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Acute myelogenous leukemia relapsing as granulocytic sarcoma of the cervix. A case report.

BACKGROUND: Granulocytic sarcoma of the uterine cervix is an unusual manifestation of acute myeloid leukemia, representing soft tissue masses of leukemic myeloblasts. An often misdiagnosed entity, it is often confused with other inflammatory or neoplastic conditions, including large cell lymphoma. CASE: A 67-year-old female presented with acute myelogenous leukemia and a normal karyotype. After eight years in complete remission, abdominal pain and an ulcerated mass in the uterine cervix developed, with a normal peripheral blood smear. Vaginal cytology examination revealed myeloid blasts, which, on subsequent cervical biopsy, stained positive for leukocyte common antigen, Kp-1 (CD68), antimyeloperoxidase, lysozyme and chloroacetate esterase, confirming the cytologic diagnosis. K-ras was not mutated at codon 12 or 13. Chemotherapy induced a complete remission, followed nine months later by central nervous system and then systemic relapse. The patient died 13 months after being diagnosed with granulocytic sarcoma of the cervix. CONCLUSION: This case illustrates the value of vaginal cytology and histologic biopsy evaluation in patients with acute myelogenous leukemia, including those without evidence of systemic disease. The characteristic cytologic features of granulocytic sarcoma led to the correct diagnosis. Histologic biopsy evaluation, including immunohistochemistry for myeloid markers, proved of value in confirming the diagnosis.

Aged↗

Histochemical comparison of mast cells obtained from the airways of mongrel dogs and Basenji-Greyhound dogs by bronchoalveolar lavage.

An abnormally large number of mast cells in the airway lumen may be an important factor in the pathogenesis of bronchial hyperreactivity. However, it is unclear just how many mast cells are present in the lumen of normal or hyperreactive airways, in part because of differences in the histochemical techniques that have been used to identify mast cells and questions about the heterogeneity of mast cells. The present study was done (1) to compare the effectiveness of six techniques in the identification of mast cells obtained from dogs by bronchoalveolar lavage (BAL), (2) to compare the mast cells in the airways of normal mongrel dogs with those from a breed of dog (Basenji-Greyhound) known to have bronchial hyperreactivity, and (3) to determine whether the two-type histochemical classification used for rodent mast cells (formaldehyde-resistant or typical and formaldehyde-sensitive or atypical) applies meaningfully to the mast cells in BAL fluid from dogs. Cells obtained by BAL were fixed with Mota's basic lead acetate or formaldehyde. Mast cells were identified by metachromatic staining with toluidine blue or methylene blue, staining of highly sulfated proteoglycans with Alcian blue or berberine sulfate, and a histochemical reaction for chloroacetate esterase (mast cell chymase). After Mota's fixation, the methods were relatively similar in their effectiveness in determining the number of mast cells in lavage fluid from the mongrel dogs, in that all of the values fell within a narrow range: 0.53 to 0.96% of the total number of cells.(ABSTRACT TRUNCATED AT 250 WORDS)

Animals↗

Histochemical and immunohistochemical characteristics of mast cells in nasal polyps.

In surgically excised nasal polyps, most epithelial mast cells were formalin sensitive, chloroacetate esterase (CAE) negative, and chymase negative. Thus, this represents a population of mast cells not identified by staining for CAE. On the other hand, most stromal mast cells were formalin resistant and CAE positive, and although there was some polyp-to-polyp variability in their content of neutral protease, most of these cells were positive for both tryptase and chymase. The percentage of metachromatic cells in the epithelium and the number of metachromatic cells per unit area of polyp tissue did not correlate with an index of allergy skin test reactivity or the serum IgE concentration. The percentage of mast cells surrounded by pericellular tryptase, suggesting activation/degranulation, was significantly higher in the stroma than in the epithelium. The findings demonstrate differences between the stroma and the epithelium in phenotype and state of activation of mast cells; these are postulated to be due to distinct microenvironmental factors that affect mast cells at these sites.

