Comparative analysis of the expression of proliferating cell nuclear antigen, p53, bax, and bcl-2 in oral lichen planus and oral squamous cell carcinoma

Several epidermiologic studies have shown the malignant transformation potential of oral lichen planus; however, this potential is subject of much controversy. To evaluate the expression of proteins related to the cell proliferation and apoptosis processes in oral lichen planus, we compared oral lichen planus with oral squamous cell carcinoma. Twenty-four cases of each lesion were submitted according to streptavidin-biotin technique to evaluate the immunohistochemical expression of proliferating cell nuclear antigen, p53, bax, and bcl-2 proteins. X 2 test showed no statistically significant differences between the expression of p53, bax, and bcl-2 in oral lichen planus and oral squamous cell carcinoma (P > .05). However, the expression of proliferating cell nuclear antigen was significantly lower in oral lichen planus than in oral squamous cell carcinoma (P < .05). No statistically significant differences between the expression of p53, bax, and bcl-2 in oral lichen planus and oral squamous cell carcinoma were observed, which may be an evidence of the potential of malignant transformation of oral lichen planus. (C) 2009 Elsevier B.V. All rights reserved.

Leiomioma vascular bucal: relato de dois casos, revisão da literatura e estudo imuno-histoquímico

INTRODUÇÃO: Os leiomiomas são neoplasias benignas do músculo liso que ocorrem mais comumente nos tratos geniturinário e gastrintestinal, entretanto são raros na cavidade bucal, na qual provavelmente a maioria desses tumores tem sua origem a partir de músculo liso vascular. OBJETIVOS: Relatar dois casos com história clínica semelhante, confrontando seus aspectos com aqueles encontrados na literatura. RELATO DOS CASOS: Dois casos de mulheres na quinta e sexta décadas de vida apresentando nódulo submucoso em ventre lingual. Após exame microscópico que revelou numerosos vasos sangüíneos entremeados a células fusiformes, com núcleos ovalados ou alongados, e exame imuno-histoquímico com positividade para actina de músculo liso nos dois casos, o diagnóstico final foi leiomioma vascular. CONCLUSÃO: O estudo do leiomioma vascular bucal é de extrema importância devido à raridade e semelhança desse com outras lesões da cavidade bucal. A análise imuno-histoquímica é importante para o diagnóstico final do leiomioma.

Immunolocalization of bone morphogenetic protein 2 during the early healing events after guided bone regeneration

Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)

Evaluation of epithelial cell proliferating activity and fibroblast nuclear kariometry in recurrent pterygium treated with mitomycin C

OBJETIVO: Avaliar a eficácia da mitomicina C (MMC) na prevenção da recorrência quando previamente utilizada no transplante autólogo de conjuntiva (TAC). A avaliação da proliferação celular epitelial pelo antígeno Ki-67 e a cariometria do núcleo dos fibroblastos foram usados como auxiliares na avaliação do tratamento. MÉTODOS: Vinte e nove pacientes com pterígio recidivado foram divididos em três grupos: Grupo (G) 1-TAC e colírio placebo (PLA); G2-TAC, MMC 0,015% subconjuntival e PLA; G3-TAC e colírio de MMC 0,02%. A imuno-histoquímica foi realizada no tecido excisado para o antígeno Ki-67, como a cariometria dos núcleos dos fibroblastos (divididos em lado nasal e temporal). A cariometria dos núcleos dos fibroblastos foi avaliada de acordo com os seguintes parâmetros: volume (Vl) e área (Ar) em pelos menos 50 células por paciente. RESULTADOS: A porcentagem das células epiteliais positivas para o antígeno Ki-67 no lado nasal e temporal após o tratamento dos três grupos estudados foi: nasal (3,30% G1, 4,49% G2 e 3,38% G3) e temporal (3,30% G1, 4,46% G2 e 4,14% G3) não mostrando diferença significativa. A cariometria do núcleo dos fibroblastos foi: Vl nasal (792,1 µ3 G1, 605,1 µ3 G2, e 549,9 µ3 G3) e a Ar (100,58 µ2 G1, 83,13 µ2 G2, e 78,41 µ2 G3). Os três grupos mostraram uma diferença significativa p=0,039 e p=0,035, respectivamente do Vl e da Ar no lado nasal. Após seis meses de tratamento, os três grupos apresentaram a seguinte taxa de recidiva: 22,22% G1, 18,18%, G2 e 33,33% G3 respectivamente. CONCLUSÃO: O uso da MMC não interferiu nas células epiteliais positivas para o antígeno Ki-67 no pterígio recidivado, mas acarretou diminuição do volume e área dos núcleos dos fibroblastos no lado nasal do pterígio. As células epiteliais positivas para o antígeno Ki-67 parecem não ter relação com a recidiva do pterígio após seis meses da cirurgia. Outros estudos devem ser realizados para avaliar o papel da proliferação das células epiteliais na recorrência do pterígio.

