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PubMed1981

Bacterial infection stimulating granulocytic sarcoma of the small bowel.

A 57-year-old white man who had abdominal pain and distension, died after a short hospitalization for increasing ascites, anorexia, and deteriorating mental status. At autopsy, the principal gross finding was a dilated, hyperemic, thickened proximal jejunum that by light microscopy consisted of a transmural infiltrate of large mononuclear cells. Intense naphthol AS-D chloroacetate esterase (NASD) positivity was observed within most of the cells, suggesting granulocytic sarcoma. However, bacterial strains and electron-microscopic examination revealed that the massive jejunal infiltrate was composed of macrophages containing numerous phagocytosed bacteria. Although occasional cells had primary and secondary granules characteristic of myeloid precursors present within their cytoplasm, most cells lacked specific granules. Attempts to reproduce this markedly enhanced NASD result experimentally in peritoneal macrophages of mice were unsuccessful. This case shows that intense NASD cytoplasmic staining may occasionally occur macrophages that have phagocytosed large numbers of bacteria.

Animals↗
PubMed1981

Enzyme histochemistry on normal and pathologic paraffin-embedded lymphoid tissues.

So far, enzyme histochemical examination has been applied, with few exceptions, either to tissue imprints or to cryostat sections of freshly collected samples. This procedure is not easily applicable in routine histopathologic examination. In this study, a simplified tissue embedding procedure is presented which can be performed using an automatic tissue changer. The paraffin embedded samples can be used for both conventional histopathologic examination and for demonstrating enzymes in sections. The enzymes studied (alkaline phosphatase, alpha-naphthyl acetate esterase, acid phosphatase, tartrate resistant acid phosphatase, ATPase, peroxidase, and chloroacetate esterase) gave comparable results in formalin-fixed cryostat sections and paraffin sections in both normal and pathologic lymphoid samples. The only exception was ATPase, which could not be demonstrated on paraffin-embedded material. The technic described has broad application in the analysis of lymphoid diseases.

Acid Phosphatase↗
PubMed1983

Granular acute lymphoblastic leukemia.

Granules in blasts are most typical of acute myeloblastic leukemia. However, there have been scattered reports of patients with acute lymphoblastic leukemia (ALL) that have lymphoblasts with azurophilic cytoplasmic granules. These reports do not describe immunologic markers or cytogenetics. We report five additional cases with detailed cytologic, immunologic, and cytogenetic studies. At diagnosis one of these patients had central nervous system disease, while the others had no unusual features. Four of the five patients attained remission. The blasts of all five patients contained distinct cytoplasmic azurophilic granules. The granules were negative for peroxidase and chloroacetate esterase and positive for PAS, acid phosphatase, alpha-naphthyl acetate esterase incompletely fluoride inhibited, alpha-naphthyl butyrate esterase, and in one case, Sudan black B. In the one patient studied by electron microscopy, characteristic lymphoblasts contained membrane-bound electron lucent inclusions, which stained positively with non-specific esterase. Immunologic markers showed a common ALL phenotype. Different cytogenetic abnormalities were seen in all cases. It is important to recognize the characteristics of this morphologic subtype of ALL in order to avoid a misdiagnosis of acute nonlymphocytic leukemia.

Adolescent↗
PubMed1983

Morphologic and cytochemical observations on the overt leukemic phase of therapy-related leukemia.

