ABSTRACT Objective: this research objective was to develop a new peritoneal adhesion animal model that would lead to adhesions formation in all operated animals, simple and reproducible, associated with maintenance the animal’s health. Methods: eighteen adult male Wistar rats (Rattus norvegicus) were randomly distributed into three groups: Control Group (anatomical and clinical parameters), Sham Group (delicate manipulation of the stomach and exposure of the peritoneal cavity to ambient air) and Surgery Group (gastrotomy followed by gastrorrhaphy). The animals were analyzed and classificated macroscopically according to two adhesion classification models and differences between groups were considered significant when p<0.05. Results: the six animals in the control group had no peritoneal adhesions, three of the six animals in the sham group had focal peritoneal adhesions, and all animals in the surgery group (gastrotomy followed by gastrorraphy) had firm peritoneal adhesions. All adhesions found were macroscopically quantified and microscopically confirmed, without carrying out a microscopic classification of the adhesions. Conclusion: the new model developed of gastrotomy followed by gastrorrhaphy, proved to be safe and efficient to induce and study peritoneal adhesions.

RESUMO Objetivo: o objetivo deste estudo foi criar um novo modelo animal de indução de aderências peritoneais capaz de levar à formação de aderências em todos os animais operados, simples e reprodutível, associado a manutenção da saúde dos animais. Métodos: Dezoito ratos machos, adultos, da linhagem Wistar (Rattus norvegicus) foram distribuídos aleatoriamente em três grupos: Grupo Controle (parâmetro anatômico e clínico), Grupo Sham (manipulação delicada do estômago e exposição de cavidade peritoneal ao ar ambiente) e Grupo Cirurgia (gastrotomia seguida de gastrorrafia). Os animais foram submetidos à análise e classificação macroscópicas, seguindo dois modelos de classificação de aderências. As diferenças entre os grupos foram consideradas estatisticamente significantes se p<0,05. Resultados: os seis animais do grupo controle não apresentavam aderências peritoneais, três dos seis animais do grupo sham apresentavam aderências peritoneais focais e todos os seis animais do grupo cirurgia (gastrotomia seguida de gastrorrafia) apresentavam aderências peritoneais firmes. Todas as aderências encontradas foram quantificadas macroscopicamente e confirmadas microscopicamente, sem a realização de classificação microscópica das aderências. Conclusão: o novo modelo desenvolvido, de gastrotomia seguida de gastrorrafia, mostrou-se seguro e eficiente para induzir e estudar aderências peritoneais.

Abstract Purpose: To evaluate the effects and mechanisms of andiroba-based emulsion (ABE) topical treatment on full-thickness cutaneous wounds in rats. Methods: The wounds were harvested on days 3, 7, 15, and 20 post-surgery. Wound contraction rate, quantitative immunohistochemistry [macrophages, myofibroblasts, capillaries, collagens (col) I and III, transforming growth factor β3β (TGFβ3)], and tensile strength were assessed. Results: Treated wounds were smaller, contracted earlier and had increased angiogenesis, fewer CD68+ and M2 macrophages on days 7 and 15, but higher on day 20. Myofibroblasts appeared on days 3 to 7 in untreated wounds and on days 7 to 15 in treated wounds. TGFβ3 levels were higher in the treated wounds, less dense collagen fibers, lower col I/III ratios and a higher tensile strength. Conclusion: These results demonstrate the important anti-inflammatory role of treatment and the associated modulation of macrophages, myofibroblasts, and TGFβ3 levels. Collagen fibers in the treated wounds were more organized and less dense, similar to unwounded skin, which likely contributed to the higher tensile strength.

