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Clinico-pathological study on denture stomatitis.

The clinical and pathological study was performed in order to determine the histopathological and cytoimmunological characteristics of denture stomatitis. All specimens were biopsy materials from seventeen patients with denture stomatitis. Normal palatal mucosae from ten patients served as the control. In addition to the usual staining methods, naphtol AS-D chloroacetate esterase stain and peroxidase-antiperoxidase method were used to detect mast cells and plasma cells. Denture stomatitis could be divided into atrophic and hyperplastic types. The former showed a smooth and atrophic mucosa. The latter showed a large number of exophytic projections which were composed of marked acanthosis and submucosal fibrosis, and was further subdivided into granular and papillary subtype according to the size of projections. In the present study, there were six cases of the atrophic type, and eleven cases of the hyperplastic type (consisting of seven granular and four papillary subtypes). The hyperplastic type was more frequently observed in patients with partial dentures compared with complete dentures and was associated frequently with ill fitting of the denture base as well as agglutination of denture plaque. Cytoimmunological study revealed that there was a pronounced increase of plasma cells, especially IgG- and IgA-producing cells, and a moderate increase of lymphocytes as well as mast cells in both types of denture stomatitis. Mast cells were always noted in the area with marked plasma cell infiltration, suggesting an intimate relation between both cells. These findings suggest that the immunological reactions play some role in the pathogenesis of denture stomatitis.

Adolescent↗

Cytochemical analysis of human peripheral blood lymphocyte subsets defined by monoclonal antibodies.

Human blood lymphocyte subpopulations, revealed by a panel of commercially available monoclonal antibodies by means of a rosetting technique, were submitted to direct cytochemical analysis and were shown to have distinguishing characteristics. T cells reactive with OKT3 antibody (T3+) displayed higher beta-glucuronidase and dot-like alpha-naphthyl acetate acid esterase (ANAE) activity than T3- cells. Helper/inducer cells (T4+) were characterized by a high level of dot-like ANAE activity, whereas cytotoxic/suppressor cells (T8+) displayed selective naphthol-ASD-chloroacetate esterase activity. These results provide evidence for association of some cytoplasmic enzyme activities with the expression of membrane differentiation markers defined by monoclonal antibodies.

Acid Phosphatase↗

Concurrent Sweet's syndrome and leukemia cutis in patients with myeloid disorders.

A 74-year-old woman with chronic auricular fibrillation, arterial hypertension, hypercholesterolemia, ischemic cardiopathy, and peripheral arteriopathy presented with purpuric lesions on the lower limbs (Fig. 1) and, to a lesser extent, on the anterior area of the chest. The mucous membranes were not affected. In 1989, she was diagnosed with anemia that evolved until 1998, when a bone marrow biopsy revealed a myelodysplastic syndrome unclassified in French-American-British Group (FAB). The patient has required periodic transfusions since February 1999. A skin biopsy of the purpuric lesions revealed a leukocytoclastic vasculitis; the lesions cleared with topical corticosteroid treatment. In May 1999, the patient presented with inflammatory and painful lesions localized on the vulva (Fig. 2), which had evolved over several days, without fever. No lesions were observed in other locations. A cutaneous biopsy showed an intense dermal edema and a diffuse and polymorphous dermal infiltrate involving the follicular structures. Exocytosis, spongiosis, and mucin deposits, demonstrated by Alcian blue stain, were observed in the follicular epithelium. Mature neutrophils were predominant in the dermal infiltrate, but a small number of eosinophils and immature cells were also present (Fig. 3). The myelogenous origin of the immature lining cells was further confirmed by positive staining of intracytoplasmic granules with naphthol-ASD chloroacetate sterase (Leder's stain). Vasculitis was not observed. Routine laboratory tests revealed 3030 leukocytes/mm(3) (60% neutrophils), a hemoglobin level of 8.4 g/dL, and 92,000 platelets/mm(3). Treatment with 30 mg/day of prednisone was started, and the lesions cleared slowly within 4 weeks. A new bone marrow biopsy in September 1999 showed a similar appearance to that taken in 1998. The patient died in January 2000 as a result of pneumonia with cardiac and respiratory failure. A 66-year-old man presented with a febrile syndrome that had evolved over 5 days, and painful and pruritic cutaneous lesions on the face and posterior neck (Fig. 4). Three months before, the patient was diagnosed with chronic myelogenous leukemia in acceleration phase. Examination revealed an edematous and erythematous face with pustular lesions on the surface, also involving the neck and the upper part of the back. The histopathologic examination revealed an intense edema and abscesses in the dermis. The infiltrate of these lesions was composed of mature neutrophils with the presence of abundant immature cells with a myelogenous aspect (Fig. 5). Analytical studies revealed 26,130 leukocytes/mm(3) (42% blasts). No specific treatment for Sweet's syndrome was administered and the lesions showed an improvement within 5 days. Eight days after admission, the patient died as a result of acute hemorrhage, before treatment for leukemia was initiated.

