Atypical gladular cells and adenocarcinoma in situ according to the Bethesda 2001 classification : cytohistological correlation

Orientadores: Luiz Carlos Zeferino, Silvia Helena Rabelo dos SantosDissertação (mestrado) - Universidade Estadual de Campinas, Faculdade de Ciencias MedicasResumo: Introdução: Há evidências de que a incidência do adenocarcinoma do colo do útero tem aumentado, apesar dos programas de rastreamento. Foi a classificação do Sistema de Bethesda, em 1988, que incluiu o diagnóstico citológico relativo às células glandulares atípicas, que não estava presente nas classificações anteriores. Em sua revisão de 2001, o Sistema de Bethesda propôs a subclassificação do diagnóstico citológico de Anormalidades em Células Glandulares em: células glandulares atípicas sem outras especificações (AGC-SOE), células glandulares atípicas favorecendo neoplasia (AGC-FN) e adenocarcinoma in situ (AIS). Os diagnósticos citológicos de AGC constituem um problema clínico em consequência da falta de critérios citomorfológicos bem definidos para sua interpretação, do alto grau de variabilidade interobservador e da falta de achados colposcópicos característicos. O diagnóstico citológico de AIS, embora preditivo de neoplasia glandular, não pode diferenciar de modo preciso entre as formas in situ e invasiva, as quais só podem ser confirmadas pela histologia. Há, na literatura, divergentes opiniões quanto à utilidade desta subclassificação e sua análise é de grande relevância para prática clínica. Objetivo: Analisar a associação entre a classificação citológica do Sistema de Bethesda 2001 para as anormalidades do epitélio glandular cervical e o resultado histológico. Métodos: O estudo foi do tipo corte transversal analítico, prospectivo, realizado no Centro de Atenção Integral à Saúde da Mulher (CAISM), UNICAMP, entre 2002 e 2005, e incluiu uma série consecutiva de 155 mulheres com anormalidades glandulares endocervicais no exame citológico. Destas, 91 mulheres foram incluídas por AGC-SOE, 15 por AGC-FN; 14 por AIS e 35 por diagnóstico combinado de AGC associada à lesão escamosa intraepitelial de alto grau (HSIL). Os diagnósticos de AGC e AIS foram confirmados por dois observadores, de acordo com Sistema de Bethesda 2001. As mulheres com alterações glandulares,de origens outras que não endocervical, não foram incluídas no estudo. No primeiro atendimento foi colhido um segundo esfregaço cervical e realizada colposcopia em todas as mulheres. Foi realizada biópsia ou conização naquelas pacientes que tinham indicação, de acordo com a rotina assistencial do serviço. Todas as mulheres incluídas foram submetidas à ecografia pélvica. A magnitude da associação entre o diagnóstico citológico das anormalidades glandulares e o diagnóstico histológico foi estimada utilizando-se Odds ratio (OR), com respectivo intervalo de confiança (IC) de 95%. Resultados: Das 155 mulheres admitidas no estudo, 126 tiveram avaliação histológica. Tomando-se como referência AGC-SOE, os diagnósticos citológicos de AGC-FN e AIS foram significativamente associados com os resultados histológicos de neoplasia escamosa (NIC 2 ou pior diagnóstico) ou adenocarcinoma (in situ ou invasivo) com valores de odds ratio respectivamente de 10,50 (95% IC: 2,94- 37,55) e 156,00 (95% IC: 17,72-1373,46). Tomando-se como referência AGC-FN, o diagnóstico citológico de AIS foi significativamente associado com os resultados histológicos de neoplasia escamosa ou adenocarcinoma (OR=14,86; 95% IC: 1,53-144,23). Uma associação similar foi observada considerando-se os resultados histológicos de adenocarcinoma (in situ ou invasivo). Nenhuma associação foi observada para neoplasia escamosa analisada isoladamente. Os resultados histológicos de NIC 2 ou pior diagnóstico foram fortemente associados com AGC quando a lesão de alto grau (HSIL) também estava presente (OR=57,60; 95%IC: 18,25-181,76), mas nenhuma associação foi observada com resultado histológico de adenocarcinoma. Conclusões: Há uma associação significante entre as subclassificações da categoria de anormalidades em células glandulares de origem endocervical, propostas pelo Sistema de Bethesda 2001, e diagnósticos histológicos significativos, pois