Improving the efficiency of aerosolized insecticide testing against mosquitoes.

Developing robust and standardised approaches for testing mosquito populations against insecticides is vital for understanding the effectiveness of new active ingredients or formulations. Methods for testing mosquito susceptibility against contact insecticides or products, such as those delivered through public health programmes, are well-established and standardised. Nevertheless, approaches for testing volatile or aerosolized insecticides used in household products can be challenging to implement efficiently. We adapted WHO guidelines for household insecticides to develop a standardised and higher-throughput methodology for testing aerosolized products in a Peet Grady test chamber (PG-chamber) using caged mosquitoes and an efficient decontamination method. The new approach was validated using insecticide resistant and susceptible Aedes and Anopheles mosquito colonies. An added feature is the inclusion of cage-facing cameras to allow real-time quantification of knockdown following insecticide exposure. The wipe-based decontamination method was highly effective for removing pyrethroids' aerosolized oil-based residues from chamber surfaces, with < 2% mortality recorded for susceptible mosquitoes tested directly on the surfaces. There was no spatial heterogeneity for knockdown or mortality of caged mosquitoes within the PG chamber. The dual-cage approach we implement yields eight-times the throughput compared to a free-flight protocol, allows simultaneous testing of different mosquito strains and effectively discriminates susceptible and resistant mosquito colonies tested side-by-side.

Psychometric validation of the Brazilian Portuguese version of the Body Image Scale ­ HIV for people living with HIV/AIDS / Validação psicométrica da versão em português brasileiro da Body Image Scale ­ HIV para pessoas vivendo com HIV/AIDS

Introduction: Body Image can be defined as the mental representation of body identity. The surroundings, the gaze of the other, they emotions, they integrity, and our own fragility are elements of this representation. The social stigma caused by body changes in people living with HIV / AIDS may be more intense than the fear of death itself, being important to assess the impact of these changes in the course of treatment. Objective: To perform the psychometric validation of the Brazilian Portuguese version of the Body Image Scale - HIV for a sample of Brazilians, of both sexes, living with HIV / AIDS. Methods: The total of 450 patients were recruited from the São Bernardo do Campo HIV/AIDS program, aged between 18 and 78 years. Confirmatory factor analysis using the Unweighted Least Square and listwise deletion was used to determine the adherence of the data to the models tested. Results: Adequate coefficients of internal reliability and evidence of construct validity were established for the Brazilian version of BIS-HIV in the unifactorial model. Conclusion:This scale, with evidenced psychometric qualities, provides a useful tool for quantitative investigations of body image in people with HIV/AIDS in the Brazilian context and extends the possibility of future cross-cultural research.

Introdução: a Imagem Corporal pode ser definida como a representação mental da identidade corporal. O meio que cerca o sujeito, o olhar do outro, suas emoções, a integridade e a própria fragilidade do corpo são elementos próprios desta representação. O estigma social provocado pelas alterações corporais nas pessoas que vivem com HIV/AIDS pode ser mais intenso que o próprio medo da morte, sendo importante acompanhar o impacto dessas alterações do decurso do tratamento.Objetivo: realizar a validação psicométrica da versão em português brasileiro da Body Image Scale ­ HIV para uma amostra de brasileiros, de ambos os sexos, vivendo com HIV/AIDS.Método: foram recrutados 450 pacientes de um ambulatório de HIV/AIDS de São Bernardo do Campo, São Paulo, Brasil, com idades entre 18 e 78 anos. A análise fatorial confirmatória, utilizando o Unweighted Least Square e deleção listwise, foi usada para determinar a aderência dos dados aos modelos testados.Resultados: coeficientes adequados de confiabilidade interna e evidência de validade de construto foram estabelecidos para a versão brasileira da BIS-HIV no modelo unifatorial.Conclusão: a escala foi validada e com suas qualidades psicométricas evidenciadas, fornece uma ferramenta útil para investigações quantitativas da imagem corporal em pessoas com HIV/AIDS no contexto brasileiro e amplia a possibilidade de futuras pesquisas transculturais.

Malignant adnexal cutaneous tumor of the scalp: a case report of difficult differential diagnosis between metastatic breast cancer and primary sweat gland tumor / Neoplasia maligna de glândula anexial cutânea em couro cabeludo: um caso de dúvida diagnóstica entre carcinoma metastático de mama e tumor primário de glândula sudoríparas

This paper describes a case of a 71-year-old female who initially went to a dermatologist to assess a scalp skin tumor, which performed an incisional biopsy. Anatomopathological and immunohistochemical study revealed a preliminary diagnosis of breast carcinoma metastasis. Although the patient had no medical history of breast cancer, due to this result, she was referred to a mastologist, who investigated her breast nodules for the possible primary focus of the carcinoma. Despite an active investigation through imaging tests, biopsies, and mammotomy, without finding any possible primary focus on the breasts. Finally, the lesion on the scalp was entirely removed by a plastic surgeon. New anatomopathological and immunohistochemical exams confirmed the diagnosis of breast carcinoma metastasis. Given these results, the authors discuss the difficulty in diagnosing differentiation from a primary or metastatic neoplasm of the scalp, with the resources currently available, until the conclusion that it was a primary carcinoma of the sweat gland.

