Comparative analyses of whole genome sequences of Leishmania infantum isolates from humans and dogs in northeastern Brazil.

The genomic sequences of 20 Leishmania infantum isolates collected in northeastern Brazil were compared with each other and with the available genomic sequences of 29 L. infantum/donovani isolates from Nepal and Turkey. The Brazilian isolates were obtained in the early 1990s or since 2009 from patients with visceral or non-ulcerating cutaneous leishmaniasis, asymptomatic humans, or dogs with visceral leishmaniasis. Two isolates were from the blood and bone marrow of the same visceral leishmaniasis patient. All 20 genomic sequences display 99.95% identity with each other and slightly less identity with a reference L. infantum genome from a Spanish isolate. Despite the high identity, analysis of individual differences among the 32 million base pair genomes showed sufficient variation to allow the isolates to be clustered based on the primary sequence. A major source of variation detected was in chromosome somy, with only four of the 36 chromosomes being predominantly disomic in all 49 isolates examined. In contrast, chromosome 31 was predominantly tetrasomic/pentasomic, consistent with its regions of synteny on two different disomic chromosomes of Trypanosoma brucei. In the Brazilian isolates, evidence for recombination was detected in 27 of the 36 chromosomes. Clustering analyses suggested two populations, in which two of the five older isolates from the 1990s clustered with a majority of recent isolates. Overall the analyses do not suggest individual sequence variants account for differences in clinical outcome or adaptation to different hosts. For the first known time, DNA of isolates from asymptomatic subjects were sequenced. Of interest, these displayed lower diversity than isolates from symptomatic subjects, an observation that deserves further investigation with additional isolates from asymptomatic subjects.

Exposure to Mycobacterium avium decreases the protective effect of the DNA vaccine pVAXhsp65 against Mycobacterium tuberculosis-induced inflammation of the pulmonary parenchyma.

This work investigated the effect of previous Mycobacterium avium exposure on the protective ability of the DNA vaccine pVAXhsp65 against inflammation in the pulmonary parenchyma. BALB/c mice were presensitized with heat-killed M. avium and then immunized with three doses of pVAXhsp65 prior to challenge with Mycobacterium tuberculosis. M. avium sensitization induced high levels of spontaneous IL-5 production that were concomitant with a positive delayed-type hypersensitivity reaction; antigen-specific IFN-γ production was also observed upon splenocyte stimulation. Prior exposure to M. avium resulted in altered cytokine and antibody production induced by immunization with pVAXhsp65; instead of a Th1 response, vaccinated mice previously exposed to M. avium developed a strong Th2 response. This switch to a Th2 response coincided with the loss of the anti-inflammatory effect of pVAXhsp65 vaccination previously observed in the pulmonary parenchyma of mice infected with M. tuberculosis. These results suggest that exposure to environmental mycobacteria can modulate immune responses induced by mycobacterial vaccines other than bacillus Calmette-Guérin.

Neonatal BCG immunization followed by DNAhsp65 boosters: highly immunogenic but not protective against tuberculosis - a paradoxical effect of the vector?

A new tuberculosis vaccine is urgently needed. Prime-boost strategies are considered very promising and the inclusion of BCG is highly desirable. In this investigation, we tested the protective efficacy of BCG delivered in the neonatal period followed by boosters in the adult phase with a DNA vaccine containing the hsp65 gene from Mycobacterium leprae (pVAXhsp65). Immune responses were characterized by serum anti-hsp65 antibody levels and IFN-gamma and IL-5 production by the spleen. Amounts of these cytokines were also determined in lung homogenates. Protective efficacy was established by the number of colony-forming units (CFU) and histopathological analysis of the lungs after challenge with Mycobacterium tuberculosis. Immunization with BCG alone triggered a significant reduction of CFU in the lungs and also clearly preserved the pulmonary parenchyma. BCG priming also increased the immunogenicity of pVAXhsp65. However, boosters with pVAXhsp65 or the empty vector abolished the protective efficacy of BCG. Also, higher IL-5 levels were produced by spleen and lungs after DNA boosters. These results demonstrated that neonatal BCG immunization followed by DNAhsp65 boosters is highly immunogenic but is not protective against tuberculosis.

Genes at human chromosome 5q31.1 regulate delayed-type hypersensitivity responses associated with Leishmania chagasi infection.

Visceral leishmaniasis (VL) caused by Leishmania chagasi is endemic to northeast Brazil. A positive delayed-type hypersensitivity skin test response (DTH+) is a marker for acquired resistance to disease, clusters in families and may be genetically controlled. Twenty-three single nucleotide polymorphisms (SNPs) were genotyped in the cytokine 5q23.3-q31.1 region IRF1-IL5-IL13-IL4-IL9-LECT2-TGFBI in 102 families (323 DTH+; 190 DTH-; 123 VL individuals) from a VL endemic region in northeast Brazil. Data from 20 SNPs were analyzed for association with DTH+/- status and VL using family-based, stepwise conditional logistic regression analysis. Independent associations were observed between the DTH+ phenotype and markers in separate linkage disequilibrium blocks in LECT2 (OR 2.25; P=0.005; 95% CI=1.28-3.97) and TGFBI (OR 1.94; P=0.003; 95% CI=1.24-3.03). VL child/parent trios gave no evidence of association, but the DTH- phenotype was associated with SNP rs2070874 at IL4 (OR 3.14; P=0.006; 95% CI=1.38-7.14), and SNP rs30740 between LECT2 and TGFBI (OR 3.00; P=0.042; 95% CI=1.04-8.65). These results indicate several genes in the immune response gene cluster at 5q23.3-q31.1 influence outcomes of L. chagasi infection in this region of Brazil.

