Host-Parasite Interactions in Chagas Disease: Genetically Unidentical Isolates of a Single Trypanosoma cruzi Strain Identified In Vitro via LSSP-PCR

The present study aims at establishing whether the diversity in pathogenesis within a genetically diverse host population infected with a single polyclonal strain of Trypanosoma cruzi is due to selection of specific subpopulations within the strain. For this purpose we infected Swiss mice, a genetically diverse population, with the polyclonal strain of Trypanosoma cruzi Berenice-78 and characterized via LSSP-PCR the kinetoplast DNA of subpopulations isolated from blood samples collected from the animals at various times after inoculation (3, 6 and 12 months after inoculation). We examined the biological behavior of the isolates in acellular medium and in vitro profiles of infectivity in Vero cell medium. We compared the characteristics of the isolates with the inoculating strain and with another strain, Berenice 62, isolated from the same patient 16 years earlier. We found that one of the isolates had intermediate behavior in comparison with Berenice-78 and Berenice-62 and a significantly different genetic profile by LSSP-PCR in comparison with the inoculating strain. We hereby demonstrate that genetically distinct Trypanosoma cruzi isolates may be obtained upon experimental murine infection with a single polyclonal Trypanosoma cruzi strain.     

A simple,reliable method to determine the mean gait speed using heel markers on a treadmill

Gait speed is an essential parameter of gait analysis. Our study proposed a simple and accurate method to extract a mean gait speed during walking on a treadmill using only kinematic data from markers placed on the heels of the participants’ feet. This method provided an attractive, simple method that remains resistant to errors in treadmill calibration. In addition, this method required only two markers, since heel markers are essential to gait analysis, and the proposed method is robust enough to differentiate among various gait speeds (mean error <1%).     

Principles,techniques, and applications of biocatalyst immobilization for industrial application

Immobilization is one of the most effective and powerful tools used in industry, which has been studied and improved since the last century. Various immobilization techniques and support materials have been used on both laboratory and industrial scale. Each immobilization technique is applicable for a specific production mostly depending on the cost and sensibility of process. Compared to free biocatalyst systems, immobilization techniques often offer better stability, increased activity and selectivity, higher resistance, improved separation and purification, reuse of enzymes, and consequently more efficient process. Recently, many reviews have been published about immobilization systems; however, most of them have focused on a specific application or not emphasized in details. This review focuses on most commonly used techniques in industry with many recent applications including using bioreactor systems for industrial production. It is also aimed to emphasize the advantages and disadvantages of the immobilization techniques and how these systems improve process productivity compared to non-immobilized systems.     

Photosynthesis and water relations of almond tree cultivars grafted on two rootstocks

Five cultivars of Prunus amygdalus Batsch (Ferragnes, Ferrastar, Marcona, Garrigues, and Non Pareil) grafted on two different rootstocks (Garrigues and GF677), and two cultivars (Ferraduel and Casa Nova) grafted on GF677, were grown for three years under rainfed conditions in an orchard in northeast Portugal. Net photosynthetic rate (PN), leaf conductance for water vapour (gs), leaf water potential (Ψ), instantaneous water use efficiency (WUE), and internal CO2 concentration (Ci) were measured at three periods of the growing season: spring, summer (June or July) and late summer (September) over two years. Ferraduel, Ferrastar, and Marcona presented the best performance in the periods when environmental conditions were not very hard (May or September). Casa Nova and Non Pareil were well adapted to high air evaporative demand, preventing the increase of leaf temperature (T1). Ferrastar, although having a good performance in May and September, did well adapt to hard climatic conditions in June 1994. In the following year, although it presented the highest T1, the values were not limiting (30.6 ± 2.1 °C), and PN was only decreased from May to July. Marcona was highly dependent on T1, but prevented its increasing. Garrigues showed lower PN in most measurement periods. GF677 frequently induced the highest PN, WUE, and Ψ. PN was mainly dependent on T1, radiation, Ci, month, and year. WUE depended on the same factors. Ψ depended mainly on gs, air temperature, month, and year. This revised version was published online in June 2006 with corrections to the Cover Date.     

An Examination of Morphometric Variations in a Neotropical Toad Population (Proceratophrys cristiceps,Amphibia, Anura,Cycloramphidae)

The species Proceratophrys cristiceps belongs to the genus Proceratophrys within the family Cycloramphidae. These amphibians are found exclusively in South America in the morphoclimatic domain of the semi-arid depression zones in northeastern Brazil known as the Caatinga. We examined intrapopulational variation using univariate and multivariate statistics with traditional and geometric morphometrics, which supported the existence of two morphotypes of this species. Our results indicated significant degrees of variation in skeletal characteristics between some natural populations of this species. Careful analyses of variability levels are fundamental to avoid taxonomic errors, principally in populations that demonstrate characteristics intimately associated with their area of occurrence, as is the case of Proceratophrys cristiceps.     

Negative potential level in the outer layer of the toad skin.

