Imunomarcação não homogênea das membranas hialinas na sindrome da angustia respiratório do adulto pulmonar, extrapulmonar e idiopática

PURPOSE: To determine the nature of hyaline membranes in different manifestations of diffuse alveolar damage, [pulmonary and extrapulmonary acute respiratory distress syndrome], and idiopathic [acute interstitial pneumonia]. MATERIALS AND METHODS: Pulmonary specimens were obtained from 17 patients with acute respiratory distress syndrome and 9 patients with acute interstitial pneumonia. They were separated into 3 different groups: (a) pulmonary diffuse alveolar damage (pDAD) (n = 8), consisting only of pneumonia cases; (b) extrapulmonary diffuse alveolar damage (expDAI) (n = 9), consisting of sepsis and septic shock cases; and (c) idiopathic diffuse alveolar damage (iDAD) (n = 9), consisting of idiopathic cases (acute interstitial pneumonia). Hyaline membranes, the hallmark of the diffuse alveolar damage histological pattern, were examined using various kinds of antibodies. The antibodies used were against surfactant apoprotein-A (SP-A), cytokeratin 7 (CK7), cytokeratin 8 (CK8), alpha smooth muscle actin (a-SMA), cytokeratin AE1/AE3 (AE1/AE3), and factor VIII-related antigen (factor VIII). RESULTS: Pulmonary diffuse alveolar damage showed the largest quantity of hyaline membranes (12.65% ± 3.24%), while extrapulmonary diffuse alveolar damage (9.52% ± 3.64%) and idiopathic diffuse alveolar damage (7.34% ± 2.11%) showed intermediate and lower amounts, respectively, with the difference being statistically significant between pulmonary and idiopathic diffuse alveolar damage (P < 0.05). No significant difference was found for hyaline membranes Sp-A immunostaining among pulmonary (15.36% ± 3.12%), extrapulmonary (16.12% ± 4.58%), and idiopathic (13.74 ± 4.20%) diffuse alveolar damage groups. Regarding factor VIII, we found that idiopathic diffuse alveolar damage presented larger amounts of immunostained hyaline membranes (14.12% ± 6.25%) than extrapulmonary diffuse alveolar damage (3.93% ± 2.86%), with this difference being statistically significant (P < 0.001). Equally significant was the difference for progressive decrease of cytokeratin AE1/AE3 immunostaining in hyaline membranes present in the extrapulmonary diffuse alveolar damage (5.42% ± 2.80%) and idiopathic diffuse alveolar damage (0.47% ± 0.81%) groups (P < 0.001). None of the groups stained for cytokeratin CK-7, CK-8, vimentin, or a anti-smooth muscle actin. CONCLUSIONS: This study showed that only the epithelial/endothelial components (SP-A, factor VIII, and AE1/AE3) of the alveolar/capillary barrier are present in hyaline membranes formation in the 3 groups of patients with diffuse alveolar damage. The significant difference in the expression of factor VIII-related antigen and cytokeratin AE1/AE3 in the expDA versus iDAD groups as well as the significant difference in the amount of hyaline membranes present in the pDAD versus iDAD groups are suggestive of a local and specific lesion with different pathways (direct, indirect, or idiopathic), depending on the type of diffuse alveolar damage.OBJETIVO: Determinar a natureza da membrana hialina nas diferentes manifestações do dano alveolar difuso [pulmonar e extrapulmonar síndrome do desconforto respiratório] e idiopático [pneumonia intersticial aguda]. MATERIAIS E MÉTODOS: Espécimes pulmonares foram obtidos de 17 pacientes com SDRA e 9 pacientes com pneumonia intersticial aguda e separados em três diferentes grupos: (a) dano alveolar difuso pulmonar (DADp) (n=8) constituído por casos de pneumonia, (b) dano alveolar difuso extrapulmonar (DADexp) (n=9) constituído por casos de sepse e choque séptico e (c) dano alveolar difuso idiopático (DADi) (n=9) constituído por casos idopáticos (ou pneumonia intersticial aguda). As características das membranas hialinas do padrão histológico de dano alveolar difuso foram examinadas usando vários tipos de anticorpos. Os anticorpos usados foram surfactante apoproteina A (SP-A), anti-citokeratina 7 (CK7), citokeratina 8 (CK8), alfa actina de músculo liso (a-SMA), citokeratina AE1/AE3 (AE1/AE3) e antígeno relacionado ao fator VIII (Fator VIII). RESULTADOS: Observaram-se aumentos maiores da quantidade de membrana hialina no dano alveolar difuso pulmonar (12.65 ± 3.24%), intermediários no dano alveolar difuso extrapulmonar (9.52 ± 3.64%) e baixos no dano alveolar difuso idiopático (7.34 ± 2.11%) respectivamente, esta diferencia foi estatística significante entre o dano alveolar difuso pulmonar e o dano alveolar difuso idiopático (

