ABSTRACT
BACKGROUND: Paracoccidioidomycosis (PCM) is a systemic fungal infection caused by Paracoccidioides brasiliensis (Pb) and associated with deficient cellular immune response, which is modulated by inflammatory cells, mainly macrophages, and cytokines. Recently, the comprehension of the macrophage polarization mediated by Th1 and Th2 cytokines has contributed to elucidate the immune response that takes part in some diseases. Thus, the aim of this study was to assess the presence of Th1- and Th2-immune response and also Pb counting in oral lesions of chronic PCM. METHODS: Forty-eight cases of chronic PCM oral lesions were included. All cases were classified as loose or dense granulomas. S100 protein, IL-1ß, IL-6, TNF-α, CD163 and CD68 immunoexpressions, and Pb localization were evaluated. The fungi present in the tissue were quantified by anti-Pb antibody. RESULTS: Most patients were white men with mean age of 47 years old and showed higher incidence of multiple lesions. Loose granulomas were predominant and exhibited a great amount of M2 macrophages, which were visualized with anti-CD163 antibody. The expression for CD163 and CD68 was similar (P = 0.05), highlighting the predominance of M2 macrophages in PCM. IL-1ß, IL-6, and TNF-α immunoexpression did not significantly change with CD163, CD68, and S100 protein. The number of fungi was significantly higher in cases with intense IL-1ß immunoexpression (P = 0.003). CONCLUSIONS: M2-activated macrophages were the majority among inflammatory cells in chronic PCM, characterizing the action of a Th2-immune response. Nevertheless, Th1 cytokines were also found; mainly IL-1ß, which was associated with fungi counting in oral lesions.
Subject(s)
Macrophages/immunology , Mouth Diseases/immunology , Mouth Diseases/microbiology , Paracoccidioides/immunology , Paracoccidioidomycosis/immunology , Adult , Aged , Aged, 80 and over , Antigens, CD/immunology , Antigens, Differentiation, Myelomonocytic/immunology , Chronic Disease , Cytokines/immunology , Cytokines/metabolism , Female , Granuloma/immunology , Granuloma/microbiology , Granuloma/pathology , Humans , Interleukin-1beta/immunology , Interleukin-6/immunology , Macrophages/metabolism , Macrophages/pathology , Male , Middle Aged , Mouth Diseases/pathology , Paracoccidioidomycosis/microbiology , Paracoccidioidomycosis/pathology , Receptors, Cell Surface/immunology , S100 Proteins/immunology , Tumor Necrosis Factor-alpha/immunology , Young AdultABSTRACT
No Brasil, as leishmanioses representam um importante problema de saúde pública pela sua elevada incidência, ampla distribuição geográfica e marcante aumento na transmissão associados à urbanização da doença. Cães são considerados os principais reservatórios da leishmaniose visceral (LV) no ambiente urbano, tornando-se fundamental o aprimoramento do diagnóstico dadoença nessa espécie. Este estudo objetivou a avaliação da técnica de imuno-histoquímica (IHQ) para o diagnóstico de Leishmania spp. em amostras de diferentes tecidos de cães recebidas no Núcleo de Patologia Quantitativa do Instituto Adolfo Lutz (NPQ-IAL). Amostras de tecidos coletadas de 134 cães, provenientes de municípios do Estado de São Paulo, positivos para LV por teste rápido(TR DPP®-Bio-Manguinhos), ensaio imunoenzimático-EIE/Elisa (Bio-Manguinhos) e pela reação em cadeia de polimerase, foram submetidas a marcação IHQ específica, em duplicata. A reação de IHQ apresentou sensibilidade de 98,51 %, especificidade de 100,00 % e acurácia de 98,61 %. A maior positividade foi detectada nas amostras de baço. O índice Kappa foi de 0,84 entre os resultados da análise dos tecidos em duplicata. A técnica de IHQ pode ser utilizada como uma técnica rotineirapara o diagnóstico das leishmanioses caninas, sendo padronizada e implantada no NPQ-IAL.
In Brazil, leishmaniasis represents an important public health problem due to its high incidence, wide geographic distribution and marked increase in transmission associated to the urbanization of the disease. Dogs are considered the main reservoirs of visceral leishmaniasis (VL) in the urban environment, making it fundamental to improve the diagnosis of this species. This study aimed at the evaluation of the immunohistochemical technique (IHC) for the diagnosis of Leishmania spp. In samples of different dog tissues received at the Adolfo Lutz Institute's Quantitative Pathology Center (NPQ-IAL). Tissue specimens collected from 134 dogs, from municipalities in the State of São Paulo, positive for LV by rapid test (TR DPP®-Bio-Manguinhos), enzyme-linked immunosorbent assay (EIE / Elisa (Bio-Manguinhos) and the chain reaction of Polymerase, were subjected to specific IHQ labeling, in duplicate. The IHQ reaction showed sensitivity of 98.51%, specificity of 100.00% and accuracy of 98.61%. The highest positivity was detected in spleen samples. The Kappa index was 0.84 between the results of the tissue analysis in duplicate. The IHC technique can be used as a routine technique for the diagnosis of canine leishmaniasis, being standardized and implanted in the NPQ-IAL.