Comparison of morphologic and cytochemical features of the leukemic cells of patients with overt acute nonlymphocytic leukemia following therapy for primary malignancies (2 degrees-ANLL) to those of patients with ANLL de novo revealed several differences. No Auer rods were seen in cells of patients with 2 degrees-ANLL, but they were seen in 54 (42%) of 129 cases of ANLL de novo. Only one (4.7%) of 21 patients with 2 degrees-ANLL was classified as M-4 by FAB criteria, compared to 46 (36%) of 129 patients with ANLL de novo classified as M-4. Only one (10%) of 10 patients with 2 degrees-ANLL had more than 10% peroxidase-positive leukemic cells, whereas 19 (95%) of 20 of a control group of patients with ANLL de novo of similar FAB subtypes had more than 10% positive cells. Two of 11 (18%) 2 degrees-ANLL patients had 10% or more naphthol-AS-D chloroacetate esterase-positive cells, in contrast to nine (47%) of 19 patients with ANLL de novo. Nonspecific esterase activity was present in 10% or more of cells in only one (10%) of 10 patients with 2 degrees-ANLL studied, as compared to seven (35%) of 20 of the control group. Previous reports indicate that patients with 2 degrees-ANLL have cytogenetic abnormalities and therapeutic responses that differ from those in ANLL de novo; our findings indicate that there are morphologic and cytochemical differences as well.

Acute Disease↗
PubMed1983

Cytochemistry of Reed-Sternberg cells in lymph node imprints.

Cytochemical reactions were examined in lymph node imprints from a group of 53 previously untreated patients with histologically proven Hodgkin's disease. In 40 of 51 cases investigated, Reed-Sternberg (R-S) cells, irrespective of the cytologic appearances and the histologic types, showed moderate to strong reactions with acid phosphatase (ACP). In 12 cases ACP activity was present in more than 25% of the R-S cells. The reaction consisted of formation of small- to medium-sized granules, which were located close to the nuclei on a diffusely positive background or irregularly distributed throughout the cytoplasm. In three cases, a coarse granular reaction product with periodic acid-Schiff was present. R-S cells were positive to the naphthol-AS acetate esterase and beta-glucuronidase reactions in four and two cases, respectively. Alkaline phosphatase and naphthol-AS-D-chloroacetate esterase reactions were completely negative. Our results have revealed a pattern of staining in the diagnostic R-S cells similar to that in its morphologic variants; this supports the view that these cells may derive from a common primitive cell. Moreover, the quality and quantity of the ACP reaction product shows that R-S cells differ from both neoplastic histiocytes of malignant histiocytosis and neoplastic lymphocytes of T-cell lymphomas. This study confirms that R-S cells lack definite cytochemical characteristics of each of supposed progenitor cells: histiocytes and T-lymphocytes.

Acid Phosphatase↗
PubMed1983

A cytochemical and ultrastructural study of acute myelomonocytic leukemia exhibiting the pseudo-Chediak-Higashi anomaly of leukemia and "splinter-type" Auer rods.

An unusual case of acute myelomonocytic leukemia (AMML) in an 81-year-old man is described. Ultrastructural examination revealed the presence of distinct Auer bodies, which commonly are found in acute promyelocytic leukemia (APL). The Auer bodies had a splinterlike appearance with a distinct tubular substructure. The morphologic appearance of the leukemic cells coupled with the results of cytochemical staining reactions (alpha-naphthyl AS-D chloroacetate esterase, alpha-naphthyl butyrate esterase, Sudan black B, and periodic acid-Schiff reaction) identified this leukemia as myelomonocytic in origin. APL-like Auer rods in AMML now can be included in the spectrum of morphologic forms commonly seen in myelogenous leukemia and lends support to the hypothesis of an aberrant stem cell. In addition to the APL-like Auer rods, the leukemic blasts contained giant pink-staining granules on Wright's stain, some of which had a tubular substructure similar to the Auer rods under the electron microscope. The presence of these megagranules is a morphologic aberration termed "the pseudo-Chediak-Higashi anomaly of acute leukemia," and now has been described for the first time in AMML at the ultrastructural level.

Aged↗
PubMed1983

Systemic angioendotheliomatosis presenting with hemolytic anemia.

