Characterization of a Trypanosoma cruzi genomic fragment complementary to several species-specific mRNAs but different from the spliced leader sequence

We have isolated a clone of Trypanosoma cruzi genimic DNA, lambda 3b2-5, which contains sequences that are reiterated in the genome. Northtern blot analysis showed that clone 3b2-5 hybridizes to 1,200-5,000 bases different mRNA species. The number of mRNAs species hybridized to clone 3b2-5 exceeds its coding capacity showing that this clone carries sequences that are common to several mRNAs species and conserved in the poly A(+) RNA. These sequences are not homologous to the T. cruzi spliced leader sequence, since clone 3b2-5 hybridize to a synthetic 20 nucleotice complementary to the spliced leader sequence. Clone 3b2-5 does not hybridize to DNA and RNA from several genera of Trypanosomatidae and other Trypanosoma species indicating that it carries T. cruzi species-specific sequences.

Characterization of soil chemical properties of strawberry fields using principal component analysis

One of the largest strawberry-producing municipalities of Rio Grande do Sul (RS) is Turuçu, in the South of the State. The strawberry production system adopted by farmers is similar to that used in other regions in Brazil and in the world. The main difference is related to the soil management, which can change the soil chemical properties during the strawberry cycle. This study had the objective of assessing the spatial and temporal distribution of soil fertility parameters using principal component analysis (PCA). Soil sampling was based on topography, dividing the field in three thirds: upper, middle and lower. From each of these thirds, five soil samples were randomly collected in the 0-0.20 m layer, to form a composite sample for each third. Four samples were taken during the strawberry cycle and the following properties were determined: soil organic matter (OM), soil total nitrogen (N), available phosphorus (P) and potassium (K), exchangeable calcium (Ca) and magnesium (Mg), soil pH (pH), cation exchange capacity (CEC) at pH 7.0, soil base (V%) and soil aluminum saturation(m%). No spatial variation was observed for any of the studied soil fertility parameters in the strawberry fields and temporal variation was only detected for available K. Phosphorus and K contents were always high or very high from the beginning of the strawberry cycle, while pH values ranged from very low to very high. Principal component analysis allowed the clustering of all strawberry fields based on variables related to soil acidity and organic matter content.

SYNTHESIS AND CHARACTERIZATION OF HDTMA-ORGANOCLAYS: INSIGHTS INTO THEIR STRUCTURAL PROPERTIES

This study aims to synthesize and characterize organoclays developed from an Argentinian montmorillonite (Bent) using hexadecyltrimethylammonium bromide (HDTMA-Br) as the intercalation agent. Subsequently, an adsorption mechanism is proposed. The obtained organoclays were more hydrophobic than the starting clay. Surfactant molecules were adsorbed initially through cation exchange in sites placed in the interlayer space of the clay. Adsorption in such sites continued until the interlayer space was saturated. Depending on the surfactant loading introduced during the intercalation process, different organizations of surfactant in the interlayer were obtained. Further adsorption of surfactant occurred in the mesopores generated by tactoids in the "house of cards" organization. This process kept surfactant molecules relatively free and out of the interlayer space.

Biheterocyclic ligands: synthesis, characterization and coordinating properties of bis(4-amino-5-mercapto-1,2,4-triazol-3-yl) alkanes with transition metal ions and their thermokinetic and biological studies

The Co(II), Ni(II) and Cu(II) metal ions complexes of Bis(4-amino-5-mercapto-1,2,4-triazol-3-yl) alkanes (BATs) have been prepared and characterized by elemental analysis, conductivity measurements infrared, magnetic susceptibility, the electronic spectral data and thermal studies. Based on spectral and magnetic results, the ligands are tetradentate coordinating through the N and S-atoms of BATs; six-coordinated octahedral or distorted octahedral and some times four-coordinated square planar were proposed for these complexes. Activation energies computed for the thermal decomposition steps were compared. The ligands and their metal complexes were tested in vitro for their biological effects. Their activities against two gram-positive, two gram-negative bacteria and two fungal species were found to vary from moderate to very strong.

