Sealing of pulp chamber dentin in endodontics: Influence of bond strategy and time-point application.

Context: Seal the dentin of the pulp chamber during endodontic treatment to avoid interfering with the restorative treatment performed afterward. Aims: The aim was to evaluate the effect of three adhesive systems applied in different bonding strategies (etch-and-rinse, self-etch, and universal adhesive) and time-point application (immediately after the cavity access preparation or after endodontic obturation) on the hybrid layer formation and dentinal penetrability. Materials and Methods: Forty-eight sound molars were randomly distributed into six groups (n = 10) according to the adhesive system used: Forty-eight sound molars were randomly distributed into six groups (n = 10) according to the adhesive system used and the time-point application: Adper Scotchbond Multi-purpose (AS), Clearfil SE (CF) and Scotchbond Universal (SU) in strategy of immediate endodontic sealing (IES) or delayed endodontic sealing (DES). In IES-AS, IES-CF, and IES-SU groups, dentin sealing was performed immediately after the cavity access, while in DES-AS, DES-CF, and DES-SU, after root canal obturation. The specimens were sectioned in the long axis, in a buccal-lingual direction, and the dentinal penetrability of the adhesive systems was evaluated using confocal microscopy images. Hybrid layer formation was analyzed by scanning electron microscopy images. Statistical Analysis Used: Dentinal penetrability data were analyzed with the ANOVA test and the Kruskal-Wallis test was performed for hybrid layer data (α = 0.05). Results: IES-CF showed the lowest dentinal penetrability (P < 0.05), while the other protocols were similar to each other (P > 0.05). No significant differences were found between groups regarding the hybrid layer formation (P > 0.05). Immediate and DES protocols do not influence the hybrid layer formation, regardless of the bond strategy used. Conclusions: Sealing the pulp chamber dentin before endodontic treatment can improve the bond strength of the final restoration but the formation of the hybrid layer was not influenced by the bond strategy.

Solução estética conservadora em paciente jovem com fluorose: clareamento e microabrasão: relato de caso / Conservative aesthetic solution in a young patient with fluorosis: whitening and microabrasion: case report

Introdução: a fluorose constitui uma patologia que afeta os dentes desencadeada pelo excesso de ingestão de flúor. Sua manifestação ocorre a nível de esmalte na presença de manchas ou defeitos anatômicos. Objetivo: relatar o manejo clínico de um paciente com fluorose tratado com procedimento minimamente invasivo. Relato do caso: paciente do sexo feminino, 18 anos, apresentando queixas estéticas clinicamente observadas na vestibular dos dentes, com diagnóstico de lesões fluoróticas. Foi executado protocolo de dessensibilização com Ultra EZ por 5 min e aplicação do Verniz (Enamelast Fluoride) previamente ao tratamento clareador. Foram realizados duas sessões de clareamento com peróxido de hidrogenio 35% (DMC) por 45 min com intervalo de sete dias entre elas. Imediatamente após a segunda sessão de clareamento, foi realizado microabrasão com pasta abrasiva Whitness RM (FGM) e taça de borracha com fricção por 20 segundos. Resultados: o tratamento clareador associado a técnica da microabrasão do esmalte demonstrou resultado estético favorável, microinvasivo e eficaz no tratamento da fluorose. Conclusão: a associação dos tratamentos resolveu o problema estético da paciente de forma rápida e segura, conservando a estrutura dentária.

Introduction: fluorosis is a pathology that affects teeth triggered by excess fluoride intake. Its manifestation occurs at the enamel level in the presence of stains or anatomical defects. Objective: to report the clinical management of a patient with fluorosis treated with a minimally invasive procedure. Case report: female patient, 18 years old, presenting aesthetic complaints clinically observed in the buccal of the teeth, with a diagnosis of fluorotic lesions. A desensitization protocol was performed with Ultra EZ for 5 min and Varnish (Enamelast Fluoride) was applied prior to the bleaching treatment. Two bleaching sessions were performed with 35% hydrogen peroxide (DMC) for 45 min, with an interval of seven days between them. Immediately after the second bleaching session, microabrasion was performed with Whitness RM abrasive paste (FGM) and a rubber cup with friction for 20 seconds. Results: The bleaching treatment associated with the enamel microabrasion technique demonstrated a favorable, microinvasive and effective esthetic result in the treatment of fluorosis. Conclusion: the combination of treatments solved the patient's aesthetic problem quickly and safely, preserving the dental structure.