Subject(s)
Animals , Dogs , Immunohistochemistry , Leishmania , Leishmaniasis, Visceral/diagnosis , Leishmaniasis, Visceral/pathologyABSTRACT
In Brazil, leishmaniasis represents a major public health problem due to its high incidence, wide distribution and remarkable increase in transmission associated with the disease urbanization. Dogs are considered as the main reservoirs of visceral leishmaniasis (VL) in the urban environment and diagnosis improvement was fundamental in this species. This study aimed at evaluating the immunohistochemical technique (IHC) for diagnosing Leishmania spp. in differents tissues samples collected from dogs. This investigation was performed at the Quantitative Pathology Center of the Institute Adolfo Lutz (NPQ-IAL). Tissue samples collected from 134 dogs, derived from municipalities of São Paulo State, positive for VL by rapid testing (TR DPP® - Bio-Manguinhos), enzyme-linked immunosorbent assay (EIA/ELISA - Bio-Manguinhos), and polymerase chain reaction, were analyzed by specific IHC in duplicate. These dogs were previously diagnosed as VL. IHC showed sensitivity of 98.51 %, specificity of 100.00 %, positive predictive value of 100.00 %, negative predictive value of 83.33 % and accuracy of 98.61 %. The major positivity was detected in spleen samples. Kappa index between the tissues duplicate results was of 0.84. IHC technique, standardized and implemented at NPQ-IAL, showed to be suitable to be used as routine test for diagnosing canine leishmaniasis effectivelly.
No Brasil, as leishmanioses representam um importante problema de saúde pública pela sua elevada incidência, ampla distribuição geográfica e marcante aumento na transmissão associados à urbanização da doença. Cães são considerados os principais reservatórios da leishmaniose visceral (LV) no ambiente urbano, tornando-se fundamental o aprimoramento do diagnóstico da doença nessa espécie. Este estudo objetivou a avaliação da técnica de imuno-histoquímica (IHQ) para o diagnóstico de Leishmania spp. em amostras de diferentes tecidos de cães recebidas no Núcleo de Patologia Quantitativa do Instituto Adolfo Lutz (NPQ-IAL). Amostras de tecidos coletadas de 134 cães, provenientes de municípios do Estado de São Paulo, positivos para LV por teste rápido (TR DPP®-Bio-Manguinhos), ensaio imunoenzimático-EIE/Elisa (Bio-Manguinhos) e pela reação em cadeia de polimerase, foram submetidas a marcação IHQ específica, em duplicata. A reação de IHQ apresentou sensibilidade de 98,51 %, especificidade de 100,00 % e acurácia de 98,61 %. A maior positividade foi detectada nas amostras de baço. O índice Kappa foi de 0,84 entre os resultados da análise dos tecidos em duplicata. A técnica de IHQ pode ser utilizada como uma técnica rotineira para o diagnóstico das leishmanioses caninas, sendo padronizada e implantada no NPQ-IAL.
Subject(s)
Animals , Dogs , Immunohistochemistry/veterinary , Leishmania/isolation & purification , Leishmaniasis, Visceral/diagnosis , Leishmaniasis, Visceral/veterinary , Enzyme-Linked Immunosorbent Assay/veterinary , Polymerase Chain Reaction/veterinary , Clinical Laboratory Techniques/veterinaryABSTRACT
Os avanços tecnológicos das últimas décadas catalisaram transformações sociais e econômicas, que influenciaram decisivamente nos padrões da morbimortalidade populacional. No Brasil, a heterogeneidade deste padrão é muito visível e complexa, em função de sua grande extensão territorial, do significativo número de habitantes e das diferenças socioeconômicas e culturais. Com o aumento da expectativa devida e o envelhecimento da população brasileira, observa-se o aparecimento cada vez mais frequente de doenças crônicas não transmissíveis. As mudanças climáticas e as condições higiênico-sanitárias, ainda deficientes em algumas regiões, podem propiciar o recrudescimento das doenças infectocontagiosas. Em face dessas transformações, faz-se necessário que o sistema de vigilância epidemiológica seja reestruturado para adequar aos novos cenários epidemiológicos, identificar novos riscos, prever e conter a expansão de áreas com riscos preexistentes de disseminação, propagação e redução das doenças. Um dos pontos cruciais desta reestruturação é o armazenamento correto e adequado das amostras biológicas, por meio da criação de biorrepositórios e biobancos, que possibilitará a aplicação de novas tecnologias para detecção, investigação e respostas às situações de surtos, epidemias e pandemias, com benefícios à pesquisa e assistência na área de saúde. Esta revisão demonstra a importância dos biobancos e biorrepositórios em Saúde Pública.
