Effect of culture conditions on the biomass determination by ergosterol of <Emphasis Type="Italic">Lentinus crinitus</Emphasis> and <Emphasis Type="Italic">Psilocybe castanella</Emphasis>

Estimating fungal growth is important in processes of soil bioremediation. It has been demonstrated that ergosterol is a good indicator of fungal biomass in solid substrata. In the present study were evaluated the effects upon the ergosterol rate of Lentinus crinitus Berk. and Psilocybe castanella Peck through the culture conditions of these fungi, which are evaluated for the bioremediation of soils contaminated by organochlorates. A good correlation between fungal biomass and ergosterol was observed for both species. The culture conditions did not influence the ergosterol rate of L. crinitus. Yet the ergosterol rate of P. castanella was influenced from 35 days of culture and when grown in the presence of 15.00 g hexachlorobenzene l⁻¹ of culture medium. So it is possible to estimate growth of both species using ergosterol as indicator in processes of soil bioremediation since the influences observed in the ergosterol rate of P. castanella are considered.     

Subsarcolemmal and intermyofibrillar mitochondria proteome differences disclose functional specializations in skeletal muscle

Skeletal muscle is a highly specialized tissue that contains two distinct mitochondria subpopulations, the subsarcolemmal (SS) and the intermyofibrillar (IMF) mitochondria. Although it is established that these mitochondrial subpopulations differ functionally in several ways, limited information exists about the proteomic differences underlying these functional differences. Therefore, the objective of this study was to biochemically characterize the SS and IMF mitochondria isolated from rat red gastrocnemius skeletal muscle. We separated the two mitochondrial subpopulations from skeletal muscle using a refined method that provides an excellent division of these unique mitochondrial subpopulations. Using proteomics of mitochondria and its subfractions (intermembrane space, matrix and inner membrane), a total of 325 distinct proteins were identified, most of which belong to the functional clusters of oxidative phosphorylation, metabolism and signal transduction. Although more gel spots were observed in SS mitochondria, 38 of the identified proteins were differentially expressed between the SS and IMF subpopulations. Compared to the SS mitochondrial, IMF mitochondria expressed a higher level of proteins associated with oxidative phosphorylation. This observation, coupled with the finding of a higher respiratory chain complex activity in IMF mitochondria, suggests a specialization of IMF mitochondria toward energy production for contractile activity.     

Analysis of membrane protein genes in a Brazilian isolate of Anaplasma marginale

The sequencing of the complete genome of Anaplasma marginale has enabled the identification of several genes that encode membrane proteins, thereby increasing the chances of identifying candidate immunogens. Little is known regarding the genetic variability of genes that encode membrane proteins in A. marginale isolates. The aim of the present study was to determine the degree of conservation of the predicted amino acid sequences of OMP1, OMP4, OMP5, OMP7, OMP8, OMP10, OMP14, OMP15, SODb, OPAG1, OPAG3, VirB3, VirB9-1, PepA, EF-Tu and AM854 proteins in a Brazilian isolate of A. marginale compared to other isolates. Hence, primers were used to amplify these genes: omp1, omp4, omp5, omp7, omp8, omp10, omp14, omp15, sodb, opag1, opag3, virb3, VirB9-1, pepA, ef-tu and am854. After polimerase chain reaction amplification, the products were cloned and sequenced using the Sanger method and the predicted amino acid sequence were multi-aligned using the CLUSTALW and MEGA 4 programs, comparing the predicted sequences between the Brazilian, Saint Maries, Florida and A. marginale centrale isolates. With the exception of outer membrane protein (OMP) 7, all proteins exhibited 92-100% homology to the other A. marginale isolates. However, only OMP1, OMP5, EF-Tu, VirB3, SODb and VirB9-1 were selected as potential immunogens capable of promoting cross-protection between isolates due to the high degree of homology (over 72%) also found with A. (centrale) marginale.     

