miRNAs and NFKB1 and TRAF6 target genes: The initial functional study in CD14+ monocytes in rheumatoid arthritis patients.

We predicted miRNAs with regulatory impact on NFKB1 and TRAF6 gene expression and selected the miR-194-5p, miR-124-3p, miR-9-5p, and miR-340-5p and their target genes for expression analyses on CD14+ monocytes from rheumatoid arthritis (RA) patients and healthy controls. Additionally, we evaluated the influence of genes and miRNA expression on RA patients' cytokine levels. No difference was observed in genes or miRNAs expression when compared to healthy controls and RA patients or clinical parameters. However, we found a significant difference between miR-194-5p and miR-9-5p levels (FC=-2.31; p=0.031; FC=-3.05;p=0.031, respectively) and non-prednisone users as compared to prednisone using patients. We conducted correlation analyses to identify the strength of the relationship between expression data and cytokine plasma levels. We observed a moderate positive correlation between miR-124-3p expression and IL-6 plasma levels (r=0.46; p=0.033). In addition, overexpression of miRNAs was concomitant to TRAF6 and NFKB1 genes as indicated by correlation analyses: TRAF6 and miR-194-5p (r=0.60;p<0.001) and miR-9-5p (r=0.63;p<0.001) and NFKB1 and miR-194-5p (r=0.72;p<0.001), miR-9-5p (r=0.72;p<0.001) and miR-340-5p (r=0.61;p<0.001). NFKB1 and TRAF6 genes and miRNAs monocyte expression do not appear to be related to RA but showed a significant difference in different groups of RA therapy. In addition, increased levels of miRNAs can be linked to concomitant overexpression of TRAF6 and NFKB1 in monocytes and act as its regulators.

Influence of IL-6, IL-8, and TGF-ß1 gene polymorphisms on the risk of human papillomavirus-infection in women from Pernambuco, Brazil.

Human papillomavirus (HPV) infections are strongly associated with the development of cervical intraepithelial neoplasias and invasive cervical cancer. Polymorphisms in cytokine-encoding genes and behavioural cofactors could play an important role in protecting an individual against viral infections and cancer. Here, we investigated whether IL-6 -174 G>C, IL-8 +396 G>T, and TGF-ß1 +869 G>C and +915 G>C polymorphisms were associated with susceptibility to HPV infection in women from north-east (Pernambuco) Brazil. We analysed 108 healthy uninfected women (HC) and 108 HPV-positive women with cervical lesions. Genetic polymorphisms were assessed using Sanger sequencing and polymerase chain reaction-restriction fragment length polymorphism. Comparison of the distribution of the genotypic and allelic frequencies of the IL-18 +396 T>G polymorphism between HPV infected woman an uninfected controls showed that the GG genotype and G allele were both more frequent in the HC group, and were associated with protection from HPV infection (p = 0.0015; OR = 0.29 CI95% = 0.13-0.61; p = 0.0005; OR = 0.45 CI95% 0.29-0.7, respectively). Individuals from the control group could have previously had HPV infection that was spontaneously eliminated; however, it was undetectable at the time of sample collection. Based on our findings, we hypothesize that the IL-8 +396 G>T polymorphism could interfere with susceptibility to HPV infection, by modulating the ability of immune system to fight the virus.

CCR2 and CCR5 genes polymorphisms in women with cervical lesions from Pernambuco, Northeast Region of Brazil: a case-control study.

Polymorphisms in chemokine receptors play an important role in the progression of cervical intraepithelial neoplasia (CIN) to cervical cancer (CC). Our study examined the association of CCR2-64I (rs1799864) andCCR5-Δ32 (rs333) polymorphisms with susceptibility to develop cervical lesion (CIN and CC) in a Brazilian population. The genotyping of 139 women with cervical lesions and 151 women without cervical lesions for the CCR2-64I and CCR5-Δ32 polymorphisms were performed using polymerase chain reaction-restriction fragment length polymorphism. The individuals carrying heterozygous or homozygous genotypes (GA+AA) for CCR2-64I polymorphisms seem to be at lower risk for cervical lesion [odds ratio (OR) = 0.37, p = 0.0008)]. The same was observed for the A allele (OR = 0.39, p = 0.0002), while no association was detected (p > 0.05) with CCR5-Δ32 polymorphism. Regarding the human papillomavirus (HPV) type, patients carrying the CCR2-64Ipolymorphism were protected against infection by HPV type 16 (OR = 0.35, p = 0.0184). In summary, our study showed a protective effect ofCCR2-64I rs1799864 polymorphism against the development of cervical lesions (CIN and CC) and in the susceptibility of HPV 16 infection.