Abstract Purpose: To evaluate the most frequent surgical techniques of high-risk colorectal anastomoses in rats. Methods: Wistar rats were enrolled in three different models comprising inflammatory (TNBS enema), vascular (portal vein occlusion) or obstructive (a non-ischemic constricting ring) mechanisms associated with colonic anastomosis that had accomplished after these former lesions. Histological analyses (Hematoxylin and eosin and Picrosirius red) were performed. Results: All anastomoses techniques were associated with risk factors and had complications, mainly anastomotic leakage. In Study 1, the use of a pharmacological agent, trinitrobenzene sulfonic acid (TNBS) mimicked an inflammatory bowel disease such as Crohn’s disease with 50% of anastomosis leakage, the higher percentage among all models tested. In Study 2, after portal ischemia followed by reperfusion it was observed a dense neutrophil infiltrate in the midst of necrotic tissue and fibrin at the anastomotic site and 5 days after the anastomosis, no collagen was produced. In Study 3, 5 days after the mechanical obstruction some denuded areas of epithelium with marked oedema of mucosa and submucosa were seen, at the anastomotic site and anastomosis group showed some reduction of collagen density when compared with Control/Sham group. Conclusion: All the experimental surgical techniques tested in rats were associated with high-risk colorectal anastomoses and were useful to study colonic anastomotic healing and intestinal leakage.

PURPOSE:Implement a laceration protocol of the rat lateral gastrocnemius (LG) and following-up its repair with ultrasound biomicroscopy (UBM), contractility tests and histology.METHODS: Sixty-three male Wistar rats were distributed into two groups. One, with sub-groups GI, GII and GIII (n=12), each containing right LG lacerated (n=6), control and sham (n=3) animals. LG muscles in GI, GII and GIII were inspected by UBM (40 MHz) immediately after, 14 and 28 days post-surgery and thereafter excised with four (GI), 14 (GII) and 28 (GIII) days post-surgery for histology. Animals in second group were distributed into right LG lacerated and control sub-groups. LG muscles in lacerated sub-group were submitted to contractility tests at four (n=8), 14 (n=8) and 28 (n=8) days post-surgery, while in the control sub-group (n=5) were submitted to contractility tests along the course of the experiments.RESULTS: Descriptive findings agreed between the lesion model, muscle repair, UBM images and histology. Contractility results for right LG were different (p<0.05) between control and injured muscle with four and 14 days post-surgery, at tetanic stimulating frequencies (50 and 70 Hz).CONCLUSION: A laceration protocol of the lateral gastrocnemius was implemented and ultrasound biomicroscopy, contractility and histology findings agreed regarding the following-up of injured muscle repair.

Macrófagos associados a tumores (MAT) representam o componente principal do estroma de muitos tumores, além de participar da angiogênese tumoral. Este estudo comparou a microdensidade vascular (MDV) e densidade de macrófagos infiltrando o tumor (DMIT) em carcinoma escamocelular da boca (CEC) com diferentes graus histológicos de malignidade. Análise histomorfométrica foi empregada após técnica imuno-histoquímica para os anticorpos fator von-Willebrand e CD68. Uma diferença significante entre MDV e carcinomas bem e moderadamente diferenciados foi observada (p<0,05). MAT estavam fortemente presentes em todos os tumores estudados e a DMIT não foi diferente entre os diferentes graus histológicos de malignidade do CEC (p=0,381). Correlação significante entre MDV e DMIT não foi observada (p=0,870). Em conclusão, os resultados desse estudo sugerem a influência de MAT e angiogênese nos diferentes graus histológicos de malignidade do CEC. Entretanto, a ausência de correlação entre MDV e DMIT sugere que a angiogênese não depende do número de macrófagos presentes neste tipo de câncer, mas do fenótipo predominante. Outros estudos devem ser realizados a fim de contribuir para melhor compreensão da participação de MAT na angiogênese tumoral.

Tumor-associated macrophages (TAM) are the main cellular component in stroma of many tumors and participate in tumor angiogenesis. The aim of present study was to compare the microvascular density (MVD) and infiltrating macrophage density (IMD) in oral squamous cell carcinomas (OSCCs) with different histological grades. A histomorphometric analysis was performed after immunohistochemistry using antibodies such as von-Willebrand factor and CD68. A significant difference in MVD was found between well and moderately differentiated OSCCs (p<0.05). TAM were largely present in all studied tumors and the IMD was not different among OSCCs with different histological grades (p=0.381). Significant correlation between MVD and IMD was not observed (p=0.870). In conclusion, these results suggest that TAM and angiogenesis have an influence at different histological grades of OSCC. However, the lack of correlation between MVD and IMD could suggest that angiogenesis does not depend on the number of macrophages present in OSCC, but their predominant phenotype. Further studies involving distinct phenotypes of macrophages should be done to better understand the influence of TAM on the tumor angiogenesis.

