Protein recovery, separation and purification: selection of optimal techniques using an expert system

The paper discusses the utilization of new techniques ot select processes for protein recovery, separation and purification. It describesa rational approach that uses fundamental databases of proteins molecules to simplify the complex problem of choosing high resolution separation methods for multi component mixtures. It examines the role of modern computer techniques to help solving these questions.

Comparative chromatin analysis of Trypanosoma congolense

The chromatin of Trypanosoma congolense was analyzed by electron microscopy. The chromatin is organized as nucleosome filaments but does not form a 30 nm fiber. There are five groups of histones, including a histone H1-like protein, which has a molecular weight within the range of the core histones, and is extremely hydrophilic. Weak histone-histone interaction, a typical feature of trypanosoma chromatin, was found. These results are similar to those for T. cruzi and T. b. brucei, but differ significantly from those for higher eukaryotes. The results confirm the model of trypanosome chromatin, and support the theory of their early separation from the other eukaryotes during the evolution. T. congolensis is an excellent model for chromatin research on trypanosomes, because it is easy to cultivate and its chromatin has, a relatively high stability, compared to that of other trypanosomes.

Separation and Mapping of Chromosomes of Parasitic Protozoa

Many protozoan parasites represent an important group of human pathogens. Pulsed Field Gradient Gel Electrophoresis (PFGE) analysis has been an important tool for fundamental genetic studies of parasites like Trypanosoma, Leishmania, Giardia or the human malaria parasite Plasmodium falciparum. We present PFGE conditions allowing a high resolution separation of chromosomes ranging from 500 to 4000 kb within a two day electrophoresis run. In addition, we present conditions for separating large chromosomes (2000-6000 kb) within 36 hr. We demontrate that the application of two dimentional PFGE (2D-PFGE) technique to parasite karyotypes is a very useful method for the analysis of dispersed gene families and comparative studies of the intrachomosomal genome organization

Seroprevalence of Trypanosoma cruzi Infection in Students at the Seven-Fourteen Age Range, Londrina, PR, Brazil, in 1995

Seropositivity for Chagas disease was evaluated in 834 children aged between 7 and 14 from the Municipal Teaching System in the district of Londrina, State of Paraná. A seroprevalence rate of 0.1% was found through the use of an indirect immunofluorescent test and an enzyme-linked immunosorbent assay. This low rate of seroprevalence provides evidence that the vectorial transmission of Chagas disease has been eliminated in Londrina. The main reason for the elimination of vectorial transmission of Trypanosoma cruzi infection, as evaluated by serological tests, may be a remarkable change in the economic structure of the northern region of Paraná in the 1960's. At that time coffee production was almost completely replaced by soy beans, wheat and grazing in the rural areas. This change deeply affected the rural ecology and caused an exodus of the population from rural to urban areas as well as a decrease in the total number of the population of that region. The measures introduced for controlling the disease through the Program of Chagas Disease Control established by the Fundação Nacional de Saúde of the Brazilian Ministry of Health, certainly, had a positive impact on the reduction of American trypanosomiasis prevalence in the area under study. However, it does not seem that this was the most relevant factor responsible for the elimination of vectorial transmission of Chagas disease in Londrina.

Small todents fleas from the bubonic plague focus located in the Serra dos Órgãos Mountain Range, State of Rio de Janeiro, Brazil

Eleven species of fleas were collected from 601 small rodents, from November 1995 to October 1997, in areas of natural focus of bubonic plague, including the municipalities of Nova Friburgo, Sumidouro and Teresópolis, State of Rio de Janeiro, Brazil. Among 924 fleas collected, Polygenis (Polygenis) rimatus (Rhopalopsyllidae) was the predominant species regarding the frequency, representing 41.3% (N:382), followed by P. (Neopolygenis) pradoi, representing 20% (N:185) and Craneopsylla minervaminerva (Stephanocircidae), representing 18.9% (N:175). The host Akodon cursor harbored 47.9% of these fleas. Other six host species were infested by 52.1% of the remaining fleas. Fleas were found on hosts and in places within the focus not previously reported by the literature.

