Distribution of collagen types I, III, and IV in gastric tissue of marmosets (Callithrix spp., Callitrichidae: Primates)

Extracellular matrix (ECM) components such as fibrillar collagens play a fundamental role in wound repair and have also been studied in association with the gastric ulcer healing process in gastroenterology. Nevertheless, there have been no studies in the literature to date regarding the description and characterization of ECM components, neither in normal nor in injured gastric tissue of primate species. The objective of this study was to investigate the expression of gastric collagen types I, III, and IV in marmosets (Callithrix sp.). Histological specimens from the stomach of 6 Callithrix jacchus, 12 C. kuhli, and 12 C. geoffroyi were evaluated. The specimens were immunostained with anti-types I and III collagen polyclonal antibodies and anti-type IV collagen monoclonal antibody. Collagen types I and III were detected in the submucosa and lamina propria between the mucosal glands while collagen type IV was detected in the muscularis mucosae, muscular layers, blood vessels, and gastric mucosa between the mucosal glands. It is hoped that these findings can contribute to future studies on the gastric extracellular matrix components in primates and to comparative studies in the area of gastroenterology.

Aplicação, em pré-colheita, de dessecante em duas cultivares de soja (Glycine max (L. ) Merril) III - Efeitos sobre a composição química (proteína, óleo e cinzas) e resíduos nas sementes

A aplicação pré-colheita do dessecante paraquat, quando real izada a partir das primeiras épocas (75 e 72 dias após o início do florescimento, para a Santa Rosa e IAC-2, respectivamente) não modificaram os teores com que ocorreram normalmente, proteína, extrato etéreo e cinzas nos grãos. As análises do resíduo de paraquat nos grãos colhidos, mostram claramente que não se deve recomendar tal prática às lavouras de soja, cujo objetivo final seja o fornecimento de grãos para a alimentação humana e animal. Entretanto pode ser indicada, sem maiores restrições, àquelas cuja finalidade é a produção de sementes comerciais.

Injúria potencial de herbicidas de solo ao girassol: III - imazaquin e imazethapyr

Conduziu-se um ensaio em campo na Estação Experimental Agronômica da Universidade Federal do Rio Grande do Sul, em Eldorado do Sul, RS, em 1989/90. O objetivo do trabalho foi determinar o efeito residual potencial dos herbicidas imazaquin e imazethapyr, aplicados em três doses à superfície do solo (PRE) ou incorporados no mesmo (PPI), na cultura de girassol. Num sistema de rotação soja-girassol, o efeito residual potencial de imazaquin foi mais acentuado que o de imazethapyr. A atividade herbicida de imazaquin e imazethapyr não foi dependente do método de aplicação. Imazaquin e imazethapyr, nas maiores doses, promoveram danos iniciais acentuados. As plantas de girassol recuperaram-se dos sintomas de intoxicação de imazethapyr, os quais diminuíram com o decorrer do tempo, mas não se recuperaram das injúrias causadas pelas maiores doses de imazaquin.

Estudos anatômicos de folhas de espécies de plantas daninhas de grande ocorrência no Brasil: III - Galinsoga parviflora, Crotalaria incana, Conyza bonariensis e Ipomoea cairica

O objetivo deste trabalho foi estudar a anatomia das folhas das espécies de plantas daninhas de grande ocorrência no Brasil: Galinsoga parviflora, Crotalaria incana, Conyza bonariensis e Ipomoea cairica, visando melhor entendimento sobre as barreiras que cada espécie impõe à penetração dos herbicidas e, assim, fornecer subsídios à busca de estratégias para superar esses obstáculos. As folhas completamente expandidas do terceiro ao quinto nó foram coletadas de plantas de ocorrência espontânea no campo. Das folhas de cada espécie foram obtidas três amostras da região central, com aproximadamente 1 cm², as quais foram utilizadas em estudos da estrutura e clarificação e em observações em microscópio eletrônico de varredura (MEV). Todas as espécies avaliadas são anfiestomáticas. A principal barreira foliar potencial à penetração de herbicidas observada na planta daninha G. parviflora foi a baixa densidade estomática na face adaxial. C. incana apresentou como possível principal obstáculo foliar à penetração de herbicidas o alto teor de cera epicuticular. Já em relação a C. bonariensis, alta densidade tricomática, grande espessura da cutícula da face adaxial e baixa densidade estomática na face adaxial foram as principais barreiras detectadas. Grande espessura da cutícula da face adaxial e baixa densidade estomática na face adaxial foram os possíveis obstáculos constatados nas folhas de I. cairica.