Effect of different adhesive strategies and storage time on bond strength of bi-functional monomers to simulated endodontically-treated dentin.

Adhesive strategies were evaluated on the bond strength of bi-functional monomers bonded to endodontically-treated-dentin (ETD). Superficial dentin was removed on human molars. Teeth were immersed in 5 mL 2.5% NaOCl, followed by immersion in 5 mL 17% EDTA. Dentin surface impregnated with epoxy resin-based sealer was then divided four groups (n=10): Scotchbond Multi-Purpose (SBMP); Single Bond Universal (SBU); Optibond All-in-One (OPB); and Tetric-N-Bond Universal (TBU). After 24-h or 1-year-of-storage specimens were submitted to microtensile bond strength (µTBS) and failure classification. A confocal laser scanning microscope (CLSM) evaluated the hybrid layer formation. Two-way ANOVA and Tukey-HSD test were performed (α=5%). The µTBS did not present statistical differences among adhesive strategies after 24-h. Significant differences were found after 1-year-of-storage. CLSM analysis showed water infiltration and consequently degradation of the hybrid layer after 1-year-of-storage. The use of SBU universal adhesive on the self-etching mode on ETD produced more stable bond over the 1-year-of-storage.

Analysis of enamel/restoration interface submitted cariogenic challenge and fluoride release.

The treatment of high-risk patients still is a challenge. The understanding and development non-invasive, non-destructive, and non-ionizing techniques, can help to guide the treatment and the diagnosis of primary and recurrent caries. The present study evaluated the behavior of enamel/restoration interface after a cariogenic challenge by Fourier domain optical coherence tomography (FD-OCT), scanning electron microscopy (SEM) examination, and the fluoride release of the different restorative materials. Cavities (1.5 × 0.5 mm) were performed in enamel surface and divided into groups (n = 8): glass ionomer cement (GIC), resin-modified glass ionomer cement (RMGIC), and resin composite (RC). The samples were submitted to pH-cycling, and the solutions analyzed for cumulative fluoride by ion-analyzer. The morphology was analyzed by SEM through replicas. The optical attenuation coefficient (OAC) was calculated through exponential decay from the images generated by FD-OCT. Data were analyzed considering α = 0.05. OAC values increased for all groups after pH-cycling indicating demineralization (p < .05). Considering the remineralizing solution, RMGIC presented higher fluoride release rate, followed by GIC, while RC did not release any fluoride. Yet for the demineralizing solution, RMGIC and GIC released similar fluoride rates, overcoming RC (p < .05). Micrographs revealed no changes on the restorations margins, although enamel detachment was observed for RC and GIC after pH-cycling.

Irrigação ultrassônica passiva proporciona menor incidência de debris após preparo do espaço para o pino / Passive ultrasonic irrigation provides less debris after post space preparation

Objetivo: esse estudo avaliou a incidência de resíduos (debris) na dentina, no espaço preparado para pino de fibra, submetida a um dos seguintes protocolos de irrigação: água destilada (AD); hipoclorito de sódio a 2,5% (HS); hipoclorito de sódio a 2,5% mais EDTA a 17% (HSE); hipoclorito de sódio a 2,5% agitado com irrigação ultrassônica passiva (HSUS). Métodos: os canais radiculares de quarenta incisivos bovinos, com comprimento radicular de 17,0 mm, foram obturados pela técnica do cone único. Após sete dias, um espaço para o pino de fibra foi preparado com brocas de Largo #1 e #2 e broca DC2 (White Post; FGM). Os espécimes foram aleatoriamente distribuídos em quatro grupos (n = 10), de acordo com o protocolo de irrigação usado: AD; HS, HSE ou HSUS. As raízes foram clivadas longitudinalmente e obtidas imagens, em microscopia eletrônica de varredura (500X), da superfície dentinária dos segmentos radiculares cervical e apical do espaço preparado para pino de fibra. Escores foram atribuídos de acordo com a presença de debris. Resultados: no segmento cervical, HSE e HSUS apresentaram menor incidência de debris do que AD e HS (p < 0,05). No segmento apical, HSUS proporcionou a menor incidência de debris na dentina radicular (p < 0,05). Não houve diferença entre os demais protocolos de irrigação, tanto no segmento cervical quanto no apical (p > 0,05). Conclusões: o protocolo HSUS proporcionou menor incidência de debris sobre a superfície dentinária do espaço preparado para pino de fibra (AU).