Two patients with systemic angioendotheliomatosis had prominent constitutional symptoms such as fever, loss of weight, and general weakness, and had multiple organ dysfunctions, including bizarre neurologic findings and dementia. Severe anemia that required frequent blood transfusions also was present. One patient developed severe hemolysis and hypersplenism that required splenectomy for relief; the other patient had intravascular hemolysis and autoimmune hemolytic anemia, which were treated unsuccessfully with conservative measures. In both cases, postmortem examination showed many large, noncohesive malignant cells within the lumen of the blood vessels in many of the organs. There was no infiltration or replacement of the normal tissues by the tumor cells. Histochemical studies showed that the tumor cells were pyroninophilic but did not have cytoplasmic immunoglobulins or activity of chloroacetate esterase and muramidase. The cells showed factor VIII antigen in their cytoplasm. Systemic angioendotheliomatosis may represent a true neoplastic process of the vascular endothelial cells.

Aged↗
PubMed1984

Granulocytic sarcoma presenting as a mediastinal tumor. Report of a case and cytological and cytochemical studies of tumor cells in vivo and in vitro.

An unusual case of granulocytic sarcoma in a 23-year-old man is reported. The patient initially presented with mediastinal tumor and was diagnosed clinically as having thymoma. The patient was treated by radiotherapy and surgical removal of the tumor. Histology of the excised tumor had been nondiagnostic because of extensive fibrous changes. Eight months later, the patient developed pleural effusion on the right, which soon was followed by blood and bone marrow pictures consistent with acute promyelocytic leukemia. In vitro culture of pleural effusion cells unexpectedly gave rise to a continuously growing peroxidase-positive myeloid cell line. Autopsy revealed the recurrent mediastinal tumor to be positive for intracytoplasmic naphthol AS-D chloroacetate esterase and lysozyme activity. From these findings, the patient retrospectively was diagnosed as having mediastinal granulocytic sarcoma, which terminated in pleural effusion and acute promyelocytic leukemia.

Adult↗
PubMed1985

Fluorescent microsphere detection of surface antigens and simultaneous cytochemistries in individual hematopoietic cells.

A method for simultaneous analysis of cell surface antigens, cell morphology, and endogenous cytochemical staining on the same hematopoietic cell has been developed. Individual cells in heterogeneous populations can be tested for cell surface antigens using monoclonal antibodies conjugated to fluorescein-containing microspheres and subsequently stained for myeloperoxidase, Sudan black B, chloroacetate esterase, or alpha naphthyl butyrate. This technic, when applied to mixed cell populations such as normal peripheral blood and leukemic bone marrow samples, provides simultaneous analysis of cell surface antigens and specific cytoplasmic cytochemistries within individual cells. Application of this technic may increase the body of knowledge about hematopoiesis by allowing individual cells to be analyzed simultaneously for multiple lineage-associated characteristics.

Antibodies, Monoclonal↗
PubMed1986

Acute leukemias with both myeloid and lymphoid surface markers. Cytoplasmic alpha-1-anti-chymotrypsin and alpha-1-anti-trypsin as possible indicators of early granulocytic differentiation.

Blast cells from ten patients (seven adults, three children) with acute lymphoblastic leukemias (ALL) contained immunoreactive cytoplasmic alpha-1-anti-trypsin (alpha-1-AT) and alpha-1-antichymotrypsin (alpha-1-ACT). Cytochemically positive reactions for block-like periodic acid-Schiff and a localized acid phosphatase suggested that the cells were of lymphoid origin rather than myeloid origin: negative for sudan black-B, nonspecific esterase, chloroacetate esterase, and myeloperoxidase. By surface phenotype, the leukemia showed positive reactions for both lymphoid (common acute lymphoblastic leukemia antigen, Ia, OKT-10) and myeloid (OKM-1, Leu M-1) antigens. Three of three patients tested portrayed the Philadelphia chromosome. Nine patients were Mexican-American and one was Japanese: all were of Asian ethnic derivation. Both myeloid and lymphoid treatment regimens were employed, with survival less than expected. Early granulocytic differentiation detectable by cytoplasmic alpha-1-AT and alpha-1-ACT in lymphoid blasts is discussed.