A cicatrização e remodelação do cólon resultam das modificações do colágeno na matriz extracelular. Algumas condições desequilibram sua renovação, enfraquecendo a resistência mecânica a cicatriz, como resultado da atividade elevada das metaloproteinases locais, e levando a um alto risco de deiscência. As metaloproteinases da matriz extracelular (matrix metalloproteinases, MMPs) constituem uma família de endopeptidases zinco-dependentes - metzincinas. São reconhecidos atualmente, em humanos, cerca de 24 genes responsáveis por cada uma delas. A colagenase (MMP-1) foi identificada por Gross e Lapière (1962) na cauda do girino da rã-touro americana. No câncer as MMPs tem ocupado um lugar especial. Evidências de que a célula neoplásica é capaz de interferir na modulação desta enzima - um co-fator associado à invasividade local e disseminação metastática. As MMP-2 e -7 são observadas com frequência no câncer de cólon, a MMP-12 parece exercer um efeito protetor (melhor prognóstico) e, ao contrário, a MMP-3 o torna pior. A associação entre alta atividade de MMPs, o pior prognóstico do câncer e o maior risco de deiscência de anastomose intestinal já vem sendo considerada, sugerindo uma trilogia consistente. A terapia farmacológica (inibidores MMPs) tem sido investigada, também para o controle do câncer. O artigo discute as informações mais relevantes e atualizadas sobre o assunto.

Colon healing and remodeling depends on the collagen changes in extracellular matrix. Some conditions, disrupt its turnover, causing strength weakening of the scar, as a result of high activity of local matrix metalloproteinases, causing a high risk of dehiscence. The extracellular matrix metalloproteinases are a family of zinc-dependent endopeptidases, or metzincines, and have been currently recognized in humans about 24 genes responsible for each one. The first MMP, colagenase (MMP-1), was described by Gross and Lapière (1962), while studying tadpole resorption of the American bullfrog. Metalloproteinases activity in cancer research, has taken a special place. Currently, evidences points to the cancer cell ability to interfere with enzymatic activity modulation - an co-factor which affects local invasiveness and metastatic dissemination. Both MMPs-2 and -7 have been frequently observed in colon cancer. Moreover, MMP-12 seems to counteract MMP-7 effect therefore considered as a protector and associated with better prognosis, in contrast to MMP-3, which may be responsible for a worse outcome. Association between high activity of MMPs, the prognosis of cancer and increased risk of intestinal anastomotic leakage has been highlighted, suggesting a consistent trilogy. Pharmacological therapy using MMPs inhibitors has been extensively studied, especially targeted for cancer control. The article discusses the most relevant information and updated information on the subject.

INTRODUÇÃO: A medicina regenerativa tem ganho grande importância nos últimos anos em decorrência da possibilidade de certas células se diferenciarem em linhagens celulares distintas e, assim, reconstruírem o tecido lesado. As células-tronco têm despontado como forma alternativa de tratamento para doenças pela sua capacidade de diferenciação nos mais de 100 tipos de tecido. A medula óssea contém células-tronco adultas, hematopoéticas e mesenquimais, que auxiliam na limitação do remodelamento cardíaco. MÉTODO: Foram utilizados 9 cães com peso entre 25 kg e 30 kg, divididos em três grupos: intracoronária, intramiocárdica-transendocárdica e retrógrada venosa. Células mononucleares da medula óssea foram coletadas por densidade Ficoll, marcadas com fluorocromo Hoechst e infundidas nas diferentes vias citadas anteriormente. RESULTADOS: Observou-se que, na via intracoronária, a distribuição das células foi homogênea do epicárdio ao endocárdio, em átrios e ventrículos direito e esquerdo; na via intramiocárdica-transendocárdica, as células ficaram mais restritas ao local da infusão, com consequente maior número em ventrículo esquerdo; e na via retrógrada venosa, as células encontravam-se com padrão de distribuição semelhante ao da via intramiocárdica, contudo sem que as mesmas chegassem à região endocárdica. Com relação à quantidade de células distribuídas pelo tecido, foi observado que havia diferença significativa (P < 0,01), quando analisada a interação via de infusão vs. região. CONCLUSÃO: Podemos concluir que existe a possibilidade de empregar técnicas distintas de entrega de células para diferentes doenças cardíacas.