Este artigo descreve o caso de uma mulher de 71 anos que inicialmente foi ao dermatologista para avaliação de tumor de pele no couro cabeludo, e que realizou biópsia incisional desta lesão. O estudo anatomopatológico e imuno-histoquímico revelou um diagnóstico preliminar de metástase de carcinoma de mama. Embora a paciente não tivesse história clínica de câncer de mama, devido a esse resultado, foi encaminhada à mastologista, que investigou seus nódulos mamários para localizar o possível foco primário do carcinoma. Apesar de uma investigação ativa por meio de exames de imagem, biópsias e mamotomia, não foi encontrado nenhum possível foco primário nas mamas. Por fim, a lesão no couro cabeludo também foi totalmente removida por um cirurgião plástico. Novos exames anatomopatológicos e imuno-histoquímicos confirmaram o diagnóstico de metástase de carcinoma de mama. Diante desses resultados, os autores discutem a dificuldade em diagnosticar a diferenciação de uma neoplasia primária ou metastática do couro cabeludo, com os recursos disponíveis atualmente, até a conclusão de que se tratava de um carcinoma primário da glândula sudorípara.

A completely digital workflow for an interim implant-supported crown: A clinical report.

This article describes a completely digital workflow for the diagnostic phase, surgical and prosthetic planning, extraction, immediate single implant placement by guided surgery, and interim implant-supported crown. From a virtual impression, zirconia abutments and a polymethylmethacrylate (PMMA) interim crown was planned in a computer-aided design (CAD) software program. This workflow shortened the time required for chairside placement of an interim restoration with enhanced function and esthetics while restoring an anterior mandibular tooth lost after trauma.

Psychometric Validation of the Brazilian Portuguese Version of the Derriford Appearance Scale-24 (DAS-24) for People Living with HIV/AIDS.

The changes in appearance of people living with HIV/AIDS (PLHA) interferes with how people around them react to their body, how social interactions take place, and how each person perceives and accepts their body. The definition of itself can be severely challenged when the body changes as a result of illness and the person does not look healthier anymore. People living with HIV/AIDS (PLHA) are an especially vulnerable group when it comes to "distress" and the psychosocial impact of appearance, yet the assessment of body image changes in these people was subjective in Brazil. The aim of this paper was to assess the psychometric properties of the Brazilian version of Derriford Appearance Scale 24 (DAS-24) for a sample of Brazilians living with HIV/AIDS. A sample of 400 patients were recruited from an HIV/AIDS ambulatory, aged between 18 and 78 years, of both sexes. The psychometric properties of DAS-24 were investigated while using confirmatory factor analysis (CFA), with unweighted least square estimation and listwise deletion for missing data. The adjustment of three structural models previously established for DAS-24 (single-factor, two-factor, and three-factor) was investigated. Evidences of construct validity-convergent and discriminant-and internal consistency-Cronbach's alpha and construct reliability-were also generated for the measure model. The results showed that the one-factor model had the best adjustment, after eliminating items 8, 17, and 20, and accepting the covariance of errors between items 4 and 10; 9 and 23; 11 and 14; and, 14 and 22. Additionally, validity and reliability evidence were satisfactory for the model. The Brazilian Portuguese version of DAS-24 seems to be a psychometrically sound scale for measuring body image distress for people living with HIV/AIDS (PLHA).

Transcriptomic analysis of insecticide resistance in the lymphatic filariasis vector Culex quinquefasciatus.

Culex quinquefasciatus plays an important role in transmission of vector-borne diseases of public health importance, including lymphatic filariasis (LF), as well as many arboviral diseases. Currently, efforts to tackle C. quinquefasciatus vectored diseases are based on either mass drug administration (MDA) for LF, or insecticide-based interventions. Widespread and intensive insecticide usage has resulted in increased resistance in mosquito vectors, including C. quinquefasciatus. Herein, the transcriptome profile of Ugandan bendiocarb-resistant C. quinquefasciatus was explored to identify candidate genes associated with insecticide resistance. High levels of insecticide resistance were observed for five out of six insecticides tested, with the lowest mortality (0.97%) reported to permethrin, while for DDT, lambdacyhalothrin, bendiocarb and deltamethrin the mortality rate ranged from 1.63-3.29%. Resistance to bendiocarb in exposed mosquitoes was marked, with 2.04% mortality following 1 h exposure and 58.02% after 4 h. Genotyping of the G119S Ace-1 target site mutation detected a highly significant association (p < 0.0001; OR = 25) between resistance and Ace1-119S. However, synergist assays using the P450 inhibitor PBO, or the esterase inhibitor TPP resulted in markedly increased mortality (to ≈80%), suggesting a role of metabolic resistance in the resistance phenotype. Using a novel, custom 60 K whole-transcriptome microarray 16 genes significantly overexpressed in resistant mosquitoes were detected, with the P450 Cyp6z18 showing the highest differential gene expression (>8-fold increase vs unexposed controls). These results provide evidence that bendiocarb resistance in Ugandan C. quinquefasciatus is mediated by both target-site mechanisms and over-expression of detoxification enzymes.

