Direct type I IFN but not MDA5/TLR3 activation of dendritic cells is required for maturation and metabolic shift to glycolysis after poly IC stimulation.

Type I interferons (IFNs) play an important role in direct antiviral defense as well as linking the innate and adaptive immune responses. On dendritic cells (DCs), IFNs facilitate their activation and contribute to CD8(+) and CD4(+) T cell priming. However, the precise molecular mechanism by which IFNs regulate maturation and immunogenicity of DCs in vivo has not been studied in depth. Here we show that, after in vivo stimulation with the TLR ligand poly IC, IFNs dominate transcriptional changes in DCs. In contrast to direct TLR3/mda5 signaling, IFNs are required for upregulation of all pathways associated with DC immunogenicity. In addition, metabolic pathways, particularly the switch from oxidative phosphorylation to glycolysis, are also regulated by IFNs and required for DC maturation. These data provide evidence for a metabolic reprogramming concomitant with DC maturation and offer a novel mechanism by which IFNs modulate DC maturation.

Enterocytozoon bieneusi detected by molecular methods in raw sewage and treated effluent from a combined system in Brazil.

BACKGROUND: Enterocytozoon bieneusi are the most common microsporidia associated with different clinical manifestations such as diarrhoea, respiratory tract inflammation and acalculous cholecystitis, especially in immunocompromised patients. Infection usually occurs by ingestion of food and water contaminated with spores, but can also result from direct contact with spores through broken skin, eye lesions, and sexual transmission, depending on the microsporidian species. Although there are reports of E. bieneusi found in humans and animals in Brazil, there are no published studies of environmental samples examined by molecular methods. OBJECTIVES: The purpose of this study was to verify the presence of E. bieneusi in raw sewage and treated effluent from a combined system by molecular methods. METHODS: Raw sewage and treated effluent samples collected from a combined system were analysed for the presence of E. bieneusi using the internal transcriber spacer (ITS) region of E. bieneusi by nested polymerase chain reaction. FINDINGS: The analysis revealed E. bieneusi presence and a novel genotype (EbRB) in one raw sewage sample and one treated effluent. MAIN CONCLUSIONS: The presence of E. bieneusi in final effluent indicates that the combined system may not remove microsporidian spores. This study is the first report of E. bieneusi in environmental samples in Brazil.

A new synthetic TLR4 agonist, GLA, allows dendritic cells targeted with antigen to elicit Th1 T-cell immunity in vivo.

Protein-based vaccines offer safety and cost advantages but require adjuvants to induce immunity. Here we examined the adjuvant capacity of glucopyranosyl lipid A (GLA), a new synthetic non-toxic analogue of lipopolysaccharide. In mice, in comparison with non-formulated LPS and monophosphoryl lipid A, formulated GLA induced higher antibody titers and generated Type 1 T-cell responses to HIV gag-p24 protein in spleen and lymph nodes, which was dependent on TLR4 expression. Immunization was greatly improved by targeting HIV gag p24 to DCs with an antibody to DEC-205, a DC receptor for antigen uptake and processing. Subcutaneous immunization induced antigen-specific T-cell responses in the intestinal lamina propria. Immunity did not develop in mice transiently depleted of DCs. To understand how GLA works, we studied DCs directly from vaccinated mice. Within 4 h, GLA caused DCs to upregulate CD86 and CD40 and produce cytokines including IL-12p70 in vivo. Importantly, DCs removed from mice 4 h after vaccination became immunogenic, capable of inducing T-cell immunity upon injection into naïve mice. These data indicate that a synthetic and clinically feasible TLR4 agonist rapidly stimulates full maturation of DCs in vivo, allowing for adaptive immunity to develop many weeks to months later.

In vitro and in vivo evaluations of glass-ionomer cement containing chlorhexidine for Atraumatic Restorative Treatment.