Períodos de interferência das plantas daninhas na cultura da cana-de-açúcar: III - capim-braquiária (Brachiaria decumbens) e capim-colonião (Panicum maximum)

Um experimento foi conduzido em São João da Boa Vista-SP, com o objetivo de determinar o período anterior à interferência (PAI) e o período total de prevenção à interferência (PTPI) das plantas daninhas na cultura da cana-de-açúcar. A cana foi plantada em abril de 1995, na época de plantio caracterizada por maior deficiência hídrica. A comunidade infestante presente foi variada, sendo Brachiaria decumbens e Panicum maximum as espécies mais importantes. Essa comunidade tendeu a apresentar acúmulo crescente de matéria seca durante todo o período de avaliação e reduziu em até 40% a produtividade de colmos da cana-de-açúcar. A cultura conviveu com a comunidade infestante até 74 dias após o plantio, sem sofrer redução significativa na produtividade (PAI). O período mínimo de controle para garantir a produtividade foi de 127 DAP (PTPI). Dessa forma, o controle das plantas daninhas foi crítico no período compreendido entre 74 e 127 dias após o plantio.

Interferências mútuas entre a cultura do milho, espécies forrageiras e plantas daninhas em um sistema de consórcio: III - implicações sobre as plantas daninhas

O objetivo desta pesquisa foi avaliar as conseqüências da adoção de um sistema de consórcio com culturas forrageiras sobre a infestação e a produção de massa seca e área foliar de três espécies daninhas, na cultura do milho. Os tratamentos resultaram da combinação entre quatro níveis do fator plantas forrageiras (Brachiaria decumbens, Brachiaria brizantha, Panicum maximum e ausência de forrageira) e três níveis do fator plantas daninhas (Ipomoea grandifolia corda-de-viola, Amaranthus hybridus caruru-roxo e Digitaria horizontalis capim-colchão). Durante a condução do experimento foram avaliadas a área foliar, a massa seca e a densidade das plantas daninhas quando submetidas à convivência com a associação do milho e espécies forrageiras. De forma geral, o sistema de produção que envolve a associação de uma cultura forrageira com a cultura do milho reduz a infestação e suprime o crescimento de plantas daninhas no sistema; B. decumbens foi a forrageira que menos reduziu a infestação de plantas daninhas; a espécie B. brizantha foi a forrageira mais eficiente em reduzir a infestação de corda-de-viola, mas não suprimiu o crescimento do caruru-roxo; e a forrageira P. maximum foi a espécie que mais reduziu o crescimento do caruru-roxo e a área foliar do capim-colchão.

Seletividade de herbicidas: III - aplicação de herbicidas em pós emergência inicial e tardia da cana-de-açúcar na época da estiagem

Este trabalho objetivou avaliar a seletividade dos herbicidas diuron+hexazinone, azafenidin+hexazinone, metribuzin e isoxaflutole aplicados em pós-emergência inicial e tardia das plantas de cana-de-açúcar, variedade RB835089, na época da estiagem, em soqueira de quarto corte, após colheita com queima prévia do canavial, em Araras, SP. O delineamento experimental foi o de blocos casualizados com testemunhas pareadas, com cinco tratamentos e quatro repetições, existindo uma testemunha capinada para cada tratamento químico dentro de cada bloco. Os tratamentos foram: diuron+hexazinone (1.170+330 g ha-1), azafenidin+ hexazinone (192,5+247,5 g ha-1), metribuzin (1.920 g ha-1), isoxaflutole (127,5 g ha¹) e testemunha capinada. Concluiu-se que os herbicidas foram mais fitotóxicos quando aplicados na pós-emergência tardia. Em pós-emergência inicial, as plantas recuperaram-se totalmente dos efeitos fitotóxicos dos herbicidas. Na pós-emergência tardia, azafenidin+hexazinone, isoxaflutole e diuron+hexazinone prejudicaram o índice de fluorescência e a produtividade agrícola.