Development and application of a tri-allelic PCR assay for screening Vgsc-L1014F kdr mutations associated with pyrethroid and organochlorine resistance in the mosquito Culex quinquefasciatus.

BACKGROUND: Culex quinquefasciatus has a widespread distribution across tropical and sub-tropical regions, and plays an important role in the transmission of vector-borne diseases of public health importance, including lymphatic filariasis (LF) and multiple arboviruses. Increased resistance to insecticides threatens the efficacy and sustainability of insecticide-based anti-vector interventions which mitigate the burden of mosquito transmitted diseases in endemic regions. In C. quinquefasciatus two non-synonymous voltage gated sodium channel (Vgsc) variants, both resulting in a leucine to phenylalanine change at codon 1014, are associated with resistance to pyrethroids and DDT. This tri-allelic variation has compromised the ability to perform high-throughput single-assay screening. To facilitate the detection and monitoring of the Vgsc-1014 locus in field-caught mosquitoes, an Engineered-Tail Allele-Specific-PCR (ETAS-PCR) diagnostic assay was developed and applied to wild mosquitoes from Brazil, Tanzania and Uganda. RESULTS: This new cost-effective, single-tube assay was compared to two, well-established, genotyping approaches, pyrosequencing and TaqMan. The ETAS-PCR assay showed high specificity for discriminating the three alleles at Vgsc-L1014F, with genotyping results strongly correlated with pyrosequencing and TaqMan results (98.64% and 100% respectively). CONCLUSIONS: Our results support the utility of the ETAS-PCR/Vgsc-1014 diagnostic assay, which stands as an effective alternative for genotyping tri-allelic variants.

Cross Cultural adaptation into Brazilian Portuguese language of Derriford Appearance Scale 24 (DAS-24) for people living with HIV/AIDS / Adaptação cultural em Português Brasileiro da Derriford Appearance Scale - 24 (DAS-24) para pessoas vivendo com HIV/AIDS

INTRODUCTION: Body image can be defined as the representation of beliefs, emotions and perceptions about the body itself, manifested in behaviors directed to the body. When the body changes because of a disease and does not seem healthy, the self-concept may be severely challenged. People living with HIV/AIDS (PLHA) are particularly vulnerable to the distress and psychosocial impact of appearance, but in Brazil the assessment of those body image changes was subjective because there was not an available scale in Brazilian Portuguese to assess body image changes in clinical practice or research. OBJECTIVE: To carry out the cross-cultural adaptation to the Brazilian Portuguese of the Derriford Appearance Scale 24 (DAS-24), with the verification of the linguistic, semantic, conceptual and cultural equivalence of the people living with HIV/AIDS in Brazil METHODS: We followed the five stages of culturally sensitive translation: direct translations, synthesis of translations, back-translations, expert committee meeting and pre-tests. The process of cultural adaptation was presented in a descriptive and analytical way, following patterns of methodological studies. The minimum, maximum and median values of the responses of each item were calculated from the pool of data from the third pretest group of 50 participants. The median of the item scores, the correlation on each item with the total score and the internal reliability, were calculated using the Cronbach alpha test. RESULTS: The analysis of the responses of the last pre-test group indicated that attention must be given to items A, H, T and V in a future psychometric study. The present study is not enough for this scale to be used in clinical practice. To ensure that the culturally adapted instrument generates valid and reliable data, a subsequent study investigating its psychometric properties should be conducted. CONCLUSION: The cross-cultural adaptation of the Derriford Appearance Scale 24 (DAS-24) in its components of linguistic, semantic, conceptual and cultural equivalence to Brazilian Portuguese for the population of people living with HIV/AIDS was fully carried out. Despite this achievement, it is emphasized that the use of the Brazilian version of DAS-24 in research and clinical routine is advised only after a psychometric study with this instrument.