OBJECTIVES: Addition of chlorhexidine has enhanced the antimicrobial effect of glass ionomer cement (GIC) indicated to Atraumatic Restorative Treatment (ART); however, the impact of this mixture on the properties of these materials and on the longevity of restorations must be investigated. The aim of this study was to evaluate the effects of incorporating chlorhexidine (CHX) in the in vitro biological and chemical-mechanical properties of GIC and in vivo clinical/ microbiological follow-up of the ART with GIC containing or not CHX. MATERIAL AND METHODS: For in vitro studies, groups were divided into GIC, GIC with 1.25% CHX, and GIC with 2.5% CHX. Antimicrobial activity of GIC was analyzed using agar diffusion and anti-biofilm assays. Cytotoxic effects, compressive tensile strength, microhardness and fluoride (F) release were also evaluated. A randomized controlled trial was conducted on 36 children that received ART either with GIC or GIC with CHX. Saliva and biofilm were collected for mutans streptococci (MS) counts and the survival rate of restorations was checked after 7 days, 3 months and one year after ART. ANOVA/Tukey or Kruskal-Wallis/ Mann-Whitney tests were performed for in vitro tests and in vivo microbiological analysis. The Kaplan-Meier method and Log rank tests were applied to estimate survival percentages of restorations (p<0.05). RESULTS: Incorporation of 1.25% and 2.5% CHX improved the antimicrobial/anti-biofilm activity of GIC, without affecting F release and mechanical characteristics, but 2.5% CHX was cytotoxic. Survival rate of restorations using GIC with 1.25% CHX was similar to GIC. A significant reduction of MS levels was observed for KM+CHX group in children saliva and biofilm 7 days after treatment. CONCLUSIONS: The incorporation of 1.25% CHX increased the in vitro antimicrobial activity, without changing chemical-mechanical properties of GIC and odontoblast-like cell viability. This combination improved the in vivo short-term microbiological effect without affecting clinical performance of ART restorations.

Clinical evaluation of composite and compomer restorations in primary teeth: 24-month results.

PURPOSE: This split-mouth, blind study evaluated the clinical performance of Dyract AP, F2000, and Heliomolar placed in primary molars of 30 children (mean age, 6 years and 2 months). MATERIALS AND METHODS: From a total of 79 restorations accomplished, 27 were built with Heliomolar (18 Class I, and 9 Class II), 30 were with F2000 (21 Class I and 9 Class II), and 22 were built with Dyract AP (14 Class I and 8 Class II). All of teeth restored had primary caries lesions. At 6, 12, 18 and 24 months, 60 restorations (75%) were evaluated using USPHS criteria for: color match (CM), marginal adaptation (MA), marginal discoloration (MD), anatomic form (AF) and secondary caries (SC) by three calibrated operators. The Alpha+Bravo score percentage was considered as clinical success. The data were subjected to statistical analysis by Mann-Whitney and Kruskal-Wallis tests (p<0.05). RESULTS: Heliomolar showed the smallest success clinical at 12 months for marginal adaptation and secondary caries, and at 18 months for marginal discoloration. Regarding color match and anatomic form, no significant differences were found among the groups at each evaluation period. When materials were compared, Heliomolar did not show a significant difference among the evaluated periods for any criteria, remaining with the lowest scores. Significant differences were observed at 12 months for F2000 (marginal adaptation), and at 24 months for Dyract AP (marginal discoloration and secondary caries) and for F2000 (color match and marginal discoloration). CONCLUSIONS: It was concluded that Dyract AP and F2000 showed the best clinical performance over 24 month-evaluations for marginal discoloration and secondary caries, and color match and marginal adaptation, respectively. The use of the resin composite Heliomolar in Class I/II restorations in primary molars should be carefully considered.

[A comparative study of the parasitological techniques: Kato-Katz and coprotest]. / Estudo comparativo de técnicas parasitológicas: Kato-Katz e coprotest.

Parasitological diagnosis should be appropriately conducted with high sensitivity and specificity so that intestinal parasite can be detected as the specific treatment depends on this result. A comparative study was developed to evaluate the concordance between the Kato-Katz and coprotest methods in the detection of helminthes in 332 individuals in Pedro de Toledo. A significant difference was found in the case of the Trichuris trichiura - 16.2% with the Kato-Katz method and 7.5% with the coprotest method. Therefore, the positive and negative samples of the coprotest method were compared with the number of eggs per gram of feces (epg) obtained by the Kato-Katz method. When the coprotest method was negative, 65 epg of Trichuris trichiura were found by the Kato-Katz method and when the coprotest was positive the epg according to the Kato-Katz were far higher, 199 epg, which demonstrated that the coprotest was inferior to the Kato-Katz method regarding low intensity infections.