Characterization and oxygen binding properties of des-Arg human hemoglobin

The role of chloride in the stabilization of the deoxy conformation of hemoglobin (Hb), the low oxygen affinity state, has been studied in order to identify the nature of this binding. Previous studies have shown that arginines 141α could be involved in the binding of this ion to the protein. Thus, des-Arg Hb, human hemoglobin modified by removal of the α-chain C-terminal residue Arg141α, is a possible model for studies of these interactions. The loss of Arg141α and all the salt bridges in which it participates is associated with subtle structural perturbations of the α-chains, which include an increase in the conformational flexibility and further shift to the oxy state, increasing oxygen affinity. Thus, this Hb has been the target of many studies of structural and functional behavior along with medical applications. In the present study, we describe the biochemical characterization of des-Arg Hb by electrophoresis, high-performance liquid chromatography and mass spectroscopy. The effects of chloride binding on the oxygen affinity and on the cooperativity to des-Arg Hb and to native human hemoglobin, HbA, were measured and compared. We confirm that des-Arg Hb presents high oxygen affinity and low cooperativity in the presence of bound chloride and show that the binding of chloride to des-Arg does not change its functional characteristics as observed with HbA. These results indicate that Arg141α may be involved in the chloride effect on Hb oxygenation. Moreover, they show that these residues contribute to lower Hb oxygen affinity to a level compatible with its biological function.

Serological, electrophoretic and biological properties of Fasciola hepatica antigens

Fasciola hepatica somatic antigen, its partially purified fractions and excretion-secretion products were investigated as to serological, electrophoretic and biological properties. In a Sephadex G-100 column (SG-100), Fasciola hepatica total antigen (FhTA) gave 5 fractions, and SDS-PAGE analysis showed they were glycoproteins ranging from 14 to 94 kDa molecular weight (MW). When these fractions were analyzed by enzyme linked immunotransfer blot (EITB) and immunodiffusion in gel (ID) with serum from immunized rats with FhTA, the presence of different antigenic components was revealed. In the SDS-PAGE of excretor-secretor antigen (ESA), it was possible to observe peptides from 12 to 22 kDa, which were also present in FhTA. When the FhTA, its fractions and the ESA were analyzed by EITB with the immune rat serum (IRS), it was observed that only some fractions of the SG-100 shared antigens with the FhTA and ESA. Moreover, DTH and ITH responses were studied in FhTA immunized rats challenged with these different antigen components, revealing that the protein/carbohydrate ratio is important for inducing DTH response. The ESA was the most active component in the DTH and ITH response.

Biological characterization of Trypanosoma cruzi strains from different zymodemes and schizodemes

The development in C3H mice of thirteen strains of Trypanosoma cruzi belonging to different zymodemes ans schizodemes was studied. Host mortality, virulence, histiotropism, parasitemia and polymorphism of the parasites were recorded. The strains were grouped into: a) high virulence - causing 100% mortality and characterized by predominance of bery broad trypomastigotes in the bloodstream at the end of infection; b) medium virulence - causing no mortality and with a predominance of broad trypomastigotes; c) low virulence - causing no mortality with blood forms not described due to the very low parasitemia. During 18 months maintenance the parasitemia curves were kept constant for all strains except one. A direct correlation between either zymodeme or schizodeme and experimental biological properties of T. cruzi strains was not found. However, the parasitemia was subpatent and patent for strains from zymodeme C and the others respectively. Furthermore the high virulence seems to be related to one of two shizodemes found within zymodeme B strains. All strains presenting patent parasitemia independent of shizodeme and ymodeme showed a myotropism towards heart and skeletal muscle with varible inflammatory intensity. The present study confirmed the heterogeneity found by isoenzyme and K-DNA patterns among the strains of T. cruzi isolated from chagasic patients in Bambuí, Minas Gerais State, Brasil.