Acetates↗

Monocytic leukemia in a horse.

On clinical examination, a six-year-old Hassian gray gelding with a history of impaired performance, slight cough, colic, and edema of the ventral abdomen, prepuce and the legs had reduced skin turgor, pale mucous membranes, forced costoabdominal breathing, reduced venous return, enlarged lymph nodes, and splenomegaly. Hematologic findings revealed anemia, leukocytosis and a high percentage of monocytoid leukemic cells. Generalized lymphadenopathy, splenomegaly, ascites, hydrothorax, and a diffusely thickened gut wall were found at necropsy. Massive infiltration with monocytoid leukemic cells was detected in lymph nodes, spleen, bone marrow, liver, gut wall, kidneys, and choroid plexus. Incubation of living cells obtained from a leukocyte concentrate with latex particles revealed phagocytosis in the leukemic cells on light and electron microscopy. The leukemic cells also had a marked alpha-naphthyl-acetate and naphthol-AS-acetate esterase activity, but were only weakly positive to naphthol-AS-D-chloroacetate esterase. A very weak alkaline phosphatase activity only was demonstrated in a few leukemic cells. On scanning electron microscopy, the leukemic cells had prominent ruffles and ridge-like profiles. These features of the leukemic cells excluded lymphocytic and granulocytic leukemia, and monocytic leukemia was diagnosed.

Alkaline Phosphatase↗

Cytochemical characterization of leukemic cells from 20 dogs.

The hematopoietic cells in blood and/or bone marrow from 20 leukemic dogs and 22 control dogs were characterized using a battery of cytochemical stains. The results of cytochemical staining led to modification of the diagnoses based on clinical, hematologic and histologic findings in seven (35%) of the leukemias. Sudan black B and chloroacetate esterase served as granulocytic markers in both the control and leukemic groups. Peroxidase activity was present in the granulocytes and monocytes of control animals but not the blasts of leukemic dogs. Alkaline phosphatase-positive staining of granulocytic precursors was a consistent finding in granulocytic and myelomonocytic leukemia, and alkaline phosphatase-positive lymphoblasts were seen in 38% of lymphocytic leukemias. Diffuse alpha naphthyl butyrate esterase-positive staining marked monocytes in both control and leukemic dogs. Cytochemical staining was found to be a valuable diagnostic aid in the classification of leukemias in the dog.

Acid Phosphatase↗

Cytochemical characterization of feline leukemic cells.

Hematopoietic cells in blood and/or bone marrow from 23 leukemic cats and ten control cats were characterized using a battery of cytochemical enzyme stains. The results of cytochemical staining led to modification of diagnosis based on clinical, hematologic and histopathologic findings in four (17%) of the leukemias. Sudan black-B and chloroacetate esterase served as granulocytic markers in both the control and leukemic groups. Peroxidase activity was seen in the granulocytes and monocytes of control animals but not in the blasts of leukemic cats. Diffuse alpha naphthyl butyrate esterase staining marked monocytes in both control and leukemic animals. Cytochemical staining was found to be a valuable aid in the classification of leukemias in the cat.

Acid Phosphatase↗

Acute megakaryoblastic leukemia in one cat and two dogs.

The clinical, hematologic, and histologic features of acute megakaryoblastic leukemia are described for an 8-year-old female Domestic Shorthair cat, a 3-year-old female mixed-breed dog, and a 3-year-old male German Shepherd Dog. The neoplastic cells were characterized as belonging to the megakaryocytic lineage. The following techniques were used: electron microscopy; detection of antibodies against human von Willebrand factor (vWF) and human platelet glycoprotein GP IIIa using a modified avidin biotin peroxidase complex technique on formalin-fixed paraffin sections; and enzyme histochemical methods on plastic sections for alkaline phosphatase, acid phosphatase, myeloperoxidase, alpha-naphthyl acetate esterase, alpha-naphthyl butyrate esterase, naphthol AS acetate esterase, and naphthol AS-D chloroacetate esterase. In addition, benign megakaryocytic cells, platelets, and neoplastic cells were labeled with lectins that have partially been shown to bind to platelet glycoproteins of other species. In healthy cats and dogs, the megakaryocytes and platelets reacted with lectins PSA, LCA, PHA-L, and WGA. Megakaryocytes and platelets from healthy cats were also labeled by lectin PNA. The lectins PHA-L and WGA reacted with neoplastic cells from the cat and both dogs. Lectin PNA bound to neoplastic cells from the cat, and lectins PSA, LCA, and SBA bound to neoplastic cells from both dogs. For the retrospective examination of paraffin-embedded material, the detection of vWF and GP IIIa appears to be the most reliable method for the identification of megakaryocytic cells.