Objective: The present study evaluated the residues (debris) incidence in dentin fiber post space submitted to irrigation protocols using distilled water (DW), 2.5% sodium hypochlorite (HS), 2.5 % sodium hypochlorite and 17% EDTA (HSE), or 2.5% sodium hypochlorite energized bypassive ultrasonic irrigation (HSUS). Methods: Forty bovine incisors root canals with standardized root length (17.0mm) were obturated using single cone technique and epoxyresin sealer. After 7 days, the fiber post space was prepared using # 1 and # 2 Largo burs, and DC2 (White Post; FGM).The specimens were randomly divided into four groups (n= 10), according to the irrigation protocol: AD; HS, HSE or HSUS. After 48 hours, the roots were sectioned. Cervical and apical segment images were obtained using scanning electron microscopy (500X). Scores were assessed according to debris presence. Results: In cervical segment, HSE and HSUS presented debris incidence lower than DW and HS (P <0.05). DW and HS or HSE and HSUS were similar to each other (P > 0.05). Conclusions: HSUS provided the lower debris incidence in dentin surface of the fiber post space (P < 0.05) (AU).

Phylogenetic relationships of Leishmania species based on trypanosomatid barcode (SSU rDNA) and gGAPDH genes: Taxonomic revision of Leishmania (L.) infantum chagasi in South America.

Phylogenetic studies on trypanosomatid barcode using V7V8 SSU rRNA and gGAPDH gene sequences have provided support for redefining some trypanosomatid species and positioning new isolates. The genus Leishmania is a slow evolving monophyletic group and including important human pathogens. The phylogenetic relationships of this genus have been determined by the natural history of its vertebrate hosts, vector specificity, clinical manifestations, geographical distribution and molecular approaches using different markers. Thus, in an attempt to better understand the phylogenetic relationships of Leishmania species, we performed phylogenetic analysis on trypanosomatid barcode using V7V8 SSU rRNA and gGAPDH gene sequences among a large number of Leishmania species and also several Brazilian visceral Leishmania infantum chagasi isolates obtained from dogs and humans. Our phylogenetic analysis strongly suggested that Leishmania hertigi and Leishmania equatoriensis should be taxonomically revised so as to include them in the genus Endotrypanum; and supported ancient divergence of Leishmania enriettii. This, together with recent data in the literature, throws light on the discussion about the evolutionary southern supercontinent hypothesis for the origin of Leishmania ssp. and validates L. infantum chagasi from Brazil, thus clearly differentiating it from L. infantum, for the first time.

Occurrence of Ehrlichia canis and Anaplasma platys in household dogs from northern Parana.

Canine monocytic ehrlichiosis caused primarily by Ehrlichia canis and canine thrombocytic anaplasmosis induced by Anaplasma platys are important emerging zoonotic tick-borne diseases of dogs. There is evidence that these pathogens can also affect humans. This study evaluated the presence of E. canis and A. platys in blood samples collected from 256 domiciled dogs in the municipality of Jataizinho, located in north region of the State of Parana, Brazil, by PCR assay. The occurrence of E. canis and A. platys was 16.4% (42/256) and 19.4% (49/256), respectively; while 5.47% (14/256) of the dogs evaluated were co-infected by these two organisms. The presence of E. canis and A. platys was not significantly associated with the variables evaluated (sex, age, outdoor access, and presence of ticks during blood collection). Infection of dogs by E. canis was associated with anemia and thrombocytopenia, while infection induced by A. platys was related only to thrombocytopenia. Canine monocytic ehrlichiosis and canine thrombocytic anaplasmosis should be included in the differential diagnoses when these hematological alterations are observed during routine laboratory evaluation of dogs.

Occurrence of Ehrlichia canis and Anaplasma platys in household dogs from northern Parana / Ocorrência de Ehrlichia canis e Anaplasma platys em cães domiciliados da região norte do Paraná

Canine monocytic ehrlichiosis caused primarily by Ehrlichia canis and canine thrombocytic anaplasmosis induced by Anaplasma platys are important emerging zoonotic tick-borne diseases of dogs. There is evidence that these pathogens can also affect humans. This study evaluated the presence of E. canis and A. platys in blood samples collected from 256 domiciled dogs in the municipality of Jataizinho, located in north region of the State of Parana, Brazil, by PCR assay. The occurrence of E. canis and A. platys was 16.4% (42/256) and 19.4% (49/256), respectively; while 5.47% (14/256) of the dogs evaluated were co-infected by these two organisms. The presence of E. canis and A. platys was not significantly associated with the variables evaluated (sex, age, outdoor access, and presence of ticks during blood collection). Infection of dogs by E. canis was associated with anemia and thrombocytopenia, while infection induced by A. platys was related only to thrombocytopenia. Canine monocytic ehrlichiosis and canine thrombocytic anaplasmosis should be included in the differential diagnoses when these hematological alterations are observed during routine laboratory evaluation of dogs.(AU)

