Variação ao longo do dia da atividade de enzimas do catabolismo de sacarose em plântulas de Hymenaea courbaril L. durante a mobilização do xiloglucano de reserva

Sementes de Hymenaea courbaril L. possuem um polissacarídeo de reserva que é mobilizado após a germinação, quando a primeira folha da planta já é fotossinteticamente ativa. No momento da mobilização das reservas, a plântula precisa coordenar duas fontes de carboidratos: a fotossíntese e a mobilização das reservas. Ambos geram sacarose como forma de exportação de carbono. Para entender a alocação de recursos na plântula, portanto, é necessário avaliar o catabolismo de sacarose nos órgãos. Neste trabalho foram analisados os carboidratos de baixo peso, quantificada a atividade da sacarose sintase e das três isoformas de invertase nos diferentes órgãos de plântulas de H. courbaril ao longo de um dia. As dosagens foram feitas no período de mobilização do xiloglucano, sendo as plântulas coletadas em intervalos de 6 horas, com uma coleta extra às 2 horas da manhã. Cada uma das enzimas apresentou um padrão característico de variação ao longo do dia, sugerindo funções distintas e independentes em cada órgão. A análise dos carboidratos mostrou altas concentrações de sacarose nos órgãos-dreno, enquanto os cotilédones apresentaram altas concentrações de monossacarídeos livres. A existência de isoformas com propriedades e distribuição celular distintas variando de forma independente ao longo do dia sugere que as isoformas podem ter funções fisiológicas distintas dentro da planta.

Effect of epithelial debridement on human cornea proteoglycans

Corneal transparency is attributed to the regular spacing and diameter of collagen fibrils, and proteoglycans may play a role in fibrillogenesis and matrix assembly. Corneal scar tissue is opaque and this opacity is explained by decreased ultrastructural order that may be related to proteoglycan composition. Thus, the objectives of the present study were to characterize the proteoglycans synthesized by human corneal explants and to investigate the effect of mechanical epithelial debridement. Human corneas unsuitable for transplants were immersed in F-12 culture medium and maintained under tissue culture conditions. The proteoglycans synthesized in 24 h were labeled metabolically by the addition of 35S-sulfate to the medium. These compounds were extracted by 4 M GuHCl and identified by a combination of agarose gel electrophoresis, enzymatic degradation with protease and mucopolysaccharidases, and immunoblotting. Decorin was identified as the main dermatan sulfate proteoglycan and keratan sulfate proteoglycans were also prominent components. When the glycosaminoglycan side chains were analyzed, only keratan sulfate and dermatan sulfate were detected (~50% each). Nevertheless, when these compounds were 35S-labeled metabolically, the label in dermatan sulfate was greater than in keratan sulfate, suggesting a lower synthesis rate for keratan sulfate. 35S-Heparan sulfate also appeared. The removal of the epithelial layer caused a decrease in heparan sulfate labeling and induced the synthesis of dermatan sulfate by the stroma. The increased deposit of dermatan sulfate proteoglycans in the stroma suggests a functional relationship between epithelium and stroma that could be related to the corneal opacity that may appear after epithelial cell debridement.

Detection of immunoglobulin G in the lung and liver of hamsters with visceral leishmaniasis

Several organs are affected in visceral leishmaniasis, not only those rich in mononuclear phagocytes. Hypergammaglobulinemia occurs during visceral leishmaniasis; anti-Leishmania antibodies are not primarily important for protection but might be involved in the pathogenesis of tissue lesions. The glomerulonephritis occurring in visceral leishmaniasis has been attributed to immune complex deposition but in other organs the mechanism has not been studied. In the current study we demonstrated the presence of IgG in the lung and liver of hamsters with visceral leishmaniasis. Hamsters were injected intraperitoneally with 2 x 10(7) amastigotes of Leishmania (Leishmania) chagasi and the presence of IgG in the liver and lung was evaluated at 7, 15, 30, 45, 80 and 102 days postinfection (PI) by immunohistochemistry. The parasite burden in the spleen and liver increased progressively during infection. We observed a deposit of IgG from day 7 PI that increased progressively until it reached highest intensity around 30 and 45 days PI, declining at later times. The IgG deposits outlined the sinusoids. In the lung a deposit of IgG was observed in the capillary walls that was moderate at day 7 PI, but the intensity increased remarkably at day 30 PI and declined at later times of infection. No significant C3 deposits were observed in the lung or in the liver. We conclude that IgG may participate in the pathogenesis of the inflammatory process of the lung and liver occurring in experimental visceral leishmaniasis and we discuss an alternative mechanism other than immune complex deposition.