Agreement between methods for diagnosing HPV-induced anal lesions in women with cervical neoplasia.

OBJECTIVE: To evaluate agreement between 3 methods for screening anal intraepithelial lesions: anal cytology, anoscopy and human papillomavirus (HPV) detection by PCR. STUDY DESIGN: This prospective, cross-sectional study screened 324 women with cervical neoplasia for anal neoplasia. Agreement between methods was calculated using the κ coefficient. RESULTS: Of 324 anal cytologies performed, 31.5% (n = 102) were found to be abnormal: low-grade anal lesions were detected in 19.1% (n = 62) of cases, high-grade lesions in 3.1% (n = 10) and atypical squamous cells of undetermined significance in 9.3% (n = 30). With respect to the biopsies, 25.7% (n = 20) were positive, consisting of 7 cases of HPV infection, 5 anal intraepithelial neoplasia (AIN) grade 1, 6 AIN grade 2, and 2 AIN grade 3. Twenty-one samples (6.5%) were inadequate for HPV analysis. Of the 303 adequate samples, 84.2% (n = 255) tested positive for HPV. Agreement between cytology and anoscopy was fair (κ = 0.31). Agreement between PCR for HPV and cytology was slight (κ = 0.08) and no agreement was found between PCR for HPV and anoscopy (κ = 0.00). CONCLUSION: Agreement between the different methods of diagnosing HPV-induced anal lesions is slight to fair; however, anal cytology permits identification of cases in which lesions are present, allowing them to be referred for anoscopy and biopsy.

MYD88, IRAK3 and Rheumatoid Arthritis pathogenesis: Analysis of differential gene expression in CD14 + monocytes and the inflammatory cytokine levels.

Rheumatoid arthritis (RA) is a well-known chronic inflammatory disorder. Two molecular players act in the inflammation balance of the disease: MyD88 (Myeloid differentiation primary response 88) is related to TLR (Toll-like receptors) response and promotes the formation of myddosome complex resulting in increased inflammation; IRAK3 (Interleukin-1 receptor associated kinase 3) acts suppressing the myddosome complex thus decreasing inflammation. In this scenario, MYD88 and IRAK3 gene expression profile in RA patients and its correlation with clinical features is still partially known. So, we evaluated the MYD88 and IRAK3 gene expressions in CD14 + monocytes from RA patients and healthy controls and its relation with patients' clinical features and cytokine plasma levels. CD14 + monocytes were isolated using positive selection by magnetic cell separation. The MYD88 and IRAK3 gene expressions were measured through real time relative quantitative PCR with specific primers; relative quantification was normalized to ACTB, GAPDH, 18S and RPLP0 reference genes. Cytokine levels were analyzed by CBA (cytokine beads assays). CD14 + monocytes from RA patients showed lower IRAK3 expression level compared to controls although with a borderline statistical significance (Fold change (FC) = -1.63; p = 0.054). Furthermore, RA patients with high disease activity had lower levels of IRAK3 when compared to patients with low/moderate activity measured by the CDAI index (FC = -1.78; p = 0.030). No significant differences were observed for MYD88 gene expression (FC = 1.20; p = 0.294) between patients and controls analyzed. Additionally, we did not we did not observe correlation between IRAK3 and MYD88 gene expression and TNF-α, IL-6, IL-2 and IL-10 levels. We suggested that IRAK3 gene expression in CD14 + monocytes appears to be relevant to the RA etiology and clinical activity, whereas, in this study, MYD88 does not play a role in RA onset and development.

Absence of influence of polymorphisms of the MBL2 gene in oral infections by HSV-1 in individuals with HIV.

Polymorphisms in the structural gene MBL-2 (mannose-binding lectin-2) may result in low MBL serum concentration, associated with greater susceptibility to infection. The study evaluated the effects of MBL-2 polymorphisms with the oral manifestations of the HSV in human immunodeficiency virus (HIV)-infected patients. An observational case-control study was carried out, with the sample comprising 64 HIV+ and 65 healthy individuals. The signs and symptoms of HSV oral infection were evaluated, and oral mucosa buccal smears were collected. Polymorphisms of the MBL-2 gene and HSV-1 DNA were amplified through real-time PCR. The data revealed that of 64 HIV+, 29.6% presented signs and symptoms of HSV oral infection. Of these, the HSV-1 DNA was detected through real-time PCR in 21% of cases, and in 13.3% of asymptomatic individuals. There was no statistically significant difference between the symptomatic (p = 1) and the asymptomatic (p = 0.52) individuals, HIV+ and HIV-. Different genotypes (AA, A0, or 00) did not contribute to the oral manifestation of HSV in the HIV+ patients (p = 0.81) or HIV- (p = 0.45). There was no statistically significant difference in either group (p = 0.52). No significant association was identified between the MBL-2 gene polymorphisms in the oral manifestation of HSV infection. However, further studies are recommended with larger population groups before discarding this interrelationship.