Subject(s)
Biological Specimen Banks , Tissue Banks , Molecular Biology , Communicable Disease Control , Public Health , Epidemiological MonitoringABSTRACT
Neste trabalho foram identificadas as associações multivariadas entre as alterações citológicas (AC) do colo uterino e o comportamento de risco das profissionais do sexo (PS) atendidas no Centro de Referência em Saúde Sexual e Reprodutiva (CRESSER) Sumaré, SP. Das 90 PS participantes, foram coletados dados sociocomportamentais por meio de questionário e amostras cérvico-vaginais para realização do exame de Papanicolaou. Para o teste estatístico foi utilizada a Análise Hierárquica de Agrupamentos (AHA). AC foram detectadas em 33 PS (36,7 por cento), sendo 18 (20,0 por cento) atipias de significado indeterminado possivelmente não neoplásica (ASC-US), quatro (4,4 por cento) atipias de significado indeterminado sem excluir lesão de alto grau (ASC-H), sete (7,8 por cento) lesão intraepitelial de baixo grau (LSIL) e quatro (4,4 por cento) lesão intraepitelial de alto grau (HSIL). Comparando os dados das PS com citologia normal e alterada, houve diferenças estatisticamente significativas quanto ao tabagismo, início da atividade sexual, relações com pessoas do mesmo sexo e detecção de Gardnerella vaginalis. AHA classificou as PS em quatro grupos distintos, em que as PS com idade média superior apresentaram maior proporção de DST, HIV, AC e pouco uso do preservativo. As PS com menor idade média mostraram menor frequência de AC, DST, atividade sexual precoce e HIV negativo.
Subject(s)
Humans , Female , Risk-Taking , Cell Biology , Cervix Uteri/cytology , Sexually Transmitted Diseases , Sex WorkersABSTRACT
O objetivo deste estudo foi analisar os dados preliminares da detecção do Epstein-Barr Virus (EBV) pela reação de hibridização in situ cromogênica (CISH) nas amostras encaminhadas ao Núcleo de Patologia Quantitativa/Instituto Adolfo Lutz. Os 12 casos com suspeita da presença do EBV foram recebidos no período de outubro de 2011 a julhode 2012, totalizando 14 amostras, sendo que quatro casos apresentavam resultados prévios pelo estudo imuno-histoquímico (IHQ): três negativos e um positivo para o EBV LMP-1. Foi utilizado o Zytofast EBV Probe e o ZytoFast Plus CISH Implementation Kit HRP DAB adaptado às condições do laboratório. Apositividade para EBER (Epstein-Barr Encoded Early RNAs) do EBV foi observada em 10 amostras, inclusive nos três casos previamente negativos pelo exame IHQ. Concluímos, neste estudo preliminar, que a reação de CISH permitiu detectar sequências gênicas do EBV, mesmo em amostras negativas para proteína latente de membrana (LMP-1) pela reação IHQ
Subject(s)
In Situ Hybridization , Immunohistochemistry , LymphomaABSTRACT
Introdução: O xilol é necessário para a diafanização de amostras citopatológicas. Pode trazer problemas de ordem ocupacional, proteção ao meio ambiente e de custos. Objetivo:Avaliar o desempenho de várias concentrações da mistura verniz/xilol em relação à diafanização e conservação em amostras de citologia coradas com a técnica dePapanicolaou (CP). Material e métodos: Foram avaliadas 75 lâminas de raspado bucal (RB) e 8.773 esfregaços cérvico-vaginais (CV). As lâminas foram coradas pela CP, com a retirada das etapas de álcool/xilol e xilol e foram secas à temperatura ambiente porvinte minutos antes da montagem com lamínula. A montagem foi realizada com soluções de verniz/xilol de concentrações 75/25%, 70/30%, 60/40%, 50/50% e 40/60%. As lâminas foram distribuídas de forma aleatória aos profissionais para avaliação da técnica, sendo conceituadas como boas, regulares ou ruins e reavaliadas após nove meses dearquivamento. A concentração de verniz/xilol que apresentou melhor qualidade final nas lâminas de RB foi aplicada também nos CV. Resultados: A concentração de 75/25% deverniz/xilol apresentou melhor desempenho para as duas situações, em RB e CV. Após nove meses, a qualidade foi mantida. Conclusão: A concentração de 75/25% apresentou melhor resultado.
