Histochemical demonstration of mouse submandibular esterproteases with a new chromogenic substrate

Lexow, Ullrich; Grossarth, C; O, von Deimling

doi:10.1007/bf00500661

Cited by 16 publications

(7 citation statements)

References 14 publications

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“…Chymase‐positive MCs were stained by using the substrate N ‐acetyl‐l‐methionine alpha‐naphtyl ester (u–N–O–Met) as previously described (16). Briefly, sections were deparaffinized, rehydrated, and washed in 0.15 M phosphate buffer, pH 7.1.…”

Section: Methodsmentioning

confidence: 99%

“…Enzymehistochemical staining of chymase-positive mast cells Chymase-positive MCs were stained by using the substrate N-acetyl-l-methionine alpha-naphtyl ester (u-N-O-Met) as previously described (16 using a Nikon Diaphot 300 microscope (Nikon, Melville, NY, USA) equipped with an OC-M calibrated eyepiece micrometer and connected to an IMAGEPRO analysis program 4.0.1 (Media Cibernetics, Atlanta, GA, USA). MCs were counted in 30 counting fields in normal lip sections (including epithelium/connective tissue junction, connective, and submucous zones) and in 20 counting fields per lip SCC section at 40· magnification (counting field area ¼ 0.4 mm 2 ).…”

Section: Lip Biopsiesmentioning

confidence: 99%

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Characterization of mast cell subpopulations in lip cancer

Rojas

Spencer

Martínez

et al. 2005

J Oral Pathology Medicine

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Section: Methodsmentioning

confidence: 99%

Section: Lip Biopsiesmentioning

confidence: 99%

Characterization of mast cell subpopulations in lip cancer

Rojas

Spencer

Martínez

et al. 2005

J Oral Pathology Medicine

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“…At pH 6.8 the proteinase splits Z-AlaOnap and C1Ac-OnapAS-D (nonspecific substrates for human neutrophil leukocyte elastase; Starkey and Barrett 1976) but shows no elastinolytic activity at the pH range of 7.2-10.0. It cleaves Ac-Met-Onap (a substrate for esterproteinases from the mouse submandibular gland; Lexow et al 1979;Schaller and von Deimling 1979) but is resistant to 1 mM pNPP (an inhibitor of these enzymes; Schaller and von Deimling 1979). Under histochemical conditions the proteinase is incapable of hydrolyzing Nblocked DL-phenylalanine-2-naphthylester (a substrate for chymotrypsin; Ravin et al 1954) as well as N-blocked naphthylamides of L-trialanine (a relatively specific substrate for elastase; Sweetman and Ornstein 1974;Smith and van Frank 1975), L-arginine and L-diarginine (substrates for trypsin; Smith and van Frank 1975), and of L-leucine, L-phenylalanine, L-phenylalanyl-L-arginine, diglycyl-L-arginine, glycyl-L-proline, and L-proline.…”

Section: Discussionmentioning

confidence: 99%

A serine proteinase in the penetration glands of the cercariae of Plagiorchis elegans (trematoda, plagiorchiidae)