Arterial diffuse intimal thickening associated with enzootic calcinosis of sheep

Foram feitos estudos morfométrico, imunoistoquímico e ultra-estrutural do espessamento intimal difuso (DIT) das artérias de 7 ovinos com sinais clínicos de calcinose enzoótica espontânea causada pela ingestão da planta Nierembergia veitchii. As lesões caracterizavam-se por deposição de sais de cálcio na média como placas e estrias que, com frequência faziam saliência para a luz, criando rugosidades e irregularidades da íntima. Não foram observadas calcificações na artéria pulmonar e no sistema venoso. Microscopicamente, as calcificações das artérias restringiam-se quase que exclusivamente à média. Na imunoistoquimica, foi verificada a-actina nas células da média e nas do espessamento intimal. Receptores para 1,25(OH)2D3 foram evidenciados nos núcleos das células musculares da média, da íntima e das células endoteliais. As análises morfométricas em microscopia ótica revelaram, nas artérias, DIT irregularmente distribuido sem relação com a intensidade dos processos de calcificação subjacente nem com a espessura da média remanescente. A morfometria das alterações ultra-estruturais das células musculares lisas da média e da íntima espessada, mostrou que nessas últimas foi verificado aumento de até 318% na fração volumétrica das organelas de síntese em detrimento dos elementos contráteis, quando comparados com os valores das células da média. Essas modificações indicam a transformação das células musculares lisas de um tipo contrátil para células de tipo sintético. Entre essses dois extremos foram evidenciadas diversas formas intermediárias. Foram observadas evidências histológicas e ultra-estruturais da transformação e migração de células da média através das fenestras da lâmina elástica interna, para a íntima, onde proliferam e formam o DIT. Foi concluído que o DIT é um componente constante nas lesões arterioscleróticas nas calcinoses dos ovinos induzidas pela planta N. veitchii e que as células predominantes são células musculares lisas provenientes de predecessoras da média. Foi constatado que o DIT só excepcionalmente apresenta calcificação, vascularização, presença de células espumosas e fendas de colesterol. Não foram vistas áreas de necrose. Sugere-se que o fator indutor das alterações arteriais é o 1,25(OH)2D3 contido em N. veitchii.

Profile of thyroid hormones in breast cancer patients

Estrogen involvement in breast cancer has been established; however, the association between breast cancer and thyroid diseases is controversial. Estrogen-like effects of thyroid hormone on breast cancer cell growth in culture have been reported. The objective of the present study was to determine the profile of thyroid hormones in breast cancer patients. Serum aliquots from 26 patients with breast cancer ranging in age from 30 to 85 years and age-matched normal controls (N = 22) were analyzed for free triiodothyronine (T3F), free thyroxine (T4F), thyroid-stimulating hormone (TSH), antiperoxidase antibody (TPO), and estradiol (E2). Estrogen receptor ß (ERß) was determined in tumor tissues by immunohistochemistry. Thyroid disease incidence was higher in patients than in controls (58 vs 18%, P < 0.05). Subclinical hyperthyroidism was the most frequent disorder in patients (31%); hypothyroidism (8%) and positive anti-TPO antibodies (19%) were also found. Subclinical hypothyroidism was the only dysfunction (18%) found in controls. Hyperthyroidism was associated with postmenopausal patients, as shown by significantly higher mean T3 and T4 values and lower TSH levels in this group of breast cancer patients than in controls. The majority of positive ERß tumors were clustered in the postmenopausal patients and all cases presenting subclinical hyperthyroidism in this subgroup concomitantly exhibited Erß-positive tumors. Subclinical hyperthyroidism was present in only one of 6 premenopausal patients. We show here that postmenopausal breast cancer patients have a significantly increased thyroid hormone/E2 ratio (P < 0.05), suggesting a possible tumor growth-promoting effect caused by this misbalance.