Subject(s)
Humans , Female , Cell Biology , Industrial Oils , Staining and Labeling , Solvents/adverse effects , Solvents/toxicity , Transillumination , Vaginal Smears , Environment , Occupational HealthABSTRACT
No presente estudo foi avaliada a história de realização dos exames citológicos (EC) prévios e posteriores com diagnóstico citológico de células escamosas atípicas (ASC) e sua correlação com os resultados de exames de biópsias. Para tanto, foi realizado o estudo retrospectivo, de 60 casos de EC cérvico-vaginais com diagnóstico de ASC e biópsias correlatas, os exames prévios e posteriores de ASC e a periodicidade de exames realizados. Das 60 análises selecionadas, 57 apresentaram diagnóstico citológico anterior de ASC-US (possivelmente não neoplásico), 30 (52,6%) foram negativos no exame de biópsia, 16 (28,1%) de NIC 1, 11 (19,3%) com diagnósticos discrepantes acima de 2 graus; 3 (100%) casos de ASC-H (não se pode afastar lesão de alto grau) foram NIC 1 na biópsia. Dentre os 60 casos, 44 (73,3%) que tiveram exames anteriores, 14 (23,3%) eram positivos; 53 (88,3%) que fizeram EC posteriores à biópsia, 10 (16,7%) eram positivos. Quanto à periodicidade dos exames, 26 (43,3%) realizaram o exame com intervalo de até 1 ano, 29 (48,3%) de 1 a 3 anos, 1 (1,7%) em intervalo maior que 3 anos. De acordo com os achados observados neste estudo, os diagnósticos citológicos de ASC, quando comparado com os exames da biópsia, estavam mais frequentemente associados às alterações morfológicas reacionais. Estes dados mostram o valioso papel da correlação cito- histológica, uma vez que a execução apenas do seguimento citológico pode ser insuficiente para obter o diagnóstico morfológico. É salientada, ainda, a necessidade de efetuar o exame de acompanhamento de seis em seis meses, para realizar o controle evolutivo dessas atipias, bem como para auxiliar na conduta terapêutica das pacientes.
Subject(s)
Uterine Cervical Dysplasia , Vaginal Smears , Retrospective Studies , Cytological Techniques , Histocytological Preparation TechniquesABSTRACT
No presente estudo foi realizado o levantamento retrospectivo de diagnóstico citopatológico das lesões pré-neoplásicas e neoplásicas pela método de Papanicolaou e sua distribuição por faixa etária das mulheres atendidas no período de 2003 a 2008. Das 222.024 amostras analisadas, 3.674 (1,65%) foram classificadas como insatisfatórias, 206.439 (92,98%) foram negativas e 11.911 (5,36%) apresentaram algum tipo de atipias nucleares. Dentre as alterações epiteliais atípicas, 6.437 (54,04%) foram de ASC-US e 210 (1,76%) de ASC-H; 3.264 (27,40%) de LSIL; 1.279 (10,74%) de HSIL; 131 (1,10%) de SCC; 552 (4,63%) por AGC-US e 23 (0,19%) de AGC-H; 7 (0,06%) de AIS e 8 (0,07%) de ADENOCA. Quanto à ocorrência e distribuição dos diagnósticos de lesões intraepiteliais escamosas e glandulares, de acordo com a faixa etária em intervalos de 5 anos, foi observada maior frequência de diagnóstico de LSIL entre as mulheres mais jovens (15-25 anos) e os diagnósticos de ASC-US, ASC-H, AGC-US, HSIL, SCC e ADENOCA nas mulheres >50 anos de idade. As alterações glandulares, como AGC-H e AIS, ocorreram em mulheres na faixa etária entre 30-34 e 45-49 anos. Foram detectados dois (0,16%) casos de HSIL em pacientes <14 anos e em 51 (3,99%) pacientes na faixa entre 15-19 anos. Os dados da presente avaliação enfatiza a importância de campanhas orientativas, rastreamento e seguimento das pacientes para garantir a melhoria e a efetividade dos programas de prevenção.
Subject(s)
Humans , Female , Vaginal Smears , Uterine Cervical Neoplasms/diagnosis , Mass Screening , Cytological Techniques , Age Distribution , Women's HealthABSTRACT
OBJECTIVE: To evaluate performance and viability of internal quality control (QC) strategies in a public health laboratory of the state of São Paulo. STUDY DESIGN: A retrospective study was performed with 3 QC strategies to improve internal cytologic diagnoses: morphologic guided-list criteria (MGLC), 100% rapid-rescreening (100% RR) of negative slides ("turret" method) and 10% rescreening (10% R) of negative slides. Cases were examined at Adolfo Lutz Institute, São Paulo, Brazil, from 2002 to 2004. Histopathologic results, when available, were considered gold standard; cytologic consensus diagnosis was by 2 pathologists when histologic results were unavailable. RESULTS: MGLC selected 20.7% samples with cytologic atypias, 10% R selected 0.6% and RR selected 2.5%. Cytologic/histologic initial concordance was 57.4%, low-grade squamous intra-epithelial lesion false negative rate was 34.9% and high-grade squamous intraepithelial lesion false negative rate was 12.2%. After diagnosis, consensus concordance was 97.2%. CONCLUSION: The 100% RR and 10% R QC strategies detected more false negative cases in liquid-based cytology than in conventional Pap smears. The 100% RR strategy reduced the false negative results and allowed evaluation of individual staff performance. The 10% R strategy did not offer significant results. We concluded that association of MGLC and 100% RR strategies might improve cytologic diagnostic quality.