Aged↗

Quantitative significance of IgE-producing plasma cells and tissue distribution of mast cells in apical periodontitis.

A total of 30 dental periapical lesions were used for estimating the frequency of IgE-producing plasma cells by the unlabelled antibody peroxidase antiperoxidase method. Mast cells were detected in adjacent sections by the naphthol-AS-D-chloroacetate esterase technique. IgE-producing plasma cells were present in the central granulation tissue in 26 of the 30 specimens. The average ratio of IgE-containing plasma cells was 0.22% as related to all plasma cells. The central granulation tissue was devoid of mast cells, but numerous mast cells were found in the fibrous capsule of periapical lesions.

Cell Count↗

Systemic mastocytosis without cutaneous involvement.

An autopsy case of systemic mastocytosis without cutaneous involvement in a 76-year-old woman was described. The patient presented with general malaise, chest and epigastric discomfort, flushing of the face and progressive hepatosplenomegaly, and she terminated in hemorrhagic complications of DIC within 2 months. There was neither rash nor urticaria pigmentosa recognizable in the entire course. The diagnosis was made by the histologic identification of abnormal aggregates of mast cells in a bone marrow aspirate. These mast cell granules were chloroacetate esterase-positive, peroxidase-negative, and electronmicroscopically they were composed of fine granular materials containing variable numbers of lamellar structures. At autopsy, diffuse infiltration of the mast cells was found in the liver, spleen, bone marrow, lymph nodes, lungs, kidneys, stomach, and adrenal glands.

Aged↗

An unusual form of chronic myeloproliferative disorder. Aleukemic basophilic leukemia.

A 52-year-old Japanese man manifested various clinical signs and symptoms such as vomiting, high fever, dyspnea, cough, sweating, palpitation, eosinophilic leukocytosis and hepatosplenomegaly. These histamine-related clinical manifestations showed a dramatic response to steroid therapy. After 10 months of hospitalization, he suddenly succumbed to candidal septicemia at the end of the third cycle of steroid therapy. Autopsy revealed neoplastic proliferation of immature basophils in various internal organs without involvement of the skin. The neoplastic cells, positive immunohistochemically for leukocyte common antigen, possessed lobulated nuclei and weakly metachromatic cytoplasmic granules, predominantly of the basophil type, which exhibited weak naphthol ASD-chloroacetate esterase activity. Mast cell-type granules were also observed ultrastructurally. The neoplastic infiltration was associated with fibrosis in the liver, spleen and bone marrow and with extramedullary hematopoiesis in the liver, spleen, lymph nodes and perihypophyseal tissue. The bone marrow showed uneven and multifocal involvement. Despite the lack of leukemic manifestations and the results of chromosomal analysis, the most suitable diagnosis was aleukemic basophilic leukemia within the category of chronic myeloproliferative disorder. Kinship of this neoplasia to systemic mastocytosis is discussed.

Basophils↗

Systemic mast cell disease with splenic infarction: a case report.

An autopsy case of systemic mast cell disease (SMCD) without primary skin lesions in a 57-year-old Japanese male is described. Initially the patient was suspected of having liver cirrhosis or malignant lymphoma because of hepatomegaly and lymph node enlargement on admission. However, a lymph node biopsy and bone marrow aspiration conducted on his third admission indicated a SMCD because of the existence of metachromatic cell aggregates stained with toluidine blue. At autopsy, the diagnosis was confirmed because the proliferating cells were histochemically proven to be mast cells by naphthol AS.D chloroacetate esterase, Giemsa and alcian blue, in addition to toluidine blue staining. The intra-abdominal and retroperitoneal lymph nodes were replaced by mast cell aggregates, which caused the splenic infarction and bilateral hydronephrosis, with infiltration of mast cells into the spleen and kidneys also being apparent. Mast cell infiltration was similarly found in the bone marrow, liver, ileum and ascending colon. Immunohistochemically, the mast cells were positive for antibodies of alpha 1-antichymotrypsin, CD45 (LCA), CD43 (MT-1), CD45R (MB-1) and the oncoprotein c-kit. Electron microscopic examination using formalin-fixed tissue gave supportive evidence of a mast cell origin for the lesions.