Animals↗

The equine endometrial mast cell during the puerperal period: evaluation of mast cell numbers and types in comparison to other inflammatory changes.

Endometrial biopsies of 44 broodmares were histologically examined on days 3, 6, and 9 postpartum. The mares were subdivided into three groups according to the course of the puerperal period. In 29 mares, parturition and expulsion of the placenta was normal, six mares showed dystocia, and in nine mares, the placenta was retained for > 2 hours. Tissue samples were evaluated histologically, and the average numbers of granulocytes, lymphocytes, macrophages, siderophages, and mast cells was determined. Protease content of mast cells was examined with a double-enzyme immunohistochemical staining technique, using a histochemical reaction for chloroacetate esterase and fast blue to detect chymase activity and an immunohistochemical staining method with a polyclonal antibody and fast red for the detection of tryptase. Analyzing the cell numbers using the statistical software Statistica, a marked inflammatory reaction was observed in the endometrium postpartum. Although the number of granulocytes decreased during the first 9 days postpartum, the number of lymphocytes, macrophages, and siderophages increased. No significant difference in the number of any of these cell types could be demonstrated in the three different courses of the puerperal period, although the numbers of these cells seemed to be lower in mares with dystocia. In contrast with other cells, no change in the number of endometrial mast cells was observed during the puerperal period, but a significantly lower number were found in the endometrium of mares with retained placenta. The enzyme immunohistochemical double-labeling technique could demonstrate only tryptase-positive mast cells; no chymase activity was detectable in any endometrial mast cells. The number of mast cells detected with the metachromatic staining technique was significantly higher than that detected with double labeling. These results support the hypothesis that a sufficient number of mast cells may be necessary for a normal postnatal period and suggest a mast cell subtype in the equine endometrium that is tryptase and chymase negative.

Animals↗

Large granular lymphocyte leukemia/lymphoma in six cats.

This report describes six cases of feline large granular lymphocyte lymphoma identified by light microscopy on the basis of their characteristic azurophilic granulation in Giemsa-stained plastic sections and by electron microscopy on the basis of their typical granules. Although the granules of all the tumor cells were negative for peroxidase activity, they all demonstrated chloroacetate-esterase and acid phosphatase activity. All the tumors reacted with cross-reacting antibodies against the CD3 antigen (epsilon chain) and did not react with a cross-reacting monoclonal antibody directed against epitopes on cytoplasmic domains of the CD20 antigen. Three tumors had a positive reaction with a monoclonal human CD57-like antibody. This is highly suggestive of either a cytotoxic T cell or a natural killer cell origin of the neoplasias. In three cats, although other abdominal organs were affected to a variable extent, the main neoplastic lesions were localized in the gastrointestinal tract and the jejunal lymph nodes. In contrast, in the other three cats, organ involvement was more widespread, affecting the lung (two), myocardium (two), precardiac mediastinum (one), salivary gland (one), and spinal cord (one); in addition, leukemia was present in two of these cats. The data presented indicate that tumors made up of large granular lymphocytes occur more frequently in cats than previously assumed and that they share many characteristic features with specific subtypes of clonal disorders of large granular lymphocytes in humans.

