Sintesis de poliuretanos a partir de polioles obtenidos a partir del aceite de higuerilla modificado por transesterificación con pentaeritritol

Castor oil was reacted by transesterification with various percentages in mass of pentaerythritol to obtain different esters of pentaerythritol. Alternatively, glycerol was also used instead of pentaerythritol for the same reaction in order to establish comparative reference products. The products of the reactions were characterized by matrix-assisted laser desorption/ionization time-of-flight mass spectroscopy in order to detect and quantify (in terms of the molecular mass and structural information) the components of the products obtained. Analysis for hydroxyl value, acid value, viscosity and specific gravity were used to complete the characterization of the polyols obtained and also of the original castor oil. The polymer characterization was accomplished by tensile stress-strain tests, Shore A hardness, thermogravimetric analysis and chemical resistance to solvents.

Caracterização de contaminantes presentes em sistemas de tratamento de esgotos, por cromatografia líquida acoplada à espectrometria de massas tandem em alta resolução

This work shows results on the characterization, by liquid chromatography coupled to high resolution tandem mass spectrometry (LC-IT-TOF-MS) with electrospray ionization, of organic compounds present in raw and treated effluents from a combined sewage treatment systems (upflow anaerobic sludge blanket-trickling filter). The sewage samples were prepared by C18 solid phase extraction and the spectra obtained from the various extracts were submitted to principal component analysis to evaluate their pattern and identify the major deprotonated species. Some target compounds were submitted to semiquantitative analysis, using phenolphtalein as internal standard. The results showed the anaerobic step had little impact on the removal of anionic surfactants (LAS), fatty acids, and some contaminantes such as bisphenol A and bezafibrate, whereas the aerobic post-treatment was very efficient in removing these organics.

COMPARISON BETWEEN ASPHALTENES (SUB)FRACTIONS EXTRACTED FROM TWO DIFFERENT ASPHALTIC RESIDUES: CHEMICAL CHARACTERIZATION AND PHASE BEHAVIOR

Asphaltenes are blamed for various problems in the petroleum industry, especially formation of solid deposits and stabilization of water-in-oil emulsions. Many studies have been conducted to characterize chemical structures of asphaltenes and assess their phase behavior in crude oil or in model-systems of asphaltenes extracted from oil or asphaltic residues from refineries. However, due to the diversity and complexity of these structures, there is still much to be investigated. In this study, asphaltene (sub)fractions were extracted from an asphaltic residue (AR02), characterized by NMR, elemental analysis, X-ray fluorescence and MS-TOF, and compared to asphaltene subfractions obtained from another asphaltic residue (AR01) described in a previous article. The (sub)fractions obtained from the two residues were used to prepare model-systems containing 1 wt% of asphaltenes in toluene and their phase behavior was evaluated by measuring asphaltene precipitation onset using optical microscopy. The results obtained indicated minor differences between the asphaltene fractions obtained from the asphaltic residues of distinct origins, with respect to aromaticity, elemental composition (CHN), presence and content of heteroelements and average molar mass. Regarding stability, minor differences in molecule polarity appear to promote major differences in the phase behavior of each of the asphaltene fractions isolated.

Umidade do solo na zona radicular do maracujazeiro cultivado sob irrigação localizada

O objetivo deste trabalho foi avaliar o efeito de níveis de irrigação aplicados por gotejamento no perfil de umidade do solo na zona de concentração radicular do maracujazeiro-amarelo (Passiflora edulis Sims f. flavicarpa Deg). O experimento foi realizado no período de julho de 1999 a janeiro de 2000, no Campo Experimental da Fazenda Areão, em Piracicaba - SP (22º42'30"S e 47º38'00"W). Os níveis de irrigação, aplicados por um sistema de irrigação por gotejamento, foram: L1 = 0,25 L; L2 = 0,50 L; L3 = 0,75 L, e L4 = 1,00 L, em que L é a lâmina de água média medida em lisímetros. Os níveis de irrigação influenciaram nos perfis de distribuição temporal do potencial matricial, apresentando comportamento diferenciado na oscilação e na manutenção dos teores de água na zona radicular do maracujazeiro. A aplicação de volume de água entre 1.528 L planta-1 ano-1 (L2) e 2.117 L planta-1 ano-1 (L3) favoreceu perfis médios de umidade na zona radicular do maracujazeiro mais adequados, enquanto L1 e L4 tiveram valores de umidade abaixo e acima do valor referente à capacidade de campo, respectivamente. Valores de potencial matricial abaixo de -0,01445 MPa e acima de -0,00357 MPa afetam negativamente os parâmetros de produção do maracujazeiro-amarelo.