An alternative aesthetic restoration for extensive coronal destruction in primary molars: indirect restorative technique with composite resin.

The indirect restorative technique with composite resin has presented efficient results when used in primary teeth. This technique enables the oral rehabilitation. Based on the results presented in the literature concerning clinical performance, this technique is presented as an alternative treatment to the primary teeth with extensive decay by showing a case of rehabilitation of primary molars. The oral rehabilitation using indirect resin composite restoration showed an important concept to be applied in primary dentition due to feasible, low cost, and lower chairside time.

Management of severe partial hypodontia: case report.

Hypodontia is characterized by partial or total congenital missing of one or more teeth, on one or both dentitions. Heredity is the main etiological factor and the principal clinical features are reduction on number, size and form of teeth, and late eruption. Removable partial prosthesis, fixed prosthesis, overdentures and adhesive prosthesis are alternative treatments; and the indication is type dependent. The aim of this study was to describe the clinical case of an eleven-year-old child with eight missing permanent teeth of idiopathic etiology. The patient had facial and skeletal symmetry, normal development and was not related to any syndrome. Clinical characteristics: permanent teeth with good periodontal conditions (16, 12, 11, 21, 22, 26, 36, 31, 42, 46), primary teeth (53, 63, 64, 73, 83); overbite and microdontia on teeth 12 and 22. The treatment plan was done initially by documenting of the case for teeth analysis (study casting models, periapicals and panoramic x-rays, and photographs), and followed by the exodontics of teeth 73 and 83. A removable appliance in autocured acrylic resin, using teeth in acrylic for maintenance of functional space and occlusion was planned and carried out. An anterior track for vertical dimension gain was used because of his accentuated overbite. The patient will be monitored until the end of the craniofacial growth, when it will be again evaluated and forwarded for the final oral rehabilitation. Hypodontia diagnosis and management should be performed as early as possible not to interfere with the craniofacial development of the child.

Classical Flt3L-dependent dendritic cells control immunity to protein vaccine.

DCs are critical for initiating immunity. The current paradigm in vaccine biology is that DCs migrating from peripheral tissue and classical lymphoid-resident DCs (cDCs) cooperate in the draining LNs to initiate priming and proliferation of T cells. Here, we observe subcutaneous immunity is Fms-like tyrosine kinase 3 ligand (Flt3L) dependent. Flt3L is rapidly secreted after immunization; Flt3 deletion reduces T cell responses by 50%. Flt3L enhances global T cell and humoral immunity as well as both the numbers and antigen capture capacity of migratory DCs (migDCs) and LN-resident cDCs. Surprisingly, however, we find immunity is controlled by cDCs and actively tempered in vivo by migDCs. Deletion of Langerin(+) DC or blockade of DC migration improves immunity. Consistent with an immune-regulatory role, transcriptomic analyses reveals different skin migDC subsets in both mouse and human cluster together, and share immune-suppressing gene expression and regulatory pathways. These data reveal that protective immunity to protein vaccines is controlled by Flt3L-dependent, LN-resident cDCs.

Proteomics analysis of the non-muscle myosin heavy chain IIa-enriched actin-myosin complex reveals multiple functions within the podocyte.

MYH9 encodes non-muscle myosin heavy chain IIA (NMMHCIIA), the predominant force-generating ATPase in non-muscle cells. Several lines of evidence implicate a role for MYH9 in podocytopathies. However, NMMHCIIA's function in podocytes remains unknown. To better understand this function, we performed immuno-precipitation followed by mass-spectrometry proteomics to identify proteins interacting with the NMMHCIIA-enriched actin-myosin complexes. Computational analyses revealed that these proteins belong to functional networks including regulators of cytoskeletal organization, metabolism and networks regulated by the HIV-1 gene nef. We further characterized the subcellular localization of NMMHCIIA within podocytes in vivo, and found it to be present within the podocyte major foot processes. Finally, we tested the effect of loss of MYH9 expression in podocytes in vitro, and found that it was necessary for cytoskeletal organization. Our results provide the first survey of NMMHCIIA-enriched actin-myosin-interacting proteins within the podocyte, demonstrating the important role of NMMHCIIA in organizing the elaborate cytoskeleton structure of podocytes. Our characterization of NMMHCIIA's functions goes beyond the podocyte, providing important insights into its general molecular role.