Angiogênese como indicador do potencial de metástase no carcinoma papilífero tireóideo

Angiogenesis is new blood vessel formation, a process that can lead to tumor development. Microvessel count has been correlated to metastasis in some neoplasias. PURPOSE: To determine if measurement of microvessel density is useful in predicting metastasis to the cervical lymph node and prognosis in patients with papillary thyroid carcinoma. METHODS: A retrospective analysis was performed in 30 patients that had undergone total thyroidectomy. They were divided in 2 groups of 15 patients - with and without metastatic disease. Immunohistochemistry was used to detect expression of CD34 in archival paraffin-embedded papillary thyroid tumors, and microvessel density was calculated based on it. Association between microvessel density and the presence of metastasis, according to histological subtype, disease recurrence, and AMES prognostic index groups was determined through statistical analysis. RESULTS: The median microvessel density for the patient group without metastasis (200.0 microvessels/mm²) was apparently, but not significantly, less than that observed among metastatic disease patients (254.4 microvessels/mm²) (P = .20). When papillary carcinoma subtypes were analyzed, this difference became significant (P =.02). The follicular variant exhibited a greater microvessel density than the other subtypes, independent of metastasis presence. There was an apparent, but not significant, tendency for a larger median microvessel density in the group of patients that presented recurrence (294.4 microvessels/mm² vs 249.6 microvessels/mm², P = .11). There was no relationship between risk level and microvessel density: in the low- and high-risk groups, the median MVD was 304.0 microvessels/mm² and 229.6 microvessels/mm², respectively (P = .27). CONCLUSIONS: The results suggest that angiogenesis is more intense among metastatic tumors in the classic and the tall cell variants, indicating that microvessel count can be an indicator of the potential for metastasis in these subtypes of papillary thyroid carcinoma. Patients that developed recurrent disease had a tendency to exhibit higher angiogenesis; however, there was no association between microvessel density and the AMES prognostic index.O desenvolvimento dos tumores depende da formação de neovasos, a angiogênese. Em algumas neoplasias, a alta densidade de microvasos tumorais correlaciona-se com a presença de metástase. OBJETIVO: Determinar se a medida da angiogênese pode indicar o potencial de metástase e o prognóstico do carcinoma papilífero tireóideo. MÉTODO: Foi feita análise retrospectiva de 30 tireoidectomizados, divididos em dois grupos de 15 indivíduos cada, respectivamente com e sem metástase. A partir dos blocos de parafina, foi calculada a densidade de microvasos no tecido tumoral por meio da quantificação da expressão do anticorpo CD34 pela imunohistoquímica. A associação da densidade de microvasos com a presença de metástase, ocorrência de recidiva e os grupos de risco do índice prognóstico AMES foi determinada por análise estatística. RESULTADOS: A mediana da densidade de microvasos no grupo de doentes sem metástase (200,0 microvasos/mm²) foi inferior àquela dos portadores de metástase (254,4 microvasos/mm²) (p = .2), sem atingir significância estatística. Ao considerar apenas os subtipos histológicos clássico e de células altas, essa diferença tornou-se significante (p = .02), uma vez que a variante folicular exibiu maior DMV que os demais subtipos, independente da presença de metástase. Houve tendência não significativa à maior densidade de microvasos entre aqueles que apresentaram recidiva (294,4 microvasos/mm² contra 249,6 microvasos/mm², p = .11). Nos grupos de baixo e alto risco, a mediana da densidade de microvasos foi de 304,0 microvasos/mm² e 229,6 microvasos/mm² respectivamente (p = .27). CONCLUSÃO: A angiogênese foi mais intensa nos tumores com metástase nos subtipos clássico e de células altas, sugerindo que a contagem de microvasos pode ser um indicador do potencial de metástase nestes subtipos histológicos do carcinoma papilífero tireóideo. Doentes que evoluíram com recidiva tenderam a exibir maior angiogênese, porém não houve associação da densidade de microvasos e o índice prognóstico