Subject(s)
Laboratories/standards , Papanicolaou Test , Quality Assurance, Health Care , Uterine Cervical Dysplasia/pathology , Uterine Cervical Neoplasms/pathology , Vaginal Smears/standards , Brazil , False Negative Reactions , Female , Humans , Predictive Value of Tests , Quality Control , Retrospective StudiesABSTRACT
We compared the performance of the Whole, Turret and Step techniques of 100% rapid rescreening (RR) in detection of false-negatives in cervical cytology. We tested RR performance with cytologists trained and among those without training. We revised 1,000 consecutive slides from women participating in an ongoing international screening trial. Two teams of experienced cytologists performed the RR techniques: one trained in RR procedures and the other not trained. The sensitivities in the trained group were Whole 46.6%, Turret 47.4% and Step 50.9%; and in the non-trained group were 38.6, 31.6 and 47.4%, respectively. The kappa coefficient showed a weak agreement between the two groups of cytologists and between the three RR techniques. The RR techniques are more valuable if used by trained cytologists. In the trained group, we did not observe significant differences between the RR techniques used, whereas in the non-trained group, the Step technique had the best sensitivity.
Subject(s)
Mass Screening/methods , Task Performance and Analysis , Uterine Cervical Diseases/diagnosis , Vaginal Smears/methods , False Negative Reactions , Female , Humans , Reproducibility of Results , Sensitivity and Specificity , Vaginal Smears/standardsABSTRACT
OBJECTIVE: To evaluate the discriminating potential of AgNOR area measurement and count in thyroid tumors using static cytometry equipment. STUDY DESIGN: Slides were analyzed by a computerized system for image analysis, CAS 200 (Becton & Dickinson, U.S.A.), using the Cell Measurement computer program (CAS 200, Becton & Dickinson). The argyrophilic reaction (NORs) was evaluated with a 400-fold amplification directly from the computer monitor. RESULTS: Thirty-three cases were analyzed for AgNOR staining. The cases studied included 3 goiters, 10 follicular adenomas, 6 Hürthle adenomas, 4 follicular carcinomas, 7 papillary carcinomas, and 3 Hürthle carcinomas. A total of 6,600 nuclei were evaluated. For statistical purposes, lesions were classified as benign and malignant, and both the number and the area of counted NORs showed very similar values. The NORs median among 19 benign tumors was 1.484 (SD +/- 0.265) and of 14 malignant tumors was 1.436 (SD +/- 0.414); the NORs areas were 2.6584 (SD +/- 1.0653) and 2.3643 (SD +/- 0.6320), respectively. CONCLUSION: Our results showed that AgNOR evaluation was not a significant parameter to discriminate between malignant and benign thyroid lesions.
Subject(s)
Image Processing, Computer-Assisted , Nucleolus Organizer Region/pathology , Thyroid Neoplasms/pathology , Humans , Image Cytometry , Silver Staining , Thyroid Neoplasms/classificationABSTRACT
OBJECTIVE: To correlate the subjective AgNOR counting method and DNA content with histologic diagnoses of thyroid cancer and invasion. STUDY DESIGN: Eighty-one consecutive cases of thyroid carcinoma were selected for DNA and AgNOR analysis. The diagnoses were: papillary carcinoma (n = 40), follicular carcinoma (n = 31), Hürthle cell adenocarcinoma (n = 4), and undifferentiated carcinoma (n = 6). Seven normal thyroids were used as controls. DNA quantitative measurement was performed with Vidas 2.0 software (Kontron Bildanalyse, Munich, Germany) connected to an MPM 210 photometer microscope (Carl Zeiss, Oberkochen, Germany). The DNA index was obtained using histograms. Counting the NORs was performed by subjectively counting the NORs in 200 malignant cells. RESULTS: DNA ploidy analysis showed all Hürthle cell adenocarcinomas, 21 (67%)follicular tumors, 23 (57%) papillary tumors and 4 (67%) undifferentiated carcinomas to be aneuploid. DNA analysis correlated with histologic type of the tumor (p = 0.032). There was no statistical significance to the AgNOR counting variables studied. Statistical analysis showed correlation between ploidy and histologic diagnosis, but not AgNOR counting, to have prognostic value. CONCLUSION: DNA ploidy is more useful than subjective counting of NORs as an adjunct method for thyroid lesion analysis.