os diagnósticos citológicos de AGC-SOE, AGC-FN e AIS indicam um aumento progressivo de risco para adenocarcinoma. A presença de HSIL associada à AGC representa maior probabilidade de neoplasia escamosa, mas não de adenocarcinomaAbstract: Introduction: There is evidence indicating that incidence of cervical adenocarcinoma has been increasing, despite the screening programs. The cytological diagnosis of atypical glandular cells, absent from previous versions, was included in the 1988 edition of the Bethesda System (TBS). In its review of 2001, the TBS has proposed the sub-classification of the cytological diagnosis of glandular cells abnormalities as: atypical glandular cells not otherwise specified (AGC-NOS), atypical glandular cells favor neoplastic (AGC-FN) and adenocarcinoma in situ "(AIS). The usefulness of the sub-classification of cytological diagnosis as atypical glandular cells not otherwise specified (AGC-NOS), atypical glandular cells favor neoplastic (AGC-FN) and adenocarcinoma in situ (AIS), as proposed in the 2001 TBS, is a controversial issue, but of great importance in clinical practice. AGC in cervical smears constitutes a clinical problem due to the lack of well-defined cytomorphological criteria for the interpretation of this finding, the high degree of interobserver variability and the lack of characteristic colposcopic features. Moreover, cytological AIS cannot specifically differentiate between AIS and invasive adenocarcinoma, which can only be confirmed by histology. Objective: to analyze the association between the 2001 TBS classification of glandular abnormalities and the histological outcome. Method: This cross-sectional and prospective study was conducted at the Center of Health Care of Women (CAISM), UNICAMP, between 2002 and 2005. The sample comprises a series of 155 women with glandular abnormalities in cervical smear. Of those, 91 women were included due to AGC-SOE, 15 due to AGC-FN, 14 due to AIS and 35 for AGC combined with high grade squamous intraepithelial lesions (HSIL). The diagnoses of AGC and AIS were confirmed by two observers according to the TBS 2001. Women with changes of glandular origin other than endocervical were excluded. In their first visit, a second cervical sample was collected from all women, and colposcopy was performed. A biopsy or a conization was performed when necessary, according to CAISM's protocols. All women underwent pelvic ultrassonography. Odds ratios (OR) with 95% confidence interval (95% CI) were used to evaluate the magnitude of the association between the cytological diagnosis of the glandular abnormalities and the significant histologic diagnosis. Results: Of the 155 women admitted to the study, 126 were submitted to histological examination. Taking as reference AGC-NOS, the cytological diagnosis of AGC-FN and AIS were significantly associated with the histological outcome of squamous neoplasia (CIN or worse) or adenocarcinoma (in situ or invasive) with OR values of 10.50 (95% CI: 2.94-37.55) and 156.00 (95% CI: 17.72-1373.46), respectively. Taking as reference AGC-FN, the cytological diagnosis of AIS was significantly associated with the histological outcome of squamous neoplasia or adenocarcinoma (OR=14.86; 95%CI: 1.53-144.23). Similar associations were observed for the histological outcome of adenocarcinoma, but no association was observed for only squamous neoplasia. Histological outcome of CIN2 or worse was strongly associated with AGC when HSIL was also present (OR = 57.60; 95% CI: 18.25- 181.76), but no association was observed with adenocarcinoma as the stand-alone histological outcome. Conclusions: There is an association between the subclassification of glandular cells abnormalities of endocervical origin proposed by 2001 TBS with significant histological outcomes. The cytological diagnoses of AGC-NOS, AGC-FN and AIS were progressively associated with adenocarcinoma. The histological outcome of squamous neoplasia is frequent but does not differ from these cytological interpretations. The presence of HSIL associated with AGC was associated with a greater probability of squamous neoplasia, but notMestradoCiencias BiomedicasMestre em Tocoginecologi