Fator de crescimento fibroblástico básico e seus receptores em relação à atividade proliferativa na placenta bubalina em diferentes fases da gestação

Estudou-se a distribuição espaço-temporal do fator de crescimento fibroblástico básico (bFGF), do receptor 1 do fator de crescimento fibroblástico (FGFR1) e do receptor 2 do fator de crescimento fibroblástico (FGFR2) na placenta bubalina, correlacionando-a à proliferação celular. Para a detecção do bFGF, FGFR1, FGFR2 e antígeno Ki-67, colheram-se 12 placentas de búfalas nos terços inicial, médio e final da gestação, em abatedouros, e realizaram-se testes de imunoistoquímica. Detectou-se e avaliou-se a expressão do bFGF, do FGFR1, do FGFR2 e do antígeno Ki-67 ao longo da gestação. No compartimento fetal da placenta, observaram-se correlações positivas entre a expressão do bFGF e Ki-67, entre FGFR1 e Ki-67 e entre FGFR2 com Ki-67 (r=0,313, 0,358 e 0,384, respectivamente). No epitélio e estroma maternos observaram-se altas correlações entre FGFR1 e Ki-67 (r=0,739 e r=0,511, respectivamente). Os resultados sugerem envolvimento do bFGF, FGFR1 e FGFR2 na proliferação do trofoblasto enquanto no compartimento materno da placenta bubalina apenas o FGFR1 atuaria como modulador dessa atividade.

Histopathological findings and detection of parasites in the eyes of dogs infected naturally with Leishmania chagasi

Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)

Estudo comparativo morfofuncional de melanócitos em lesões de melasma

FUNDAMENTOS - Melasma é hipermelanose comum caracterizada por máculas acastanhadas em áreas fotoexpostas, cuja fisiopatogenia não é totalmente esclarecida. OBJETIVOS - Caracterizar e comparar morfologica e funcionalmente os melanócitos da epiderme sã com os da pele afetada por melasma. MÉTODOS - Avaliaram-se 12 pacientes portadores de melasma facial, sendo realizadas biópsias da pele lesada e pele sã adjacente. Os cortes foram corados por hematoxilina-eosina, Fontana-Masson, marcados pelo Melan-A e submetidos à microscopia eletrônica. A quantificação epidérmica de melanina e melanócitos foi estimada a partir de análise citomorfométrica digital. RESULTADOS - Todas as pacientes eram mulheres com média de idade 41,3±2,8 anos. Ao Fontana-Masson evidenciou-se importante aumento da melanina epidérmica na pele lesada em relação à pele sã. A marcação pelo Melan-A demonstrou melanócitos maiores com dendritos proeminentes na pele lesada. Observou-se maior densidade de melanina epidérmica na pele lesada, e a análise digital do número de melanócitos da epiderme não demonstrou diferença significativa entre pele lesada e sã. À microscopia eletrônica, observaram-se número aumentado de melanossomas maduros nos ceratinócitos e melanócitos com organelas citoplasmáticas proeminentes na pele lesada. CONCLUSÕES - Melanogênese aumentada na epiderme com melasma em relação à epiderme normal adjacente.