Erliquiose monocítica canina, causada principalmente por Ehrlichia canis, e anaplasmose trombocítica canina, devida a infecção com Anaplasma platys, são importantes doenças transmitidas por carrapatos que acometem os cães, com evidências que podem também acometer o homem. O presente estudo avaliou a ocorrência desses agentes em amostras de sangue de 256 cães domiciliados na cidade de Jataizinho, na região Norte do Paraná, Brasil, utilizando a técnica da Reação em Cadeia da Polimerase (PCR). A ocorrência de E. canis e A. platys foi de 16,4% (42/256) e 19,4% (49/256), respectivamente, com 5,47% (14/256) dos animais apresentando coinfecção. Não foi observada associação significativa com as variáveis sexo, idade, acesso à rua e presença de carrapatos no momento da coleta de sangue. A infecção por E. canis teve relação com anemia e com trombocitopenia, enquanto a infecção por A. platys apresentou relação apenas com trombocitopenia. Com base nos resultados obtidos, reforçou-se a necessidade de que erliquiose e anaplasmose canina devem estar entre os diagnósticos diferenciais, quando da detecção de anemia e trombocitopenia em exames laboratoriais.(AU)

Molecular detection and occurrence of 'Candidatus Mycoplasma haemobos' in dairy cattle of Southern Brazil.

Bovine hemoplasmas are bacteria found on the erythrocyte surface or free in the plasma of cattle. The aim of the present study was to evaluate the occurrence of 'Candidatus Mycoplasma haemobos' ('C. M. haemobos') in Holstein and Jersey cattle raised in Londrina and surroundings, northern region of the State of Parana, Southern Brazil. PCR testing directed to 16S rRNA gene fragment was performed to investigate the occurrence and characterize the molecular identity of 'C. M. haemobos'. A total of 264/433 (60.97%) blood samples were positive by PCR. Further alignment of 500-bp amplicons to available sequences at the GenBank database showed high identity (100%) to 'C. M. haemobos'. To the author's knowledge, this is the first molecular confirmation of the hemoplasma 'C. M. haemobos' in cattle from Brazil. Moreover, 'C. M. haemobos' was observed in high occurrence in dairy cattle, and may have significant impact in livestock production.

Molecular detection and occurrence of 'Candidatus Mycoplasma haemobos' in dairy cattle of Southern Brazil / Detecção molecular e ocorrência de 'Candidatus Mycoplasma haemobos' em bovinos de leite do Sul do Brasil

Bovine hemoplasmas are bacteria found on the erythrocyte surface or free in the plasma of cattle. The aim of the present study was to evaluate the occurrence of 'Candidatus Mycoplasma haemobos' ('C. M. haemobos') in Holstein and Jersey cattle raised in Londrina and surroundings, northern region of the State of Parana, Southern Brazil. PCR testing directed to 16S rRNA gene fragment was performed to investigate the occurrence and characterize the molecular identity of 'C. M. haemobos'. A total of 264/433 (60.97%) blood samples were positive by PCR. Further alignment of 500-bp amplicons to available sequences at the GenBank database showed high identity (100%) to 'C. M. haemobos'. To the author's knowledge, this is the first molecular confirmation of the hemoplasma 'C. M. haemobos' in cattle from Brazil. Moreover, 'C. M. haemobos' was observed in high occurrence in dairy cattle, and may have significant impact in livestock production.

Hemoplasmas de bovinos são bactérias encontradas na superfície de hemácias, ou livre no plasma de bovinos. O objetivo do presente estudo foi avaliar a ocorrência de Candidatus Mycoplasma haemobos' ('C. M. haemobos') em bovinos das raças Holandesa e Jersey da região de Londrina, norte do Paraná, sul do Brasil. Para investigar a ocorrência e caracterizar a identidade molecular do 'C. M haemobos' uma PCR baseada no fragmento do gene 16S rRNA foi realizada. A PCR identificou como positivas 264/433 (61%) amostras de sangue testadas. O alinhamento deste fragmento de 500 pb com seqüências disponíveis no GenBank mostrou 100% de identidade 'C. M. haemobos'. Pela bibliografia consultada, esta é a primeira confirmação molecular do hemoplasma 'C. M. haemobos' em bovinos no Brasil. Além disso, foi observada uma alta prevalência deste hemoplasma em bovinos de leite, que pode ter um impacto importante na pecuária bovina.