Purification and biochemical characterization of four iron superoxide dismutases in Trypanosoma cruzi

Four superoxide dismutase (SOD) activities (SOD I, II, III, and IV) have been characterized in the epimastigote form of Trypanosoma cruzi. The total extract was subjected to two successive ammonium sulphate additions between 35 and 85%, and the resulting fraction was purified using two continuous chromatography processes (ion exchange and filtration). Enzymes were insensitive to cyanide but sensitive to hydrogen peroxide, properties characteristic of iron-containing SODs. The molecular masses of the different SODs were 20 kDa (SOD I), 60 kDa (SOD II), 50 kDa (SOD III) and 25 kDa (SOD IV), whereas the isoelectric points were 6.9, 6.8, 5.2 and 3.8, respectively. Subcellular location and digitonin experiments have shown that these SODs are mainly cytosolic, with small amounts in the low-mass organelles (SOD II and SOD I) and the mitochondrion (SOD III), where these enzymes play an important role in minimizing oxidative damage.

Heterologous expression and biochemical characterization of an α1,2-mannosidase encoded by the Candida albicans MNS1 gene

Protein glycosylation pathways, commonly found in fungal pathogens, offer an attractive new area of study for the discovery of antifungal targets. In particular, these post-translational modifications are required for virulence and proper cell wall assembly in Candida albicans, an opportunistic human pathogen. The C. albicans MNS1 gene is predicted to encode a member of the glycosyl hydrolase family 47, with 1,2-mannosidase activity. In order to characterise its activity, we first cloned the C. albicans MNS1 gene into Escherichia coli, then expressed and purified the enzyme. The recombinant Mns1 was capable of converting a Man9GlcNAc2 N-glycan core into Man8GlcNAc2 isomer B, but failed to process a Man5GlcNAc2-Asn N-oligosaccharide. These properties are similar to those displayed by Mns1 purified from C. albicansmembranes and strongly suggest that the enzyme is an ±1,2-mannosidase that is localised to the endoplasmic reticulum and involved in the processing of N-linked mannans. Polyclonal antibodies specifically raised against recombinant Mns1 also immunoreacted with the soluble ±1,2-mannosidases E-I and E-II, indicating that Mns1 could share structural similarities with both soluble enzymes. Due to the high degree of similarity between the members of family 47, it is conceivable that these antibodies may recognise ±1,2-mannosidases in other biological systems as well.

Chemical and physical properties of organic residues

Due to human activity, large amounts of organic residue are generated daily. Therefore, an adequate use in agricultural activities requires the characterization of the main properties. The chemical and physical characterization is important when planning the use and management of organic residue. In this study, chemical and physical properties of charcoal, coffee husk, pine-bark, cattle manure, chicken manure, coconut fiber, sewage sludge, peat, and vermiculite were determined. The following properties were analyzed: N-NH4+, N-N0(3)-, and total concentrations of N, P, S, K, Ca, Mg, Mn, Zn, Cu, and B, as well as pH, Electrical Conductivity (EC) and bulk density. Coffee husk, sewage sludge, chicken manure and cattle manure were generally richer in nutrients. The EC values of these residues were also the highest (0.08 - 40.6 dS m-1). Peat and sewage sludge had the highest bulky density. Sodium contents varied from 0 to 4.75 g kg-1, with the highest levels in chicken manure, cattle manure and sewage sludge. Great care must be taken when establishing proportions of organic residues in the production of substrates with coffee husk, cattle or chicken manure or sewage sludge in the calculation of the applied fertilizer quantity in crop fertilization programs.

Highlands of the upper Jequitinhonha valley, Brazil: I - characterization and classification