EFFECTS OF CYCLOSPORINE-A IMMUNOSUPPRESSION ON PANCREATICODUODENAL TRANSPLANTATION IN ALLOXAN-DIABETIC RATS

1. Forty-five outbred Wistar rats were randomly assigned to three experimental groups: GI, 10 non-diabetic control rats; GII, 10 alloxan-diabetic control rats; GIII, 25 alloxan-diabetic tats which received pancreaticoduodenal transplantation (PDT) from normal Wistar donor rats and were immunosuppressed with cyclosporin A (Cy-A), 10 mg kg body weight-1 day-1, administered intraperitoneally for 30 days.2. In parallel, 15 alloxan-diabetic inbred Wistar rats received isogeneic PDT from normal Wistar donor rats.3. Cy-A prevented graft rejection in the 15 surviving animals in group III. These observations were confirmed by clinical and biochemical parameters (body weight, urine output, water and food intake, blood and urinary glucose and plasma insulin) and by histology and immunohistochemistry of the pancreas.4. However, Cy-A was associated with 60% of the infectious complications in transplanted rats leading to 40% mortality. Pulmonary infections were the main cause of death. There were no side effects of immunosuppression on the pancreas. Infections were not significant in inbred rats submitted to PDT.

Epithelial-stromal transition of MMP-7 immunolocalization in the rat ventral prostate following bilateral orchiectorny

Epithelial cells from involuting rat ventral prostate (VP) express Matrilysin (MMP-7) mRNA. Herein, we investigated by immunohistochemistry the NIMP-7 protein location and its association with tissue changes following castration in the VP. Normal and castrated adult male Wistar rats were sacrificed at different times after surgery. VP was examined by immunocytochemistry and immunoprecipitation. Castration promoted a shrinking of prostate ducts with an extensive stromal remodeling. In the VP from normal rats, MMP-7 immunoreactivity was found in epithelial secretory granules. Three days after castration, immunostaining for MMP-7 was found in both the epithelial secretory granules and in the stroma just below the epithelium, mainly at the distal ductal tips. At seven and 21 days after castration, the immunostaining for MMP-7 was found only in the stromal space. Immunoprecipitation confirmed the specificity of the primary antibody by rescuing a pro-enzyme form (28 kDa) in the prostate extracts. The present results suggest that MMP-7 participates in the epithelial-stromal interface remodeling of the ventral prostate during the involution achieved by castration, probably in the degradation of components of the epithelial basement membrane. (c) 2007 International Federation for Cell Biology. Published by Elsevier Ltd. All rights reserved.

DUCTAL CARCINOMA IN-SITU OF THE BREAST - HISTOLOGIC CATEGORIZATION AND ITS RELATIONSHIP TO PLOIDY AND IMMUNOHISTOCHEMICAL EXPRESSION OF HORMONE RECEPTORS, P53, AND C-ERBB-2 PROTEIN

Background. Ductal carcinoma in situ (DCIS) of the breast has been diagnosed increasingly since the advent of mammography. However, the natural history of these lesions remains uncertain. Ductal carcinoma in situ of the breast does not represent a single entity but a heterogeneous group with histologic and clinical differences. The histologic subtype of DCIS seems to have an influence on its biologic behavior, but there are few studies correlating subtype with biologic markers.Methods. The authors studied a consecutive series of 40 cases of DCIS and after its histologic categorization verified its relationship with ploidy using image analysis and analyzing estrogen receptor (ER), progesterone receptor (PR), p53 and c-erbB-2 expression using immunohistochemistry.Results. The three groups proposed according to the grade of malignancy were correlated significantly with some of the additional parameters studied, including aneuploidy and c-erB-2 expression. Aneuploidy was detected in 77.5% of cases of DCIS mainly in high and intermediate grade subtypes (100% and 80% vs. 35.7% in low grade) whereas immunoreactivity for c-erbB-2 was detected in 45% of cases of DCIS mainly in the high grade group. Expression of ER and PR were observed frequently in this study (63.9% and 65.7% respectively), but without correlation with the histologic subtype of DCIS, although we found a somewhat significant association between high grade DCIS and lack of ER. p53 protein expression was detected in 36.8% of these cases, but no relationship between this expression and histologic subtype or grading of DCIS was found.Conclusions. These results provide further evidence for the morphologic and biologic heterogeneity of DCIS. Besides histologic classification and nuclear grading, some biologic markers such as aneuploidy and c-erbB-2 expression constitute additional criteria of high grade of malignancy.