Unusual remodeling of the hyalinization band in vulval lichen sclerosus by type V collagen and ECM 1 protein

OBJECTIVES: The vulva is the primary site affected in lichen sclerosus, a chronic dermatosis in women that is histologically characterized by a zone of collagen remodeling in the superior dermis. The normal physiological properties of the vulva depend on the assembly of collagen types I (COLI), III (COLIII) and V (COLV), which form heterotypic fibers, and extracellular matrix protein interactions. COLV regulates the heterotypic fiber diameter, and the preservation of its properties is important for maintaining normal tissue architecture and function. In the current work, we analyzed the expression of COLV and its relationship with COLI, COLIII, elastic fibers and extracellular matrix protein 1 in vulvar biopsies from patients with lichen sclerosus. METHODS: Skin biopsies from 21 patients with lichen sclerosus, classified according to Hewitt histological criteria, were studied and compared to clinically normal vulvar tissue (N=21). Morphology, immunohistochemistry, immunofluorescence, 3D reconstruction and morphometric analysis of COLI, COLIII, COLV deposition, elastic fibers and extracellular matrix 1 expression in a zone of collagen remodeling in the superior dermis were performed. RESULTS: A significant decrease of elastic fibers and extracellular matrix 1 protein was present in the hyalinization zone of lichen sclerosus compared to healthy controls. The non-homogeneous distribution of collagen fibers visualized under immunofluorescence in the hyalinization zone of lichen sclerosus and control skin was confirmed by histomorphometry. Lichen sclerosus dermis shows a significant increase of COLI, COLIII and COLV expression compared to the healthy controls. Significant inverse associations were found between elastic fibers and COLV and between COLV and extracellular matrix 1 expression. A direct association was found between elastic fiber content and extracellular matrix 1 expression. Tridimensional reconstruction of the heterotypic fibers of the lichen sclerosus zone of collagen remodeling confirmed the presence of densely clustered COLV. CONCLUSIONS: Increased deposition of abnormal COLV and its correlation with extracellular matrix 1 and elastic fibers suggest that COLV may be a trigger in the pathogenesis of lichen sclerosus

Nonhomogeneous immunostaining of hyaline membranes in different manifestations of diffuse alveolar damage Imunomarcação não homogênea das membranas hialinas na sindrome da angustia respiratório do adulto pulmonar, extrapulmonar e idiopática