Uncertainties regarding dengue modeling in Rio de Janeiro, Brazil

Dengue fever is currently the most important arthropod-borne viral disease in Brazil. Mathematical modeling of disease dynamics is a very useful tool for the evaluation of control measures. To be used in decision-making, however, a mathematical model must be carefully parameterized and validated with epidemiological and entomological data. In this work, we developed a simple dengue model to answer three questions: (i) which parameters are worth pursuing in the field in order to develop a dengue transmission model for Brazilian cities; (ii) how vector density spatial heterogeneity influences control efforts; (iii) with a degree of uncertainty, what is the invasion potential of dengue virus type 4 (DEN-4) in Rio de Janeiro city. Our model consists of an expression for the basic reproductive number (R0) that incorporates vector density spatial heterogeneity. To deal with the uncertainty regarding parameter values, we parameterized the model using a priori probability density functions covering a range of plausible values for each parameter. Using the Latin Hypercube Sampling procedure, values for the parameters were generated. We conclude that, even in the presence of vector spatial heterogeneity, the two most important entomological parameters to be estimated in the field are the mortality rate and the extrinsic incubation period. The spatial heterogeneity of the vector population increases the risk of epidemics and makes the control strategies more complex. At last, we conclude that Rio de Janeiro is at risk of a DEN-4 invasion. Finally, we stress the point that epidemiologists, mathematicians, and entomologists need to interact more to find better approaches to the measuring and interpretation of the transmission dynamics of arthropod-borne diseases.

Expansion of host range as a driving force in the evolution of Toxoplasma

The apicomplexan parasite Toxoplasma gondii is unusual in being able to infect almost any cell from almost any warm-blooded animal it encounters. This extraordinary host-range contrasts with its far more particular cousins such as the various species of the malaria parasite Plasmodium where each species of parasite has a single genus or even species of host that it can infect. Genetic and genomic studies have revealed a key role for a number of gene families in how Toxoplasma invades a host cell, modulates gene expression of that cell and successfully evades the resulting immune response. In this review, I will explore the hypothesis that a combination of sexual recombination and expansion of host range may be the major driving forces in the evolution of some of these gene families and the specific genes they encompass. These ideas stem from results and thoughts published by several labs in the last few years but especially recent papers on the role of different forms of rhoptry proteins in the relative virulence of F1 Toxoplasma progeny in a particular host species (mice).

Sandflies (Diptera: Psychodidae) from caves of the quartzite Espinhaço Range, Minas Gerais, Brazil

This study investigated the sandfly fauna of two quartzite caves in the Espinhaço Mountain Range, located in the municipality of Diamantina, state Minas Gerais. From August 2010-July 2011, entomological sampling was performed in the caves of Salitre and Monte Cristo with two HP light traps exposed in the photic and aphotic zones of each cave. The sandfly fauna consisted of 17 species, among which Lutzomyia cipoensis was predominant (54.76%). The male/female ratio in the total captures was 1:2.5. The aphotic zone showed the highest frequency of specimens captured (65%). A greater density of sandflies occurred during the summer (January-February), coinciding with the period of higher temperatures, humidity and rainfall. The presence of Lutzomyia longipalpis, Lutzomyia whitmani and Lutzomyia pessoai, proven or suspected vectors of leishmaniasis, is of concern because the area is visited by many tourists.

Updating the geographical distribution and frequency of Aedes albopictus in Brazil with remarks regarding its range in the Americas

The geographical distribution of Aedes albopictus in Brazil was updated according to the data recorded across the country over the last eight years. Countrywide house indexes (HI) for Ae. albopictus in urban and suburban areas were described for the first time using a sample of Brazilian municipalities. This mosquito is currently present in at least 59% of the Brazilian municipalities and in 24 of the 27 federal units (i.e., 26 states and the Federal District). In 34 Brazilian municipalities, the HI values for Ae. albopictus were higher than those recorded for Ae. aegypti, reaching figures as high as HI = 7.72 in the Southeast Region. Remarks regarding the current range of this mosquito species in the Americas are also presented. Nineteen American countries are currently infested and few mainland American countries have not confirmed the occurrence of Ae. albopictus. The large distribution and high frequency of Ae. albopictus in the Americas may become a critical factor in the spread of arboviruses like chikungunya in the new world.