Acid Phosphatase↗

Correlation analysis of alveolar macrophage cytochemical parameters in smoking and pulmonary oncology.

Cytochemical examination of alveolar macrophages (AM) obtained by bronchoalveolar lavage (BAL) was performed in healthy volunteers (11 non-smokers and 11 smokers) and in 9 patients with squamous lung carcinoma (all of them smokers or ex-smokers) in order to analyze its peculiarities related to the smoking habit and to lung malignancy. Assessment of non-specific esterases: alpha-naphthyl acetate esterase (ANAE) and butyrate esterase (BUT), chloroacetate esterase (CHL), acid phosphatase (AcP), intracellular glycogen (PAS reaction), lipids (Sudan black B reaction-SBB) and iron (Perl's reaction) was performed by a semiquantitative cytochemical method (1). A significant correlation was obtained between BUT and stage of squamous lung carcinoma (varying between I and IV) (r = 0.52, p < 0.05). There was a correlation between BUT and Perl's in healthy controls (r = 0.76, p < 0.05). The same type of correlation was observed in control smokers (r = 0.64, p < 0.05), in addition to a correlation between CHL and AcP (r = 0.69, p < 0.05). There was no significant BUT/Perl's correlation in patients with squamous cell lung carcinoma (r = 0.23, p > 0.05), but significant AcP/CHL correlation as was observed in control smokers (r = 0.73, p < 0.05), and a "new" type of correlation was shown to exist between ANAE and SBB (r = 0.77, p < 0.05). In spite of the unresolved nature of lung cancer, correlation analysis of cytochemical parameters in AM might have an important part in the analysis of their relative contribution to the development of smoking-related disorders and lung malignancies.

Acid Phosphatase↗

Cytochemical reactions of human hematopoietic cells in liquid culture.

Growth of human bone marrow in liquid suspension cultures has been used to study normal hematopoietic cell differentiation and abnormalities in blood diseases. A variety of cytochemical stains were applied to human marrow cells cultured in vitro for up to 14 days. AS-D- CHLOROACETATE ESTERASE AND ALPHA-NAPHYHYL BUTYrate esterase were most useful in distinguishing different cell lines in culture. Peroxidase activity disappeared with mononuclear phagocyte morphogenesis and diminished with culture in intermediate and mature granulocytes. Acid phosphatase activity and methyl greed pyronin staining intensity increased with macrophage maturation.

Adult↗

Cytologic characterization of pulmonary alveolar macrophages by enzyme histochemistry in plastic.

Quantitative morphometric analyses of macrophages should facilitate a more precise definition of the role of macrophages in neoplastic diseases and inflammatory diseases of lymphoid and hemopoietic tissues. The usefulness of the available phenotypic markers is affected by the extent to which they are expressed by macrophages in tissue sections. For our study of phenotypic markers for macrophages in human tissue sections, we selected lung, since pulmonary alveolar macrophages are more readily identified morphologically in tissue sections than macrophages in most other tissues. In 1-2 micron sections of freshly fixed lung embedded in methacrylate, 89.7% of pulmonary alveolar macrophages had histochemically demonstrable acid phosphatase; 86.2%, nonspecific esterase with naphthol ASD chloroacetate as the substrate; 86.2%, nonspecific esterase with alpha-naphthyl butyrate as the substrate; 80.5%, peroxidase; and 75.8%, iron. With respect to their expression of markers, the observed heterogeneity among pulmonary alveolar macrophages is interesting; this heterogeneity may reflect the degree to which they have been activated, the different periods of time since they arrived in the lung, and differences in their local environments. Except for peroxidase, all examined markers were as well demonstrated when tissues were fixed after storage over liquid nitrogen as when fixation was carried out with fresh tissue. Acid phosphatase and nonspecific esterase with the chloroactetate substrate gave bright colors that would facilitate morphometric analyses. The storage of tissue over liquid nitrogen will be equally satisfactory for the characterization of macrophages with histochemical markers and monoclonal antibodies.