INTRODUÇÃO: A imagem corporal pode ser definida como a representação das crenças, emoções e percepções a respeito do próprio corpo, manifesta em comportamentos voltados ao corpo. Quando o corpo muda como consequência de doença e não parece mais saudável, a definição de si mesmo pode ser severamente desafiada. As pessoas vivendo com HIV/AIDS (PVHA) são um público especialmente vulnerável quando se trata do "distress" e do impacto psicossocial da aparência, mas a avaliação destas alterações de imagem corporal era subjetiva porque não havia nenhuma escala em Português Brasileiro para avaliar alterações da imagem disponível para uso clínico ou para pesquisa. OBJETIVO: Realizar a adaptação transcultural para o português Brasileiro da Derriford Appearance Scale 24 (DAS-24), com a verificação da equivalência idiomática, semântica, conceitual e cultural, para o público-alvo pessoas vivendo com HIV/AIDS (PVHA) no Brasil MÉTODO: Seguiu-se guia de cinco etapas para adaptação de escala transcultural: traduções, síntese de traduções, retrotraduções, reunião de comitê de especialistas e pré-testes. O processo de adaptação cultural foi apresentado de forma descritiva e analítica, seguindo padrões de estudos metodológicos. Os valores mínimo, máximo e mediano das respostas de cada item foram calculados a partir do pool de dados do terceiro grupo de pré-teste de 50 participantes. A mediana dos escores dos itens, a correlação de cada item com o escore total e a confiabilidade interna foram calculados pelo teste alfa de Cronbach. RESULTADO: A análise das respostas do último grupo pré-teste indicou que deve ser dada atenção aos itens A, B, G, H e K em um futuro estudo psicométrico. O presente estudo não é suficiente para que essa escala seja utilizada na prática clínica. Para garantir que o instrumento culturalmente adaptado gere dados válidos e confiáveis, um estudo subsequente que investigue suas propriedades psicométricas deve ser conduzido. CONCLUSÃO: A adaptação transcultural da Derriford Appearance Scale 24 (DAS-24), em seus componentes de equivalência linguística, semântica, conceitual e cultural para o português brasileiro para a população de pessoas vivendo com HIV/AIDS foi plenamente realizada. Apesar dessa conquista, ressalta-se que o uso da versão brasileira do DAS-24 em pesquisa e rotina clínica é aconselhado somente após um estudo psicométrico com este instrumento.

CCR5Δ32 (rs333) polymorphism is associated with decreased risk of chronic and aggressive periodontitis: A case-control analysis based in disease resistance and susceptibility phenotypes.

Chronic and aggressive periodontitis are infectious diseases characterized by the irreversible destruction of periodontal tissues, which is mediated by the host inflammatory immune response triggered by periodontal infection. The chemokine receptor CCR5 play an important role in disease pathogenesis, contributing to pro-inflammatory response and osteoclastogenesis. CCR5Δ32 (rs333) is a loss-of-function mutation in the CCR5 gene, which can potentially modulate the host response and, consequently periodontitis outcome. Thus, we investigated the effect of the CCR5Δ32 mutation over the risk to suffer periodontitis in a cohort of Brazilian patients (total N=699), representative of disease susceptibility (chronic periodontitis, N=197; and aggressive periodontitis, N=91) or resistance (chronic gingivitis, N=193) phenotypes, and healthy subjects (N=218). Additionally, we assayed the influence of CCR5Δ32 in the expression of the biomarkers TNFα, IL-1ß, IL-10, IL-6, IFN-γ and T-bet, and key periodontal pathogens P. gingivalis, T. forsythia, and T. denticola. In the association analysis of resistant versus susceptible subjects, CCR5Δ32 mutant allele-carriers proved significantly protected against chronic (OR 0.49; 95% CI 0.29-0.83; p-value 0.01) and aggressive (OR 0.46; 95% CI 0.22-0.94; p-value 0.03) periodontitis. Further, heterozygous subjects exhibited significantly decreased expression of TNFα in periodontal tissues, pointing to a functional effect of the mutation in periodontal tissues during the progression of the disease. Conversely, no significant changes were observed in the presence or quantity of the periodontal pathogens P. gingivalis, T. forsythia, and T. denticola in the subgingival biofilm that could be attributable to the mutant genotype.

Local selection in the presence of high levels of gene flow: Evidence of heterogeneous insecticide selection pressure across Ugandan Culex quinquefasciatus populations.

BACKGROUND: Culex quinquefasciatus collected in Uganda, where no vector control interventions directly targeting this species have been conducted, was used as a model to determine if it is possible to detect heterogeneities in selection pressure driven by insecticide application targeting other insect species. METHODOLOGY/PRINCIPAL FINDINGS: Population genetic structure was assessed through microsatellite analysis, and the impact of insecticide pressure by genotyping two target-site mutations, Vgsc-1014F of the voltage-gated sodium channel target of pyrethroid and DDT insecticides, and Ace1-119S of the acetylcholinesterase gene, target of carbamate and organophosphate insecticides. No significant differences in genetic diversity were observed among populations by microsatellite markers with HE ranging from 0.597 to 0.612 and low, but significant, genetic differentiation among populations (FST = 0.019, P = 0.001). By contrast, the insecticide-resistance markers display heterogeneous allelic distributions with significant differences detected between Central Ugandan (urban) populations relative to Eastern and Southwestern (rural) populations. In the central region, a frequency of 62% for Vgsc-1014F, and 32% for the Ace1-119S resistant allele were observed. Conversely, in both Eastern and Southwestern regions the Vgsc-1014F alleles were close to fixation, whilst Ace1-119S allele frequency was 12% (although frequencies may be underestimated due to copy number variation at both loci). CONCLUSIONS/SIGNIFICANCE: Taken together, the microsatellite and both insecticide resistance target-site markers provide evidence that in the face of intense gene flow among populations, disjunction in resistance frequencies arise due to intense local selection pressures despite an absence of insecticidal control interventions targeting Culex.