Expression of the proteins MMP-2, MMP-9, MMP-14, TIMP-1, TIMP-2 and VEGF-A in the CIN 3 and cervical cancer

Orientadores: Luiz Carlos Zeferino, Silvia Helena Rabelo dos SantosTexto em português e inglêsTese (doutorado) - Universidade Estadual de Campinas, Faculdade de Ciências MédicasResumo: Introdução: O carcinoma escamoso do colo uterino é precedido pela neoplasia intraepitelial cervical grau 3 (NIC 3). A invasão tumoral envolve a degradação da matriz extracelular e membrana basal do epitélio por enzimas proteolíticas denominadas metaloproteinases (MMPs). Os inibidores teciduais das metaloproteinases (TIMPs) também interferem no processo de invasão. Angiogênese é condição indispensável para a progressão tumoral. Objetivo: Analisar a expressão de MMP-2, MMP-9, MMP-14, TIMP-1, TIMP-2 e VEGF-A na NIC 3 e carcinoma do colo uterino. Sujeito e Métodos: Estudo do tipo comparativo observacional constituído de três grupos:- Grupo 1: 55 casos com diagnóstico de NIC 3, Grupo 2: 30 casos com NIC 3 e carcinoma associados e Grupo 3: 46 casos com carcinoma. A expressão protéica foi pesquisada separadamente nas células tumorais e estromais por reação imunoistoquímica. Para estabelecer a porcentagem de células imunopositivas utilizou-se software morfométrico. Análise Estatística: Aplicou-se o Teste T-pareado ou de Mann-Whitney ou Wilcoxon Signed Rank. Resultados: Em todos os grupos, a expressão tumoral de MMP-14 foi maior que a estromal. Inversamente, a expressão de TIMP-2 foi maior nas células estromais que nas tumorais, em cada grupo diagnóstico. A expressão de MMP-9 foi maior nas células estromais que nas tumorais, com exceção do componente invasor do Grupo 2. A expressão estromal de TIMP-1 foi maior que a tumoral no carcinoma e, ao contrário, sua expressão foi maior nas células tumorais da NIC 3. A expressão de VEGF-A foi maior apenas nas células tumorais da NIC 3. Comparando a expressão dos marcadores entre os grupos, foram encontradas as maiores diferenças entre grupos extremos, ou seja, entre NIC 3 e carcinoma. A expressão de MMP-2 nas células estromais foi maior no componente NIC 3 do Grupo 2 que no NIC 3 do Grupo 1. A expressão de VEGF-A nas células estromais do carcinoma foi maior que nas células estromais da NIC 3. Conclusões: Os resultados deste estudo sugerem que a expressão de TIMP-1 aumenta nas células do estroma e diminui nas células tumorais quando a NIC 3 progride para carcinoma invasor. MMP-9 e TIMP-2 tiveram expressão similar na NIC 3 e no carcinoma, o que limita inferências sobre seu papel na progressão neoplásica. O padrão imunoistoquímico da expressão das MMPs, TIMPs e VEGF-A na NIC 3 e no carcinoma invasivo, quando estas lesões estavam associadas, foi semelhante. A expressão do VEGF-A foi maior nas células tumorais do que nas estromais da NIC 3, porém quando esta lesão progride para carcinoma invasivo sua expressão aumenta nas células do estroma e não se altera nas tumorais. A expressão de MMP-14, MMP-2, TIMP-1 e VEGF-A aumentou com a gravidade da neoplasiaAbstract: Introduction: Squamous cell carcinoma of the cervix is preceded by cervical intraepithelial neoplasia grade 3 (CIN 3). Tumor invasion involves degradation of extracellular matrix and epithelium basement membrane by proteolytic enzymes called metalloproteinases (MMPs). Tissue inhibitors of metalloproteinases (TIMPs) are also involved in the invasion process. Angiogenesis is a prerequisite for tumor progression. Objective: To analyze the expression of MMP-2, MMP-9 and MMP-14, TIMP-1, TIMP-2 and VEGF-A in CIN 3 and invasive carcinoma. Subject and Methods: This comparative observational study was consists of three groups: Group 1: 55 cases diagnosed with CIN 3, Group 2: 30 cases with CIN 3 associated with invasive carcinoma and Group 3: 46 cases with invasive carcinoma. Protein expression was investigated separately in tumor and stromal cells by immunohistochemistry and evaluated by the percentage of cells positive for immunostaining using morphometric software. Statistical Analysis: Was performed applying paired t-test or Mann-Whitney or Wilcoxon Signed Rank. Results: In each diagnostic group, expression markers were significantly higher: MMP-14 in tumor cells, and TIMP-2 in stromal cells; also MMP-9 expression was significantly higher in stromal cells, except in invasive component of group 2, and TIMP-1 had significantly higher expression in stromal cells of invasive carcinoma and in tumor cells of CIN 3. VEGF-A expression was significantly higher only in tumor cells CIN 3. Comparing the expression of markers between groups, two by two, we find the greatest differences between the extreme groups, i.e. between invasive carcinoma and CIN 3. The expression of MMP-2 was significantly greater in the stromal component CIN 3 in group 2 than in CIN 3 only. The expression of VEGF-A was significantly higher in the group stromal cell carcinoma when compared to stromal cells CIN 3. Conclusions: The results of this study suggest that the expression of TIMP-1 increases in the stromal cells and decreases in tumor cells when CIN 3 progresses to invasive carcinoma. MMP-9 and TIMP-2 had similar expression in CIN 3 and invasive carcinoma, which limits inferences about its role in neoplastic progression. The immunohistochemical pattern of expression of MMPs, TIMPs and VEGF-A in CIN 3 and invasive carcinoma, as these lesions were associated, was similar. The expression of VEGF-A was higher in tumor cells than in stromal cells in CIN 3, but when the lesion progresses to invasive carcinoma its expression increases in the stromal cells and the tumor cells does not change. The expression of MMP-14, MMP-2, TIMP-1 and VEGF-A was increased with the severity of the neoplasiaDoutoradoOncologia Ginecológica e MamáriaDoutora em Ciências da Saúd