In the upper Jequitinhonha valley, state of Minas Gerais, Brazi, there are large plane areas known as "chapadas", which are separated by areas dissected by tributaries of the Jequitinhonha and Araçuaí rivers. These dissected areas have a surface drainage system with tree, shrub, and grass vegetation, more commonly known as "veredas", i.e., palm swamps. The main purpose of this study was to characterize soil physical, chemical and morphological properties of a representative toposequence in the watershed of the Vereda Lagoa do Leandro, a swamp near Minas Novas, MG, on "chapadas", the highlands of the Alto Jequitinhonha region Different soil types are observed in the landscape: at the top - Typic Haplustox (LVA), in the middle slope - Xanthic Haplustox (LA), at the footslope - Xanthic Haplustox, gray color, here called "Gray Haplustox" ("LAC") and, at the bottom of the palm swamp - Typic Albaquult (GXbd). These soils were first morphologically described; samples of disturbed and undisturbed soils were collected from all horizons and subhorizons, to evaluate their essential physical and chemical properties, by means of standard determination of Fe, Al, Mn, Ti and Si oxides after sulfuric extraction. The contents of Fe, Al and Mn, extracted with dithionite-citrate-bicarbonate and oxalate treatments, were also determined. In the well-drained soils of the slope positions, the typical morphological, physical and chemical properties of Oxisols were found. The GXbd sample, from the bottom of the palm swamp, is grayish and has high texture gradient (B/A) and massive structure. The reduction of the proportion of crystalline iron compounds and the low crystallinity along the slope confirmed the loss of iron during pedogenesis, which is reflected in the current soil color. The Si and Al contents were lowest in the "LAC" soil. There was a decrease of the Fe2O3/TiO2 ratio downhill, indicating progressive drainage restriction along the toposequence. The genesis and all physical and chemical properties of the soils at the footslope and the bottom of the palm swamp of the "chapadas" of the Alto Jequitinhonha region are strongly influenced by the occurrence of ground water on the surface or near the surface all year long, at present and/or in the past. Total concentrations of iron oxides, Fe d and Fe o in soils of the toposequence studied are related to the past and/or present soil colors and drainage conditions.

Characterization and classification of soils in the Taquari river basin - Pantanal region, state of Mato Grosso do Sul, Brazil

Among the soils in the Mato Grosso do Sul, stand out in the Pantanal biome, the Spodosols. Despite being recorded in considerable extensions, few studies aiming to characterize and classify these soils were performed. The purpose of this study was to characterize and classify soils in three areas of two physiographic types in the Taquari river basin: bay and flooded fields. Two trenches were opened in the bay area (P1 and P2) and two in the flooded field (P3 and P4). The third area (saline) with high sodium levels was sampled for further studies. In the soils in both areas the sand fraction was predominant and the texture from sand to sandy loam, with the main constituent quartz. In the bay area, the soil organic carbon in the surface layer (P1) was (OC) > 80 g kg-1, being diagnosed as Histic epipedon. In the other profiles the surface horizons had low OC levels which, associated with other properties, classified them as Ochric epipedons. In the soils of the bay area (P1 and P2), the pH ranged from 5.0 to 7.5, associated with dominance of Ca2+ and Mg2+, with base saturation above 50 % in some horizons. In the flooded fields (P3 and P4) the soil pH ranged from 4.9 to 5.9, H+ contents were high in the surface horizons (0.8-10.5 cmol c kg-1 ), Ca2+ and Mg² contents ranged from 0.4 to 0.8 cmol c kg-1 and base saturation was < 50 %. In the soils of the bay area (P1 and P2) iron was accumulated (extracted by dithionite - Fed) and OC in the spodic horizon; in the P3 and P4 soils only Fed was accumulated (in the subsurface layers). According to the criteria adopted by the Brazilian System of Soil Classification (SiBCS) at the subgroup level, the soils were classified as: P1: Organic Hydromorphic Ferrohumiluvic Spodosol. P2: Typical Orthic Ferrohumiluvic Spodosol. P3: Typical Hydromorphic Ferroluvic Spodosol. P4: Arenic Orthic Ferroluvic Spodosol.