PURPOSE: To determine the nature of hyaline membranes in different manifestations of diffuse alveolar damage, [pulmonary and extrapulmonary acute respiratory distress syndrome], and idiopathic [acute interstitial pneumonia]. MATERIALS AND METHODS: Pulmonary specimens were obtained from 17 patients with acute respiratory distress syndrome and 9 patients with acute interstitial pneumonia. They were separated into 3 different groups: (a) pulmonary diffuse alveolar damage (pDAD) (n = 8), consisting only of pneumonia cases; (b) extrapulmonary diffuse alveolar damage (expDAI) (n = 9), consisting of sepsis and septic shock cases; and (c) idiopathic diffuse alveolar damage (iDAD) (n = 9), consisting of idiopathic cases (acute interstitial pneumonia). Hyaline membranes, the hallmark of the diffuse alveolar damage histological pattern, were examined using various kinds of antibodies. The antibodies used were against surfactant apoprotein-A (SP-A), cytokeratin 7 (CK7), cytokeratin 8 (CK8), alpha smooth muscle actin (a-SMA), cytokeratin AE1/AE3 (AE1/AE3), and factor VIII-related antigen (factor VIII). RESULTS: Pulmonary diffuse alveolar damage showed the largest quantity of hyaline membranes (12.65% ± 3.24%), while extrapulmonary diffuse alveolar damage (9.52% ± 3.64%) and idiopathic diffuse alveolar damage (7.34% ± 2.11%) showed intermediate and lower amounts, respectively, with the difference being statistically significant between pulmonary and idiopathic diffuse alveolar damage (P < 0.05). No significant difference was found for hyaline membranes Sp-A immunostaining among pulmonary (15.36% ± 3.12%), extrapulmonary (16.12% ± 4.58%), and idiopathic (13.74 ± 4.20%) diffuse alveolar damage groups. Regarding factor VIII, we found that idiopathic diffuse alveolar damage presented larger amounts of immunostained hyaline membranes (14.12% ± 6.25%) than extrapulmonary diffuse alveolar damage (3.93% ± 2.86%), with this difference being statistically significant (P < 0.001). Equally significant was the difference for progressive decrease of cytokeratin AE1/AE3 immunostaining in hyaline membranes present in the extrapulmonary diffuse alveolar damage (5.42% ± 2.80%) and idiopathic diffuse alveolar damage (0.47% ± 0.81%) groups (P < 0.001). None of the groups stained for cytokeratin CK-7, CK-8, vimentin, or a anti-smooth muscle actin. CONCLUSIONS: This study showed that only the epithelial/endothelial components (SP-A, factor VIII, and AE1/AE3) of the alveolar/capillary barrier are present in hyaline membranes formation in the 3 groups of patients with diffuse alveolar damage. The significant difference in the expression of factor VIII-related antigen and cytokeratin AE1/AE3 in the expDA versus iDAD groups as well as the significant difference in the amount of hyaline membranes present in the pDAD versus iDAD groups are suggestive of a local and specific lesion with different pathways (direct, indirect, or idiopathic), depending on the type of diffuse alveolar damage.<br>OBJETIVO: Determinar a natureza da membrana hialina nas diferentes manifestações do dano alveolar difuso [pulmonar e extrapulmonar síndrome do desconforto respiratório] e idiopático [pneumonia intersticial aguda]. MATERIAIS E MÉTODOS: Espécimes pulmonares foram obtidos de 17 pacientes com SDRA e 9 pacientes com pneumonia intersticial aguda e separados em três diferentes grupos: (a) dano alveolar difuso pulmonar (DADp) (n=8) constituído por casos de pneumonia, (b) dano alveolar difuso extrapulmonar (DADexp) (n=9) constituído por casos de sepse e choque séptico e (c) dano alveolar difuso idiopático (DADi) (n=9) constituído por casos idopáticos (ou pneumonia intersticial aguda). As características das membranas hialinas do padrão histológico de dano alveolar difuso foram examinadas usando vários tipos de anticorpos. Os anticorpos usados foram surfactante apoproteina A (SP-A), anti-citokeratina 7 (CK7), citokeratina 8 (CK8), alfa actina de músculo liso (a-SMA), citokeratina AE1/AE3 (AE1/AE3) e antígeno relacionado ao fator VIII (Fator VIII). RESULTADOS: Observaram-se aumentos maiores da quantidade de membrana hialina no dano alveolar difuso pulmonar (12.65 ± 3.24%), intermediários no dano alveolar difuso extrapulmonar (9.52 ± 3.64%) e baixos no dano alveolar difuso idiopático (7.34 ± 2.11%) respectivamente, esta diferencia foi estatística significante entre o dano alveolar difuso pulmonar e o dano alveolar difuso idiopático (p<0.05). Não se encontrou significância estatística para a quantidade de imunomarcação de Sp-A nos grupos de dano alveolar difuso pulmonar (15.36 ± 3.12%), extrapulmonar (16.12 ± 4.58%) e idiopático (13.74 ± 4.20%). Com relação ao Fator VIII, nós encontramos maiores aumentos da imunomarcação da membrana hialina no grupo dano alveolar difuso idiopático (14.12 ± 6.25%) do que no dano alveolar difuso extrapulmonar (3.93 ± 2.86%), com significância estatística (p<0.001). Da mesma forma houve um aumento progressivo da imunomarcação da membrana hialina com citokeratina AE1/AE3 nos grupos de dano alveolar difuso extrapulmonar (5.42 ± 2.80%) e dano alveolar difuso idiopático (0.47 ± 0.81%) (p<0.001). Nenhuns dos grupos marcou para as citokeratina CK-7, CK-8 ou para a vimentina ou actina de músculo liso. CONCLUSÕES: Este estudo só mostra os componentes epitelio/endotelio (SP-A, Fator VIII e AE1/AE) da barreira alvéolo/capilar que estão presentes na formação da membrana hialina nos três diferentes grupos, sugerindo que as diferenças na sua patogênese dependem da via do ferimento (direto, indireto, ou idiopático); que dependem da causa do dano alveolar difuso. As diferenças na expressão do antígeno relacionado ao Fator VIII e nas citoqueratinas AE1/AE3 no grupo DADexp versus DADi, bem como as diferenças entre HM presente no grupo DADp versus DADi sugerem a ocorrência de lesões específicas com vias diferentes (direta, indireta ou idiopática) dependentes do tipo de DAD