Detection and molecular characterization of a canine piroplasm from Brazil.

In the beginning of the 20th century, a new canine disease was reported in Brazil under the name "nambiuvú", whose etiological agent was called Rangelia vitalii, a distinct piroplasm that was shown to parasitize not only erythrocytes, but also leucocytes and endothelial cells. In this new century, more publications on R. vitalii were reported from Brazil, including an extensive study on its ultrastructural analysis, in addition to clinical, pathological, and epidemiological data on nambiuvú. However, a molecular analysis of R. vitalii has not been performed to date. In the present study, we performed molecular phylogenetic analyses of R. vitalii based on fragments of the genes 18S rRNA and the heat shock protein 70 (hsp70), amplified by PCR performed on blood samples derived from five clinical cases of dogs presumably infected with R. vitalii in southern Brazil. In addition, we examined Giemsa-stained thin blood smears from these same dogs. DNA sequences (604-bp) of the 18S rRNA gene obtained from the five dogs were identical to each other, and by Blast analysis, this sequence shared the highest degree of sequence identity (95%) with Babesia sp. China-BQ1. DNA sequences (1056-bp) of the hsp70 gene obtained from the five dogs were identical to each other, and by Blast analysis, this sequence shared the highest degree of sequence identity (87%) with Babesia bigemina. Phylogenetic analyses inferred from either of the two genes resulted in the newly genotype being placed in the Babesia spp. sensu stricto clade with very high bootstrap support (95-100%) in three analyses (Neighbor-Joining, Maximum parsimony, and Maximum likelihood). Giemsa-stained thin blood smears from the dogs were shown to contain piroplasm organisms within erythrocytes, monocytes and neutrophils (individual forms), and schizont-like forms within neutrophils, in accordance with literature reports of R. vitalii. Based on these results, we conclude that R. vitalii, the etiological agent of "nambiuvú" in southern Brazil, is a valid species of piroplasm. Further studies are required to evaluate the validity of the genus Rangelia.

Detecção de peptídeos importantes para o diagnóstico da cisticercose bovina no immunoblot / Detection of important peptides for bovine cysticercosis diagnosis in immunoblot

A dificuldade de se localizar cisticercos em bovinos com infecção discreta torna o diagnóstico tradicional da cisticercose bovina em matadouros um recurso de baixa sensibilidade, o que gera uma demanda de diagnóstico por métodos alternativos como o immunoblot. Para tanto, neste trabalho, foram realizados ensaios de immunoblot a fim de analisar os peptídeos responsáveis pela reação do antígeno vesicular de Taenia crassiceps. Foram utilizadas 28 amostras de soro de bovinos comprovadamente negativos e 28 amostras coletadas de bovinos infectados experimentalmente com ovos de Taenia saginata. Os resultados dos ensaios de immunoblot com as amostras de soros-controle mencionadas indicaram que os peptídeos de 4-6, 14 e 18kDa destacaram-se entre os demais, mostrando altas taxas de desempenho no diagnóstico da cisticercose bovina e uma aparência diferenciada, com área e largura maiores, ao contrário dos peptídeos de média e alta massa molecular, que se apresentaram sob a forma de linha e com reações inespecíficas. O immunoblot mostrou ter um potencial como uma alternativa de diagnóstico da cisticercose bovina, incluindo sua aplicação em animais vivos, como método auxiliar de diagnóstico em estudos epidemiológicos da doença.

The diagnosis of cysticercosis in bovines that presents discreet infection is committed by the conventional methodology of detection employed in slaughterhouses. This deficiency leads to the development of alternative methods, such as immunoblot, in order to improve the detection of this disease. An immunoblot assay was developed to detect peptides from Taenia crassiceps antigenand it was used to test 56 samples of bovine serum (28 confirmed as negative and 28 from experimentally infected animals with Taenia saginata eggs). The peptides 4-6, 14 and 18 kDa showed better performance in bovine cysticercosis diagnosis due to an enhanced appearance, once they showed higher area and wide, contrasting with medium and high molecular weight peptides, that showed unspecific reactions and discreet appearance. The obtained results indicated that immunoblot can represent a potential alternative in bovine cysticercosis diagnosis, including its use in live animals, improving the detection of this disease even in epidemiological studies.