Detection and quantitation of copy number variation in the voltage-gated sodium channel gene of the mosquito Culex quinquefasciatus.

Insecticide resistance is typically associated with alterations to the insecticidal target-site or with gene expression variation at loci involved in insecticide detoxification. In some species copy number variation (CNV) of target site loci (e.g. the Ace-1 target site of carbamate insecticides) or detoxification genes has been implicated in the resistance phenotype. We show that field-collected Ugandan Culex quinquefasciatus display CNV for the voltage-gated sodium channel gene (Vgsc), target-site of pyrethroid and organochlorine insecticides. In order to develop field-applicable diagnostics for Vgsc CN, and as a prelude to investigating the possible association of CN with insecticide resistance, three assays were compared for their accuracy in CN estimation in this species. The gold standard method is droplet digital PCR (ddPCR), however, the hardware is prohibitively expensive for widespread utility. Here, ddPCR was compared to quantitative PCR (qPCR) and pyrosequencing. Across all platforms, CNV was detected in ≈10% of mosquitoes, corresponding to three or four copies (per diploid genome). ddPCR and qPCR-Std-curve yielded similar predictions for Vgsc CN, indicating that the qPCR protocol developed here can be applied as a diagnostic assay, facilitating monitoring of Vgsc CN in wild populations and the elucidation of association between the Vgsc CN and insecticide resistance.

Genetic variability and spatial distribution in small geographic scale of Aedes aegypti (Diptera: Culicidae) under different climatic conditions in Northeastern Brazil.

BACKGROUND: The study of the genetic structure of Aedes aegypti is essential to understanding their population dynamics as well as for the analysis of factors responsible for their resistance and ecological adaptation. The use of molecular markers in identifying differences amongst populations of Ae. aegypti in different geographical areas as well as the temporal variation of the vector populations has contributed to the improvement of vector control strategies. The present study aims to determine the genetic variability of Ae. aegypti populations in a small geographical area (state of Sergipe, Northeastern Brazil) by means of inter-simple sequence repeat (ISSR) and single nucleotide polymorphism (SNP) molecular markers. RESULTS: ISSR markers revealed a more heterogeneous pattern of genetic diversity among the populations with an expected heterozygosity (H E) ranging from 0.261 ± 0.03 to 0.120 ± 0.032, while a similar trend was detected through SNPs across populations with an H E between 0.375 ± 0.054 and 0.269 ± 0.042. The population's genetic differentiation assessed with ISSR and SNP markers indicated a very low structuring among the populations with the highest diversity observed within the populations 72 % (ISSR) and 92 % (SNP). Clustering analysis also suggested little variation among populations: the seven populations were grouped into only three ISSR clusters and a single panmictic group based on SNP markers. The present study identified a close relationship between the populations, which probably results mainly from passive gene flow between mosquitoes from distinct geographic regions, influenced by humans commuting along roads. CONCLUSIONS: There was an intense migration of mosquitos across municipalities, leading to a potential increase in risk of arbovirus and insecticide resistance associated-alleles spreading between mosquito populations.

Preenchimento facial com Polimetilmetacrilato em pacientes que vivem com a síndrome da imunodeficiência adquirida (AIDS) / Facial filling with polymethylmethacrylate in patients with acquired immunodeficiency syndrome

INTRODUÇÃO: Pacientes que vivem com síndrome da imunodeficiência adquirida (AIDS) em uso da Terapia Antirretroviral de Alta Potência (TARV) são suscetíveis a desenvolver síndrome lipodistrófica. O preenchimento facial com polimetilmetacrilato é opção de tratamento. O objetivo é analisar o procedimento de preenchimento facial e avaliar os pacientes em relação à percepção, incômodo, revelação do diagnóstico, expectativa quanto ao preenchimento e a satisfação e impacto em suas vidas. MÉTODOS: Análise em 63 pacientes submetidos ao preenchimento facial. Foram realizados procedimentos, analisados prontuários dos pacientes e o Protocolo do Ambulatório de Lipodistrofia do Programa Municipal de doenças sexualmente transmissíveis (DST)/AIDS e Hepatites Virais de São Bernardo do Campo, atendidos no período de janeiro a julho de 2009. RESULTADOS: Todos os 63 pacientes iniciais que concordaram em participar da pesquisa permaneceram até o término deste trabalho. Apenas seis pacientes (9,5%) eram de origem de outros municípios, enquanto 57 pacientes (90,5%) eram moradores de São Bernardo. 68,2% eram homens e 100% brancos. A média das idades foi 49,7 anos. Em média, o Vírus da Imunodeficiência Humana (HIV) foi diagnosticado há 11,5 anos, com tempo médio de uso de TARV por 10 anos e tempo médio de lipoatrofia facial de 3,8 anos. A maioria fez uso de Estavudina e/ou Efavirenz. Quem ficava mais desconfortável com as alterações na face eram os próprios pacientes. 85,7% não revelaram o diagnóstico para terceiros. 100% dos pacientes ficaram satisfeitos ou muito satisfeitos com o resultado obtido. CONCLUSÃO: 100% dos pacientes ficaram satisfeitos ou muito satisfeitos com o resultado obtido. Em 100% dos casos houve um impacto favorável na vida. Não houve efeitos adversos ao procedimento cirúrgico de preenchimento.

