The present study evaluated the glass fibre membrane (GFM)-polymerase chain reaction (PCR)-enzyme-linked immunosorbent assay (ELISA) technique for genotyping the Plasmodium vivax variants, to verify the distribution of P. vivax variants (VK210, VK247 and P. vivax-like) in parts of Brazil and their correlation with levels of parasitaemia, previous malaria experience and clearance of parasitaemia linked to different treatment schedules. The samples were taken from individuals living in Macapá, Porto Velho and Belém, all of which are endemic areas of vivax malaria in the Amazon region of Brazil. Blood samples were collected on GFMs. The gene that codes for the circumsporozoite proteins of P. vivax variants was amplified by PCR and the amplified fragments were hybridized to variant-specific, digoxigenin-labelled oligonucleotide probes by ELISA. The GFM-PCR-ELISA technique was shown to be accurate for epidemiological surveys of the vivax complex. All variants were detected in all 3 areas, but only P. vivax VK210 was found as a single agent of infection, while the other 2 occurred as mixed infections. The P. vivax-like variant was found to be associated with low parasitaemia and VK210 with the highest parasitaemia levels; none of the P. vivax variants was linked with a previous malaria experience. In all cases parasitaemia clearance was identical regarding the type of treatment and consequently it is not possible to confirm the previously reported correlation between P. vivax genotype and response to chloroquine.

The purpose of this study was to compare the efficacy of different chelating solutions (17% EDTA and 10% citric acid) on the smear layer removal, and their effect on tubular dentin sealer penetration. Sixty root canals were prepared and distributed into four groups (n = 15) according to the final irrigation protocol: G1, final irrigation with 2.5 mL of distilled water; G2, final irrigation with 2.5 mL of 2.5% sodium hypochlorite solution; G3, final irrigation with 2.5 mL of 17% EDTA; and G4, final irrigation with 2.5 mL of 10% citric acid. Five specimens from each group were not filled to assess smear layer removal by scanning electron microscopy. Ten specimens from each group were filled for analysis of sealer penetration into dentinal tubules by confocal laser scanning microscopy. Smear layer removal (Kruskal-Wallis and Dunn's tests) and sealer penetration (F and Tukey's tests) were statistically analyzed with 95% of significance level. G3 and G4 had greater smear layer removal rates in the cervical and middle thirds, in comparison with G1 and G2 (p < .05). G3 and G4 had the highest percentages of sealer penetration in all thirds, in comparison with G1 and G2 (p < .05). Smear layer removal was effective only at the cervical and middle thirds when the chelating solutions were used. Sealer penetration into the dentinal tubules significantly increased in all root thirds when the specimens were treated with both chelating solutions.

African trypanosomes (Trypanosoma) are vector-borne haemoparasites that survive in the vertebrate bloodstream through antigenic variation of their Variant Surface Glycoprotein (VSG). Recombination, or rather segmented gene conversion, is fundamental in Trypanosoma brucei for both VSG gene switching and for generating antigenic diversity during infections. Trypanosoma vivax is a related, livestock pathogen whose VSG lack structures that facilitate gene conversion in T. brucei and mechanisms underlying its antigenic diversity are poorly understood. Here we show that species-wide VSG repertoire is broadly conserved across diverse T. vivax clinical strains and has limited antigenic repertoire. We use variant antigen profiling, coalescent approaches and experimental infections to show that recombination plays little role in diversifying T. vivax VSG sequences. These results have immediate consequences for both the current mechanistic model of antigenic variation in African trypanosomes and species differences in virulence and transmission, requiring reconsideration of the wider epidemiology of animal African trypanosomiasis.

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The circumsporozoite protein (CSP) of the Plasmodium vivax infective sporozoite is considered to be a major target for the development of recombinant malaria vaccines. The Duffy blood group molecule acts as the red blood cell receptor for P. vivax. We review the frequency of P. vivax CSP variants and report their association with the Duffy blood group genotypes from Brazilian Amazon patients carrying P. vivax malaria. Peripheral blood samples were collected from 155 P. vivax-infected individuals from five Brazilian malaria-endemic areas. The P. vivax CSP variants and the Duffy blood group genotypes were assessed using PCR/RFLP. In single infections, the VK210 variant was the commonest followed by the P. vivax-like variant. The typing of P. vivax indicated that the frequency of variants among the study areas was significantly different from one to another. This is the first detection of the VK247 and P. vivax-like variant in single infections in endemic areas of Brazil. Association of the CSP P. vivax variants with the heterozygous Duffy blood group system genotype was significant for VK210 single infection. These observations provide additional data on the Plasmodium-host interactions concerning the Duffy blood group and P. vivax capability of causing human malaria.

ObjectiveTo analyze possible correlations among tubular dentine cement penetration, adhesiveness and apical leakage in fillings performed with gutta percha and an endodontic cement based on epoxy amine resin. Material and MethodsSixty similar, extracted human mandibular central incisors were irrigated, instrumented and filled following the same protocol. First, apical leakage was quantified by fluid filtration tests. Then, these same specimens were sectioned for analysis of tubular dentine cement penetration and the middle thirds were submitted to push-out tests to analyze the adhesiveness of the fillings. ResultsIn brief, the means and standard deviations with a confidence interval of 95% were as follows: tubular dentine cement penetration (8.875±4.540), adhesiveness (4.441±2.683) and apical leakage (0.318±0.215). The data were confronted using the Pearson's test (P>0.05), and it was possible to prove that there was no correlation between (1) tubular dentine cement penetration and apical leakage (r2: 0.08276), (2) tubular dentine cement penetration and adhesiveness (r2: -0.2412) and (3) adhesiveness and apical leakage (r2: 0.1340). ConclusionAfter analysis of these data, it could be observed that there exists no correlation among the variables analyzed in this study.

Five adult donkeys were experimentally infected with Brazilian strain of Trypanosoma evansi originally isolated from a naturally infected dog to study the hematological biochemical and histopathological alterations during the evolution of the disease. The course of the experimental infection was followed up to 145 days. Hematological analyses of the infected donkeys revealed a marked decline in hemoglobin, packed-cell volume, and erythrocyte count. Anemia was observed after successive peaks of parasitemia. Biochemical analyses showed increased levels of icterus index, serum globulins and decreased serum albumin and glucose values. All infected donkeys revealed enlargement of spleen and its white pulp, enlargement of mediastinal lymph nodes and lungs congestion. The main histopathological features consisted of meningoencephalitis. Demyelination in some areas of the cerebellum pediculus and neuropil vacuolization were observed. This study showed that donkeys infected with a Brazilian strain of T. evansi developed a chronic disease.