The Impact Of A Community Intervention To Improve Cervical Cancer Screening Uptake In The Amazon Region Of Brazil.

In the northern region of Brazil, cervical cancer is the most important cause of cancer-related deaths among women. There is considerable likelihood, however, that official incidence and mortality figures are greatly underestimated. The aim of this study was to estimate the repercussions from improvement in cervical cancer screening programs on the incidence of pre-invasive and invasive cervical lesions in a municipality in this region. This was a quasi-experimental study that assessed process dimensions relevant to the program objectives. The study comprised a sample of 2,226 women seen at primary healthcare units in Cruzeiro do Sul, a small city in the Brazilian Amazon region, from April 2003 to July 2004. Women were recruited through local radio advertisements and by oral communication from the investigators. The women answered a structured questionnaire and underwent pelvic examination, which included Papanicolaou (Pap) smears and naked-eye inspection of the cervix after applying diluted acetic acid. Women with positive Pap smears or abnormal gynecological examination were referred for colposcopy and possible biopsy, diathermic large loop excision of the transformation zone or conization. The results obtained were compared with historical official data retrieved from the Brazilian Ministry of Health's database. Intervention resulted in a 40% increase in positive Pap smears and detection of cancer was nine times higher than had been observed in routine screening. Detection of pre-invasive and invasive cervical lesions in the intervention group was remarkably higher than among women seen during routine screening.12542-