Doses de composto orgânico comercial na composição de substratos para a produção de mudas de maracujazeiro em diferentes tipos de cultivo protegido

Doses de composto orgânico comercial misturado ao solo foram utilizadas como substrato na produção de mudas de maracujazeiro amarelo. O experimento foi conduzido no Campus de Aquidauana, da Universidade Estadual de Mato Grosso do Sul, de setembro a dezembro de 2007. Utilizaram-se seis ambientes protegidos: estufa plástica com pé-direito de 2,5m; viveiro telado com Sombrite® 50%, com pé-direito de 2,5m; viveiro telado de tela termo-refletora Aluminet® 50%, com pé-direito de 2,5m; viveiro coberto com palha de coqueiro, com pé-direito de 1,8m; estufa plástica com pé-direito de 4,0m, possuindo abertura zenital e tela termo-refletora 50% sob o plástico e, viveiro telado de tela de sombreamento 50%, com pé-direito de 3,5m. Foram utilizados cinco substratos com doses de 0, 7, 14, 21 e 28% de composto orgânico misturado ao solo. Utilizou-se o delineamento em parcelas subdivididas, com dez repetições. As parcelas principais foram os ambientes de cultivo e as subparcelas foram os substratos. Conclui-se que a estufa agrícola com altura de 4,0m, o viveiro agrícola com altura de 3,5m e o viveiro coberto com palha de coqueiro nativo propiciaram melhor desenvolvimento das mudas. As doses de 7%, 14% e 21% de composto orgânico se mostraram viáveis para composição de substratos com solo da região.

Desenvolvimento e produtividade do maracujazeiro-amarelo irrigado sob diferentes lâminas de irrigação em ambiente protegido e natural

A irrigação é uma prática que, além de aumentar a produtividade, pode proporcionar a obtenção de um produto com melhor qualidade. O cultivo do maracujazeiro em ambiente protegido não é uma atividade comum, sendo necessária a pesquisa sob esta condição, especialmente sobre a irrigação. Este trabalho foi conduzido com o objetivo de avaliar o efeito das diferentes tensões de água no solo sobre o comportamento da cultura em ambiente protegido e natural, na região de Lavras - MG. O experimento foi instalado em ambiente protegido e natural na área experimental da Engenharia -UFLA, com delineamento em blocos casualizados e quatro repetições. Os tratamentos foram constituídos de quatro tensões de água no solo, preestabelecidas em 15; 30; 45 e 60 kPa, à profundidade de 0,20 m. Os resultados obtidos permitiram concluir que: a irrigação do maracujazeiro, sob tensões de até 60 kPa, tanto em ambiente protegido quanto em condições naturais de cultivo, não afetou, significativamente, a produções total, comercial e não comercial; o cultivo em ambiente protegido promoveu crescimento mais rápido das plantas (altura e diâmetro do caule) e, também, a antecipação da colheita em relação ao ambiente natural; a irrigação poderá ser realizada para tensão de água no solo de 60 kPa em ambiente protegido e natural sem comprometer a produtividade da cultura; a tensão de água no solo afetou o crescimento das plantas, entretanto não afetou o diâmetro do caule.

Anatomia de escapos de Tillandsia L. (Bromeliaceae) dos Campos Gerais do Paraná, Brasil