The role of complement in the early phase of Leishmania (Leishmania) amazonensis infection in BALB/c mice

Complement-depleted and -non-depleted BALB/c mice were inoculated with Leishmania (Leishmania) amazonensis promastigotes into the hind footpad to study the role of the complement system in cutaneous leishmaniasis. Total serum complement activity was measured by hemolytic assay and C3 fragment deposit at the inoculation site was determined by direct immunofluorescence in the early period of infection, i.e., at 3, 24, 48 h and 7 days post-infection. The inflammatory reaction and the parasite burden were evaluated in the skin lesion at 7 and 30 days post-infection. Total serum complement activity decreased in the early phase of infection, from 3 to 24 h, in non-depleted mice compared to non-infected and non-depleted mice. C3 fragment deposit at the site of parasite inoculation was present throughout the period of infection in non-depleted mice. In contrast, no C3 fragment deposit was observed at the inoculation site in complement-depleted mice. Complement-depleted mice showed a significant decrease in the inflammatory response and a significant increase in the number of parasites (70.0 ± 5.3 vs 5.3 ± 1.5) at 7 days of infection (P < 0.05). A higher number of parasites were also present at 30 days of infection at the inoculation site of complement-depleted mice (78.5 ± 24.9 vs 6.3 ± 5.7). These experiments indicate that complement has an important role at the beginning of experimental cutaneous leishmaniasis caused by L. (L.) amazonensis by controlling the number of parasites in the lesion.

Protein turnover, amino acid requirements and recommendations for athletes and active populations

Skeletal muscle is the major deposit of protein molecules. As for any cell or tissue, total muscle protein reflects a dynamic turnover between net protein synthesis and degradation. Noninvasive and invasive techniques have been applied to determine amino acid catabolism and muscle protein building at rest, during exercise and during the recovery period after a single experiment or training sessions. Stable isotopic tracers (13C-lysine, 15N-glycine, ²H5-phenylalanine) and arteriovenous differences have been used in studies of skeletal muscle and collagen tissues under resting and exercise conditions. There are different fractional synthesis rates in skeletal muscle and tendon tissues, but there is no major difference between collagen and myofibrillar protein synthesis. Strenuous exercise provokes increased proteolysis and decreased protein synthesis, the opposite occurring during the recovery period. Individuals who exercise respond differently when resistance and endurance types of contractions are compared. Endurance exercise induces a greater oxidative capacity (enzymes) compared to resistance exercise, which induces fiber hypertrophy (myofibrils). Nitrogen balance (difference between protein intake and protein degradation) for athletes is usually balanced when the intake of protein reaches 1.2 g·kg-1·day-1 compared to 0.8 g·kg-1·day-1 in resting individuals. Muscular activities promote a cascade of signals leading to the stimulation of eukaryotic initiation of myofibrillar protein synthesis. As suggested in several publications, a bolus of 15-20 g protein (from skimmed milk or whey proteins) and carbohydrate (± 30 g maltodextrine) drinks is needed immediately after stopping exercise to stimulate muscle protein and tendon collagen turnover within 1 h.

Carbohydrate composition of ripe pineapple (cv. perola) and the glycemic response in humans

Brazil is the third largest producer of pineapple (Ananas comosus) and the market for fresh pineapple is sustained by the Hawaii and Perola cultivars. In this work the Perola cultivar was divided into three main parts, shell, core and pulp, for characterization. Moisture in the pulp was higher (between 10 and 15%) than in the shell and core. The amount of protein was higher in the core (35%) than in the pulp and shell. Perola contained relatively low concentrations of total ascorbic acid in the edible parts, although higher levels of ascorbic acid in the shell. Citric acid corresponded to almost 60% of the total organic acids. The total soluble sugars [~7-12% (FW)] were predominantly sucrose, fructose and glucose. The core had almost twice as much total sugar (12%) than the pulp (6.8%). The amount of insoluble dietary fiber was around 1%, and the soluble fiber was less than 0.1%. The pulp showed the highest concentration of polyphenols (0.49%) and antioxidant activity (33 µmol.g-1) out of the parts. The consumption of the pineapple pulp or core produced a high glycemic index (~93%), but considering the glycemic load, this fruit can be considered as low dietary.