Subject(s)
Aneuploidy , Antigens, Nuclear , Nuclear Proteins , Thyroid Neoplasms/diagnosis , Adenocarcinoma, Follicular/diagnosis , Adenocarcinoma, Follicular/genetics , Adenocarcinoma, Follicular/metabolism , Adenoma, Oxyphilic/diagnosis , Adenoma, Oxyphilic/genetics , Adenoma, Oxyphilic/metabolism , Adolescent , Adult , Aged , Antigens, Nuclear/analysis , Carcinoma, Papillary/diagnosis , Carcinoma, Papillary/genetics , Carcinoma, Papillary/metabolism , Child , Data Interpretation, Statistical , Humans , Image Cytometry , Karyotyping , Middle Aged , Nuclear Proteins/analysis , Nucleolus Organizer Region/genetics , Nucleolus Organizer Region/metabolism , Ploidies , Thyroid Neoplasms/genetics , Thyroid Neoplasms/metabolismABSTRACT
INTRODUÇAO: A exposição da tireóide à irradiação está associada à alteração do componente cromossômico nessa glândula. OBJETIVO: Avaliar o conteúdo de DNA (ploidia) em glândula tireóide submetida à radiação. MATERIAL E MÉTODOS: Foram irradiadas tireóides de 75 ratos; 25 foram usados como grupo controle e não sofreram irradiação. A exposição à irradiação foi realizada com o uso de Cobalt-60 na região cervical anterior direita com espaço de 3-30cm, com anel traqueal de 2º e 6º e dose de 600 centigrays (cGY). O conteúdo de DNA (ploidia) foi obtido com o método de Feulgen-Thionin e quantificado com o aparelho CAS 200 de citometria estática. RESULTADO: Dos 92 casos, 88 (95,7 por cento) foram diplóides, independente do tempo de exposição; aneuploidia foi observada em quatro casos (4,3 por cento), somente no grupo sacrificado aos 33 dias. Oito casos foram excluídos por problemas técnicos. CONCLUSAO: A irradiação afeta muito precocemente o conteúdo de DNA nuclear levando a aneuploidia.
Subject(s)
Animals , Rats , Animals , Diploidy , DNA Mutational Analysis , Thyroid Gland/pathology , Neoplasms, Radiation-Induced/genetics , Neoplasms, Radiation-Induced/chemistry , Thyroid Neoplasms/genetics , Thyroid Neoplasms/chemistry , Rats, Wistar , X-RaysABSTRACT
This study was designed to analyze the cross-sectional comparison of the p16 and Ki-67 immunocytochemical expression in negative and equivocal (atypical squamous cells of undetermined significance (ASC-US)) liquid-based cytology (LBC) samples testing positive for high-risk human papillomavirus (HPV) types with HC2 assay or polymerase-chain reaction (PCR). A series of 199 consecutive LBC specimens derived from the same number of women participating in the ongoing Latin American Screening Study at Leonor Mendes de Barros Hospital, Sao Paulo, were analyzed using immunocytochemistry for expression of p16 and Ki-67 in negative and equivocal LBC samples testing positive for high-risk HPV types with hybrid capture II test (HC2) or PCR. All patients with at least one test positive (cytology, PCR, and/or HC2) were followed each 6 months for 3 years. The follow-up procedure consisted of visual examination, colposcopic inspection, cytology, and HC2 assay. Among the negative cytologic samples, 101 were HPV-positive and 55 HPV-negative. Of the HPV-positive group, 59 of 101 cases (58.4%) were positive for both p16 and Ki67 immunostaining, and 17 of 101 (16.8%) were negative for both. The proportion of Ki-67-positivity increased almost in parallel with the increasing grade of p16-positivity (p = 0.0001 for linear trend). In the HPV-negative group, both markers were negative in 41 of 55 cases (74.5%), and no statistical relationship was observed between the two markers (Pearson, p = 0.595). HPV-positive ASC-US samples demonstrated a simultaneous positive immunoreaction for p16 and Ki67 in 11 of 16 cases (68.7%), whereas 3 (18.7%) were concurrently negative. The relationship between the two markers was of borderline significance (Pearson, p = 0.053), but no linear relationship was found between the graded p16 and Ki-67 expression (p = 0.065 for linear trend). In the HPV-negative ASC-US group, there was no statistical association between the graded p16 and Ki-67 positivity (Pearson, p = 0.281). After 36 months of follow-up of the ASC-US patients, 6 women still displayed ASC-US smear, of which 4 of 6 were HPV-positive and expressed both p16 and Ki-67 markers. Two of 43 ASC-US smears had high-grade squamous intraepithelial lesions diagnosed (4.6%), and 1 had low-grade squamous intraepithelial lesion (2.3%). All of those were positive for HPV, p16 and Ki-67. Patients with ASC-US diagnosis and positive high-risk HPV status and positive for p16 Ki67 should be carefully observed to exclude occurrence of a squamous intraepithelial lesion. The combination of these two markers can be a useful implement for management of women with equivocal cytology.