Fatal Outcome↗

Distinguishing between proliferating nodal lymphoid blasts in chronic myelogenous leukemia and non-Hodgkin lymphoma: report of three cases and detection of a bcr/abl fusion signal by single-cell analysis.

Lymph node biopsies were analyzed from three patients with chronic myelogenous leukemia (CML) showing nodal blast proliferation. Immunohistochemically, the blasts from all three patients had an immature marker profile with a T-blast population (cCD3+, CD4-, CD7+, CD8-, CD99+, terminal deoxynucleotidyl transferase +) and a hematopoietic progenitor cell marker (CD34). In two patients, the blasts also expressed myeloid lineage specificity (naphthol AS-D chloroacetate esterase activity and myeloperoxidase positivity). However, it was difficult to distinguish between blast proliferation in CML and non-Hodgkin lymphoma from these immunohistopathological findings alone. Subsequently, bcr gene rearrangement and bcr/abl mRNA expression were detected by Southern blot and reverse transcription-polymerase chain reaction analysis of the lymph nodes. Fluorescence in situ hybridization (FISH) analysis of lymph node touch smears also disclosed bcr/abl gene fusion signals in the blasts of all patients, confirming that the blasts were derived from Philadelphia chromosome-positive CML. Accurate discrimination between the proliferating nodal blasts of CML and non-Hodgkin lymphoma is essential for determining subsequent therapy. FISH analysis of bcr/abl in single-cell blast preparations is an efficient tool that allows rapid, accurate cytopathological diagnosis of extramedullary blast-phase CML and its discrimination from non-Hodgkin lymphoma.

12E7 Antigen↗

A novel bacterium that utilizes monochlorobiphenyls and 4-chlorobenzoate as growth substrates.

A bacterial isolate, tentatively named SK-3, was isolated from tertiary lagoon sludge contaminated with polychlorinated biphenyls for over 25 years. SK-3 was able to grow on 2-chlorobiphenyl, 3-chlorobiphenyl, 4-chlorobiphenyl (4-CB), chloroacetate and 4-chlorobenzoic acid (4-CBA) as sole carbon and energy sources. During growth on 4-CB, a stoichiometric amount of chloride ion was produced and growth yields were comparable to those observed during growth on biphenyl. The production of 4-CBA as a metabolite was not observed. Protein yields during growth on 4-CB and biphenyl suggested that the presence of the chlorine did not impede use of carbons on both aromatic rings. Growth on 4-CBA also resulted in nearly stoichiometric production of chloride ion. Benzoate-grown SK-3 was also able to degrade several Aroclor 1242 congeners without the need for a primary substrate of previous growth on biphenyl.

Aroclors↗

Value of immunohistochemistry in the diagnosis of leukemia cutis: study of 54 cases using paraffin-section markers.

A grave prognosis is usually associated with leukemic skin infiltrates (leukemia cutis). However, some leukemic skin infiltrates are clinically similar to reactive non-leukemic infiltrates in patients with leukemia; thus it is of great importance to distinguish them. Fifty-four cases which were thought clinically to be leukemia cutis underwent immunophenotyping with a panel of nine T, B, monocytic, and macrophage markers using paraffin sections. Immunohistochemistry helped identify 44 cases with leukemia cutis and 10 with reactive infiltrates. In all cases of leukemia cutis, the staining patterns of skin infiltrates were concordant with cell type in the bone marrow. Furthermore, the panel of markers was usually helpful in distinguishing reactive from leukemia infiltrates, especially in cases with chronic lymphatic leukemia. Immunohistochemistry is a valuable adjunct in histopathologic differentiation of skin infiltrates in most cases of leukemia. With formalin-fixed, paraffin-embedded biopsies, we recommend that CD45 (LCA), CD45RO (UCHL-1), CD3, CD20 (L-26), CD43 (Leu-22), CD68 (KP-1), lysozyme, and chloroacetate esterase be considered in cases of systemic leukemia with cutaneous papules and nodules that prove difficult to interpret with routine section.

Acute Disease↗

Cutaneous myelofibrosis.