Influence of IL-6, IL-8, and TGF-β 1 gene polymorphisms on the risk of human papillomavirus-infection in women from Pernambuco, Brazil

Human papillomavirus (HPV) infections are strongly associated with the development of cervical intraepithelial neoplasias and invasive cervical cancer. Polymorphisms in cytokine-encoding genes and behavioural cofactors could play an important role in protecting an individual against viral infections and cancer. Here, we investigated whether IL-6 -174 G>C, IL-8 +396 G>T, and TGF-β1 +869 G>C and +915 G>C polymorphisms were associated with susceptibility to HPV infection in women from north-east (Pernambuco) Brazil. We analysed 108 healthy uninfected women (HC) and 108 HPV-positive women with cervical lesions. Genetic polymorphisms were assessed using Sanger sequencing and polymerase chain reaction-restriction fragment length polymorphism. Comparison of the distribution of the genotypic and allelic frequencies of the IL-18 +396 T>G polymorphism between HPV infected woman an uninfected controls showed that the GG genotype and G allele were both more frequent in the HC group, and were associated with protection from HPV infection (p = 0.0015; OR = 0.29 CI95% = 0.13-0.61; p = 0.0005; OR = 0.45 CI95% 0.29-0.7, respectively). Individuals from the control group could have previously had HPV infection that was spontaneously eliminated; however, it was undetectable at the time of sample collection. Based on our findings, we hypothesize that the IL-8 +396 G>T polymorphism could interfere with susceptibility to HPV infection, by modulating the ability of immune system to fight the virus.

CCR2 and CCR5 genes polymorphisms in women with cervical lesions from Pernambuco, Northeast Region of Brazil: a case-control study

Polymorphisms in chemokine receptors play an important role in the progression of cervical intraepithelial neoplasia (CIN) to cervical cancer (CC). Our study examined the association of CCR2-64I (rs1799864) andCCR5-Δ32 (rs333) polymorphisms with susceptibility to develop cervical lesion (CIN and CC) in a Brazilian population. The genotyping of 139 women with cervical lesions and 151 women without cervical lesions for the CCR2-64I and CCR5-Δ32 polymorphisms were performed using polymerase chain reaction-restriction fragment length polymorphism. The individuals carrying heterozygous or homozygous genotypes (GA+AA) for CCR2-64I polymorphisms seem to be at lower risk for cervical lesion [odds ratio (OR) = 0.37, p = 0.0008)]. The same was observed for the A allele (OR = 0.39, p = 0.0002), while no association was detected (p > 0.05) with CCR5-Δ32 polymorphism. Regarding the human papillomavirus (HPV) type, patients carrying the CCR2-64Ipolymorphism were protected against infection by HPV type 16 (OR = 0.35, p = 0.0184). In summary, our study showed a protective effect ofCCR2-64I rs1799864 polymorphism against the development of cervical lesions (CIN and CC) and in the susceptibility of HPV 16 infection.

Chlamydia infection in patients with and without cervical intra-epithelial lesions tested by real-time PCR vs. direct immunofluorescence.

This study compares the detection rates of Chlamydia trachomatis by two techniques, direct immunofluorescence (IMF) and real time polymerase chain reaction (PCR), in patients with and without intra-epithelial cervical lesions (SIL) in Recife. We conducted a transversal study involving 35 women with SIL and 35 without SIL attended at Ambulatório Especializado da Mulher, Recife, Brazil. They were tested for Chlamydia trachomatis using two techniques, direct IMF or real time PCR. The rates of Chlamydia trachomatis detection were compared and the association with intra-epithelial cervical lesions was determined using the chi-square test at a 5% level of significance. Concordance between the tests was evaluated using kappa. The global prevalence of Chlamydia infection was 47.1% by direct IMF and 58.6% by real time PCR. A significant association was observed between Chlamydia diagnosis and presence of intra-epithelial cervical lesions, with about 80% positive results by direct IMF and 77.1% by real time PCR. However, the detected rate of infection with Chlamydia trachomatis was significantly greater in patients without intra-epithelial cervical lesions tested by real time PCR (40%) when compared to direct IMF (14.3%). The concordance between the tests was weak, with a kappa coefficient of 0.4. Both real time PCR and direct IMF detected elevated rates of Chlamydia infection in patients with intra-epithelial cervical lesions (80%) but the tests were discordant when patients without cervical lesions were tested, possibly because sensitivity of real time PCR is greater.