Acid Phosphatase↗

Distribution of chymase-containing mast cells in human bronchi.

Mast cell chymase stimulates secretion from cultured airway gland serous cells and hydrolyzes bronchoactive peptides in vitro. To explore the likelihood of these interactions occurring in situ, we examined the distribution and concentration of chymase-containing mast cells near glands and smooth muscle of major human bronchi from eight individuals without known airway disease. Total airway mast cells and the subset of mast cells containing chymase were detected by staining for methylene blue metachromasia and chloroacetate esterase activity, respectively. The percentage of chymase-containing mast cells was found to differ strikingly among bronchial tissue compartments. Near glands, for example, the concentration of chymase-positive mast cells (640 +/- 120 cells/mm3) was 73 +/- 9% that of total mast cells (910 +/- 130 cells/mm3), whereas in smooth muscle the concentration of chymase-positive mast cells (450 +/- 200 cells/mm3) was only 14 +/- 4% that of total mast cells (2920 +/- 620 cells/mm3). Of all chymase-containing mast cells in the airway subepithelium, 30 +/- 4% were located within 20 microns of submucosal glands. Although the percentage of chymase-containing cells varied, the absolute concentration of chymase-containing mast cells was similar in all compartments. These results reveal a differential distribution of mast cell subpopulations in human airway and suggest that mast cells containing chymase are near gland and smooth muscle targets.

Bronchi↗

Demonstration of tryptase in bovine cutaneous and tumor mast cells.

We examined three tissue samples from each of four cows with non-lesional skin, tissue samples from a cow with multiple cutaneous mast cell tumors, and samples from another cow in which mast cells were infiltrating multiple lymphosarcomas of the skin, for the presence of tryptase and chymase by enzyme cytochemical and immunohistological methods. The enzyme activities of tryptase and chymase were tested using N-carbobenzoxy-glycilglycil-L-arginine-2-naphthylamide (Z-Gly-Gly-Arg-NA) and naphthol-AS-D-chloroacetate (N-AS-D-CA) as substrates, respectively. Tryptase reactivity could be demonstrated in frozen and Carnoy-fixed paraffin sections. Chymase reactivity was seen in neither frozen nor paraffin sections of formalin- or Carnoy-fixed skin tissues. Antibody linkage with a polyclonal rabbit anti-human skin tryptase antibody was highly specific in bovine normal cutaneous, infiltrating, and tumor mast cells. More than 90% of the tumor mast cells were distinctly tryptase-positive. With alcian blue, only slightly more than 10% of the mast cells stained clearly positive and with methylene blue hardly any staining of mast cell granules could be demonstrated. No antibody labeling of mast cell granules in any of the tissue sections was detected by the use of rabbit anti-dog chymase antiserum. These results indicate that there is a striking antigenic similarity of bovine tryptase to its canine and human equivalents. The demonstration of tryptase is an important tool in confirming the diagnosis of undifferentiated mast cell tumors. In contrast to other species, chymase appears to be completely absent in bovine skin mast cells.

Amino Acid Sequence↗

The in vitro production and characterization of neutrophils from embryonic stem cells.

An embryonic stem (ES) cell/OP9 coculture system for the effective production of functional neutrophils is described. A 3-step differentiation strategy was developed that uses liquid culture, enabling reliable and abundant production of neutrophils at high purity without the need of sorting for isolation of mature neutrophils. Use of the OP9 stromal cell line significantly enhances the number, percentage, and duration of differentiated neutrophils produced from embryonic stem cells. Effective and sustained differentiation of ES cells into neutrophils provides a useful model system for studying neutrophil differentiation and function and the factors that regulate them. Morphologic and functional evaluation of these ES-derived neutrophils indicates that large numbers of mature neutrophils can be produced from pluripotent ES cells in vitro. Specifically, their morphology, ability to produce superoxides, flux calcium, undergo chemotaxis in response to macrophage inflammatory protein 2 (MIP-2), stain for the granulocyte-specific marker-specific chloroacetate esterase, and contain the neutrophil-specific markers Gr-1 and the mouse neutrophil-specific antigen indicates that they are comparable with purified mouse bone marrow neutrophils. They also express gelatinase and lactoferrin granule proteins. During the differentiation of these ES-derived neutrophils, regional areas of neutrophil production can be identified that have been designated as neutrophil generating regions (NGRs).