INTRODUCTION: Patients with acquired immunodeficiency syndrome (AIDS) who use highly active antiretroviral therapy (HAART) can develop lipodystrophy syndrome, for which facial filling with polymethylmethacrylate is a treatment option. The objective is to analyze the procedure of facial filling and evaluate patients in relation to their perception, discomfort, revelation of the diagnosis to third parties, expectation concerning facial filling, and satisfaction with the treatment outcome and its impact on their lives. METHODS: Sixty-three patients who underwent facial filling were evaluated. Procedures performed between January and July 2009 were assessed, the records of the patients were analyzed, and the outpatient lipodystrophy protocol of the STD/AIDS and Viral Hepatitis Municipal Program of São Bernardo do Campo was used. RESULTS: All the 63 patients who agreed to participate in the research completed the study. Only 6 patients (9.5%) were from other municipalities, while 57 patients (90.5%) were residents of São Bernardo. Of the patients, 68.2% were men and 100% were Caucasian. The mean age of the patients was 49.7 years. Human immunodeficiency virus was diagnosed 11.5 years prior on average, with 10-year average use of HAART and 3.8-year average time of facial lipoatrophy. Most of the patients used stavudine and/or efavirenz. The patients themselves felt more uncomfortable with facial changes. Among the patients, 85.7% did not reveal the diagnosis to third parties. CONCLUSION: All of the patients were satisfied or very satisfied with the result obtained, which had a favorable impact on their lives. The filling surgical procedure had no adverse effects.

La modulación de la expresión de IL-10 por el polimorfismo IL-10-592C/A (rs1800872) es independiente de la presencia y carga bacteriana de los periodontopatógenos clásicos / Modulation of IL-10 mRNA levels in periodontal lesions by IL-10-592C/A polymorphism (rs1800872) is independent of the frequency and load of classic periodontopathic bacteria

Objetivo: Reportamos la asociación entre el polimorfismo de nucleótido simple de IL-10-592C/A (rs1800872) y la detección/abundancia relativa de los periodontopatógenos Porfiromonas gingivalis, Tenerella forsythia, Treponema denticola y Aggregatibacter actinomycetemcomitans. Además investigamos la influencia de los determinantes genéticos y microbiológicos en los niveles de expresión de IL-10 en lesiones periodontales. Metodología Fueron reclutados 117 pacientes con periodontitis crónica y 58 controles. Luego del examen clínico fueron obtenidas muestras microbiológicas y la presencia/carga bacteriana de especies de periodontopatógenos fue cuantificada por RT-PCR. El genotipo para IL-10-592C/A fue determinado mediante restriction fragment length polymorphism. Resultados La distribución alélica del SNP rs1800872 en la población investigada cumplió con el equilibrio de Hardy-Weinberg (p = 0,64). Como ya ha sido reportado, los sujetos polimórficos demostraron menor expresión de IL-10 y riesgo aumentado de sufrir periodontitis crónica. El polimorfismo IL-10-592C/A no demostró relación con la detección o carga bacteriana de ninguna de las bacterias investigadas, además los niveles de expresión de IL-10 no fueron influenciados por el perfil microbiológico, sino que se correlacionaron directamente con el genotipo para el polimorfismo IL-10-592C/A.

Objective: A study was conducted to investigate the possible influence of the single nucleotide polymorphism (SNP) IL-10-592 C/A on the occurrence and load of the periodontal pathogens: P. gingivalis, T. forsythia, T. denticolaand A. Actinomycetemcomitans; as well to investigate the influence of microbial and genetic factors on the modulation of local IL-10 mRNA levels. Methodology The study included 117 cases and 58 controls. After clinical examination microbiological samples were obtained and the detection/quantification of the target bacterial species was performed by RT-PCR. SNP rs1800872 was assayed by restriction fragment length polymorphism (RFLP). Results Allele distribution of rs1800872 was in Hardy-Weinberg equilibrium (P = 64). As previously reported, polymorphic subjects demonstrated decreased IL-10 expression and increased risk of suffering chronic periodontitis. IL-10-592C/A rs1800872 SNP was not associated with the detection or the bacterial load of the investigated pathogens. Moreover, the presence/load of bacteria at periodontal sites did not influence IL-10 expression, which was determined by the genetic background of the study subjects. IL-10-592C/A SNP was not associated with detection/bacterial load of pathogenic bacteria. IL-10 expression levels were determined by the genetic background and were independent of the bacterial microenvironment.