Analysis of the polymorphism of osteoprotegerin by polymerase chain reaction in patients with type 2 diabetes and the association with periodontal condition / Análise de polimorfismos da região promotora T/C950 da osteoprotegerina através da reação em cadeia de polimerase em pacientes diabéticos tipo 2 e com periodontite

Objective: The aim of this paper was to analyze the presence of polymorphism in the promoter region T/C950 of the osteoprotegerin gene and its distribution in diabetic patients with periodontitis, when compared to the control group. Methods: 67 patients took part in the research. The test group (n = 32) was composed of diabetic patients with periodontitis and the control group (n = 35) included patients without diabetes and without periodontitis. For the diagnosis of periodontitis, the following clinical parameters were evaluated: probing depth, bleeding on probing and clinical attachment level. The DNA to investigate the polymorphisms of osteoprotegerin, obtained through the technique of polymerase chain reaction, was obtained from the blood serum of the participants.Results: Polymorphisms of osteoprotegerin were found in promoter region -950T/C but there was no significance (p=1.000). Only the control group showed significant results for the probing depth according to the polymorphic region. Conclusion: No influence was found between genetic polymorphisms of osteoprotegerin in patients with diabetes and periodontitis.

Objetivo: Analisar a presença de polimorfismos na região promotora T/C950 do gene da osteoprotegerina, e a sua distribuição em pacientes diabéticos e com periodontite, quando comparados ao grupo controle saudável. Métodos: A pesquisa contou com a participação de 67 indivíduos distribuídos em um grupo teste (n=32), constituído por pacientes diabéticos e com periodontite, e um grupo controle (n=35) que incluía pacientes não diabéticos e sem periodontite. Para o diagnóstico da periodontite, foram avaliados os parâmetros clínicos: profundidade de sondagem, sangramento à sondagem e nível de inserção clínica. O DNA para a investigação dos polimorfismos da osteoprotegerina, através da técnica da reação em cadeia de polimerase convencional, foi obtido a partir de amostras sanguíneas dos participantes. Resultados: Polimorfismos no gene da osteoprotegerina foram encontrados na posição -950T/C da região promotora, porém sem significância estatística (p=1,000). Apenas o grupo controle apresentou resultados significativos para a profundidade de sondagem segundo a região polimórfica (p=0,017).Conclusão: Não foi observada influência entre o polimorfismo da região T/C950 do gene da osteoprotegerina em pacientes com periodontite e a diabetes mellitus.

Comparação de critérios de diagnóstico clínico de periodontite em diabéticos / Comparison of clinical diagnostic criteria of periodontitis in diabetics

Objetivo: A diversidade de critérios de diagnóstico clínico para periodontite em trabalhos científicos tem dificultado a determinação adequada da frequência desta doença e diminuído a possibilidade de comparações entre os estudos. O objetivo foi comparar cinco critérios encontrados na literatura que a associam a profundidade de sondagem (PS) e a perda de inserção clínica (PIC) na determinação da doença periodontal. Material e Métodos: 92 pacientes diabéticos foram submetidos à avaliação periodontal, sendo registrado medidas de PS e PIC em seis sítios em cada dente, e sangramento a sondagem. Com os dados registrados foi calculada a frequência da doença periodontal aplicando os diferentes critérios e posteriormente foi avaliado a concordância entre 5 critérios, utilizando o índice de Kappa (K). Resultados: A frequência de casos de periodontite para os critérios I (PIC ≥ 5 mm em 4 ou mais sítios, e pelo menos um deles com PS ≥ 4 mm), II (PS ≥ 4 mm e PIC ≥ 4 mm em pelo menos um sítio), III (4 ou mais dentes com pelo menos 1 sítio com PS ≥ 4 mm e PIC ≥ 3 mm), IV (2 ou mais dentes com pelo menos 1 sítio PS ≥ 4 mm e PIC ≥ 3mm) e V (PIC ≥ 6 mm em 2 ou mais dentes e PS ≥ 5 mm em 1 ou mais sítios) foi de 59,8%, 67,4%, 51,1%, 56,5% e 57,6%, respectivamente. A concordância entre os critérios de diagnósticos clínicos pelo Kappa variou entre 0,47 (47%) e 0,89 (89%). Conclusão: A frequência de distribuição da periodontite em pacientes diabéticos utilizando diversos critérios de diagnóstico não apresentou grande variação, e a concordância foi considerada entre razoável a boa e excelente. (AU)