Estudou-se a anatomia dos escapos de Tillandsia crocata (E. Morren) Baker, T. gardneri Lindl., T. geminiflora Brongn., T. linearis Vell., T. lorentziana Griseb., T. mallemontii Glaziou ex Mez, T. recurvata L., T. streptocarpa Baker, T. stricta Soland ex Sims, T. tenuifolia L. e Tillandsia sp., que ocorrem nos Campos Gerais do Paraná. São plantas epífitas de galhos de árvores ou de paredões rochosos. Os escapos são eretos ou recurvados, revestidos por brácteas e emergem do centro da roseta foliar. Verificaram-se dois padrões anatômicos de distribuição dos feixes vasculares. Um padrão onde os feixes vasculares ocorrem condensados perifericamente no cilindro vascular, circundados e/ou imersos num cilindro esclerótico - periciclo. Outro padrão onde os feixes vasculares ocorrem dispersos, mas concentrados na periferia do cilindro vascular, envolvidos total ou parcialmente por bainha fibrosa, sem formar um cilindro esclerótico. A grande maioria das espécies apresenta escapos com epiderme de células espessadas, hipoderme com células espessadas, e feixes vasculares colaterais envolvidos total ou parcialmente por fibras. A presença de células com paredes espessadas, de hipoderme, de idioblastos de ráfides, mucilagem e canais de ar constituem características adaptativas ao hábito epifítico. Os tipos de espessamento das células epidérmicas, da hipoderme, e do periciclo, a distribuição dos feixes vasculares, e a presença de feixes vasculares compostos possibilitam separar as espécies dentro do gênero. É apresentada uma chave de identificação das Tillandsia estudadas com base nas características anatômicas do escapo.

Anatomia foliar de Tillandsia L. (Bromeliaceae) dos Campos Gerais, Paraná, Brasil

Foi estudada a anatomia foliar de Tillandsia crocata (E. Morren) Baker, T. gardneri Lindl., T. geminiflora Brongn., T. linearis Vell., T. lorentziana Griseb., T. mallemontii Glaziou ex Mez, T. recurvata L., T. streptocarpa Baker, T. stricta Soland ex Sims, T. tenuifolia L. T. usneoides L. e Tillandsia sp., dos Campos Gerais, Paraná, Brasil. Em vista frontal a epiderme apresenta células com paredes lineares até sinuosas, corpos silicosos e escamas epidérmicas que protegem os estômatos anomocíticos. A epiderme e o primeiro estrato da hipoderme apresentam células lignificadas na maioria das espécies. Em secção transversal observa-se estômatos que ocorrem um pouco abaixo do nível das demais células da epiderme; presença de parênquima aqüífero; canais de ar longitudinais e feixes vasculares colaterais circundados por bainha dupla. Essas estruturas anatômicas são xeromórficas e usualmente consideradas como adaptações ao hábito epifítico das Tillandsia atmosféricas. Além disso, elas poderiam também ser usadas com finalidades diagnósticas para as espécies. Forma do limbo da folha em secção transversal, ornamentação da cutícula, estrutura das escamas epidérmicas, espessamento das paredes das células epidérmicas, distribuição dos estômatos, estrutura e distribuição das células do parênquima aqüífero, presença de canais de ar e tamanho de feixes vasculares são caracteres que podem auxiliar na delimitação taxonômica das espécies dentro do gênero.

Somaclonal variation: a morphogenetic and biochemical analysis of Mandevilla velutina cultured cells

Cell cultures of Mandevilla velutina have proved to be an interesting production system for biomass and secondary metabolites able to inhibit the hypotensive activity of bradykinin, a nonapeptide generated in plasma during tissue trauma. The crude ethyl acetate extract of cultured cells contains about 31- to 79-fold more potent anti-bradykinin compounds (e.g., velutinol A) than that obtained with equivalent extracts of tubers. Somaclonal variation may be an explanation for the wide range of inhibitor activity found in the cell cultures. The heterogeneity concerning morphology, differentiation, carbon dissimilation, and velutinol A production in M. velutina cell cultures is reported. Cell cultures showed an asynchronous growth and cells in distinct developmental stages. Meristematic cells were found as the major type, with several morphological variations. Cell aggregates consisting only of meristematic cells, differentiated cells containing specialized cell structures such as functional chloroplasts (cytodifferentiation) and cells with embryogenetic characteristics were observed. The time course for sucrose metabolism indicated cell populations with significant differences in growth and metabolic rates, with the highest biomass-producing cell line showing a cell cycle 60% shorter and a metabolic rate 33.6% higher than the control (F2 cell population). MALDI-TOF mass spectrometric analysis of velutinol A in selected cell lines demonstrated the existence of velutinol A producing and nonproducing somaclones. These results point to a high genetic heterogeneity in general and also in terms of secondary metabolite content.