BACKGROUND: Regenerative medicine has become increasingly important in recent years due to the possibility of certain cells to differentiate into different cell lines and thus recover the damaged tissue. The stem cell has emerged as an alternative treatment for diseases as a result of their ability to differentiate in more than 100 types of tissue. Bone marrow contains adult stem cells, hematopoietic and mesenchymal cells, which limit heart remodeling. METHODS: Nine dogs weighing between 25 and 30 kg were divided into three groups: intracoronary group, intramyocardial-transendocardial group and retrograde venous group. Mononuclear cells were collected from bone marrow by Ficoll density, stained with Hoechst fluorocrom and infused through the different routes mentioned above. RESULTS: It was observed that distribution of the cells was homogeneous from the epicardium to the endocardium in the right and left atrium and ventricle when the intracoronary route was used. With the transendocardial route the cells were more restricted to the infusion site, with a consequent larger number in the left ventricle. A similar distribution pattern was observed with retrograde venous infusion, however, cells did not reach the endocardial region. As to the number of cells distributed over the tissue, a significant difference (P < 0.01) was observed for the interaction between the infusion route versus region. CONCLUSION: We concluded that different cell delivery techniques may be used according to the different heart diseases.

FUNDAMENTOS: As neoplasias malignas da pele de grandes dimensões apresentam dificuldades de reconstrução após a excisão. OBJETIVO: O objetivo deste estudo foi avaliar a exeqüibilidade de uma nova proposta de cobertura para feridas cirúrgicas criadas após a ressecção de grandes tumores cutâneos, a combinação da derme acelular humana com epitélio autólogo cultivado. MÉTODOS: A aplicação dos substitutos de pele foi feita em quatro pacientes com área de implante variando de 33 a 120 cm2. Além da observação dos resultados clínicos, realizou-se estudo morfológico para avaliação da integração dos implantes. RESULTADOS: Ceratinócitos autólogos cultivados foram enxertados em dois pacientes e não demonstraram integração. A derme acelular foi aplicada em quatro pacientes, sendo que em um deles foram feitas duas aplicações. Dos cinco implantes de derme acelular realizados, dois não apresentaram integração, em dois a integração foi de 70%, e de 50% no último. CONCLUSÃO: A cobertura imediata e definitiva de defeitos cirúrgicos através da aplicação de derme acelular humana combinada com epitélio autólogo cultivado é exeqüível. Em oncologia cutânea apenas em situações especiais o uso de substitutos de pele pode ser conveniente no sentido de evitar reconstruções mais complexas.

BACKGROUND: Reconstruction difficulties may arise after excision of large malignant skin neoplasms.a OBJECTIVE: The objective of this study was to assess the feasibility of a new coverage for surgical wounds following resection of large skin tumors: a combination of human acellular dermis with cultured autologous epithelium. METHODS: The skin substitute was implanted in four patients, one of them received two implants and the area ranged from 33 to 120 cm2. Clinical results and morphologic studies were assessed as to implant integration. RESULTS: Cultured autologous epithelium was grafted in two patients and no integration was observed. The acellular dermis was applied to four patients. Out of five acellular dermis implants, two did not present integration, two presented 70% integration and the remaining, 50% integration. CONCLUSION: The immediate and definite coverage of surgical defects by means of application of human acellular dermis combined with cultured autologous epithelium is feasible. In skin oncology, the use of skin substitutes might be convenient only in special situations to avoid more complex reconstructions.