The three phases of the ensemble forecasting of niche models: geographic range and shifts in climatically suitable areas of Utetheisa ornatrix (Lepidoptera, Arctiidae)

Species' geographic ranges are usually considered as basic units in macroecology and biogeography, yet it is still difficult to measure them accurately for many reasons. About 20 years ago, researchers started using local data on species' occurrences to estimate broad scale ranges, thereby establishing the niche modeling approach. However, there are still many problems in model evaluation and application, and one of the solutions is to find a consensus solution among models derived from different mathematical and statistical models for niche modeling, climatic projections and variable combination, all of which are sources of uncertainty during niche modeling. In this paper, we discuss this approach of ensemble forecasting and propose that it can be divided into three phases with increasing levels of complexity. Phase I is the simple combination of maps to achieve a consensual and hopefully conservative solution. In Phase II, differences among the maps used are described by multivariate analyses, and Phase III consists of the quantitative evaluation of the relative magnitude of uncertainties from different sources and their mapping. To illustrate these developments, we analyzed the occurrence data of the tiger moth, Utetheisa ornatrix (Lepidoptera, Arctiidae), a Neotropical moth species, and modeled its geographic range in current and future climates.

Compositional and structural variabilities of Mg-rich iron oxide spinels from tuffite

Maghemite (γFe2O3) from tuffite is exceptionally rich in Mg, relatively to most of those reportedly found in other mafic lithosystems. To investigate in detail the compositional and structural variabilities of this natural magnetic iron oxide, sets of crystals were isolated from samples collected at different positions in a tuffite weathering mantle. These sets of crystal were individually powdered and studied by X-ray diffractometry, Mössbauer spectroscopy, magnetization measurements and chemical analysis. Lattice parameter of the cubic cell (a0) was found to vary from 0.834(1) to 0.8412(1) nm. Lower a0-values are characteristic of maghemite whereas higher ones are related to a magnetite precursor. FeO content ranges up to 17 mass % and spontaneous magnetization ranges from 8 to 32 J T-1 kg-1. Zero-field room temperature Mössbauer spectra are rather complex, indicating that the hyperfine field distributions due to Fe3+ and mixed valence Fe3+/2+ overlap. The structural variabilities of the (Mg, Ti)-rich iron oxide spinels is essentially related to the range of chemical composition of its precursor (Mg, Ti)-rich magnetite, and probably to the extent to which it has been oxidized during transformation in soil.

Pedological and isotopic relations of a highland tropical peatland, Mountain Range of the Espinhaço Meridional (Brazil)

The evolution of organic matter sources in soil is related to climate and vegetation dynamics in the past recorded in paleoenvironmental Quaternary deposits such as peatlands. For this reason, a Histosol of the mineralotrophic peatland from the Pau-de-Fruta Special Protection Area - SPA, Espinhaço Meridional, State of Minas Gerais, was described and characterized to evidence the soil constituent materials and properties as related to changes in environmental conditions, supported by the isotopic and elementary characterization of soil C and N and 14C ages. Samples were collected in a depression at 1,350 m asl, where Histosols are possibly more developed due to the great thickness (505 cm). Nowadays, the area is colonized by vegetation physiognomies of the Cerrado Biome, mainly rocky and wet fields (Campo Rupestre and Campo Úmido), aside from fragments of Semidecidual Seasonal Forest, called Capões forests. The results this study showed that early the genesis of the analyzed soil profile showed a high initial contribution of mostly herbaceous organic matter before 8,090 ± 30 years BP (14C age). In the lower-mid Holocene, between 8,090 ± 30 years AP (14C age) to ± 4,100 years BP (interpolated age), the vegetation gradually became more woody, with forest expansion, possibly due to increased humidity, suggesting the existence of a more woody Cerrado in the past than at present. Drier climate conditions than the current were concluded ± 2,500 years BP (interpolated age) and that after 430 years BP (14C age) the forest gave way to grassland, predominantly. After the dry season, humidity increased to the current conditions. Due to these climate fluctuations during the Holocene, three decomposition stages of organic matter were observed in the Histosols of this study, with prevalence of the most advanced (sapric), typical of a deposit in a highly advanced stage of pedogenetic evolution.