Adult↗
PubMed1990

Biphenotypic acute leukemia in adults.

Leukemias are characterized by an idiopathic proliferation of a progenitor cell that is committed to a single cell lineage. However, leukemias with dual-lineage differentiation are being described, especially within the pediatric age group. The authors reviewed 118 cases of adult acute leukemia phenotyped by immunofluorescent flow cytometry; 7 cases demonstrated mixed cell lineage. Immunophenotypically these cases were defined by early B-lymphocyte differentiation (TdT, HLA-DR, and CD19) coexpressed with a myeloid receptor (CD13, CD15, or CD33) on the same leukemic cell. Routine cytochemical evaluation demonstrated punctate positivity of the blasts with naphthol AS-D chloroacetate esterase stain in five of seven cases. Cytogenetic analysis revealed structural abnormalities of chromosome 11 in four of the seven cases. Three of these studies showed a break at 11q23-24, the location of the human proto-oncogene ets-1. Clinically, two of these leukemias represented chronic myelogenous leukemia in blast crisis, and all cases behaved aggressively. The authors' data suggest that mixed lineage leukemias are an identifiable subset of adult acute leukemias and are associated with a poor prognosis.

Acute Disease↗
PubMed1991

Granulocytic sarcoma presenting as an epidural mass with acute paraparesis in an aleukemic patient.

Granulocytic sarcomas are rare tumors composed of granulocytic precursor cells. They are most commonly encountered in patients with acute myelogenous leukemias and myeloproliferative disorders in blast crisis. Rarely, patients presenting with granulocytic sarcoma show no evidence of acute leukemia. The authors report an aleukemic patient with acute paraparesis from an epidural granulocytic sarcoma. Only five such cases have been reported previously. Immunoperoxidase stain for lysozyme and chloroacetate esterase stain were used to prove the myeloid origin of the tumor cells.

Aged↗
PubMed1991

Granulocytic sarcoma (chloroma). Presentation of an unusual case.

An unusual case of granulocytic sarcoma with a large retro-orbital tumor mass is described. The tumor had an uncommon cytomorphology and ultrastructure that mimicked a signet ring cell lymphoma. It was negative by chloroacetate esterase (CAE) stain. The patient was treated successfully with CHOP-regimen polychemotherapy and irradiation. Seventeen months after the initial diagnosis of malignant lymphoma, acute myeloid leukemia developed. Additional immunohistochemistry, including an immunoperoxidase staining for lysozyme, clearly demonstrated the early myeloid nature of the original tumor. This case emphasizes the importance of staining for lysozyme and other myeloid markers in addition to CAE staining in cases that demonstrate unusual morphological features.

Carboxylic Ester Hydrolases↗
PubMed1991

Prospective gene rearrangement studies and multiparameter analysis of acute myeloid leukemia.

Twenty-six cases of acute myeloid leukemia (AML) with cytochemical and immunophenotypic data were studied prospectively for immunoglobulin and T-cell receptor gene rearrangement. Dysmyelopoiesis was seen in 100% and Auer rods in 18%. Sudan black B was positive in 83% of the cases, peroxidase in 76%, nonspecific esterase in 74% (fluoride-inhibited in 82%), chloroacetate in 70%, acid phosphatase and PAS in 100%, and immunoperoxidase stains for platelet glycoprotein IIIa and factor VIII in 0% of the cases studied. Flow cytometry revealed myeloid phenotype in 19 of 20 cases. In four cases 5-86% of cells were TdT positive. Heavy-chain gene rearrangement was demonstrated in three cases (12%) and kappa light chain gene rearrangement in one; clinically significant rearrangement of the T-cell receptor gene was not found. Rearrangements of immunoglobulin genes are found occasionally in AML; these may represent nonspecific findings or coexistent lymphoid differentiation in AML.