Spatial distribution and preferential substrate of Neoregelia johannis (Carriére) L.B. Smith (Bromeliaceae) in a disturbed area of Atlantic Rainforest at Ilha Grande, RJ, Brazil

In bromeliad populations, amount of light and available substrates influence individuals spatial organization. In Atlantic Rainforest of Ilha Grande, the heliophylous bromeliad Neoregelia johannis is a large and abundant species. In this forest, it would be expected that N. johannis would occupy stable substrates, as large trunks, large branches, rock boulders or ground, with high sunlight, enough for the bromeliad survivor. In the present work, we analyzed the distribution and most used substrates of N. johannis in secondary forest. We analyzed the frequency of reproductive modes (sexual and vegetative) used by the bromeliad shoots, registering if the shoots were originated from seeds or by vegetative reproduction. The results indicated an aggregated distribution pattern (Ip = 0.052). The preferred substrate was boulders (91%), whereas tree trunks (6%) and the ground (3%) were rarely used. Small and fragile substrates are unstable to support large adults of this species, which may explain the predominant pattern of establishment over boulders within the secondary forest, as the presence of this substrate also results in more opened canopy cover. Approximately 50% of young individuals entered the population by vegetative reproduction. We conclude that the preferential habit and the aggregated distribution of N. johannis are due to the conjunction of preferred substrate with higher amount of light resulting from breaks in tree canopy over areas with rock blocks, and high frequency of recruitment by vegetative reproduction.

Molecular cloning of exons II and III of the a-globin major gene from Odontophrynus americanus 2n and 4n (Amphibia, Anura)

The a-globin major genes from diploid and tetraploid Odontophrynus americanus were studied using PCR-based technology. The cloned and sequenced amplified fragments were shown to contain most of the exon II sequences as well as the whole exon III sequence of the a-globin gene. Unexpectedly, intron 2 was entirely absent in the amplified fragments of both 2n and 4n origin. High conservation was observed among the obtained sequences when compared to corresponding sequences from human and Xenopus laevis origin. The possibility that these sequences might be pseudogenes is raised

The insulin receptor substrate 1 associates with phosphotyrosine phosphatase SHPTP2 in liver and muscle of rats

Insulin stimulates the tyrosine kinase activity of its receptor resulting in the phosphorylation of its cytosolic substrate, insulin receptor substrate-1 (IRS-1) which, in turn, associates with proteins containing SH2 domains. It has been shown that IRS-1 associates with the tyrosine phosphatase SHPTP2 in cell cultures. While the effect of the IRS-1/SHPTP2 association on insulin signal transduction is not completely known, this association may dephosphorylate IRS-1 and may play a critical role in the mitogenic actions of insulin. However, there is no physiological demonstration of this pathway of insulin action in animal tissues. In the present study we investigated the ability of insulin to induce association between IRS-1 and SHPTP2 in liver and muscle of intact rats, by co-immunoprecipitation with anti-IRS-1 antibody and anti-SHPTP2 antibody. In both tissues there was an increase in IRS-1 association with SHPTP2 after insulin stimulation. This association occurred when IRS-1 had the highest level of tyrosine phosphorylation and the decrease in this association was more rapid than the decrease in IRS-1 phosphorylation levels. The data provide evidence against the participation of SHPTP2 in IRS-1 dephosphorylation in rat tissues, and suggest that the insulin signal transduction pathway in rat tissues is related mainly to the mitogenic effects of the hormone.

Erythrocyte glucose-6-phosphate dehydrogenase activity assay and affinity for its substrate under "physiological" conditions

Glucose-6-phosphate dehydrogenase (G6PD) activity and the affinity for its substrate glucose-6-phosphate were investigated under conditions similar to the physiological environment in terms of ionic strength (I: 0.188), cation concentration, pH 7.34, and temperature (37oC). A 12.4, 10.4 and 21.4% decrease was observed in G6PD B, G6PD A+ and G6PD A- activities, respectively. A Km increase of 95.1, 94.4 and 95.4% was observed in G6PD B, G6PD A+ and G6PD A-, respectively, leading to a marked decrease in affinity. In conclusion, the observation of the reduced activity and affinity for its natural substrate reflects the actual pentose pathway rate. It also suggests a much lower NADPH generation, which is crucial mostly in G6PD-deficient individuals, whose NADPH availability is poor.

Reference values for lung function tests: III. Carbon monoxide diffusing capacity (transfer factor)