Objective: The variability of clinical diagnostic criteria for periodontitis in scientific papers has difficulted the proper determination of the frequency of this disease and decreases the possibility of comparisons between studies. The aim of this study was to compare five criteria found in the literature associated with probing depth (PD) and clinical attachment loss (CAL) in the determination of periodontal disease. Methods: 92 diabetic patients underwent periodontal evaluation, and measures of PD and CAL was performed at six sites on each tooth, and bleeding on probing. With the recorded data the frequency of periodontal disease was calculated by applying different criteria and was subsequently evaluated the correlation between 5 criteria, using the Kappa index (K). Results: The frequency of cases of periodontitis for the criteria I (CAL ≥ 5 mm in 4 or more sites, with at least one site PD ≥ 4 mm), II (PD ≥ 4 mm and CAL ≥ 4 mm in at least one site), III (4 or more teeth with at least one site PD ≥ 4 mm and CAL ≥ 3 mm), IV (2 or more teeth with at least one site PD ≥ 4 mm e CAL ≥ 3mm) e V (CAL ≥ 6 mm in 2 or more teeth and PD ≥ 5 mm in one or more sites) was 59.8%, 67.4%, 51.1%, 56.5% and 57.6%, respectively. The agreement between the criteria for clinical diagnosis by Kappa ranged from 0.47 (47%) and 0.89 (89%). Conclusion: The frequency distribution of periodontitis in diabetic patients using various diagnostic criteria did not show great variation, and the agreement was considered reasonable to good and excelente (AU)

Mannose binding lectin genes (MBL2) polymorpshisms and the periodontal disease in diabetic patients / Polimorfimos da lectina de ligação da manose (MBL2) e doença periodontal em pacientes diabéticos

PURPOSE: To assess the association between the polymorphism in exon-1 of the MBL2 gene and the periodontal disease in type 2 diabetic patients. METHODS: The sample comprised of 100 patients, who were submitted to a clinical periodontal examination that evaluated in six sites per tooth the probing depth (PD), bleeding on probing (BOP), clinical attachment loss (CAL), plaque index (PI) and the number of teeth present. Periodontal disease was defined as at least four sites with loss of attachment of >5 mm, with one or more of those sites having a pocket of > 4 mm. The collection of scaling cells from the oral mucosa was carried out and the detection of MBL2 polymorphism was made by real time PCR and melting temperature curve analysis. RESULTS: In a type 2 diabetic population, no significant statistical differences in MBL2 polymorphisms genotype or allele frequencies were observed among subjects with periodontal disease. CONCLUSION: This study indicates that the polymorphisms in exon-1 of the MBL2 gene are not related to periodontal disease in a type 2 diabetic population.

OBJETIVO: Avaliar a associação entre o polimorfismo no exon-1 do gene MBL2 e a doença periodontal em pacientes diabéticos tipo 2. MÉTODO: A amostra foi composta por 100 pacientes que foram submetidos a um exame clínico periodontal que avaliou seis sítios por dente a profundidade de sondagem (PS), sangramento à sondagem (SS), perda de inserção clínica (PIC), índice de placa (IP) e o número de dentes presente. A doença periodontal foi definida como pelo menos quatro sítios com perda de inserção de >5mm, com um ou mais destes sítios tendo uma bolsa de >4mm. Foram coletadas células de descamação da mucosa oral e a detecção do polimorfismo foi feita através da PCR em tempo real e análise da temperatura de melting. RESULTADOS: Em uma população de diabéticos tipo 2, não houve diferenças estatisticamente significantes nos genótipos do polimorfismo da MBL2 ou freqüência alélica observadas entre os indivíduos com doença periodontal. CONCLUSÃO: Este estudo indicou que o polimorfimos no exon-1 do gene da MBL2 não foi relacionado à doença periodontal em uma população de diabéticos tipo 2.

Condição Periodontal e Marcadores do Controle Metabólico em Pacientes Diabéticos / Periodontal Conditions and Metabolic Control Markers in Diabetic Patients

Objetivo: Avaliar a condição periodontal dos pacientes diabéticos tipo 2, relacionando com o controle de marcadores metabólicos.Método: Para este estudo foram selecionados 92 pacientes diabéticos que recebiam tratamento em dois centros de referência em Recife, Brasil. Foram realizados exames clínicos periodontais em seis sítios de cada dente, avaliando a profundidade de sondagem, sangramento à sondagem, perda de inserção, índice de placa dentária e número de dentes presentes. A doença periodontal foi definida pela presença de mais de 4 sítios com perda de inserção ≥ 5 mm, sendo um ou mais destes sítios com profundidade de sondagem de 4 mm ou mais. Foram realizados exames hematológicos para avaliar os marcadores metabólicos (hemoglobina glicosilada, glicemia de jejum, triglicérides, colesterol total, colesterol HDL e LDL). Foi realizada análise estatística bi-variada para verificar a associação entre as variáveis em estudo.Resultados: A avaliação da condição periodontal dos pacientes avaliados mostrou que 59,8% dos pacientes diabéticos eram portadores de doença periodontal, caracterizada por periodontite. A média de idade observada foi de 54,8 anos com DP = 9,3, o sangramento gengival à sondagem e o índice de placa foram de 33,8% e 61,07%, respectivamente. A maioria dos pacientes com níveis elevados de glicemia (≥ 126 mg / dL) apresentaram doença periodontal (62,3%),o perfil lipídico dos pacientes com doença periodontal se mostrou controlado.Conclusão: Não foi observada associação entre a condição periodontal e os marcadores do controle metabólico dos pacientes diabéticos