Angiogenesis as an indicator of metastatic potential in papillary thyroid carcinoma Angiogênese como indicador do potencial de metástase no carcinoma papilífero tireóideo

Angiogenesis is new blood vessel formation, a process that can lead to tumor development. Microvessel count has been correlated to metastasis in some neoplasias. PURPOSE: To determine if measurement of microvessel density is useful in predicting metastasis to the cervical lymph node and prognosis in patients with papillary thyroid carcinoma. METHODS: A retrospective analysis was performed in 30 patients that had undergone total thyroidectomy. They were divided in 2 groups of 15 patients - with and without metastatic disease. Immunohistochemistry was used to detect expression of CD34 in archival paraffin-embedded papillary thyroid tumors, and microvessel density was calculated based on it. Association between microvessel density and the presence of metastasis, according to histological subtype, disease recurrence, and AMES prognostic index groups was determined through statistical analysis. RESULTS: The median microvessel density for the patient group without metastasis (200.0 microvessels/mm²) was apparently, but not significantly, less than that observed among metastatic disease patients (254.4 microvessels/mm²) (P = .20). When papillary carcinoma subtypes were analyzed, this difference became significant (P =.02). The follicular variant exhibited a greater microvessel density than the other subtypes, independent of metastasis presence. There was an apparent, but not significant, tendency for a larger median microvessel density in the group of patients that presented recurrence (294.4 microvessels/mm² vs 249.6 microvessels/mm², P = .11). There was no relationship between risk level and microvessel density: in the low- and high-risk groups, the median MVD was 304.0 microvessels/mm² and 229.6 microvessels/mm², respectively (P = .27). CONCLUSIONS: The results suggest that angiogenesis is more intense among metastatic tumors in the classic and the tall cell variants, indicating that microvessel count can be an indicator of the potential for metastasis in these subtypes of papillary thyroid carcinoma. Patients that developed recurrent disease had a tendency to exhibit higher angiogenesis; however, there was no association between microvessel density and the AMES prognostic index.<br>O desenvolvimento dos tumores depende da formação de neovasos, a angiogênese. Em algumas neoplasias, a alta densidade de microvasos tumorais correlaciona-se com a presença de metástase. OBJETIVO: Determinar se a medida da angiogênese pode indicar o potencial de metástase e o prognóstico do carcinoma papilífero tireóideo. MÉTODO: Foi feita análise retrospectiva de 30 tireoidectomizados, divididos em dois grupos de 15 indivíduos cada, respectivamente com e sem metástase. A partir dos blocos de parafina, foi calculada a densidade de microvasos no tecido tumoral por meio da quantificação da expressão do anticorpo CD34 pela imunohistoquímica. A associação da densidade de microvasos com a presença de metástase, ocorrência de recidiva e os grupos de risco do índice prognóstico AMES foi determinada por análise estatística. RESULTADOS: A mediana da densidade de microvasos no grupo de doentes sem metástase (200,0 microvasos/mm²) foi inferior àquela dos portadores de metástase (254,4 microvasos/mm²) (p = .2), sem atingir significância estatística. Ao considerar apenas os subtipos histológicos clássico e de células altas, essa diferença tornou-se significante (p = .02), uma vez que a variante folicular exibiu maior DMV que os demais subtipos, independente da presença de metástase. Houve tendência não significativa à maior densidade de microvasos entre aqueles que apresentaram recidiva (294,4 microvasos/mm² contra 249,6 microvasos/mm², p = .11). Nos grupos de baixo e alto risco, a mediana da densidade de microvasos foi de 304,0 microvasos/mm² e 229,6 microvasos/mm² respectivamente (p = .27). CONCLUSÃO: A angiogênese foi mais intensa nos tumores com metástase nos subtipos clássico e de células altas, sugerindo que a contagem de microvasos pode ser um indicador do potencial de metástase nestes subtipos histológicos do carcinoma papilífero tireóideo. Doentes que evoluíram com recidiva tenderam a exibir maior angiogênese, porém não houve associação da densidade de microvasos e o índice prognóstico