Animals↗

Establishment and characterization of a novel human monocytic leukemia cell line (KP-1) expressing scavenger receptor.

A new monocytic leukemia cell line (KP-1) was established from a 2-y-old Japanese girl with acute monocytic leukemia. The KP-1 cells were maintained in suspension culture with a doubling time of 96 h. The cells were positively stained with alpha-naphtyl butyrate esterase, but not with naphthol AS-D chloroacetate esterase, myeloperoxidase, and periodic acid-Schiff reagent. Cell surface marker analysis revealed that the cells were CD4, CD11a, CD11c, CD13, CD14, CD18, CD33, and HLA-DR positive. Karyotype analysis revealed near diploidy (47 XX) and a translocation t(11;19) was found. When treated with 12-o-tetradecanoylphorbol 13-acetate, KP-1 cells became tightly adherent, showed the enhanced reactivity for alpha-naphtyl butyrate esterase, and produced several monokines such as IL-1 beta, tumor necrosis factor-alpha, and macrophage colony-stimulating factor. Immunoelectron microscopy demonstrated that the human macrophage scavenger receptor was expressed after 12-o-tetradecanoylphorbol-13-acetate treatment, and the cells accumulated a large amount of cholesterol esters in the presence of acetylated LDL. Compared with another human monocytic leukemia cell line, THP-1, KP-1 expressed scavenger receptor and accumulated cholesterol ester more rapidly in the presence of 12-o-tetradecanoyl phorbol-13-acetate and acetylated LDL. Scatchard analysis using 125I-labeled acetylated LDL revealed a typical saturation curve with an apparent kd of 1.7 x 10(-7) M and 3400 binding sites per cell. KP-1 retained the characteristics of monocyte-macrophage lineage cells and will facilitate the in vitro studies of the pathologic and physiologic roles of scavenger receptors.

Antigens, Neoplasm↗

Granulocytic sarcoma presenting as an epidural mass with cord compression.

Granulocytic sarcoma (GS) usually occurs during the course of, or as a presenting sign of myelogenous leukemia. Rarely it is found before peripheral blood or bone marrow evidence of leukemia is present. We describe a man who presented with low back pain and lower extremity weakness. He had spinal cord compression due to GS without evidence of leukemia. Only four such instances have been previously reported. Such aleukemic presentations of GS are frequently misdiagnosed. The chloroacetate esterase stain and electron microscopy are useful in demonstrating the myeloid origin of GS cells. GS lesions are probably best treated by localized radiation therapy and systemic chemotherapy.

Humans↗

Hypericin induces both differentiation and apoptosis in human promyelocytic leukemia HL-60 cells.

Hypericin is a unique photosensitizing plant pigment and has been separately reported to induce differentiation and apoptosis in neoplastic cells. In this study, we examined the relationship between activities to induce differentiation and apoptosis in human promyelocytic leukemia HL-60 cells, at a concentration range of 0.15 to 0.2 microM. When treated with hypericin, the cell ratio reducible of nitroblue tetrazolium was significantly increased and the cell size was enlarged by flow cytometry analysis. Hypericin also significantly increased the ratio of the cells, which were of positive alpha-naphthyl acetate esterase activity and phagocytic activity, whereas it hardly influenced the naphthol AS-D chloroacetate esterase activity in the cells, as well as 1 alpha, 25(OH)2D3 (10 nM). In addition, hypericin increased hypodiploid nuclei and caused a nucleosomal ladder. These results indicate that hypericin induces both differentiation toward monocyte/macrophage lineage and apoptosis in HL-60 cells.

Anthracenes↗