Histological and ultrastructural alterations in lung tissue of BALB/c mice infected with dengue virus serotype 2 (non-neuroadapted), by intraperitoneal and intravenous routes were analyzed. Lung tissues were processed following the standard techniques for photonic and electron transmission microscopies. Histopathological and ultrastructural studies showed interstitial pneumonia, characterized by the presence of mononuclear cells. In the mouse model, the dengue virus serotype 2 seems to led to a transient inflammatory process without extensive damage to the interalveolar septa, but caused focal alterations of the blood-exchange barrier. Endothelial cells of blood capillaries exhibited phyllopodia suggesting activation by presence of dengue virus. Morphometrical analysis of mast cells showed an expressive increase of the number of these cells in peribronchiolar spaces and adjacent areas to the interalveolar septa. Alveolar macrophages showed particles dengue virus-like inside rough endoplasmic reticulum and Golgi complex, suggesting viral replication. The tissue alterations observed in our experimental model were similar to the observed in human cases of dengue fever and dengue hemorrhagic fever. Our results show that BALB/c mice are permissive host for dengue virus serotype 2 replication and therefore provides an useful model to study of morphological aspects of dengue virus infection.

One of the main difficulties in studying dengue virus infection in humans and in developing a vaccine is the absence of a suitable animal model which develops the full spectrum of dengue fever, dengue haemorrhagic fever, and dengue shock syndrome. It is our proposal to present morphological aspects of an animal model which shows many similarities with the dengue infection in humans. BALB/c mice were intraperitoneally infected with non-neuroadapted dengue virus serotype 2 (DENV-2). Histopathological and morphometrical analyses of liver tissue revealed focal alterations along the infection, reaching wide-ranging portal and centrolobular veins congestion and sinusoidal cell death. Additional ultrastructural observations demonstrated multifocal endothelial injury, platelet recruitment, and alterated hepatocytes. Dengue virus antigen was detected in hepatocytes and in the capillar endothelium of the central lobular vein area. Liver function tests showed high levels of aspartate transaminase and alanine transaminase enzyme activity. Lung tissue showed interstitial pneumonia and mononuclear cells, interseptal oedema, hyperplasia, and hypertrophy of the bronchiolar epithelial cells. DENV-2 led to a transient inflammatory process, but caused focal alterations of the blood-exchange barrier. Viremia was observed from 2nd to 11th day p.i. by isolation of DENV-2 in C6/36 mosquito cell line inoculated with the supernatant of macerated liver, lung, kidney, and cerebellum tissues of the infected mice.

FUNDAMENTOS: O líquen escleroso (LS) apresenta zona de hialinização do colágeno na derme superior característica, que persiste pouco definida do ponto de vista morfológico e cujo significado permanece sem explicação. Já se pôde demonstrar que no LS há profundas modificações da matriz extracelular (MEC), com acúmulo de proteínas colagênicas e de glicosaminoglicanos sulfatados na região hialina. OBJETIVOS: Caracterizar morfologicamente a presença nessa zona de decorina e condroitim sulfato que, ao interagir com as fibrilas colágenas, entre outras proteínas matriciais, poderiam contribuir para essa peculiar apresentação da MEC. MÉTODOS: 31 casos de LS vulvar foram subdivididos segundo a gradação histológica de Hewitt e analisados por imuno-histoquímica utilizando anticorpos contra decorina e condroitim sulfato revelados pela diaminobenzidina. Esses resultados foram comparados aos do grupo controle constituído por fragmentos de retalhos cutâneos excisados durante cirurgias corretivas da região vulvoperineal. RESULTADOS: Ocorreu predomínio da decorina quando a matriz apresentava um aspecto frouxo/edematoso, e o condroitim sulfato foi mais evidente quando a MEC assumia um padrão compacto, parecendo que ambos contribuem para o aspecto hialino, porém em fases diferentes da patogenia dessa doença. CONCLUSÕES: A seqüência observada na síntese desses proteoglicanos/glicosaminoglicanos levou à suposição de que a decorina seja um possível marcador precoce do LS vulvar e de que o condroitim sulfato possa estar relacionado à contenção da alteração matricial no nível da derme média.

