Hepsin immunohistochemical expression in prostate cancer in relation to Gleason's grade and serum prostate specific antigen.

Background: Hepsin (HPN), a type II trans-membrane serine protease, has been reported to be one of the most up regulated genes in prostate cancer. The aim of the present study was to find out immuno-histochemistry based phenotypic expression of HPN in formalin fixed paraffin embedded sections of prostate cancer compared with that in benign prostatic hyperplasia, in a prospective clinical setting, to know the differential status of HPN expression in benign and malignant prostatic disease. Materials and Methods: Tissue biopsies of histologically proven cases of prostatic cancers (48), benign prostatic hyperplasia (42), benign prostatic hyperplasia with prostatic intraepithelial neoplasia (7) and 4 cases of benign prostatic hyperplasia with prostatitis, were subjected to immunohistochemical staining with HPN antibody by strepavidin biotin method. Results: Hepsin expression was 100% in prostate carcinoma, 11.9% (5/42) in benign prostatic hyperplasia, 57.14% (4/7) in benign prostatic hyperplasia with prostatic intraepithelial neoplasia (PIN), and none in benign prostatic hyperplasia with prostatitis. Hepsin staining showe higher expression in high grade tumor in comparison to low grade tumor. Conclusions: Positive immunohistochemical expression of hepsin in cent percent cases of prostate cancer cases is intriguing, underscoring the significance of hepsin gene expression in prostate cancer.

Subcellular localization of an intracellular serine protease of 68 kDa in Leishmania (Leishmania) amazonensis promastigotes

Here we report the subcellular localization of an intracellular serine protease of 68 kDa in axenic promastigotes of Leishmania (Leishmania) amazonensis, using subcellular fractionation, enzymatic assays, immunoblotting, and immunocytochemistry. All fractions were evaluated by transmission electron microscopy and the serine protease activity was measured during the cell fractionation procedure using a-N-r-tosyl-L-arginine methyl ester (L-TAME) as substrate, phenylmethylsulphone fluoride (PMSF) and L-1-tosylamino-2-phenylethylchloromethylketone (TPCK) as specific inhibitors. The enzymatic activity was detected mainly in a membranous vesicular fraction (6.5-fold enrichment relative to the whole homogenate), but also in a crude plasma membrane fraction (2.0-fold). Analysis by SDS-PAGE gelatin under reducing conditions demonstrated that the major proteolytic activity was found in a 68 kDa protein in all fractions studied. A protein with identical molecular weight was also recognized in immunoblots by a polyclonal antibody against serine protease (anti-SP), with higher immunoreactivity in the vesicular fraction. Electron microscopic immunolocalization using the same polyclonal antibody showed the enzyme present at the cell surface, as well as in cytoplasmic membranous compartments of the parasite. Our findings indicate that the internal location of this serine protease in L. amazonensis is mainly restricted to the membranes of intracellular compartments resembling endocytic/exocytic elements.

Inflammatory mediators in nasal lavage among school-age children from urban and rural areas in São Paulo, Brazil

CONTEXTO: Alguns estudos demonstram que o processo inflamatório nas vias aéreas nasais poderia refletir ou mesmo afetar as vias aéreas inferiores. Decidimos avaliar indiretamente o estado inflamatório das vias aéreas nasais de dois grupos de escolares com diferente sensibilização aos aeroalérgenos mais comuns. OBJETIVO: Comparar a atividade inflamatória nas vias aéreas nasais, através da determinação de mediadores inflamatórios no lavado nasal em duas populações distintas de crianças em idade escolar. TIPO DE ESTUDO: Estudo transversal. LOCAL: O estudo foi realizado em duas escolas públicas de ensino fundamental, uma em zona urbana e outra em zona rural, no Estado de São Paulo. MÉTODOS: Foram constituídos dois grupos de 40 escolares que apresentam diferentes taxas de sensibilização a aeroalérgenos comuns. Amostras do lavado nasal foram colhidas para determinação de proteína catiônica eosinofílica (ECP) e triptase. Testes não-paramétricos foram usados na análise estatística. RESULTADOS: Níveis significativamente maiores de proteína catiônica eosinofílica foram encontrados nos estudantes da área urbana (p < 0,05). Não houve diferença estatística nos níveis de triptase entre os dois grupos. Observou-se ainda que, na área urbana, as crianças sensibilizadas aos aeroalérgenos apresentaram maiores concentrações de proteína catiônica eosinofílica, o que não foi observado nas crianças da zona rural. DISCUSSAO: A ausência de atividade de mastócitos e a degranulação aumentada de eosinófilos revelaram uma inflamação crônica nas vias aéreas das crianças estudadas. A maior atividade de eosinófilos na zona urbana, coincidindo com a maior sensibilização aos aeroalérgenos, sugere que deve haver algum fator a mais na área urbana que modula a resposta das vias aéreas influenciando a ativação das células inflamatórias locais. CONCLUSAO: Nossos achados não mostraram diferenças nos níveis de triptase no lavado nasal entre os dois grupos estudados. Por outro lado, as crianças da area urbana apresentaram maiores concentrações de proteína catiônica eosinofílica do que aquelas da zona rural. Observamos ainda que, na area urbana, as crianças sensibilizadas por aeroalérgenos apresentaram maiores concentrações de proteína catiônica eosinofílica do que aquelas não sensibilizadas, enquanto esta diferença não foi observada nas crianças da area rural.

Eosinophil Inflammation of Nasal Polyp Tissue: Relationships with Matrix Metalloproteinases, Tissue Inhibitor of Metalloproteinase-1, and Transforming Growth Factor-beta1

Eosinophil and mast cell infiltrations are consistent findings in nasal polyp tissue. Previous studies have shown that matrix metalloproteinases (MMPs) may be involved in eosinophil infiltration in airway mucosa of asthmatic patients, and that transforming growth factor-beta1 (TGF-beta1) induces extracellular matrix deposition in nasal polyp tissue. The aim of this study was to evaluate the role of MMPs and tissue-inhibitor of metalloproteinase-1 (TIMP-1) in association with TGF-beta1, eosinophils and mast cell activation in nasal polyp tissue. Nasal polyp tissues from 20 patients who underwent polypectomies were collected and prepared into tissue homogenate. Eosinophil cationic protein (ECP) and tryptase levels were measured by CAP system (Pharmacia, Sweden). MMP-2, MMP-9, TIMP-1 and TGF-beta1 levels were measured by enzyme-liked immunosorbent assay. MMP-2 was the predominant form of MMPs, followed by MMP-9 and TIMP-1. There were significant correlations between ECP, and MMP-9, MMP-2, TGF-beta1 and tryptase, but not with TIMP-1. Significant correlations were noted between tryptase, and MMP-2, MMP-9, and TGF-beta1, but not with TIMP-1. Close correlations were noted between TGF-beta1, and MMP-9 and MMP-2, but not with TIMP-1. MMP-2, MMP-9, and TGF-beta1 may contribute to eosinophil and mast cell migrations into nasal polyp tissue.