Moczon

1996

Parasitol Res

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Cercariae of Plagiorchis elegans secrete a serine proteinase from their penetration glands. The enzyme hydrolyzes both azocoll and gelatin at the optimal pH of 8.4 but is incapable of hydrolyzing elastin at the pH range of 7.2-10.0 Ca2+ and Mg2+ activate it, whereas metal chelators (1 mM EGTA, 1 mM EDTA) and serine proteinase inhibitors (1 mM PMSF and 0.1 mM DFP) act as strong inhibitors. The proteinase activity is insensitive to thiol-blocking compounds and to 5 mM 1,10-phenanthroline, a relatively specific chelator of zinc. It appears, therefore, that the active proteinase represents a metalenzyme complex rather than a metalloenzyme. Being capable of hydrolyzing N-blocked L-alanine-1-naphthylester, N-blocked L-methionine-1-naphthylester, and naphthyl AS-D chloroacetate at pH 6.8, the proteinase is insensitive to 1 mM p-nitrophenyl phosphate, an inhibitor of some mammalian esterproteinases. The enzyme does not split N-blocked DL-phenylalanine-2-naphthylester, nonspecific esterase substrates (1-naphthyl acetate, 1-naphthyl butyrate, 5-bromoindoxyl acetate), or several N-blocked L-aminoacyl- and N-blocked L-peptidyl-naphthylamides bearing L-arginine, L-alanine, L-phenylalanine, L-leucine, or L-proline at the P1 subsite. At operative pH values of 4.8 and 3.8 generated during electrophoresis in a stacking and a resolving gel, respectively, the cercarial proteinase migrates toward the cathode. The separated enzyme produces three bands of proteolysis in a gelatin-containing polyacrylamide gel at the optimal pH of 8.4. The presence of the proteinase in the penetration glands, in the secretory canals, and in the secretion from the glands suggests that the enzyme facilitates penetration, thus enabling the larvae to force their way through tissues of their hosts.

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“…Chymase activity was analyzed by using an incubation mixture containing the substrate N-acetyl-Lmethionine alpha-naphthyl ester (-N-O-Met) as described previously (19) . Briefly, paraffin sections (5-m thick) were deparaffinized in xylene and rehydrated, washed in 0.15 M phosphate buffer, pH 7.1, and then incubated with the substrate mixture which consisted of 5 mg of -N-O-Met dissolved in 0.2 ml dimethyl formamide (Sigma, St. Louis, MO) and brought up to a final volume of 25 ml with 0.15 M phosphate buffer, pH 7.1, containing 3.2 mM Fast Blue B salt (Sigma) as a capture reagent.…”

Section: Enzyme-histochemical Staining Of Mast Cell Chymasementioning

confidence: 99%

Characterization of mast cells according to their content of tryptase and chymase in normal and neoplastic human uterine cervix

Cabanillas-Saez

Schalper

Nicovani

et al. 2002

Int J Gynecol Cancer

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Cabanillas-Saez A, Schalper JA, Nicovani SM, Rudolph MI. Characterization of mast cells according to their content of tryptase and chymase in normal and neoplastic human uterine cervix. Int J Gynecol Cancer 2002;12:92-98.Mast cells (MC) have been associated with diverse human cancers. The primary function of these cells is to store and release a number of biologically active mediators, including the serine proteases tryptase and chymase. These proteases have been closely related with angiogenesis and tumor invasion, two critical steps during tumor progression. In the present work we analyzed the presence of MC in human uterine cervix from both normal and neoplastic tissues by using metachromatic, immunohistochemical, and enzymohistochemical staining. Tryptasepositive (MC T )-and tryptase/chymase-positive (MC TC )-mast cells were found in both normal and neoplastic tissues. The phenotype predominantly expressed in normal tissues as well as in benign and malignant lesions of the uterine cervix was the MC T . The total number of MC remained constant through the different stages of malignant transformation (cervical intraepithelial neoplasia grade 1-3) but a significant increase in the invasive carcinoma (IC) group was observed, this increase being mainly due to the MC T phenotype. Furthermore, we detected abundant MC T but not MC TC infiltrating tumors in sections of IC. Regarding the potent angiogenic properties described for tryptase, these findings suggest that in advanced stages of malignancy the significant number of MC T distributed within the cervical tissues could provide an effective mechanism to create the abundantly vascularized microenvironment required for tumor cells to proliferate and disseminate.

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Histochemical demonstration of mouse submandibular esterproteases with a new chromogenic substrate

Cited by 16 publications

References 14 publications

Characterization of mast cell subpopulations in lip cancer

Characterization of mast cell subpopulations in lip cancer

A serine proteinase in the penetration glands of the cercariae of Plagiorchis elegans (trematoda, plagiorchiidae)

Characterization of mast cells according to their content of tryptase and chymase in normal and neoplastic human uterine cervix

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