Objective: Evaluate the periodontal condition in diabetic patients type 2, relating to markers of metabolic control.Methods: 92 diabetic patients who received treatment in two centers in Recife, Brazil, had been invited to participate in this study. It was carried through periodontal clinical examinations in six sites of each tooth, evaluating the depth probing, bleeding probing, attachment loss, dental plaque and the number of teeth present. Periodontal disease was defined as the presence of 4 + sites with attachment loss of ≥ 5 mm with one or more than these sites with depth probing of 4 + mm. Hematologic examinations were carried out to evaluate the metabolic markers (Glycosylated Hemoglobin, fasting blood glucose, triglycerides, total cholesterol, cholesterol HDL and LDL). Bi-variate analysis were used to verified the association between variables.Results: Evaluation of periodontal condition in the sample have shown 59.8% of diabetic patients had periodontal diseases, characterized as periodontitis. The average of age observed was 54.8 years with DP= 9.3, average of bleed on probing and plaque index were 33.8% and 61.07%, respectively. The majority of patients with raised glicemic levels (≥126 mg/dL) had presented DP (62,3%), in relation to the lipidic profile the patients with periodontal disease had been presented controlled. Conclusion: In the studied population there were no signs of any association between the periodontal condition and the markers of metabolic control

Prevalência dos genótipos do papilomavírus humano: comparação entre três métodos de detecção em pacientes de Pernambuco, Brasil / Prevalence of human papillomavirus genotypes: comparison between three detection methods in patients of Pernambuco, Brazil

OBJETIVO: comparar três métodos para detecção do HPV e determinar a prevalência dos genótipos encontrados. MÉTODOS: um total de 120 amostras de raspagem da região cervical de mulheres portadoras de neoplasia intraepitelial cervical foram analisadas pela reação em cadeia da polimerase convencional, usando os sistemas de primers MY09/11, GP05+/06+ e pela Nested-PCR. As amostras foram submetidas à extração de DNA e, logo após, amplificadas com os primers GH20 e PC04 (β-globina) para verificação da qualidade do DNA obtido e pela reação em cadeia da polimerase convencional e Nested-PCR. Os fragmentos amplificados foram visualizados em gel de agarose a 1,2 por cento, corados com Blue Green Loading Dye I. As amostras positivas foram sequenciadas usando o sequenciador automático de DNA "MegaBACE 1000". Para análise estatística foram utilizados os teste do Χ2 e o de Fisher com nível de significância de 5 por cento. RESULTADOS: quinze amostras não se amplificaram para os primers de β-globina, sendo eliminadas do estudo. Das amostras restantes, 40 por cento (42/105) foram positivas para os primers MY09/11, 98 por cento (103/105) para os primers GP05+/06+ e 92 por cento (97/105) para Nested-PCR. Considerado as técnicas MY09/11 e GP05+/06+, foi possível observar 100 por cento de amostras positivas para o HPV. Neste estudo, a prevalência dos genótipos foi de 58, 23, 5, 4 e 3 por cento para HPVs 16, 18, 31, 33 e 56, respectivamente. Os HPV 67 e 83 apresentaram 2 por cento e os HPV 6, 11, 58 e candHPV85, 1 por cento cada. A prevalência dos genótipos neste estudo está de acordo com o reportado em todo o mundo (IC95 por cento=0,4657-0,8976). CONCLUSÕES: para obter resultados mais confiáveis, é necessário o uso de mais que um sistema de primers para detecção do HPV. Acredita-se que as três técnicas estudadas são importantes e adequadas para o diagnóstico clínico do HPV quando apropriadamente combinadas.

