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Effect of Ovariectomy and Estrogen Treatment on Uterine Benzylamine Oxidase and Monoamine Oxidase Type A
418
MISCELLANEOUS
was done using sulfhydryl reagent-treated red blood cells as model paroxysmal nocturnal hemoglobinuria cells. These cells resemble paroxysmal nocturnal hemoglobinuria cells closely with respect to their behavior with complement, surface structure revealed by electron microscopy, and membrane proteins and lipids. Rabbit antiserum to human red blood cells was used to sensitize erythrocytes for specific antibody-dependent complement lysis sensitivity tests. For the general laboratory a modification of the sucrose lysis test is described. This test depends upon the activation of complement in an isosmolar solution of reduced ionic strength. Titrating paroxysmal nocturnal hemoglobinuria cells with serum in sucrose provides a test analogous to one using specific antibodies and allows the demonstration of complement-sensitive cells, as well as the measurement of the fractional size of the subpopulation of abnormal cells. The test has the capability to estimate the relative complement sensitivities or the subpopulations. Testing procedures for paroxysmal nocturnal hemoglobinuria should be controlled more carefully in view of reports of an association of paroxysmal nocturnal hemoglobinuria with a variety of lymphoproliferative disorders. M. G. F. 4 figures, 3 tables, 42 references
by inherent biologic variability. With a fairly new and simple plastic-embedding technique enzyme histochemistry was studied in >300 nonneoplastic lymph nodes. The morphologic appearance of the tissues was strikingly better than that in paraffin sections and enzyme localization was excellent. Cells of the mononuclear phagocyte system were distinguishable readily from each other and from lymphoid cells. Subgroups of lymphoid cells, including T cells, mantle-zone lymphocytes and plasma cells, could be identified specifically. Neutrophils and mast cells could be identified easily. Stromal cells showed specific staining, which simplified evaluation of the relative extent of T and B cell regions. The technique may be valuable in evaluating reactive and neoplastic lymph nodes. Eventually, a system may be provided that permits plastic-embedded sections to be stained directly for enzyme histochemical and immunohistochemical markers. M.G.F. 13 figures, 1 table, 28 references
Analysis of 25-Hydroxy Vitamin. Da in Plasma by HighPerformance Liquid Chromatography
K. G.
J.C. K. Loo AND R. BRIEN, Bureau of Drug Research, Health Protection Branch, Ottawa, Ontario, Canada Res. Comm. Chem. Path. Pharm., 41: 139-148 (July) 1983 Current analytical procedures for the determination of 25(0H) vitamin D3 in plasma samples are based on radioimmunological and competitive binding assays, and high performance liquid chromatographic assays. Although the former assays are sensitive and specific, tedious procedures are required to obtain the binding proteins. The latter methods generally require tedious purification of plasma samples by multiple extraction and column chromatographic steps before high performance liquid chromatography analysis. A specific high performance liquid chromatography procedure for the determination of 25-0H vitamin D3 in human plasma is described. The method simplifies the cleanup procedure and is sufficiently sensitive and specific for the determination of plasma samples containing as little as 2 mg. 25-0H vitamin D3 per ml. The reproducibility and efficiency of the extraction procedure were determined with radioactive 25-0H vitamin D3. M. G. F. 1 figure, 2 tables, 15 references
The Evaluation of Human Lymph Nodes, Using Plastic Sections and Enzyme Histochemistry J. H. BECKSTEAD, Department of Pathology, University of California School of Medicine, San Francisco, California
Effect of Ovariectomy and Estrogen Treatment on Uterine Benzylamine Oxidase and Monoamine Oxidase Type A BHANSALI, B. E. HAYES, V. R. MUKKU, G. M. STANCEL AND D. E. CLARKE, Department of Pharmacology, University