TBX21-1993T/C (rs4794067) polymorphism is associated with increased risk of chronic periodontitis and increased T-bet expression in periodontal lesions, but does not significantly impact the IFN-g transcriptional level or the pattern of periodontophatic bacterial infection.

Th1-polarized host response, mediated by IFN-γ, has been associated with increased severity of periodontal disease as well as control of periodontal infection. The functional polymorphism TBX21-1993T/C (rs4794067) increases the transcriptional activity of the TBX21 gene (essential for Th1 polarization) resulting in a predisposition to a Th-1 biased immune response. Thus, we conducted a case-control study, including a population of healthy controls (H, n = 218), chronic periodontitis (CP, n = 197), and chronic gingivitis patients (CG, n = 193), to investigate if genetic variations in TBX21 could impact the development of Th1 responses, and consequently influence the pattern of bacterial infection and periodontitis outcome. We observed that the polymorphic allele T was significantly enriched in the CP patients compared to CG subjects, while the H controls demonstrated and intermediate genotype. Also, investigating the putative functionality TBX21-1993T/C in the modulation of local response, we observed that the transcripts levels of T-bet, but not of IFN-γ, were upregulated in homozygote and heterozygote polymorphic subjects. In addition, TBX21-1993T/C did not influence the pattern of bacterial infection or the clinical parameters of disease severity, being the presence/absence of red complex bacteria the main factor associated with the disease status and the subrogate variable probing depth (PD) in the logistic regression analysis.

Spatial distribution and esterase activity in populations of Aedes (Stegomyia) aegypti (Linnaeus) (Diptera: Culicidae) resistant to temephos.

INTRODUCTION: The need for studies that describe the resistance patterns in populations of Aedes aegypti (Linnaeus) in function of their region of origin justified this research, which aimed to characterize the resistance to temephos and to obtain information on esterase activity in populations of Aedes aegypti collected in municipalities of the State of Paraíba. METHODS: Resistance to temephos was evaluated and characterized from the diagnostic dose of 0.352mg i.a./L and multiple concentrations that caused mortalities between 5% and 99%. Electrophoresis of isoenzymes was used to verify the patterns of esterase activity among populations of the vector. RESULTS: All populations of Aedes aegypti were resistant to temephos, presenting a resistance rate (RR) greater than 20. The greatest lethal dose 50% of the sample (CL50) was found for the municipality of Lagoa Seca, approximately forty-one times the value of CL50 for the Rockefeller population. The populations characterized as resistant showed two to six regions of α and ß-esterase, called EST-1 to EST-6, while the susceptible population was only seen in one region of activity. CONCLUSIONS: Aedes aegypti is widely distributed and shows a high degree of resistance to temephos in all municipalities studied. In all cases, esterases are involved in the metabolism and, consequently, in the resistance to temephos.

Spatial distribution and esterase activity in populations of Aedes (Stegomyia) aegypti (Linnaeus) (Diptera: Culicidae) resistant to temephos

INTRODUCTION: The need for studies that describe the resistance patterns in populations of Aedes aegypti (Linnaeus) in function of their region of origin justified this research, which aimed to characterize the resistance to temephos and to obtain information on esterase activity in populations of Aedes aegypti collected in municipalities of the State of Paraíba. METHODS: Resistance to temephos was evaluated and characterized from the diagnostic dose of 0.352mg i.a./L and multiple concentrations that caused mortalities between 5% and 99%. Electrophoresis of isoenzymes was used to verify the patterns of esterase activity among populations of the vector. RESULTS: All populations of Aedes aegypti were resistant to temephos, presenting a resistance rate (RR) greater than 20. The greatest lethal dose 50% of the sample (CL50) was found for the municipality of Lagoa Seca, approximately forty-one times the value of CL50 for the Rockefeller population. The populations characterized as resistant showed two to six regions of α and β-esterase, called EST-1 to EST-6, while the susceptible population was only seen in one region of activity. CONCLUSIONS: Aedes aegypti is widely distributed and shows a high degree of resistance to temephos in all municipalities studied. In all cases, esterases are involved in the metabolism and, consequently, in the resistance to temephos. .

Non-inflammatory destructive periodontal disease: a clinical, microbiological, immunological and genetic investigation.