PURPOSE: to compare three methods for the detection of HPV infection and to determine the prevalence of the genotypes found. METHODS: a total of 120 cervical scrape samples from patients with cervical intraepithelial neoplasia were analyzed by the conventional polymerase chain reaction using the MY09/11 and GP05+/06+ primers, and by the Nested polymerase chain reaction. The samples were subjected to DNA amplification with the GH20 and PC04 primers (β-globin) to verify DNA quality and also by polymerase chain reaction and Nested polymerase chain reaction. The amplicons were visualized in 1.2 percent agarose gel stained with Blue Green Loading Dye I. Positive samples also were sequenced using the automatic DNA sequencer "MegaBACE 1000". The Χ2 and Fisher tests were used for statistical analysis with the level of significance set at 5 percent. RESULTS: fifteen samples were eliminated from the study because they failed to amplify the β-globin gene. Of the remaining samples, 40 percent (42/105) were positive using primers MY09/11, 98 percent (103/105) using primers GP05+/06+, and 92 percent (97/105) using Nested-PCR. With the MY09/11 and GP05+/06+ techniques, it was possible to obtain 100 percent HPV-positive samples. In this study, the prevalence of the genotypes found was 57, 23, 5, 4 and 3 percent for HPV genotypes 16, 18, 31, 33 and 56, respectively. HPVs 67 and 83 were present in 2 percent, and genotypes 6, 11, 58 and candHPV85 were present in 1 percent each. The prevalence of the more common genotypes (HPV 16 and 18) in this study agrees with that reported worldwide (IC95 percent=0.4657-0.8976). CONCLUSIONS: to obtain more reliable results, it is necessary the use of more than one primer system to detect HPV infections. We believe that the three techniques studied are important and suitable for the clinical diagnosis of HPV, when they are appropriately combined.

Polimorfismos genéticos e o insucessodos implantes dentários: estudo piloto / Genetic polymorphism and failure ofdental implants: a pilot study

Objetivo: verificar a relação entre polimorfismos genéticos ligados às citocinas da remodelação óssea osteoprotegerina (OPG) e o ligante do ativador do receptor do fator nuclear kappa ß(RANKL), além de relacionar a presença de polimorfismos em OPG e RANKL com o insucesso clínico dos implantes dentários ao longo do tempo e determinar quais combinações de polimorfismos de OPG e RANKL estão associados ao insucesso de implantes dentários. Métodos: vinte pacientes, de ambos os sexos, maiores de 18 anos, reabilitados com 34 implantes dentários, foram avaliados por um período de 24 meses após a instalação das coroas sobre implantes. Após os exames clínico e radiográfico, os pacientes foram incluídos no grupo de insucesso dos implantes caso um ou mais do seguintes critérios fossem encontrados: mobilidade, queixas subjetivas persistentes, infecção peri-implantar recorrente com supuração, radiolucência contínua ao redor do implan- te, profundidade de sondagem ≥ 5mm e sangramento à sondagem. Foi coletado sangue periférico para a análise do polimorfismo das citocinas OPG e RANKL por meio da reação em cadeia da polimerase (PCR). Resultados: não houve diferença estatisticamente significativa entre o grupo de falha dos implantes em relação aos genótipos de OPG e RANKL. Conclusão: os polimorfismos de OPG e RANKL não influenciaram no insucesso dos implantes dentários na amostra avaliada.

Objective: To investigate the relationship between genetic polymorphisms related to cytokinesin bone remodeling osteoprotegerin (OPG) and receptor activator of nuclear factor kappa ß ligand (RANKL). It also aims to relate the presence of OPG and RANKL polymorphisms with clinicalfailure of dental implants over time and to determine which combinations of OPG and RANKL polymorphisms are associated with failure of dental implants. Methods: Twenty patients of both sexes, over eighteen years, rehabilitated with thirty-four dental implants were evaluated during 24months after implant-supported crown placement. After clinical and radiographic examination, patients were included in a failure group if one or more of the following criteria were identified: mobility, persistent subjective complaints, recurrent peri-implantitis with suppuration, continuous radiolucency around the implant, probing depth ≥ 5 mm and bleeding on probing. Peripheral blood was collected for analysis of cytokine OPG and RANKL polymorphisms by polymerase chain reaction (PCR). Results: There were no statistically significant differences between the failure group in relation to genotypes OPG and RANKL. Conclusion: OPG and RANKL polymorphisms didnot influence dental implants failure in the investigated sample.

Imunidade inata nas doenças periodontais: [revisão] / Innate immunity in the periodontal disease: [review]

Sabe-se que o fator etiológico primário das doenças periodontais é o biofilme bacteriano, e, todos os indivíduos que tiverem acúmulo de bactérias na superfície dental próxima à margem gengival, vão após algum tempo apresentar sinais de inflamação gengival. Entretanto, a magnitude da progressão da doença, isto é, o quanto e em que velocidade ela vai destruir o periodonto, é dependente da patogenicidade dos microorganismos da placa e da capacidade de defesa do hospedeiro através de seu sistema imune. O presente trabalho teve como objetivo, através de uma revisão da literatura, apresentar os aspectos envolvidos na resposta imune do hospedeiro, em particular, na resposta imune inata, relacionando-a com o desenvolvimento da doença periodontal. Pode-se concluir que a manifestação da doença peiodontal em sua forma destrutiva está intimamente relacionada aos mecanismos imunológicos do hospedeiro, determinando sua susceptibilidade e vaiações nas respostas do organismo dentro do processo patogênico.