The isolated skin of the toad Bufo marinus ictericus when impaled from the outer surface by glass microelectrodes filled with 3 M KCl shows a voltage profile which is a continuous function of the depth of impalement. The superficial intraepithelial potential difference measured with reference to the external solution (PDi) is negative with NaCl-Ringer's solution on both sides of the skin, displaying a minimum of -26.7+/-3.6 mV at 6+/-2 mum. Null value is obtained at 19+/-3 mum, with positive values for deeper impalements. Indications of cell impalements (abrupt voltage and resistance jumps) were frequently observed at sites deeper than 25 mum from the outer surface. Measurements of the electrical resistance between the microelectrode and the external solution, made with single- and double-barreled microelectrodes, showed great discrepancies, which may be attributed to distinct pathways of different resistances in the stratum corneum. PDi measured at a depth of 5 mum was a logarithmic function of Na2SO4 or K2SO4 concentration in the external solution, increasing in negativity with a reduction in concentration. Substitution of Na by K in the external solution had only minor effects on PDi. Acidification of the external solution from pH 9 is accompanied by a reduction in the negative value of PDi. At pH 3 PDi was positive. PDi was interpreted as a diffusion potential at the tip of the microelectrode due to KCl diffusion from the electrode into the matrix of the stratum corneum. Differences in K and Cl mobilities, responsible for the origin of PDi, were attributed to fixed charges in the matrix of the stratum corneum, with density and polarity determined by their degree of proponation, controlled by the hydrogen ion concentration of the external solution. Skin potential, short-circuit current and their relationship to PDI were discussed.     

Environmental and Animal Characteristics as Factors Associated with American Cutaneous Leishmaniasis in Rural Locations with Presence of Dogs,Brazil

The aim of the study was to investigate the importance of dogs, other domesticated animals and environmental characteristics as risk factors in the epidemiology of American cutaneous leishmaniasis (ACL). A retrospective survey of cases of human ACL in the last ten years and visits to homes in rural locations were carried out in the municipality of Arapongas (southern Brazil) from 2008 to 2010. ACL in humans was significantly associated with a distance of up to 25 meters from the residence to a forest area (OR 5.08; 95% CI: 1.35–21.04), undergrowth area (OR 6.80; 95% CI: 1.69–45.33) and stream (OR 5.87; 95% CI: 1.15–24.59); banana plants near the residence (OR 5.98; 95% CI: 1.49–39.84), absence of ceiling below the roof in the residence (OR 7.30; 95% CI: 1.26–158.1), the dumping of trash in the forest area (OR 26.33; 95% CI: 7.32–93.46) and presence of ACL in dogs in the surrounding area (OR 4.39; 95% CI: 1.37–13.45). In dogs, ACL was associated with a distance of 25 to 50 meters and 51 to 100 meters, respectively, from the residence to a forest area (OR 2.59; 95% CI: 1.08–5.98; OR 3.29; 95% CI: 1.64–6.62), the presence of a stream up to 25 m from the residence (OR 6.23; 95% CI: 2.34–16.54) and banana plants near the residence (OR 0.45; 95% CI: 0.25–0.80). In the locations studied in the municipality of Arapongas (Brazil), the results reveal that canine infection increases the risk of human infection by ACL and the characteristics surrounding the residence increase the risk of infection in both humans and dogs. Thus, integrated environmental management could be a useful measure to avoid contact between humans and phlebotomines.     

Acidocalcisomes are functionally linked to the contractile vacuole of Dictyostelium discoideum.

The mass-dense granules of Dictyostelium discoideum were shown to contain large amounts of phosphorus, magnesium, and calcium, as determined by x-ray microanalysis, either in situ or when purified using iodixanol gradient centrifugation. The high phosphorus content was due to the presence of pyrophosphate and polyphosphate, which were also present in the contractile vacuoles. Both organelles also possessed a vacuolar H(+)-ATPase, an H(+)-pyrophosphatase, and a Ca(2+)-ATPase, as determined by biochemical methods or by immunofluorescence microscopy. The H(+)-pyrophosphatase activity of isolated mass-dense granules was stimulated by potassium ions and inhibited by the pyrophosphate analogs aminomethylenediphosphonate and imidodiphosphate and by KF and N-ethylmaleimide in a dose-dependent manner. The mass-dense granules and the contractile vacuole appeared to contact each other when the cells were submitted to hyposmotic stress. Acetazolamide inhibited the carbonic anhydrase activity of the contractile vacuoles and prolonged their contraction cycle in a dose-dependent manner. Similar effects were observed with the anion exchanger inhibitor 4,4' -diisothiocyanatodihydrostilbene-2, 2' -disulfonic acid and the vacuolar H(+)-ATPase inhibitor bafilomycin A(1). Together, these results suggest that the mass-dense granules of D. discoideum are homologous to the acidocalcisomes described in protozoan parasites and are linked to the function of the contractile vacuole.     

Oxypetalum banksii subsp. banksii: a taxon of Asclepiadaceae with an extragynoecial compitum

 The floral morphology of seven Oxypetalum species and, in particular, the spatial relationship between the five stigmatic chambers and two separate ovaries of their flowers with respect to transmission of the pollen tube are studied. In all species, except O. banksii subsp. banksii, floral morphology is similar to that in other Asclepiadeae, and the flowers pollinated with one pollinium develop only one follicle, which means compitum absence. In O. banksii subsp. banksii flowers, the secretory interstaminal tissue lines the inner walls of the stigmatic chambers as in the other species studied, but it also reaches the upper part of the inner surface of the filament tube, where it surrounds the styles, an unprecedented feature for Asclepiadaceae. This tissue secretes nectar and mucilage; the latter acts as transmitting medium for the growth of pollen tubes from pollinia inserted and hydrated in stigmatic chambers (“hyperstigmas”). Mucilage also functions as an extragynoecial compitum: in flowers pollinated with one pollinium both carpels develop into a follicle. Received August 28, 2001; accepted April 9, 2002 Published online: October 14, 2002 Addresses of the authors: Milene Faria Vieira (e-mail: mfvieira@mail.ufv.br), Departamento de Biologia Vegetal, Universidade Federal de Vi?osa, 36571-000, Vi?osa, Minas Gerais, Brazil. George John Shepherd, Departamento de Botanica, Instituto de Biologia, Universidade Estadual de Campinas, C.P. 6109, 13083-970, Campinas, S?o Paulo, Brazil.