of Houston and Department of Pharmacology, University of Texas Medical School at Houston, Houston, Texas Res. Comm. Chem. Path. Pharm., 41: 37-49 (July) 1983 Uterine monoamine oxidase type A and benzylamine oxidase activities were assayed in rats treated with ovariectomy alone or with estrogen. Control animals were subjected to sham ovariectomy. Total and specific activities of monoamine oxidase type A were decreased markedly by ovariectomy and restored partially by estradiol treatment. Benzylamine oxidase was not influenced selectively. Total benzylamine oxidase activity followed changes in uterine weight such that the specific activity of benzylamine oxidase per mg. tissue was not statistically different among the 3 experimental groups. The specific activity of benzylamine oxidase changed significantly on the basis of changes in the protein and deoxyribonucleic acid contents of the uterus. Although monoamine oxidase type A and benzylamine oxidase activities follow a similar trend in studies of the effects of sex steroids in rat aortas, their cellular location in the human uterus differs radically. Monoamine oxidase type A is found primarily in the endometrium and, possibly, in the sympathetic innervation. Since benzylamine oxidase appears to be insensitive to direct modulation by sex steroids and may be located predominantly in uterine blood vessels, it is suggested that the enzyme be used as a marker for hormonal influences on the vascularity and structure of the uterine vascular bed. M. G. F. 4 tables, 21 references
Amer. J. Clin. Path., 80: 131-139 (Aug.) 1983 Immunologic markers recently have been used to assist in histopathologic evaluation of human lymph nodes. Although enzyme histochemical techniques have been available for many years they have been used mainly with frozen sections and touch preparations that do not permit precise correlation of enzymes with tissue morphologic characteristics. Interpretation of enzyme histochemical reactions is complicated by the variable sensitivity of the enzymes to fixation and processing, and
MISCELLANEOUS Nephrogenic Metaplasia of the Ureter M. LUGO, R. 0. PETERSEN, I. B. ELFENBEIN, B. S. STEIN AND N. J. DUKER, Departments of Pathology, Urology, and Fels
Research Institute of Temple University Hospital and School of Medicine, Philadelphia, Pennsylvania
MISCELLANEOUS
was done using sulfhydryl reagent-treated red blood cells as model paroxysmal nocturnal hemoglobinuria cells. These cells resemble paroxysmal nocturnal hemoglobinuria cells closely with respect to their behavior with complement, surface structure revealed by electron microscopy, and membrane proteins and lipids. Rabbit antiserum to human red blood cells was used to sensitize erythrocytes for specific antibody-dependent complement lysis sensitivity tests. For the general laboratory a modification of the sucrose lysis test is described. This test depends upon the activation of complement in an isosmolar solution of reduced ionic strength. Titrating paroxysmal nocturnal hemoglobinuria cells with serum in sucrose provides a test analogous to one using specific antibodies and allows the demonstration of complement-sensitive cells, as well as the measurement of the fractional size of the subpopulation of abnormal cells. The test has the capability to estimate the relative complement sensitivities or the subpopulations. Testing procedures for paroxysmal nocturnal hemoglobinuria should be controlled more carefully in view of reports of an association of paroxysmal nocturnal hemoglobinuria with a variety of lymphoproliferative disorders. M. G. F. 4 figures, 3 tables, 42 references
by inherent biologic variability. With a fairly new and simple plastic-embedding technique enzyme histochemistry was studied in >300 nonneoplastic lymph nodes. The morphologic appearance of the tissues was strikingly better than that in paraffin sections and enzyme localization was excellent. Cells of the mononuclear phagocyte system were distinguishable readily from each other and from lymphoid cells. Subgroups of lymphoid cells, including T cells, mantle-zone lymphocytes and plasma cells, could be identified specifically. Neutrophils and mast cells could be identified easily. Stromal cells showed specific staining, which simplified evaluation of the relative extent of T and B cell regions. The technique may be valuable in evaluating reactive and neoplastic lymph nodes. Eventually, a system may be provided that permits plastic-embedded sections to be stained directly for enzyme histochemical and immunohistochemical markers. M.G.F. 13 figures, 1 table, 28 references
Analysis of 25-Hydroxy Vitamin. Da in Plasma by HighPerformance Liquid Chromatography