It is known that the primary etiological factor of periodontal diseases is the bacterial biofilm and all individuals who have accumulation of bacteria in dental surface near the gingival margin will show signs of gingival inflammation after some time. However, the magnitude of progression of the disease, which is, how much and in what speed it will destroy the periodontal tissue support, it is dependent on the pathogenicity of microorganisms plaque and the ability to defend of the host through your immune system. This study aimed to, through a review of the literature, present the issues involved in the immune response of the host, in particular, the innate immune response, listing it with the developmente of the periodontal disease. It is concluded that the expression of the periodontal disease in its destructive way, is closely related to immunological mechanisms of the host, determining their susceptibility and variations in the responses of the organism in the pathogenic process.

Principles and applications of polymerase chain reaction in medical diagnostic fields: a review: [revision] / Princípios e aplicações da reação em cadeia de Polimerase em áreas médicas diagnósticas: uma revisão: [revisão]

Recent developments in molecular methods have revolutionized the detection and characterization of microorganisms in a broad range of medical diagnostic fields, including virology, mycology, parasitology, microbiology and dentistry. Among these methods, Polymerase Chain Reaction (PCR) has generated great benefits and allowed scientific advancements. PCR is an excellent technique for the rapid detection of pathogens, including those difficult to culture. Along with conventional PCR techniques, Real-Time PCR has emerged as a technological innovation and is playing an ever-increasing role in clinical diagnostics and research laboratories. Due to its capacity to generate both qualitative and quantitative results, Real-Time PCR is considered a fast and accurate platform. The aim of the present literature review is to explore the clinical usefulness and potential of both conventional PCR and Real-Time PCR assays in diverse medical fields, addressing its main uses and advances.

O advento dos métodos moleculares tem, nos últimos anos, revolucionado a detecção e caracterização dos microorganismos em diversas áreas médicas diagnósticas, tais como virologia, micologia, parasitologia, microbiologia e odontologia. Dentre as técnicas baseadas em biologia molecular, a PCR (Polymerase Chain Reaction) trouxe enormes benefícios e desenvolvimentos científicos, se mostrando como um excelente caminho para a rápida detecção de patógenos, até mesmo aqueles de difícil cultivo. Derivada da PCR convencional, a PCR em Tempo Real se mostra como uma inovação tecnológica e vem conquistando espaço nos diagnósticos clínicos e nos laboratórios de pesquisa por apresentar a capacidade de gerar, além de resultados qualitativos, resultados quantitativos, se mostrando de forma mais rápida e precisa. Este trabalho de revisão tem por objetivo explorar a utilidade clínica da técnica de PCR convencional e em Tempo real nas diversas áreas médicas supracitadas, abrangendo seus principais usos e avanços, direcionando para o cotidiano profissional.

Chlamydia infection in patients with and without cervical intra-epithelial lesions tested by real-time PCR vs. direct immunofluorescence

This study compares the detection rates of Chlamydia trachomatis by two techniques, direct immunofluorescence (IMF) and real time polymerase chain reaction (PCR), in patients with and without intra-epithelial cervical lesions (SIL) in Recife. We conducted a transversal study involving 35 women with SIL and 35 without SIL attended at Ambulatório Especializado da Mulher, Recife, Brazil. They were tested for Chlamydia trachomatis using two techniques, direct IMF or real time PCR. The rates of Chlamydia trachomatis detection were compared and the association with intra-epithelial cervical lesions was determined using the chi-square test at a 5 percent level of significance. Concordance between the tests was evaluated using kappa. The global prevalence of Chlamydia infection was 47.1 percent by direct IMF and 58.6 percent by real time PCR. A significant association was observed between Chlamydia diagnosis and presence of intra-epithelial cervical lesions, with about 80 percent positive results by direct IMF and 77.1 percent by real time PCR. However, the detected rate of infection with Chlamydia trachomatis was significantly greater in patients without intra-epithelial cervical lesions tested by real time PCR (40 percent) when compared to direct IMF (14.3 percent). The concordance between the tests was weak, with a kappa coefficient of 0.4. Both real time PCR and direct IMF detected elevated rates of Chlamydia infection in patients with intra-epithelial cervical lesions (80 percent) but the tests were discordant when patients without cervical lesions were tested, possibly because sensitivity of real time PCR is greater.