Identification of Mycobacterium bovis antigens by analysis of bovine T-cell responses after infection with a virulent strain

Purification and characterization of individual antigenic proteins are essential for the understanding of the pathogenic mechanisms of mycobacteria and the immune response against them. In the present study, we used anion-exchange chromatography to fractionate cell extracts and culture supernatant proteins from Mycobacterium bovis to identify T-cell-stimulating antigens. These fractions were incubated with peripheral blood mononuclear cells (PBMC) from M. bovis-infected cattle in lymphoproliferation assays. This procedure does not denature proteins and permits the testing of mixtures of potential antigens that could be later identified. We characterized protein fractions with high stimulation indices from both culture supernatants and cell extracts. Proteins were identified by two-dimensional gel electrophoresis followed by N-terminal sequencing or MALDI-TOF. Culture supernatant fractions containing low molecular weight proteins such as ESAT6 and CFP10 and other proteins (85B, MPB70), and the novel antigens TPX and TRB-B were associated with a high stimulation index. These results reinforce the concept that some low molecular weight proteins such as ESAT6 and CFP10 play an important role in immune responses. Also, Rv3747 and L7/L12 were identified in high stimulation index cell extract fractions. These data show that protein fractions with high lymphoproliferative activity for bovine PBMC can be characterized and antigens which have been already described and new protein antigens can also be identified in these fractions.

Proteomic inventory of myocardial proteins from patients with chronic Chagas' cardiomyopathy

Chronic Chagas' disease cardiomyopathy (CCC) is an often fatal outcome of Trypanosoma cruzi infection, with a poorer prognosis than other cardiomyopathies. CCC is refractory to heart failure treatments, and is the major indication of heart transplantation in Latin America. A diffuse myocarditis, plus intense myocardial hypertrophy, damage and fibrosis, in the presence of very few T. cruzi forms, are the histopathological hallmarks of CCC. To gain a better understanding of the pathophysiology of CCC, we analyzed the protein profile in the affected CCC myocardium. Homogenates from left ventricular myocardial samples of end-stage CCC hearts explanted during heart transplantation were subjected to two-dimensional electrophoresis with Coomassie blue staining; protein identification was performed by MALDI-ToF mass spectrometry and peptide mass fingerprinting. The identification of selected proteins was confirmed by immunoblotting. We demonstrated that 246 proteins matched in gels from two CCC patients. They corresponded to 112 distinct proteins. Along with structural/contractile and metabolism proteins, we also identified proteins involved in apoptosis (caspase 8, caspase 2), immune system (T cell receptor ß chain, granzyme A, HLA class I) and stress processes (heat shock proteins, superoxide dismutases, and other oxidative stress proteins). Proteins involved in cell signaling and transcriptional factors were also identified. The identification of caspases and oxidative stress proteins suggests the occurrence of active apoptosis and significant oxidative stress in CCC myocardium. These results generated an inventory of myocardial proteins in CCC that should contribute to the generation of hypothesis-driven experiments designed on the basis of the classes of proteins identified here.

Isolation and characterization of a serine proteinase with thrombin-like activity from the venom of the snake Bothrops asper

A serine proteinase with thrombin-like activity was isolated from the venom of the Central American pit viper Bothrops asper. Isolation was performed by a combination of affinity chromatography on aminobenzamidine-Sepharose and ion-exchange chromatography on DEAE-Sepharose. The enzyme accounts for approximately 0.13% of the venom dry weight and has a molecular mass of 32 kDa as determined by SDS-PAGE, and of 27 kDa as determined by MALDI-TOF mass spectrometry. Its partial amino acid sequence shows high identity with snake venom serine proteinases and a complete identity with a cDNA clone previously sequenced from this species. The N-terminal sequence of the enzyme is VIGGDECNINEHRSLVVLFXSSGFL CAGTLVQDEWVLTAANCDSKNFQ. The enzyme induces clotting of plasma (minimum coagulant dose = 4.1 µg) and fibrinogen (minimum coagulant dose = 4.2 µg) in vitro, and promotes defibrin(ogen)ation in vivo (minimum defibrin(ogen)ating dose = 1.0 µg). In addition, when injected intravenously in mice at doses of 5 and 10 µg, it induces a series of behavioral changes, i.e., loss of the righting reflex, opisthotonus, and intermittent rotations over the long axis of the body, which closely resemble the `gyroxin-like' effect induced by other thrombin-like enzymes from snake venoms.