Subject(s)
Biomarkers, Tumor/analysis , Cyclin-Dependent Kinase Inhibitor p16/biosynthesis , Ki-67 Antigen/biosynthesis , Papillomavirus Infections/complications , Precancerous Conditions/diagnosis , Uterine Cervical Neoplasms/diagnosis , Female , Humans , Immunohistochemistry , Papanicolaou Test , Papillomaviridae , Polymerase Chain Reaction , Precancerous Conditions/metabolism , Precancerous Conditions/virology , Tumor Virus Infections/complications , Uterine Cervical Neoplasms/metabolism , Uterine Cervical Neoplasms/virology , Vaginal Smears , Uterine Cervical Dysplasia/diagnosis , Uterine Cervical Dysplasia/metabolism , Uterine Cervical Dysplasia/virologyABSTRACT
OBJECTIVE: To verij5 the efficacy of the quality control (QC) program in a cytologic laboratwy with a rapid rescreening (RR) protocol. STUDY DESIGN: RR, according to the Turret RR method, of all samples initially screened as negative at the Laboratory of Cytology, Adolfo Lutz Institute, was performed. The slides were reviewed for 60 seconds. Suspect smears were fully checked by 2 reviewers to determine the final diagnoses. A total of 2954 sequential cytologic results were considered in this study. Of the 2954, 2568 (86.9%) were considered initially negative according to our internal QC, and these cases underwent RR. Also, 10% were randomly selected from these negative cases for full reviewing. The internal QC in our laboratory includes review of cases selected according to clinical and cytomorphologic criteria. RESULTS: Among the 2954 total cases, QC detected 386 (13%) atypias with final diagnoses reported according to The Bethesda System 2001 as follows: 82 (2.18%) low grade squamous intraepithelial lesions (LSILs), 35 (1.18%) high grade squamous intraepithelial lesions (HSILs), 2 (0.06%) squamous cell carcinomas, 105 (3.5%) atypical cells of undetermined significance (ASC-US), 4 (0.12%) atypical endocervical cells (AECs) and 158 (5.3%) unsatisfactory samples. RR of 2568 smears initially considered negative selected 194 (7.5%) slides. Of the 194, 146 (75.3%) were negative, 28 (14.4%) ASC-US, 5 (2.6%) AEC, 1 (0.5%) LSIL and 14 (7.2%) unsatisfactory. Full review of a 10% random fraction of the 2568 cases interpreted as negative did not detect lesions but did detect 5 (1.95%) unsatisfactory samples. CONCLUSION: Internal QC used in our laboratory based on clinical and cytomorphologic criteria to select cases for review proved to be an efficient method of detecting HSIL and cervical cancer. The consensus basis of this program strongly limits the false positive and false negative rates and also provides subjects with continuing education. One hundred percent RR is more efficient than 10% full reviewing in detecting cervical abnormalities.
Subject(s)
Cytodiagnosis/standards , Laboratories/standards , Mass Screening/standards , Public Health Practice/standards , Quality Assurance, Health Care/methods , Uterine Cervical Dysplasia/diagnosis , Uterine Cervical Neoplasms/diagnosis , Vaginal Smears/standards , Brazil , Female , Humans , Medical Audit , Sensitivity and SpecificityABSTRACT
OBJECTIVE: To correlate high-risk HPV (hrHPV) detection by Hybrid Capture II (HC2) (Digene, Gaithersburg, Maryland, U.S.A.) with DNA content (ploidy) of cervical biopsies analyzed by a computer-assisted system. STUDY DESIGN: Cervical biopsies from 54 women examined at Leonor Mendes de Barros Hospital, São Paulo, as part of the Latin American Screening study during 2002--2003, were tested for hrHPV with HC2. All patients had been referred for colposcopic examination due to an abnormal cervical cytology. The final diagnosis included 30 cervicitis, 14 cervical intraepithelial neoplasia (CIN) 1, 5 CIN 2, 4 CIN 3 and 1 squamous cell carcinoma (SCC). Five-micrometer sections of each biopsy were stained with Feulgen-tionine and evaluated with the CAS 200 System (Becton Dickinson, U.S.A.), using the 3.0 software (version 8.1) of the DNA Quantitative Measurement Program (Becton Dickinson). Ploidy was evaluated from histograms obtained by analyzing atypical nuclei. RESULTS: Of the 30 cervicitis cases, 28 (93.3%) were diploid, and hrHPV was detected in 8 (28.5%) of the cases. Two tetraploid cervicitis lesions were observed, 1 positive and 1 negative for hrHPV. Among the CIN 1 lesions, 8 (57.1%) were diploid and 6 (42.8%) aneuploid. Of the latter, 4 (66.6%) were negative and 2 (33.3%) positive for hrHPV. Of the 5 CIN 2 lesions, 2 were diploid, 2 aneuploid and 1 tetraploid; all were positive for hrHPV. All CIN 3 lesions and the SCC proved to be aneuploid and positive for hrHPV. CONCLUSION: The data suggest that the majority of cervicitis and CIN 1 lesions are diploid and negativef or hrHPV. This is in sharp contrast to high grade CIN 2-3 lesions, all of which were positive for hrHPV in this study and also aneuploid, consistent with their progressive potential.