Microfilariae infection in wild birds from the Brazilian cerrado

We report the occurrence of microfilariae in wild birds of a cerrado area in northern Brazil (Tocantins State). Analyses of 166 passerine birds belonging to 46 species and 17 families captured between 2006 and 2008 revealed that 11 individuals (6.6%) were hosts for microfilariae. Two bird species, Formicivora grisea and Formicivora rufa (Thamnophilidae), were identified as hosts for microfilariae for the first time, and had high intensities of microfilaremia (65 and 107 in 100 microscopic fields, respectively). The prevalence and intensity of microfilariae described in the present study are among the highest reported for wild bird communities in the neotropics.     

Evaluation of water quality of an urban stream in southeastern Brazil using Chironomidae larvae (Insecta: Diptera)

In order to estimate the water quality of S?o Pedro stream, through distribution and composition of Chironomidae larvae present in the sediment four sampling sites were selected. In each sampling site, three sediment samples were collected within a period of twelve months using the Petersen (0.0189 m2) and the van Veen (0.0518 m2) dredges. Samples were washed through a sieve with a 0.21 mm mesh and the collected organisms were sorted in transparent trays, with a light shine being reflected into the tray. The sites located in the greatest urban mesh showed high densities of the genus Chironomus and lower values for diversity, uniformity and taxa richness, in relation to sites located in a less urbanized area. A significant difference in density of Chironomidae larvae (p = 0.02; H = 5.89) was observed between the sites without domestic sewage effluents (site I) and those with the input of the effluents (sites II, III and IV). The Chironomidae larvae composition and the physical and chemical parameters were effective as indicators of the environmental alterations in S?o Pedro stream.     

Computational model for the transition from peristaltic to pulsatile flow in the embryonic heart tube

Early in development, the heart is a single muscle-wrapped tube without formed valves. Yet survival of the embryo depends on the ability of this tube to pump blood at steadily increasing rates and pressures. Developmental biologists historically have speculated that the heart tube pumps via a peristaltic mechanism, with a wave of contraction propagating from the inflow to the outflow end. Physiological measurements, however, have shown that the flow becomes pulsatile in character quite early in development, before the valves form. Here, we use a computational model for flow though the embryonic heart to explore the pumping mechanism. Results from the model show that endocardial cushions, which are valve primordia arising near the ends of the tube, induce a transition from peristaltic to pulsatile flow. Comparison of numerical results with published experimental data shows reasonably good agreement for various pressure and flow parameters. This study illustrates the interrelationship between form and function in the early embryonic heart.     

Glyceroneogenesis and the supply of glycerol-3-phosphate for glyceride-glycerol synthesis in liver slices of fasted and diabetic rats

The pathways of glycerol-3-phosphate (G3P) generation for glyceride synthesis were examined in precision-cut liver slices of fasted and diabetic rats. The incorporation of 5 mM [U-(14)C]glucose into glyceride-glycerol, used to evaluate G3P generation via glycolysis, was reduced by approximately 26-36% in liver slices of fasted and diabetic rats. The glycolytic flux was reduced by approximately 60% in both groups. The incorporation of 1.0 mM [2-(14)C]pyruvate into glyceride-glycerol (glyceroneogenesis) increased approximately 50% and approximately 36% in slices of fasted and diabetic rats, respectively, which also showed a two-fold increase in the activity phosphoenolpyruvate carboxykinase. The increased incorporation of 1.0 mM [2-(14)C]pyruvate into glyceride-glycerol by slices of fasted rats was not affected by the addition of 5 mM glucose to the incubation medium. The activity of glycerokinase and the incorporation of 1 mM [U-(14)C]glycerol into glyceride-glycerol, evaluators of G3P formation by direct glycerol phosphorylation, did not differ significantly from controls in slices of the two experimental groups. Rates of incorporation of 1 mM [2-(14)C]pyruvate and [U-(14)C]glycerol into glucose of incubation medium (gluconeogenesis) were approximately 140 and approximately 20% higher in fasted and diabetic slices than in control slices. It could be estimated that glyceroneogenesis by liver slices of fasted rats contributed with approximately 20% of G3P generated for glyceride-glycerol synthesis, the glycolytic pathway with approximately 5%, and direct phosphorylation of glycerol by glycerokinase with approximately 75%. Pyruvate contributed with 54% and glycerol with 46% of gluconeogenesis. The present data indicate that glyceroneogenesis has a significant participation in the generation of G3P needed for the increased glyceride-glycerol synthesis in liver during fasting and diabetes.