Cutaneous extramedullary hematopoiesis is rare, usually occurring in neonates following intrauterine viral infections, hereditary spherocytosis, or the twin transfusion syndrome. Only 20 cases of cutaneous extramedullary hematopoiesis have been reported in adults, all with myelofibrosis. The cutaneous infiltrates may be atypical and difficult to distinguish from leukemia cutis. We have studied a 65-year-old woman with myelofibrosis and approximately 40 violaceous, firm, non-tender cutaneous nodules measuring 1 to 4 cm in diameter, located on her abdomen near a splenectomy scar. Histologically, the lesions had a dense infiltrate of myeloid cells in all stages of maturation, atypical large cells with multilobate nuclei or multiple nuclei, resembling atypical megakaryocytes, and fibroblasts. Although the patient received erythropoietin therapy prior to the development of the nodules, erythroid progenitors were not seen. Reticulin was increased particularly surrounding the atypical megakaryocytes. The myeloid cells stained for chloroacetate esterase and with the polyclonal antibody MAC 387. Atypical megakaryocytes stained for Factor XIIIa and Factor VIII-related antigen. Dendritic Factor XIIIa positive cells were also increased. The skin lesions remain unchanged grossly one year after their development.

Aged↗

CD56+ blastic transformation of chronic myeloid leukemia involving the skin.

We report on two patients with chronic myeloid leukemia (CML) who presented blastic transformation involving the skin, with leukemic infiltrates showing unusual morphologic and immunohistologic characteristics. Both patients were elderly men with a 36-month and a 40-month history of CML, respectively. They presented with disseminated, reddish to violaceous papules and plaques (case 1), and with localized reddish nodules on the left temporal area (case 2). Concurrent features of blastic transformation in the bone marrow were observed in one patient (case 1). Histopathologic examination of skin lesions revealed similar features in both cases. There was a moderate to dense dermal infiltrate composed mainly of medium-sized atypical mononuclear myeloid precursor cells with only few relatively well-differentiated cells of the granulocytic series. Histochemical staining for naphthol-ASD-chloroacetate esterase revealed strong positivity (>50% of neoplastic cells) in case 2 and only scattered positivity (< 10% of neoplastic cells) in case 1. Immunohistologic analysis performed on paraffin-embedded sections showed in both cases variable reactivity of neoplastic cells for leucocyte common antigen (CD45), lysozyme, myeloperoxidase, CD11c, CD15, CD43, CD66, CD68, HLA-DR, and the neural cell adhesion molecule (NCAM) CD56. A negative reaction was observed for CD3, CD34, and TdT. The immunohistologic findings were remarkably similar to those reported for acute myeloid leukemia (AML) with monocytic differentiation (French-American-British [FAB] classification, subtype M4). Examination of blasts from the bone marrow performed in one patient (case 1) revealed a similar phenotype also with CD56 expression. In conclusion, our observations show that specific cutaneous infiltrates in CML may show morphologic and immunohistochemical characteristics similar to those observed in AML with monocytic differentiation. Moreover, specific cutaneous manifestations of CML may express CD56.

Aged↗

Differential effect of the serine protease inhibitor phenyl methyl sulfonyl fluoride on cytochemically detectable esterases in human leucocytes and platelets.

Esterases of human leucocytes and platelets were studied by cytochemical methods. The aim of the study was to clarify the cellular distribution and possible nature of esterases types differing in their substrate specificity and/or their inhibitor sensitivity. 3 substrates (alpha-naphthyl acetate: ANA; naphthol AS-D chloroacetate: NASDCA; and N-acetyl DL-alanine alpha-naphthyl ester: NACALA) were used and the effects of 2 inhibitors (sodium fluoride and the serine protease inhibitor phenyl methyl sulfonyl fluoride: PMSF) were evaluated. 4 enzyme types were described: Type I, present in granulocytes, was detected using NASDCA and NACALA and was resistant to fluoride but sensitive to PMSF. Other types were detected using ANA as substrate. Type II, present in monocytes, was inhibited by both fluoride and PMSF. Type III, present in platelets and plasma cells, was inhibited by fluoride but resistant to PMSF. Type IV, present in lymphocytes, was resistant to both fluoride and PMSF. The specific aims and possible areas for application of these results are discussed.

Adult↗

Cytochemical, immunological and ANAE-isoenzyme studies in acute myelomonocytic leukaemia: a study of 39 cases.

This study reports the cytochemical, electrophoretic and immunological characteristics of blasts cells from 39 cases of acute myelomonocytic leukaemia (M4). The results indicate considerable cytochemical heterogeneity, particularly with respect to esterase (alpha naphthyl acetate and chloroacetate) activities and suggest that an increased serum lysozyme concentration is a more consistent feature. Investigations with a range of monoclonal antibodies also revealed some differences in expression of monocyte-associated determinants although it is considered that immunological assessments are more consistent than cytochemistry in the detection of monocytic blast cell components. Analysis of ANAE isoenzymes by isoelectric focusing was found to be of particular value in cases where interpretation of ANAE cytochemistry was difficult.