K. G.
J.C. K. Loo AND R. BRIEN, Bureau of Drug Research, Health Protection Branch, Ottawa, Ontario, Canada Res. Comm. Chem. Path. Pharm., 41: 139-148 (July) 1983 Current analytical procedures for the determination of 25(0H) vitamin D3 in plasma samples are based on radioimmunological and competitive binding assays, and high performance liquid chromatographic assays. Although the former assays are sensitive and specific, tedious procedures are required to obtain the binding proteins. The latter methods generally require tedious purification of plasma samples by multiple extraction and column chromatographic steps before high performance liquid chromatography analysis. A specific high performance liquid chromatography procedure for the determination of 25-0H vitamin D3 in human plasma is described. The method simplifies the cleanup procedure and is sufficiently sensitive and specific for the determination of plasma samples containing as little as 2 mg. 25-0H vitamin D3 per ml. The reproducibility and efficiency of the extraction procedure were determined with radioactive 25-0H vitamin D3. M. G. F. 1 figure, 2 tables, 15 references
The Evaluation of Human Lymph Nodes, Using Plastic Sections and Enzyme Histochemistry J. H. BECKSTEAD, Department of Pathology, University of California School of Medicine, San Francisco, California
Effect of Ovariectomy and Estrogen Treatment on Uterine Benzylamine Oxidase and Monoamine Oxidase Type A BHANSALI, B. E. HAYES, V. R. MUKKU, G. M. STANCEL AND D. E. CLARKE, Department of Pharmacology, University
of Houston and Department of Pharmacology, University of Texas Medical School at Houston, Houston, Texas Res. Comm. Chem. Path. Pharm., 41: 37-49 (July) 1983 Uterine monoamine oxidase type A and benzylamine oxidase activities were assayed in rats treated with ovariectomy alone or with estrogen. Control animals were subjected to sham ovariectomy. Total and specific activities of monoamine oxidase type A were decreased markedly by ovariectomy and restored partially by estradiol treatment. Benzylamine oxidase was not influenced selectively. Total benzylamine oxidase activity followed changes in uterine weight such that the specific activity of benzylamine oxidase per mg. tissue was not statistically different among the 3 experimental groups. The specific activity of benzylamine oxidase changed significantly on the basis of changes in the protein and deoxyribonucleic acid contents of the uterus. Although monoamine oxidase type A and benzylamine oxidase activities follow a similar trend in studies of the effects of sex steroids in rat aortas, their cellular location in the human uterus differs radically. Monoamine oxidase type A is found primarily in the endometrium and, possibly, in the sympathetic innervation. Since benzylamine oxidase appears to be insensitive to direct modulation by sex steroids and may be located predominantly in uterine blood vessels, it is suggested that the enzyme be used as a marker for hormonal influences on the vascularity and structure of the uterine vascular bed. M. G. F. 4 tables, 21 references
Amer. J. Clin. Path., 80: 131-139 (Aug.) 1983 Immunologic markers recently have been used to assist in histopathologic evaluation of human lymph nodes. Although enzyme histochemical techniques have been available for many years they have been used mainly with frozen sections and touch preparations that do not permit precise correlation of enzymes with tissue morphologic characteristics. Interpretation of enzyme histochemical reactions is complicated by the variable sensitivity of the enzymes to fixation and processing, and
MISCELLANEOUS Nephrogenic Metaplasia of the Ureter M. LUGO, R. 0. PETERSEN, I. B. ELFENBEIN, B. S. STEIN AND N. J. DUKER, Departments of Pathology, Urology, and Fels
Research Institute of Temple University Hospital and School of Medicine, Philadelphia, Pennsylvania