Enterocytozoon bieneusi detected by molecular methods in raw sewage and treated effluent from a combined system in Brazil

BACKGROUND Enterocytozoon bieneusi are the most common microsporidia associated with different clinical manifestations such as diarrhoea, respiratory tract inflammation and acalculous cholecystitis, especially in immunocompromised patients. Infection usually occurs by ingestion of food and water contaminated with spores, but can also result from direct contact with spores through broken skin, eye lesions, and sexual transmission, depending on the microsporidian species. Although there are reports of E. bieneusi found in humans and animals in Brazil, there are no published studies of environmental samples examined by molecular methods. OBJECTIVES The purpose of this study was to verify the presence of E. bieneusi in raw sewage and treated effluent from a combined system by molecular methods. METHODS Raw sewage and treated effluent samples collected from a combined system were analysed for the presence of E. bieneusi using the internal transcriber spacer (ITS) region of E. bieneusi by nested polymerase chain reaction. FINDINGS The analysis revealed E. bieneusi presence and a novel genotype (EbRB) in one raw sewage sample and one treated effluent. MAIN CONCLUSIONS The presence of E. bieneusi in final effluent indicates that the combined system may not remove microsporidian spores. This study is the first report of E. bieneusi in environmental samples in Brazil.

In vitro and in vivo evaluations of glass-ionomer cement containing chlorhexidine for Atraumatic Restorative Treatment

Abstract Objectives: Addition of chlorhexidine has enhanced the antimicrobial effect of glass ionomer cement (GIC) indicated to Atraumatic Restorative Treatment (ART); however, the impact of this mixture on the properties of these materials and on the longevity of restorations must be investigated. The aim of this study was to evaluate the effects of incorporating chlorhexidine (CHX) in the in vitro biological and chemical-mechanical properties of GIC and in vivo clinical/ microbiological follow-up of the ART with GIC containing or not CHX. Material and Methods: For in vitro studies, groups were divided into GIC, GIC with 1.25% CHX, and GIC with 2.5% CHX. Antimicrobial activity of GIC was analyzed using agar diffusion and anti-biofilm assays. Cytotoxic effects, compressive tensile strength, microhardness and fluoride (F) release were also evaluated. A randomized controlled trial was conducted on 36 children that received ART either with GIC or GIC with CHX. Saliva and biofilm were collected for mutans streptococci (MS) counts and the survival rate of restorations was checked after 7 days, 3 months and one year after ART. ANOVA/Tukey or Kruskal-Wallis/ Mann-Whitney tests were performed for in vitro tests and in vivo microbiological analysis. The Kaplan-Meier method and Log rank tests were applied to estimate survival percentages of restorations (p<0.05). Results: Incorporation of 1.25% and 2.5% CHX improved the antimicrobial/anti-biofilm activity of GIC, without affecting F release and mechanical characteristics, but 2.5% CHX was cytotoxic. Survival rate of restorations using GIC with 1.25% CHX was similar to GIC. A significant reduction of MS levels was observed for KM+CHX group in children saliva and biofilm 7 days after treatment. Conclusions: The incorporation of 1.25% CHX increased the in vitro antimicrobial activity, without changing chemical-mechanical properties of GIC and odontoblast-like cell viability. This combination improved the in vivo short-term microbiological effect without affecting clinical performance of ART restorations.

[Knowledge and practices of oral health on hospitalized children with cancer]. / Conhecimentos e práticas em saúde bucal com crianças hospitalizadas com câncer.

The aim of this study was to evaluate the knowledge and practices of oral health (OH) in hospitalized children with cancer. The sample was composed by the nurse team, caretakers and children. It was applied a questionnaire concerning the general knowledge about OH, methods and instruments used for oral hygiene (OH). According to the results, the responsible of OH of children are the caretakers (90.7%) who receive instructions from the nurse team in 21.4% of cases. As for the oral cavity discomfort, the nurse team reported that all patients exhibited clinical manifestations while the caretakers reported a different number, 62.8% of cases. All participants considered important having a dentist in the oncology sector. According to the results obtained, it was possible to conclude that there is no oral health protocol for hospitalized children with cancer and that the most frequent oral manifestations among patients going through antineoplastic treatment were: mucositis, nausea, vomit, xerostomy and lack of sense of taste.