Produção deficiente de citocinas Th1 em camundongos BALB/c jovens / Deficient production of Th1 cytokines in young BALB/c mice

O objetivo do presente estudo foi comparar a produção de IFN-gama , IL-12 e IL-4 entre camundongos jovens (5, 12e 19 dias de idade) e adultos (30 dias de idade). As avaliações foram feitas por estimulação, in vitro, de células esplênicas com Concanavalina A (ConA) , Staphylococcus aureus (S. aureus) e lipopolissacarídeo (LPS). Diferentes concentrações de cada estímulo foram testadas e os sobrenadantes das culturas foram coletados após 48 horas de incubação e as concentrações de IFN-gama IL-12 e IL-4 determinadas por ELISA. Células de camundongos jovens e adultos produziram níveis igualmente elevados de IFN-gama após estímulo com ConA. Somente animais adultos produziram IFN-gama em resposta ao estímulo com S. aureus. Em culturas estimuladas com LPS, a produção desta citocina foi baixa e similar nos animais jovens e significativamente elevada nos animais adultos. Somente células de animais adultos estimuladas com S. aureus foram capazes de produzir IL-12. O único estímulo capaz de induzir níveis detectáveis de IL-4 foi ConA, sendo que estes níveis foram mais elevados nos animais com 12 e 19 dias de idade em comparação com animais neonatos e adultos. A diminuição das doses ótimas dos estímulos não mudou o perfil de produção de cada citocina nos animais jovens. Estes resultados permitem concluir que a idade afeta a produção de citocinas: ocorre maior produção de IL-4 em camundongos jovens e maior produção de IL-12 e IFN-gama em animais adultos. Estas informações são importantes devido ao papel destas citocinas na polarização das respostas imunes nos sentidos Th1 e Th2.

A radiographic comparison of apical root resorption after orthodontic treatment with 3 different fixed appliance techniques.

Apical root resorption is an undesirable, but frequent side effect of orthodontic treatment, and therefore improvements in orthodontic techniques and materials are in constant development to decrease it. One of the most recently developed orthodontic techniques is the Bioefficient Therapy that uses contemporary orthodontic materials. Therefore, the primary objective of this study was to compare the amount of root resorption after orthodontic treatment between the simplified standard edgewise technique (group 1), the edgewise straight wire system (group 2), and the Bioefficient Therapy (group 3). It was also the purpose of this investigation to evaluate the amount of root resorption in the whole sample studied and the prevalence of root resorption in the upper and lower incisors. Thus, periapical radiographs were obtained with the long cone paralleling technique for the upper and lower incisors from 30 patients for each group. Root resorption was ranked by scores by 2 examiners who had an excellent intra and interexaminer calibration by Kendall concordance coefficient. Results of the Kruskal-Wallis test demonstrated that group 3 (Bioefficient Therapy) presented less root resorption than the others. It was speculated that the factors responsible for the lesser resorption in this technique were the use of heat-activated and superelastic wires with the bracket design in this technique as well as the use of a smaller rectangular stainless steel wire (0.018 x 0.025 inch) in a 0.022 x 0.028 inch slot during incisor retraction and the finishing stages, as compared to the other techniques. Considering the whole sample, there was no root resorption in 2.25% of the analyzed teeth. There was only a slight resorption in 42.56%, a moderate resorption in 53.37%, an accentuated resorption in 1.40% and an extreme root resorption in only 0.42% of the teeth. The prevalence of resorption for each incisor indicated, in decreasing order, a greater resorption for the upper centrals, followed by the upper laterals, lower centrals, and lastly the lower lateral incisors.

Dental maturation in subjects with extreme vertical facial types.

The purpose of this research was to investigate whether there is a difference in dental maturation between skeletal open bite and skeletal deep bite subjects of the same chronological age. The material consisted of 40 lateral headfilms and 40 panoramic radiographs of 20 male and 20 female white subjects, with a mean chronological age of 9 years and 2 months (range: 7 years 6 months to 10 years 11 months). These subjects were selected on the basis of lower anterior face height as a percentage of total face height and on the amount of open or deep-bite from a total sample of 400 subjects. The persons exhibiting the most extreme values at both ends of the distribution were selected to create two groups with 20 subjects in each (10 males and 10 females). Thus, the groups represented subjects with either a large lower anterior face height associated with an open bite or a small lower anterior face height associated with a deep bite. A double blind determination of dental maturation, expressed by dental age, for each subject was performed on the panoramic radiographs using the system of Demirjian et al. (1973). A covariance analysis was used to eliminate variability introduced by the large age range of the sample. The skeletal open bite and deep bite groups presented mean dental ages of 120.48 and 114.00 months, respectively. Statistical analysis demonstrated that this difference was statistically significant at P < 0.05. Therefore, it seems that skeletal open bite subjects presented a slight tendency to have an advanced dental maturation, expressed by dental age, as compared with skeletal deep bite subjects.

[Ankylosis of deciduous teeth]. / Anquilose de dentes decíduos.

The authors reviewed the literature concerning ankylosis of deciduous teeth, with emphasis on the diagnosis, the incidence, the aetiology and the treatment of this manifestation, illustrated with representative clinical cases.

[Bilateral posterior open bite caused by idiopathic eruption failure. Clinical case]. / Mordida aberta posterior bilateral ocasionada por falta de erupção idiopática: caso clínico.

The treatment of a patient with bilateral posterior open bite due to primary failure of eruption is presented. The characteristics, differential diagnosis with posterior open bite due to other etiologies and the treatment of these problems are presented and discussed.

[Impressions in orthodontic finishing]. / Moldagem com finalidade ortodontica.

The authors review the subject and recommend a sequence to be followed during the impression taking, starting always in the lower arch, due to its comfort and less possibility of provoking nausea, this way acquiring the patient's trust.