Unusual remodeling of the hyalinization band in vulval lichen sclerosus by type V collagen and ECM 1 protein

OBJECTIVES: The vulva is the primary site affected in lichen sclerosus, a chronic dermatosis in women that is histologically characterized by a zone of collagen remodeling in the superior dermis. The normal physiological properties of the vulva depend on the assembly of collagen types I (COLI), III (COLIII) and V (COLV), which form heterotypic fibers, and extracellular matrix protein interactions. COLV regulates the heterotypic fiber diameter, and the preservation of its properties is important for maintaining normal tissue architecture and function. In the current work, we analyzed the expression of COLV and its relationship with COLI, COLIII, elastic fibers and extracellular matrix protein 1 in vulvar biopsies from patients with lichen sclerosus. METHODS: Skin biopsies from 21 patients with lichen sclerosus, classified according to Hewitt histological criteria, were studied and compared to clinically normal vulvar tissue (N=21). Morphology, immunohistochemistry, immunofluorescence, 3D reconstruction and morphometric analysis of COLI, COLIII, COLV deposition, elastic fibers and extracellular matrix 1 expression in a zone of collagen remodeling in the superior dermis were performed. RESULTS: A significant decrease of elastic fibers and extracellular matrix 1 protein was present in the hyalinization zone of lichen sclerosus compared to healthy controls. The non-homogeneous distribution of collagen fibers visualized under immunofluorescence in the hyalinization zone of lichen sclerosus and control skin was confirmed by histomorphometry. Lichen sclerosus dermis shows a significant increase of COLI, COLIII and COLV expression compared to the healthy controls. Significant inverse associations were found between elastic fibers and COLV and between COLV and extracellular matrix 1 expression. A direct association was found between elastic fiber content and extracellular matrix 1 expression. Tridimensional reconstruction of the heterotypic fibers of the lichen sclerosus zone of collagen remodeling confirmed the presence of densely clustered COLV. CONCLUSIONS: Increased deposition of abnormal COLV and its correlation with extracellular matrix 1 and elastic fibers suggest that COLV may be a trigger in the pathogenesis of lichen sclerosus