AIDS-related lymphoma in Brazil - Histopathology, immunophenotype, and association with Epstein-Barr virus

The occurrence of malignant lymphoma is an increasingly important cause of morbidity and mortality in AIDS patients. The incidence of AIDS-related lymphoma in some developing countries such as Brazil is increasing as the survival of HIV infection has improved. Although there is a clear association between several types of immunodeficiency related lymphomas and Epstein-Barr virus (EBV), the association of EBV infection in AIDS-related lymphoma in Brazil, where the incidence of AIDS is high, is unknown. Formalin-fixed, paraffin-embedded tissue from 24 cases of AIDS-related lymphoma in Brazil were analyzed for morphologic classification, immunophenotype, and EBV association using in situ hybridization studies with an EBV-EBER1 biotinylated probe. Twenty cases of AIDS-related lymphoma were classified as non-Hodgkin's lymphoma and four cases were Hodgkin's disease. Eleven non-Hodgkin's lymphomas were classified as diffuse large cell type, five cases were small non-cleaved cell, Burkitt-type, and four cases were large cell immunoblastic non-Hodgkin's lymphoma. Eighteen cases were of B-cell phenotype; one was a T-cell lymphoma, and one was classified as null. Epstein-Barr virus (EBV) was demonstrated in the majority of tumor cells of 11 of 20 (55%) of the cases non-Hodgkin's lymphomas and in 3 of 4 (75%) cases of Hodgkin's disease. AIDS-related lymphomas in Brazil are usually of large cell/immunoblastic type, but Hodgkin's disease is also seen. Both non-Hodgkin's lymphoma and Hodgkin's disease are often associated with EBV infection. The non-Hodgkin's lymphoma is predominantly of B-cell phenotype.

Immunohistochemical characterization of mononuclear cells in delayed hypersensitivity reactions to Paracoccidiodes brasiliensis (paracoccidioidin test)

The density and distribution of T cells, T helper cells, macrophages and B cells at the site of skin tests with a cytoplasmic Paracoccidioides brasiliensis antigen (paracoccidioidin) was studied at 24 and 48 h post-challenge in 10 patients with the chronic form of paracoccidioidomycosis and in 5 noninfected individuals. The in situ study was carried out using immunoperoxidase techniques and monoclonal antibodies. The controls showed negative skin test. In the patients, the great majority of the cells in the perivascular foci were T cells (CD43-positive cells) making up 47% and 48.6% of the total number of cells at 24 and 48 h respectively. Most of the T cells showed a T helper phenotype (CD45RO-positive cells). Approximately 25% of the cells were macrophages (CD68-positive cells) and there were very few B lymphocytes (CD20-positive cells). The present data on the microanatomy of paracoccidioidin skin test sites were consistent with a delayed type hypersensitivity pattern. Our results were comparable to those reported on skin tests for other granulomatous chronic diseases.

Aneuploidies, deletion, and overexpression of TP53 gene in intestinal metaplasia of patients without gastric cancer

Gastric carcinogenesis is attributable to interacting environmental and genetic factors, through a sequence of events including intestinal metaplasia. Using a fluorescence in situ hybridization technique, we investigated the occurrence of ancuploidies of chromosomes 3, 7, 8, 9, and 17, TP53 gene deletion, and expression of p53 in 21 intestinal metaplasia (IM) samples from cancer-free patients and in 20 gastric adenocarcinoma samples. Aneuploidies were found in 71% (15/21) of the IM samples. Trisomy of chromosomes 7 and 9 occurred mainly in complete-type IM; in the incomplete type, trisomy of chromosomes 7 and 8 were more commonly found. The TP53 gene deletion was observed in 60% (3/5) of the IM cases, and immunohistochemistry revealed p53 overexpression in 12% (2/17) of the analyzed IM cases. All gastric adenocarcinoma cases presented higher frequencies of trisomy or tetrasomy of chromosomes 3, 7, 8, 9, and 17. The TP53 deletion was found in all three of the gastric adenocarcinoma analyzed for it, and immunohistochemistry detected overexpression of protein p53 in 80% (12/15) of the analyzed cases. Our study revealed for the first time the presence of aneuploidies of chromosomes 7, 8, 9, and 17 and of TP53 gene deletion and overexpression in IM samples from cancer-free patients. These results suggest that IM and gastric adenocarcinoma may share the same genetic alterations. (C) 2004 Elsevier B.V. All rights reserved.