Subject(s)
Cervix Uteri/pathology , Diagnosis, Computer-Assisted/methods , Papillomaviridae/isolation & purification , Papillomavirus Infections/diagnosis , Ploidies , Carcinoma, Squamous Cell , Cell Nucleus/pathology , Diagnosis, Differential , Enzyme-Linked Immunosorbent Assay , Female , Humans , Risk , Uterine Cervicitis/diagnosis , Uterine Cervical DysplasiaABSTRACT
OBJECTIVE: To evaluate the nuclear DNA content of cervical lesions in liquid-based cytologic specimens prepared for static cytometry. STUDY DESIGN: The DNA content of cervical lesions was evaluated in cervical samples prepared with the Autocyte PREP liquid-based cytology system (TriPath Imaging Inc., Burlington, North Carolina, U.S.A.). A series of 47 samples stained with the Papanicolaou method (chronic cervicitis, n = 15; cervical intraepithelial neoplasia [CIN] 1, n = 25; CIN 2, n = 5; CIN 3, n = 2) were collected from consecutive women enrolled in an ongoing screening study at Leonor Mendes de Barros Hospital, São Paulo, Brazil, in 2002. Each residual sample was processed according to the Feulgen-thionin method (TriPath Imaging). Ploidy evaluation was performed using the CAS 200 image analysis system and Quantitative DNA Measurement software 3.0 (version 8.1) (Becton Dickinson, San Jose, Califoria, U.S.A.). Cellular ploidy was analyzed from atypical nuclei, and the DNA index was obtained using histograms for interpretation. RESULTS: All chronic cervicitis cases were diploid. Of the CIN 1 cases, 44% were diploid, 12% tetraploid, 32% aneuploid and 12% polyploid (diploid plus tetraploid). CIN 2 lesions were diploid in 60% and aneuploid in 40% of cases, whereas all CIN 3 lesions (100%) were aneuploid. CONCLUSION: The liquid-based cytologic samples proved to be suitable and highly useful for DNA analysis by image cytometry, which was capable of discriminating CIN 3 lesions from CIN 1 and 2 but not CIN 1 from 2 lesions. Aneuploidy was closely associated with CIN 3 lesions.
Subject(s)
DNA, Neoplasm/analysis , Image Cytometry/methods , Papanicolaou Test , Uterine Cervical Dysplasia/diagnosis , Uterine Cervical Neoplasms/diagnosis , Vaginal Smears/methods , DNA/analysis , Female , Humans , Ploidies , Precancerous Conditions/diagnosis , Precancerous Conditions/pathology , Uterine Cervical Diseases/diagnosis , Uterine Cervical Diseases/pathology , Uterine Cervical Neoplasms/pathology , Uterine Cervical Dysplasia/pathologyABSTRACT
Alterações citopáticas causadas pela Candida sp ou Trichomonas vaginalis são bem conhecidas em esfregaços cervicais. Atenção especial deve ser considerada nestes casos para evitar resultados falso-positivos. Objetivo deste estudo foi identificar os parâmetros morfológicos principais utilizados no diagnóstico de ASCUS ou SIL com Candida sp ou Trichomonas vaginalis. Foram selecionados 78 casos consecutivos com diagnóstico de ASCUS ou SIL que apresentavam infecção por Candida sp ou Trichomonas vaginalis. Estes casos foram revistos por três observadores em estudo cego e as alterações principais foram registradas. As alterações mais freqüentemente observadas foram: halo perinuclear e aumento do volume nuclear (100 por cento); hipercromasia discreta (70,8 por cento); binucleação (47,2 por cento); multinucleação (34,7 por cento); disqueratose (22,2 por cento). Outros achados citológicos como: células escamosas metaplásicas (84,7 por cento) e infiltrado leucocitário (61,5 por cento). Alterações citopáticas causadas pela infecção por Candida sp ou Trichomonas vaginalis podem reproduzir alguns critérios utilizados no diagnóstico de ASCUS ou SIL, e também simular algumas alterações morfológicas relacionadas ao HPV. Estas alterações celulares devem ser cuidadosamente consideradas, uma vez Palavras-chave ASCUS; SIL; Candida sp; Trichomonas vaginalis; secreção cervical.que podem ser observados resultados falso-positivos.