Characterization and nutrient release from silicate rocks and influence on chemical changes in soil

The expansion of Brazilian agriculture has led to a heavy dependence on imported fertilizers to ensure the supply of the growing food demand. This fact has contributed to a growing interest in alternative nutrient sources, such as ground silicate rocks. It is necessary, however, to know the potential of nutrient release and changes these materials can cause in soils. The purpose of this study was to characterize six silicate rocks and evaluate their effects on the chemical properties of treated soil, assessed by chemical extractants after greenhouse incubation. The experimental design consisted of completely randomized plots, in a 3 x 6 factorial scheme, with four replications. The factors were potassium levels (0-control: without silicate rock application; 200; 400; 600 kg ha-1 of K2O), supplied as six silicate rock types (breccia, biotite schist, ultramafic rock, phlogopite schist and two types of mining waste). The chemical, physical and mineralogical properties of the alternative rock fertilizers were characterized. Treatments were applied to a dystrophic Red-Yellow Oxisol (Ferralsol), which was incubated for 100 days, at 70 % (w/w) moisture in 3.7 kg/pots. The soil was evaluated for pH; calcium and magnesium were extracted with KCl 1 mol L-1; potassium, phosphorus and sodium by Mehlich 1; nickel, copper and zinc with DTPA; and the saturation of the cation exchange capacity was calculated for aluminum, calcium, magnesium, potassium, and sodium, and overall base saturation. The alternative fertilizers affected soil chemical properties. Ultramafic rock and Chapada mining byproduct (CMB) were the silicate rocks that most influenced soil pH, while the mining byproduct (MB) led to high K levels. Zinc availability was highest in the treatments with mining byproduct and Cu in soil fertilized with Chapada and mining byproduct.

Characterization of the soil fertility and root system of restinga forests

The Restinga vegetation consists of a mosaic of plant communities, which are defined by the characteristics of the substrates, resulting from the type and age of the depositional processes. This mosaic complex of vegetation types comprises restinga forest in advanced (high restinga) and medium regeneration stages (low restinga), each with particular differentiating vegetation characteristics. The climate along the coast is tropical (Köppen). Of all ecosystems of the Atlantic Forest, Restinga is the most fragile and susceptible to anthropic disturbances. Plants respond to soil characteristics with physiological and morphological modifications, resulting in changes in the architecture (spatial configuration) of the root system. The purpose of this study was to characterize the soil fertility of high and low restinga forests, by chemical and physical parameters, and its relation to the root system distribution in the soil profile. Four locations were studied: (1) Ilha Anchieta State Park, Ubatuba; (2) two Ecological Stations of Jureia-Itatins and of Chauás, in the municipality of Iguape; (3) Vila de Pedrinhas in the municipality of Ilha Comprida; and (4) Ilha do Cardoso State Park, Cananeia. The soil fertility (chemical and physical properties) was analyzed in the layers 0-5, 0-10, 0-20, 20-40 and 40-60 cm. In addition, the distribution of the root system in the soil profile was evaluated, using digital images and the Spring program. It was concluded that the root system of all vegetation types studied is restricted to the surface layers, 0-10 and 10-20 cm, but occupies mainly the 0-10 cm layer (70 %); that soil fertility is low in all environments studied, with base saturation values below 16 %, since most exchange sites are occupied by aluminum; and that restinga vegetation is edaphic.

Soil and phytosociological characterization of an area with predominance of arnica (Lychnophora pohlii sch. bip.)

Lychnophora pohlii Sch. Bip. (Asteraceae), known as "Arnica mineira", is widely used in folk medicine and very abundant in the altitude vegetation of rocky grassland. The aim of this work was to study the density of this species and its relationship with soil parameters in rocky grassland in Diamantina, in the Upper Jequitinhonha region, Minas Gerais. Ten contiguous 20 x 50 m plots were marked (total sampled area 10,000 m²) on the campus Juscelino Kubitschek of the Federal University of Jequitinhonha and Mucuri Valleys (UFVJM). The plants in these plots were evaluated for frequency, dominance and density. The relationship between the density of this species with nine soil physical and chemical properties was analyzed by means of canonical correspondence analysis (CCA). The highest plant abundance (I) of the species Lychnophora pohlii Sch. Bip. was found in the vegetation sampling areas: plot 6 with 255 plants, plot 7 with 173, plot 8 with 189, plot 9 with 159, and plot 1 with 151 plants. In these areas, the floristic soil characteristics were similar, resulting in spatial proximity in the ACC diagrams. The density of Lychnophora pohlii was higher in plots with higher pH, P-rem and base saturation, the variables most strongly correlated with the first axis of canonical correspondence analysis.