PNPLA 3 I148M genetic variant associates with insulin resistance and baseline viral load in HCV genotype 2 but not in genotype 3 infection

Peer reviewe

The GenTree Dendroecological Collection, tree-ring and wood density data from seven tree species across Europe

The dataset presented here was collected by the GenTree project (EU-Horizon 2020), which aims to improve the use of forest genetic resources across Europe by better understanding how trees adapt to their local environment. This dataset of individual tree-core characteristics including ring-width series and whole-core wood density was collected for seven ecologically and economically important European tree species: silver birch (Betula pendula), European beech (Fagus sylvatica), Norway spruce (Picea abies), European black poplar (Populus nigra), maritime pine (Pinus pinaster), Scots pine (Pinus sylvestris), and sessile oak (Quercus petraea). Tree-ring width measurements were obtained from 3600 trees in 142 populations and whole-core wood density was measured for 3098 trees in 125 populations. This dataset covers most of the geographical and climatic range occupied by the selected species. The potential use of it will be highly valuable for assessing ecological and evolutionary responses to environmental conditions as well as for model development and parameterization, to predict adaptability under climate change scenarios

The GenTree Platform: growth traits and tree-level environmental data in 12 European forest tree species

Background: Progress in the field of evolutionary forest ecology has been hampered by the huge challenge of phenotyping trees across their ranges in their natural environments, and the limitation in high-resolution environmental information. Findings: The GenTree Platform contains phenotypic and environmental data from 4,959 trees from 12 ecologically and economically important European forest tree species: Abies alba Mill. (silver fir), Betula pendula Roth. (silver birch), Fagus sylvatica L. (European beech), Picea abies (L.) H. Karst (Norway spruce), Pinus cembra L. (Swiss stone pine), Pinus halepensis Mill. (Aleppo pine), Pinus nigra Arnold (European black pine), Pinus pinaster Aiton (maritime pine), Pinus sylvestris L. (Scots pine), Populus nigra L. (European black poplar), Taxus baccata L. (English yew), and Quercus petraea (Matt.) Liebl. (sessile oak). Phenotypic (height, diameter at breast height, crown size, bark thickness, biomass, straightness, forking, branch angle, fructification), regeneration, environmental in situ measurements (soil depth, vegetation cover, competition indices), and environmental modeling data extracted by using bilinear interpolation accounting for surrounding conditions of each tree (precipitation, temperature, insolation, drought indices) were obtained from trees in 194 sites covering the species’ geographic ranges and reflecting local environmental gradients. Conclusion: The GenTree Platform is a new resource for investigating ecological and evolutionary processes in forest trees. The coherent phenotyping and environmental characterization across 12 species in their European ranges allow for a wide range of analyses from forest ecologists, conservationists, and macro-ecologists. Also, the data here presented can be linked to the GenTree Dendroecological collection, the GenTree Leaf Trait collection, and the GenTree Genomic collection presented elsewhere, which together build the largest evolutionary forest ecology data collection available

Between but not within species variation in the distribution of fitness effects

New mutations provide the raw material for evolution and adaptation. The distribution of fitness effects (DFE) describes the spectrum of effects of new mutations that can occur along a genome, and is therefore of vital interest in evolutionary biology. Recent work has uncovered striking similarities in the DFE between closely related species, prompting us to ask whether there is variation in the DFE among populations of the same species, or among species with different degrees of divergence, i.e., whether there is variation in the DFE at different levels of evolution. Using exome capture data from six tree species sampled across Europe we characterised the DFE for multiple species, and for each species, multiple populations, and investigated the factors potentially influencing the DFE, such as demography, population divergence and genetic background. We find statistical support for there being variation in the DFE at the species level, even among relatively closely related species. However, we find very little difference at the population level, suggesting that differences in the DFE are primarily driven by deep features of species biology, and that evolutionarily recent events, such as demographic changes and local adaptation, have little impact