computerized tomography, ventilation-perfusion scan, or pulmonary angiography. Patients were followed up for three months. Seventy-seven (30%) of the patients had PE. The CRP alone had a sensitivity of 84% (95% confidence interval [CI).: 74 to 92%) and a negative predictive value (NPV) of 87% (95% CI: 78 to 93%) at a cutpoint of 5 mg/l. Overall, 61 (24%) patients with a low clinical probability of PE had a CRP &lt; 5 mg/l. Due to the low prevalence of PE (9%) in this subgroup, the NPV increased to 97% (95% CI: 89 to 100%). The D-dimer (cutpoint 500 micro g/l) showed a sensitivity of 100% (95% CI: 95 to 100%) and a NPV of 100% (95% CI: 94 to 100%) irrespective of clinical probability and accurately rule out PE in 56 (22%) patients. Standard CRP tests alone or combined with clinical probability assessment cannot safely exclude PE.

Quantify this! Report on a round table discussion on quantitative mass spectrometry in proteomics.

Following the success of the first round table in 2001, the Swiss Proteomic Society has organized two additional specific events during its last two meetings: a proteomic application exercise in 2002 and a round table in 2003. Such events have as their main objective to bring together, around a challenging topic in mass spectrometry, two groups of specialists, those who develop and commercialize mass spectrometry equipment and software, and expert MS users for peptidomics and proteomics studies. The first round table (Geneva, 2001) entitled "Challenges in Mass Spectrometry" was supported by brief oral presentations that stressed critical questions in the field of MS development or applications (Stöcklin and Binz, Proteomics 2002, 2, 825-827). Topics such as (i) direct analysis of complex biological samples, (ii) status and perspectives for MS investigations of noncovalent peptide-ligant interactions; (iii) is it more appropriate to have complementary instruments rather than a universal equipment, (iv) standardization and improvement of the MS signals for protein identification, (v) what would be the new generation of equipment and finally (vi) how to keep hardware and software adapted to MS up-to-date and accessible to all. For the SPS'02 meeting (Lausanne, 2002), a full session alternative event "Proteomic Application Exercise" was proposed. Two different samples were prepared and sent to the different participants: 100 micro g of snake venom (a complex mixture of peptides and proteins) and 10-20 micro g of almost pure recombinant polypeptide derived from the shrimp Penaeus vannamei carrying an heterogeneous post-translational modification (PTM). Among the 15 participants that received the samples blind, eight returned results and most of them were asked to present their results emphasizing the strategy, the manpower and the instrumentation used during the congress (Binz et. al., Proteomics 2003, 3, 1562-1566). It appeared that for the snake venom extract, the quality of the results was not particularly dependant on the strategy used, as all approaches allowed Lication of identification of a certain number of protein families. The genus of the snake was identified in most cases, but the species was ambiguous. Surprisingly, the precise identification of the recombinant almost pure polypeptides appeared to be much more complicated than expected as only one group reported the full sequence. Finally the SPS'03 meeting reported here included a round table on the difficult and challenging task of "Quantification by Mass Spectrometry", a discussion sustained by four selected oral presentations on the use of stable isotopes, electrospray ionization versus matrix-assisted laser desorption/ionization approaches to quantify peptides and proteins in biological fluids, the handling of differential two-dimensional liquid chromatography tandem mass spectrometry data resulting from high throughput experiments, and the quantitative analysis of PTMs. During these three events at the SPS meetings, the impressive quality and quantity of exchanges between the developers and providers of mass spectrometry equipment and software, expert users and the audience, were a key element for the success of these fruitful events and will have definitively paved the way for future round tables and challenging exercises at SPS meetings.

The stereochemistry of the amino acid side chain influences the inflammatory potential of muramyl dipeptide in experimental meningitis.

Intrathecal injections of 50 to 100 micro g of (N-acetylmuramyl-L-alanyl-D-isoglutamine) muramyl dipeptide (MDP)/rabbit dose-dependently triggered tumor necrosis factor alpha (TNF-alpha) secretion (12 to 40,000 pg/ml) preceding the influx of leukocytes in the subarachnoid space of rabbits. Intrathecal instillation of heat-killed unencapsulated R6 pneumococci produced a comparable leukocyte influx but only a minimal level of preceding TNF-alpha secretion. The stereochemistry of the first amino acid (L-alanine) of the MDP played a crucial role with regard to its inflammatory potential. Isomers harboring D-alanine in first position did not induce TNF-alpha secretion and influx of leukocytes. This stereospecificity of MDPs was also confirmed by measuring TNF-alpha release from human peripheral mononuclear blood cells stimulated in vitro. These data show that the inflammatory potential of MDPs depends on the stereochemistry of the first amino acid of the peptide side chain and suggest that intact pneumococci and MDPs induce inflammation by different pathways.