Antibodies, Monoclonal↗

Granular common acute lymphoblastic leukaemia in adults: a morphological study.

2 cases of acute lymphoblastic leukaemia characterised by the presence of cytoplasmic inclusions morphologically similar to azurophil granules are described. Azurophil granulation of blasts is one of the cardinal features which differentiate acute myeloid from acute lymphoblastic leukaemia. Although such granulation of lymphoblasts has caused diagnostic confusion in the past, we found that the granules could be distinguished from myeloid azurophil granules both morphologically and by their characteristic cytochemical staining reactions. They were negative for peroxidase/sudan black and chloroacetate esterase, but gave coarse scattered granular positivity for both acid phosphatase and alpha-naphthyl acetate esterase. Both the electron microscopic appearance of the granules and their cytochemical staining reactions suggested that they were lysosomes. Granular ALL does not appear to be associated with any morphological subtype or karyotype; but is strongly associated with the common ALL phenotype. Its prognostic significance remains uncertain. It occurs more frequently than the small number of previous reports might suggest and, given the potential for misdiagnosis, should be more widely recognised.

Adult↗

Myelo-monocytic differentiation potential of the CALLA-positive Nalm 6M1 cell line.

Nalm 6M1 cells, derived from an acute lymphoblastic leukemia of the non-T, non-B type, are known to possess characteristics of early B-lymphocytes. In order to determine whether these cells are able to differentiate along the granulocyte-monocyte pathway, they were cultured in diffusion chambers, and classified according to morphological, cytochemical and immunological criteria. On the basis of this characterization, Nalm 6M1 cells positively labeling with monoclonal anti-human granulocyte or monocyte-specific antibodies were observed in all cases. These immunological changes were accompanied by the appearance of cells positive for the naphthyl-AS-D-chloroacetate esterase reaction. However, no changes in the blast-like appearance of the cultured cells were seen. From these data, we conclude that Nalm 6M1 cells already committed to the B-cell lineage retain their capacity to develop cellular markers of the granulocyte-monocyte series.

Acute Disease↗

A novel in-situ-zymography technique localizes gelatinolytic activity in human skin to mast cells.

Matrix-metallo-proteinases play a key role in cutaneous tissue remodeling and wound healing, and have been implicated as the rate-limiting factor in cutaneous tumor invasion and metastasis. We here describe a novel in-situ-zymographic method, which allows to directly localize sites of gelatinolytic activity in human skin. Gelatinolysis was detected through protein-hydrolysis in a 200 microm thick polyacrylamide gel underlying tissue sections. The lysis was substrate-dependent, demonstrated time- and temperature-dependent kinetics, and was inhibited by both EDTA and 1,10-phenanthroline. Normal and diseased skin sections demonstrated multiple focal points of gelatinolysis which co-localized with individual cells. Histochemically, these were shown to represent most likely mast cells (via AS-d-chloroacetate esterase staining and metachromasia). However, immunohistochemical staining for gelatinases A and B showed no immunoreactivity patterns that corresponded to the identified foci of gelatinolysis. The reported in-situ-zymographic technique offers a decisive advantage over immunohistochemistry, since it detects only the activated and catabolically relevant proteases, and provides further evidence for a role of mast cells in extracellular matrix remodeling.

Gelatinases↗

Cytochemical staining characteristics of chicken heterophils and eosinophils.

Cytochemistry was used to differentiate chicken heterophils and eosinophils in blood smears and cytospin preparations of isolated leukocytes. Cytochemical staining also was performed to evaluate any alteration of cell staining characteristics after heterophils had been isolated using discontinuous Ficoll-Hypaque gradients. Cytochemical staining reactions of heterophils and eosinophils were the same before and after discontinuous gradient cell separation. Characteristic positive reactions did not occur when chicken heterophils were stained with alkaline phosphatase, peroxidase, Sudan black B, acid phosphatase, alpha-naphthyl acetate esterase, naphthol AS-D chloroacetate esterase, periodic acid-Schiff, or Luna's eosinophil granule techniques. Chicken eosinophils stained positively when peroxidase, Sudan black B, acid phosphatase, and Luna methods were performed. In highly cellular cytospin preparations, peroxidase reactions readily distinguished the few contaminating eosinophils (< 1%) from the heterophils.

Journal Article↗