Carbon monoxide diffusing capacity (DLCO) or transfer factor (TLCO) is a particularly useful test of the appropriateness of gas exchange across the lung alveolocapillary membrane. With the purpose of establishing predictive equations for DLCO using a non-smoking sample of the adult Brazilian population, we prospectively evaluated 100 subjects (50 males and 50 females aged 20 to 80 years), randomly selected from more than 8,000 individuals. Gender-specific linear prediction equations were developed by multiple regression analysis with single breath (SB) absolute and volume-corrected (VA) DLCO values as dependent variables. In the prediction equations, age (years) and height (cm) had opposite effects on DLCOSB (ml min-1 mmHg-1), independent of gender (-0.13 (age) + 0.32 (height) - 13.07 in males and -0.075 (age) + 0.18 (height) + 0.20 in females). On the other hand, height had a positive effect on DLCOSB but a negative one on DLCOSB/VA (P<0.01). We found that the predictive values from the most cited studies using predominantly Caucasian samples were significantly different from the actually measured values (P<0.05). Furthermore, oxygen uptake at maximal exercise (VO2max) correlated highly to DLCOSB (R = 0.71, P<0.001); this variable, however, did not maintain an independent role to explain the VO2max variability in the multiple regression analysis (P>0.05). Our results therefore provide an original frame of reference for either DLCOSB or DLCOSB/VA in Brazilian males and females aged 20 to 80 years, obtained from the standardized single-breath technique.

The calcium-modulated proteins, S100A1 and S100B, as potential regulators of the dynamics of type III intermediate filaments

The Ca2+-modulated, dimeric proteins of the EF-hand (helix-loop-helix) type, S100A1 and S100B, that have been shown to inhibit microtubule (MT) protein assembly and to promote MT disassembly, interact with the type III intermediate filament (IF) subunits, desmin and glial fibrillary acidic protein (GFAP), with a stoichiometry of 2 mol of IF subunit/mol of S100A1 or S100B dimer and an affinity of 0.5-1.0 µM in the presence of a few micromolar concentrations of Ca2+. Binding of S100A1 and S100B results in inhibition of desmin and GFAP assemblies into IFs and stimulation of the disassembly of preformed desmin and GFAP IFs. S100A1 and S100B interact with a stretch of residues in the N-terminal (head) domain of desmin and GFAP, thereby blocking the head-to-tail process of IF elongation. The C-terminal extension of S100A1 (and, likely, S100B) represents a critical part of the site that recognizes desmin and GFAP. S100B is localized to IFs within cells, suggesting that it might have a role in remodeling IFs upon elevation of cytosolic Ca2+ concentration by avoiding excess IF assembly and/or promoting IF disassembly in vivo. S100A1, that is not localized to IFs, might also play a role in the regulation of IF dynamics by binding to and sequestering unassembled IF subunits. Together, these observations suggest that S100A1 and S100B may be regarded as Ca2+-dependent regulators of the state of assembly of two important elements of the cytoskeleton, IFs and MTs, and, potentially, of MT- and IF-based activities.

Nonconventional amide bond formation catalysis: programming enzyme specificity with substrate mimetics

This article reports on the design and characteristics of substrate mimetics in protease-catalyzed reactions. Firstly, the basis of protease-catalyzed peptide synthesis and the general advantages of substrate mimetics over common acyl donor components are described. The binding behavior of these artificial substrates and the mechanism of catalysis are further discussed on the basis of hydrolysis, acyl transfer, protein-ligand docking, and molecular dynamics studies on the trypsin model. The general validity of the substrate mimetic concept is illustrated by the expansion of this strategy to trypsin-like, glutamic acid-specific, and hydrophobic amino acid-specific proteases. Finally, opportunities for the combination of the substrate mimetic strategy with the chemical solid-phase peptide synthesis and the use of substrate mimetics for non-peptide organic amide synthesis are presented.

Development of an operational substrate for ZapA, a metalloprotease secreted by the bacterium Proteus mirabilis

The protease ZapA, secreted by Proteus mirabilis, has been considered to be a virulence factor of this opportunistic bacterium. The control of its expression requires the use of an appropriate methodology, which until now has not been developed. The present study focused on the replacement of azocasein with fluorogenic substrates, and on the definition of enzyme specificity. Eight fluorogenic substrates were tested, and the peptide Abz-Ala-Phe-Arg-Ser-Ala-Ala-Gln-EDDnp was found to be the most convenient for use as an operational substrate for ZapA. A single peptide bond (Arg-Ser) was cleaved with a Km of 4.6 µM, a k cat of 1.73 s-1, and a catalytic efficiency of 376 (mM s)-1. Another good substrate for ZapA was peptide 6 (Abz-Arg-Pro-Pro-Gly-Phe-Ser-Pro-Phe-Arg-Gln-EDDnp) which was cleaved at a single bond (Phe-Ser) with a Km of 13.6 µM, a k cat of 3.96 s-1 and a catalytic efficiency of 291 (mM s)-1. The properties of the amino acids flanking the scissile bonds were also evaluated, and no clear requirement for the amino acid residue at P1 was found, although the enzyme seems to have a preference for a hydrophobic residue at P2.