BACKGROUND: Lichen sclerosus is characterized by a collagenous hyaline/homogeneous zone at upper dermis, which remains undefined morphologically and biologically. Our previous investigation showed that there are several changes at the extracellular matrix hyaline zone, such as accumulation of collagenic proteins (types I and III) and sulfated proteoglycans/glycosaminoglycans. OBJECTIVES: Decorin and chondroitim sulfate (sulfated proteoglycans/glycosaminoglycans) immunoexpressions were the present investigation´s aim, emphasizing the hyaline zone related changes. METHODS: Thirty one vulvar LS untreated clinical lesions were biopsed and evaluated histologically according to Hewitt’s gradation and by immunohistochemical methods. Results were compared with ones of the control group, which was formed by cutaneous fragments from vulvoperineal corrective surgeries. RESULTS: We could demonstrate that decorin and chondroitin sulfate were present at the hyaline zone in different moments of matrix modulation. In all Hewitt stages chondroitin sulfate prevailed at the extracellular matrix in cases with a compact aspect of the hyaline zone while decorin was only seen in areas of less compactness. CONCLUSION: This proteoglycans/glycosaminoglycans synthesis sequence suggests that decorin may be a possible initial marker/indicator for vulvar LS. We suppose either that chondroitin sulfate is possibly a factor that limit matricial changes extension till middle dermis level.

OBJETIVO: Os autores propõem uma nova abordagem no tratamento de lesões extensas de natureza pré-cancerosa da mucosa jugal, utilizando enxerto de mucosa escamosa autógena cultivada em laboratório. MÉTODO: O enxerto é aplicado no mesmo tempo cirúrgico da ressecção da lesão original. Foram operados cinco pacientes, os quais receberam acompanhamento pós-operatório, sendo submetidos à biopsia de controle no 90º dia. A avaliação da integração do enxerto com o leito receptor foi realizada utilizando-se critérios clínicos e morfológicos, incluindo microscopia óptica e eletrônica. RESULTADOS: O estudo anatomopatológico com microscopia óptica e eletrônica dos cinco pacientes mostrou haver integração do enxerto da mucosa cultivada com o leito receptor. As células da mucosa cultivada formam camadas que se organizam e se diferenciam à semelhança da zona doadora. À microscopia eletrônica a mucosa enxertada apresentava lâmina basal com descontinuidades focais, presença de hemidesmosomas e fibrilas de ancoragem. CONCLUSÕES: Os resultados demonstraram que a técnica é oportuna e viável para o tratamento de lesões pré-cancerosas, outrora consideradas irressecáveis pela extensão e pode ser considerada uma possibilidade real para o tratamento cirúrgico definitivo das mesmas.

OBJECTIVE: The authors propose a new approach to the treatment of extensive pre-malignant lesions of the jugal mucosa, using laboratory grown autogenous squamous mucosa as graft. METHOD: The graft was implanted at the same time the original lesion was excised. Five patients underwent surgery and were followed up until a control biopsy was obtained after 90 days. Both clinical and morphological criteria, including light and electronic microscopy, were used to assess integration of the graft to the receptor bed. RESULTS: The anatomopathological study under light and electronic microscopy of the five patients showed integration of the cultivated mucosa to the receptor bed. Cells from the cultivated mucosa make up layers which organize and differentiate according to the pattern of the donor zone. The ultrastructure of the grafted mucosa showed a basal layer with discrete areas of loss of continuity, hemidesmosomes and anchorage fibrillae. CONCLUSIONS: Our results demonstrated that being a timely and feasible technique for the treatment of pre-malignant lesions, so far deemed not amenable to resection owing to their extension, the technique may be considered a real possibility for the definitive treatment.