Fluoretação da Água e seu Heterocontrole no Município de Nova Friburgo, Rio de Janeiro - Brasil / Water Fluoridation and its external control in the municipality of Nova Friburgo, Rio de Janeiro - Brazil

A fluoretação da água, apoiada pela Política Nacional de Saúde Bucal, tem sido um desafio no Brasil, devido a algumas insuficiências atuais na gestão pública para a implantação e controle desta medida. Este estudo objetivou vigiar, de forma periódica e sistemática, os níveis de fluoretos na água para consumo humano, em Nova Friburgo - RJ- Brasil (NF), por um período de 12 meses, como medida de heterocontrole, realizada por uma universidade pública. As amostras de água foram coletadas mensalmente, em duplicata, em 26 localidades em todo o município e em datas estabelecidas por sorteio. O método para medir a concentração de fluoretos nas amostras de água foi o eletrométrico, utilizando-se um potenciômetro digital com eletrodo específico para o íon flúor. A concentração de fluoretos nas amostras foi analisada de acordo com padrões fornecidos pelo Cecol/USP (2011). Os dados mostraram grande variação na concentração de fluoretos nas amostras, variando entre um mínimo de 0,00 ppmF e um máximo de 0,98 ppmF, com algumas dentro do intervalo de 0,65 e 0,94 ppmF, considerado o de melhor benefício para prevenir a cárie dentária. A concentração de fluoretos na água de abastecimento público do município foi considerada abaixo dos níveis mínimos exigidos, na maioria das amostras avaliadas, mesmo naquelas pertencentes às Estações de Tratamento de Água (ETA), cuja medida de fluoretação fora previamente implantada. Desta forma, a fluoretação da água e seu respectivo heterocontrole deveriam ser estabelecidos de forma contínua em NF, já que são medidas públicas relevantes para a promoção de saúde bucal.

Water fluoridation, supported by the Brazilian Oral Health Policy, has been a challenge as a public measure to protect Oral Health, due to some current insufficiencies in public management for the implantation of this measure in most Brazilian cities. The present study aimed to monitor, the levels of fluoride in drinking water in Nova Friburgo - RJ-Brazil (NF), on a regular and a systematic manner for a 12 months period, as an external control measurement performed by a public university. Water samples were collected, in duplicate, from 26 localities throughout the city, on a monthly basis, on randomly established dates. The fluoride concentration in the water samples was measured by using an ion-specific electrode coupled to a potentiometer, according to standards supplied by Cecol/USP (2011). Data showed a large variation of fluoride concentration in water samples, ranging from minimum 0.00 ppmF and maximum 0.98 ppmF, with few samples into the interval between 0.65 and 0.94 ppmF, which had the best benefit to prevent dental caries. Fluoride concentration in public water in this city can be considered under the minimum required levels in most of the evaluated samples, even in those from the Water Treatment System (WTS), which has water fluoridation. Thus, water fluoridation and its external control should be established on a regular basis in Nova Friburgo-RJ, Brazil because they are relevant public measures to promote oral health.

Dental caries in inland Brazilian adolescents and its relationship with social determinants

AIM: To identify the prevalence of dental caries in inland Brazilian adolescents, and to analyze the influence of socio-demographic and clinical variables, and access to dental service on caries experience. METHODS: This study had a non-probabilistic sample comprising 504 adolescents aged 12 years, attending public schools in the city of Nova Friburgo, mountain region of Rio de Janeiro, Brazil. The World Health Organization (WHO, 1997) criterion was used by previously calibrated examiners to report dental caries. Variables were obtained by means of a semi-structured questionnaire applied to the adolescents' parents. RESULTS: The D3MFT mean was 1.90 and the Significant Caries (SIC) Index was 4.54, mainly represented by the carious component. Multiple logistic regression analysis revealed that parents with over 8 years of schooling (OR=0.579), absence of pain (OR=0.396) and not visiting the dentist (OR=0.270) might suggest protective factors against the disease; the possible risk factors were male gender (OR=1.982) and pain, extraction and others were reasons for consultation (OR=2.435). CONCLUSIONS: Although the prevalence of caries was slightly below the national mean, polarization of the disease was clearly observed. Education and no access to the dentist led to protection against the disease. These results may contribute to planning of oral health actions directed towards this target population for the control of caries...