Least limiting water range in soil under crop rotations and chiseling

Soil water availability to plants is affected by soil compaction and other variables. The Least Limiting Water Range (LLWR) comprises soil physical variables affecting root growth and soil water availability, and can be managed by either mechanical or biological methods. There is evidence that effects of crop rotations could last longer than chiseling, so the objective of this study was to assess the effect of soil chiseling or growing cover crops under no-till (NT) on the LLWR. Crop rotations involving triticale (X Triticosecale) and sunflower (Helianthus annuus) in the fall-winter associated with millet (Pennisetum glaucum), sorghum (Sorghum bicolor) and sunn hemp (Crotalaria juncea) as cover crops preceding soybean (Glycine max) were repeated for three consecutive years. In the treatment with chiseling (performed only in the first year), the area was left fallow between the fall-winter and summer crops. The experiment was carried out in Botucatu, São Paulo State, Brazil, from 2003 to 2006 on a Typic Rhodudalf. The LLWR was determined in soil samples taken from the layers 0-20 cm and 20- 40 cm, after chemical desiccation of the cover crops in December of the first and third year of the experiment. Chiseling decreases soil bulk density in the 0-20 cm soil layer, increasing the LLWR magnitude by lowering the soil water content at which penetration resistance reaches 2.0 MPa; this effect is present up to the third year after chiseling and can reach to a depth of 0.40 m. Crop rotations involving sunflower + sunn hemp, triticale + millet and triticale + sunn hemp for three years prevented soil bulk density from exceeding the critical soil bulk density in the 0- 0.20 m layer. This effect was observed to a depth of 0.40 m after three years of chiseling under crop rotations involving forage sorghum. Hence, chiseling and some crop rotations under no tillage are effective in increasing soil quality assessed by the LLWR.

Structural changes in latosols of the cerrado region: I - relationships between soil physical properties and least limiting water range

The agricultural potential of Latosols of the Brazilian Cerrado region is high, but when intensively cultivated under inappropriate management systems, the porosity can be seriously reduced, leading to rapid soil degradation. Consequently, accelerated erosion and sedimentation of springs and creeks have been observed. Therefore, the objective of this study was to evaluate structural changes of Latosols in Rio Verde, Goiás, based on the Least Limiting Water Range (LLWR), and relationships between LLWR and other physical properties. Soil samples were collected from the B horizons of five oxidic Latosols representing the textural variability of the Latosols of the Cerrado biome. LLWR and other soil physical properties were determined at various soil compaction degrees induced by uniaxial compression. Soil compaction caused effects varying from enhanced plant growth due to higher water retention, to severe restriction of edaphic functions. Also, inverse relationships were observed between clay content and bulk density values (Bd) under different structural conditions. Bd values corresponding to critical soil macroporosity (BdcMAC) were more restrictive to a sustainable use of the studied Latosols than the critical Bd corresponding to LLWR (BdcLLWR). The high tolerable compression potential of these oxidic Latosols was related to the high aeration porosity associated to the granular structure.

Least limiting water range in assessing compaction in a Brazilian Cerrado latosol growing sugarcane

In the south-central region of Brazil, there is a trend toward reducing the sugarcane inter-harvest period and increasing traffic of heavy harvesting machinery on soil with high water content, which may intensify the compaction process. In this study, we assessed the structural changes of a distroferric Red Latosol (Oxisol) by monitoring soil water content as a function of the Least Limiting Water Range (LLWR) and quantified its effects on the crop yield and industrial quality of the first ratoon crop of sugarcane cultivars with different maturation cycles. Three cultivars (RB 83-5054, RB 84-5210 and RB 86-7515) were subjected to four levels of soil compaction brought about by a differing number of passes of a farm tractor (T0 = soil not trafficked, T2 = 2 passes, T10 = 10 passes, and T20 = 20 passes of the tractor in the same place) in a 3 × 4 factorial arrangement with three replications. The deleterious effects on the soil structure from the farm machinery traffic were limited to the surface layer (0-10 cm) of the inter-row area of the ratoon crop. The LLWR dropped to nearly zero after 20 tractor passes between the cane rows. We detected differences among the cultivars studied; cultivar RB 86-7515 stood out for its industrial processing quality, regardless of the level of soil compaction. Monitoring of soil moisture in the crop showed exposure to water stress conditions, although soil compaction did not affect the production variables of the sugarcane cultivars. We thus conclude that the absence of traffic on the plant row maintained suitable soil conditions for plant development and may have offset the harmful effects of soil compaction shown by the high values for bulk density between the rows of the sugarcane cultivars.