Objetivo: avaliar o valor da presença da proteína p53 nos casos de recidiva/progressão da neoplasia intra-epitelial vulvar (VIN) III. Métodos: foram selecionadas 20 pacientes com VIN III indiferenciada, seguidas semestralmente por período de até quatro anos, divididas em dois grupos: quatorze sem e seis com recidiva/progressão da lesão. Os casos de recidiva/progressão foram distribuídos da seguinte forma: em três pacientes a recidiva ocorreu uma única vez, em duas, houve dupla recorrência e apenas uma evoluiu para carcinoma escamoso. Em ambos os grupos foram avaliados o sítio vulvar acometido e a presença da proteína p53 com análise do padrão de marcação imunohistoquímica. Estudo semelhante foi realizado nos casos de recidiva/progressão além da análise do intervalo de tempo para o surgimento de recidiva/progressão. Resultados: observou-se recidiva da VIN III em 25% dos casos e, em 5%, progressão para carcinoma. O tempo médio de recidiva foi de 24,5 meses. A localização multifocal da lesão primária foi a mais freqüente (50%) em ambos os grupos. Na maioria dos casos (87,5%), a recidiva/progressão ocorreu na mesma localização da lesão vulvar primária. A presença da proteína p53 mostrou-se positiva em 50% das lesões primárias de VIN III e em 75% dos casos de recidiva/progressão. Conclusões: a presença da proteína p53 parece desempenhar papel importante na gênese e na predição do curso clínico das VIN III. As recidivas/progressão das VIN III tendem a ocorrer na mesma área da doença inicial, sugerindo a presença de campo molecular alterado.

Purpose: to evaluate p53 overexpression value in vulvar intraepithelial neoplasia (VIN) III recurrence/progression. Methods: twenty patients with undifferentiated VIN III were selected and followed up every six months for four years and divided into two groups: fourteen without and six with recurrence/progression lesion. The recurrence/progression cases were distributed as follows: in three patients recurrence occurred only once; in two, twice, and only one progressed to squamous cancer. In both groups the site of vulvar lesion and p53 overexpression and immunostaining pattern were analyzed. A similar study was performed in recurrence/progression cases, besides the analysis of the time interval to occur the arise of recurrence/progression. Results: recurrence was observed in 25% of the cases and, in 5%, progression to carcinoma. The mean time interval for recurrence was 24.5 months. Multifocal location of the initial lesion was the predominant form (50%) in both groups. In the majority of the cases (87.5%) recurrence/progression occurred at the same site of the initial vulvar lesion. p53 overexpression was observed in 50% of the VIN III primary lesions and in 75% of the recurrence/progression cases. Conclusions: p53 overexpression seems to play an important role in VIN III pathogenesis and may predict the clinical course of the lesions. VIN III recurrence/progression has a tendency to occur in the same area of the initial lesion, suggesting the presence of molecular disturbance.

Mast cells and eosinophils actively participate in tissue repair and are prominent components of Schistosoma mansoni granulomas. Since pentoxifillyne (PTX) is an immunomodulatory and antifibrotic substance, we aimed to characterize, by morphological techniques, the effect of this drug on fibrosis developed inside murine hepatic schistosomal granulomatous reaction, beyond the quantification of eosinophil and mast cell populations. The drug (1 mg/100 g animal weight) was administrated from 35 to 90 days post-infection, when the animals were killed. The intragranulomatous interstitial collagen network was analyzed by confocal laser scanning microscopy, the number of eosinophils and mast cells was quantified and the results were validated by t-student test. Treatment did not interfere on the granuloma evolution but caused a significant decrease in the total and involutive number of hepatic granulomas (p = 0.01 and 0.001, respectivelly), and in the intragranulomatous accumulation of eosinophils (p = 0.0001). Otherwise, the number of mast cells was not significantly altered (p = 0.9); however, it was positively correlated with the number of granulomatous structures (r = 0.955). In conclusion, PTX does not affect development and collagen deposition in S. mansoni murine granuloma, but decreases the intragranulomatous eosinophil accumulation possibly due to its immunomodulatory capability, interfering in cellular recruitment and/or differentiation.