Conhecimentos e práticas sobre saúde bucal em pacientes infantis com câncer / Knowledge and practice of oral health in child patients with cancer

The aim of this study was to use a qualitative approach of an exploratory nature to analyze the knowledge and practice of a nursing staff at Hospital Infantil Joana de Gusmão as regards oral health. The sample consisted of 2 nurses and 10 nursing technicians. Data collection was carried out through semi-structured interviews and the content was analysed considering 2 thematic groups: practice and oral health knowledge and meanings of health care. The results revealed a lack of attention concerning oral health care in child patients undergoing antineoplastic treatment. It could be concluded that there is a clear need to adopt measures directed towards oral health care to promote patient health and quality of life. Furthermore, training courses on oral health in oncology, as well as the inclusion of a dentist on the hospital staff, are also suggested.

Conhecimentos e práticas em saúde bucal com crianças hospitalizadas com câncer / Knowledge and practices of oral health on hospitalized children with cancer

O objetivo do estudo foi avaliar os conhecimentos e práticas em saúde bucal (SB) com crianças hospitalizadas com câncer. A amostra foi composta pela equipe de enfermagem, cuidadores e crianças. Foi aplicado um questionário relacionado ao conhecimento geral sobre SB, métodos e instrumentos utilizados para a higiene oral (HO) e dados socioeconômicos. Baseado nos resultados, quem realiza a HO das crianças são os cuidadores (90,7 por cento), que receberam orientações da equipe de enfermagem em 21,4 por cento dos casos. Com relação ao desconforto na cavidade bucal, a equipe de enfermagem reportou que todos apresentaram manifestações clínicas, enquanto apenas 62,8 por cento dos cuidadores reportaram casos. Todos os participantes consideram importante haver um CD no setor de oncologia. Pôde-se concluir que não existe um protocolo de cuidados com a higiene bucal de crianças hospitalizadas com câncer e que as manifestações bucais mais frequentes entre os pacientes em tratamento antineoplásico foram: mucosite, enjoos, vômitos, xerostomia e ausência de paladar.

The aim of this study was to evaluate the knowledge and practices of oral health (OH) in hospitalized children with cancer. The sample was composed by the nurse team, caretakers and children. It was applied a questionnaire concerning the general knowledge about OH, methods and instruments used for oral hygiene (OH). According to the results, the responsible of OH of children are the caretakers (90.7 percent) who receive instructions from the nurse team in 21.4 percent of cases. As for the oral cavity discomfort, the nurse team reported that all patients exhibited clinical manifestations while the caretakers reported a different number, 62.8 percent of cases. All participants considered important having a dentist in the oncology sector. According to the results obtained, it was possible to conclude that there is no oral health protocol for hospitalized children with cancer and that the most frequent oral manifestations among patients going through antineoplastic treatment were: mucositis, nausea, vomit, xerostomy and lack of sense of taste.

Effect of saliva contamination and re-etching time on the shear bond strength of a pit and fissure sealant.

The aim of this study was to evaluate the effect of saliva contamination (SCT) and re-etching time (RET) on the shear bond strength (SBS) of the Fluroshield sealant. Forty-five extracted third molars were sectioned and flattened until reach an enamel surface area. Then, all samples were etched for 30 sec with 35% phosphoric acid and then they were distributed into 9 groups (n=10) according to SCT and RET (seconds), respectively: G1- control (no SCT and no RET); G2- 30s and 0s; G3- 60s and 0s; G4-30s and 2s; G5- 30s and 5s; G6- 30s and 15s; G7-60s and 2s; G8- 60s and 5s; G9- 60s and 15s. The sealant was applied according to the manufacturer's instructions. The samples were stored in distilled water at 37ºC for 72h and subjected to the SBS test. The results indicated that there was no statistically significant difference between the groups (p>0.05). However, it could be noticed that: 1- the longer the SCT, the lower the SBS values; 2 - the longer the RET, the higher the SBS values. It could be concluded that there was a tendency to the shortest SCT (30s) associated to the longest RET (5 and 15s) to reach similar SBS values for the control group.