Periodontitis comprises a group of multifactorial diseases in which periodontopathogens accumulate in dental plaque and trigger host chronic inflammatory and immune responses against periodontal structures, which are determinant to the disease outcome. Although unusual cases of non-inflammatory destructive periodontal disease (NIDPD) are described, their pathogenesis remains unknown. A unique NIDPD case was investigated by clinical, microbiological, immunological and genetic tools. The patient, a non-smoking dental surgeon with excessive oral hygiene practice, presented a generalized bone resorption and tooth mobility, but not gingival inflammation or occlusion problems. No hematological, immunological or endocrine alterations were found. No periodontopathogens (A. actinomycetemcomitans, P. gingivalis, F. nucleatum and T. denticola) or viruses (HCMV, EBV-1 and HSV-1) were detected, along with levels of IL-1ß and TNF-a in GCF compatible with healthy tissues. Conversely ALP, ACP and RANKL GCF levels were similar to diseased periodontal sites. Genetic investigation demonstrated that the patient carried some SNPs, as well HLA-DR4 (*0404) and HLA-B27 alleles, considered risk factors for bone loss. Then, a less vigorous and diminished frequency of toothbrushing was recommended to the patient, resulting in the arrest of alveolar bone loss, associated with the return of ALP, ACP and RANKL in GCF to normality levels. In conclusion, the unusual case presented here is compatible with the previous description of NIDPD, and the results that a possible combination of excessive force and frequency of mechanical stimulation with a potentially bone loss prone genotype could result in the alveolar bone loss seen in NIDPD.

The use of chronic gingivitis as reference status increases the power and odds of periodontitis genetic studies: a proposal based in the exposure concept and clearer resistance and susceptibility phenotypes definition.

AIM: Current literature on chronic periodontitis genetics encompasses numerous single nucleotide polymorphisms-focused case-control studies with inconsistent and controversial results, which typically disregards the exposure concept embraced by case-control definition. Herein, we propose a case-control design reappraisal by clear phenotype selection, where chronic gingivitis represents a genetically resistant phenotype/genotype opposing the susceptible cohort. MATERIAL AND METHODS: The hypothesis was tested in healthy, chronic periodontitis and gingivitis groups through Real-time PCR-based allelic discrimination of classic variants IL1B-3954, IL6-174, TNFA-308, IL10-592 and TLR4-299. RESULTS: Observed allele/genotype frequencies characterize the healthy group with an intermediate genetic profile between periodontitis and gingivitis cohorts. When comparing genotype/allele frequencies in periodontitis versus healthy and periodontitis versus gingivitis scenarios, the number of positive associations (2-4) and the degree of association (p and odds ratio values) were significantly increased by the new approach proposed (periodontitis versus gingivitis), suggesting the association of IL1B-3954, TNFA-308, IL10-592 and TLR4-299 with periodontitis risk. Power study was also significantly improved by the new study design proposed when compared to the traditional approach. CONCLUSIONS: The data presented herein support the use of new case-control study design based on the case-control definition and clear resistance/susceptibility phenotypes selection, which can significantly impact the study power and odds of identification of genetic factors involved in PD.

Association of human T lymphotropic virus 1 amplification of periodontitis severity with altered cytokine expression in response to a standard periodontopathogen infection.

BACKGROUND: Periodontal diseases (PDs) are infectious diseases in which periodontopathogens trigger chronic inflammatory and immune responses that lead to tissue destruction. Recently, viruses have been implicated in the pathogenesis of PDs. Individuals infected with human T lymphotropic virus 1 (HTLV-1) present with abnormal oral health and a marked increased prevalence of periodontal disease. METHODS: In this study, we investigated the patterns of periodontopathogen infection and local inflammatory immune markers in HTLV-1-seropositive individuals with chronic periodontitis (CP/HTLV-1 group) compared with HTLV-1-seronegative individuals with chronic periodontitis (CP group) and periodontally healthy, HTLV-1-seronegative individuals (control group). RESULTS: Patients in the CP/HTLV-1 group had significantly higher values of bleeding on probing, mean probing depth, and attachment loss than patients in the CP group. The expression of tumor necrosis factor alpha and interleukin (IL) 4 was found to be similar in the CP and CP/HTLV-1 groups, whereas IL-12 and IL-17 levels trended toward a higher expression in the CP/HTLV-1 group. A significant increase was seen in the levels of IL-1beta and interferon gamma in the CP/HTLV-1 group compared with the CP group, whereas expression of the regulatory T cell marker FOXp3 and IL-10 was significantly decreased in the lesions from the CP/HTLV-1 group. Interestingly, similar frequency and/or load of periodontopathogens (Porphyromonas gingivalis, Tannerella forsythia, Treponema denticola, and Aggregatibacter actinomycetemcomitans) and frequency of viruses (herpes simplex virus 1, human cytomegalovirus, and Epstein-Barr virus) characteristically associated with PDs were found in the CP/HTLV and CP groups. CONCLUSIONS: HTLV-1 may play a critical role in the pathogenesis of periodontal disease through the deregulation of the local cytokine network, resulting in an exacerbated response against a standard periodontopathogen infection.