cDNA and deduced primary structure of basic phospholipase A2 with neurotoxic activity from the venom secretion of the Crotalus durissus collilineatus rattlesnake

To illustrate the construction of precursor complementary DNAs, we isolated mRNAs from whole venom samples. After reverse transcription polymerase chain reaction (RT-PCR), we amplified the cDNA coding for a neurotoxic protein, phospholipase A2 D49 (PLA2 D49), from the venom of Crotalus durissus collilineatus (Cdc PLA2). The cDNA encoding Cdc PLA2 from whole venom was sequenced. The deduced amino acid sequence of this cDNA has high overall sequence identity with the group II PLA2 protein family. Cdc PLA2 has 14 cysteine residues capable of forming seven disulfide bonds that characterize this group of PLA2 enzymes. Cdc PLA2 was isolated using conventional Sephadex G75 column chromatography and reverse-phase high performance liquid chromatography (RP-HPLC). The molecular mass was estimated using matrix-assisted laser desorption ionization-time-of-flight (MALDI-TOF) mass spectrometry. We tested the neuromuscular blocking activities on chick biventer cervicis neuromuscular tissue. Phylogenetic analysis of Cdc PLA2 showed the existence of two lines of N6-PLA2, denominated F24 and S24. Apparently, the sequences of the New World’s N6-F24-PLA2 are similar to those of the agkistrodotoxin from the Asian genus Gloydius. The sequences of N6-S24-PLA2 are similar to the sequence of trimucrotoxin from the genus Protobothrops, found in the Old World.

Induction of a gloverin-like antimicrobial polypeptide in the sugarcane borer Diatraea saccharalis challenged by septic injury

Diatraea saccharalis (Fabricius, 1794) (Lepidoptera: Crambidae) is an important pest for Brazilian sugarcane. In the present study, we detected two distinct spots in hemolymph from septic injured larvae (HDs1 and HDs2), which are separated by 2DE gel electrophoresis. Both spots were subjected to in-gel tryptic digestion and MALDI-TOF/TOF analysis, which revealed the sequence VFGTLGSDDSGLFGK present in both HDs1 and HDs2. This sequence had homology and 80% identity with specific Lepidoptera antimicrobial peptides called gloverins. Analyses using the ImageMaster 2D software showed pI 8.94 of the HDs1 spot, which is similar to that described to Hyalophora gloveri gloverin (pI 8.5). Moreover, the 14-kDa molecular mass of the spot HDs1 is compatible to that of gloverins isolated from the hemolymph of Trichoplusia ni, Helicoverpa armigera and H. gloveri. Antimicrobial assays with partially purified fractions containing the HDs1 and HDs2 polypeptides demonstrated activity against Escherichia coli. This is the first report of antimicrobial polypeptides in D. saccharalis, and the identification of these peptides may help in the generation of new strategies to control this pest.

Biotechnological richness of the northeastern semi-arid region: antioxidant activity of casein hydrolysates from Moxotó goat milk (Capra hircus Linnaeus, 1758) obtained by papain action

This study aimed to identify antioxidant peptides from caprine casein hydrolysates by papain application using MALDI-TOF mass spectrometer, and a 2² full factorial design, with 4 axial points, in order to evaluate kinetic parameters (time and pH) effects on the degree of hydrolysis as well as the antioxidant activity of Moxotó goat milk casein peptides. Degree of hydrolysis was determined by total and soluble protein ratio in casein. Antioxidant activity was measured by ABTS method with 2, 2-cation-azinobis (3-ethylbenzothiazoline-6-sulfonic acid). TROLOX was used as standard. Peptide pattern and sequence of antioxidant amino acids were obtained using MALDI-TOF/MS. The highest degree of hydrolysis (28.5%) and antioxidant activity (2329.6 mmol.L TROLOX. mg- 1 peptide) were observed in the permeate. NENLL, NPWDQVK and LLYQEPVLGPV peptides, detected in the permeate, were pointed as the responsible for antioxidant activity, suggesting their potential application as food supplement and pharmaceutical products.