Adolescent↗
PubMed1991

The transient appearance of small blastoid cells in the marrow after bone marrow transplantation.

Of 14 patients who underwent allogeneic or syngeneic bone marrow transplantation, 6 had a transient appearance of small blastoid cells in the bone marrow after transplantation. Most of these patients (11) had leukemia, although 3 had severe aplastic anemia. The cells were 8-18 micron in diameter and had scant cytoplasm and dense nuclei with smooth, homogeneous chromatin. They often had distinct nuclear clefts. These cells constituted 4.0-21.3% of the total number of bone marrow cells. They were not reactive with peroxidase, alpha-naphtyl butylate esterase, naphthol AS-D chloroacetate esterase, or periodic acid-Schiff stains. Immunocytochemical analysis revealed that the small blastoid cells expressed terminal deoxynucleotidyl transferase, Ia-like, CD19, and CD10 antigens and cytoplasmic mu heavy chains, indicating a precursor B-cell phenotype. CD20 antigen was not expressed on these cells. The data suggest that cytoplasmic mu may be expressed earlier than CD20 antigen in the differentiation of B-cell lineage. The morphologic, cytochemical, and immunophenotypic characteristics did not distinguish these nonneoplastic cells distinctly from leukemic lymphoblastic cells. The increase of small blastoid cells was a transient and self-limited phenomenon, in contrast to that of neoplastic blasts. These cells should be recognized as a common component of the bone marrow of marrow transplant recipients. The significance and role of these cells in immune recovery and hematopoiesis remain uncertain.

Bone Marrow↗
PubMed1988

Detection of leukotriene B4 in cardiac tissue and its role in infarct extension through leucocyte migration.

The main left coronary artery of rats was ligated near its origin under light ether anaesthesia and the infarction observed for 48 h. The ischaemic area was determined after an intravenous injection of pontamine sky blue dye 1 h before induction of cardiac arrest with potassium chloride. The unstained area (true ischaemic area) decreased with time to 27.1% of the left ventricle at 48 h, whereas the intensely stained area between the normal and the true ischaemic areas increased with time, suggesting that blood was flowing to the border from the normal surrounding tissue. The infarcted area, identified by its lack of triphenyltetrazolium chloride staining, became evident after 3 h and stabilised at 12 h (42% of left ventricle). The polymorphonuclear leucocyte counts in the hearts, differentiated by staining of their chloroacetate esterase, increased gradually up to 5500 cells per section at 24 h. The leukotriene B4 concentration, determined by radioimmunoassay after freezing of the beating heart in liquid nitrogen, increased to eight times that of the sham operated hearts and peaked at 8 h (9.4(0.6) ng per heart, mean(SEM) n = 5) before the leucocyte counts reached their maximum. A single oral dose of a selective 5-lipoxygenase inhibitor (AA-861, 80 mg.kg-1, 1 h before ligation) lowered the leukotriene B4 concentration to that of the sham operated hearts and decreased the leucocyte count by 49.4% and 41.2% at 12 and 24 h respectively. The inhibitor also reduced the infarct size at 48 h by 34.4%. It was concluded that leukotriene B4, generated in ischaemic cardiac tissue, may increase infarct size through migration of polymorphonuclear leucocytes.

Animals↗
PubMedDate not supplied

Human endometriosis-derived permanent cell line (FbEM-1): establishment and characterization.

A human epithelial-like cell line derived from peritoneal implants from a patient with gonadotrophin-releasing hormone (GnRH) agonist-resistant endometriosis graded as stage IVd according to the American Fertility Society classification was established in vitro. This cell line, designated FbEM-1, exhibited an epithelial-like morphology, grew in suspension and was immunoreactive for cytokeratins 8, 18, 19, vimentin and human leukocyte class I antigens. The cultured cells were negative for various haematopoietic cell markers, including the lymphoid cell antigens CD3, CD20 and CD45, von Willebrandt factor, carcinoembryonic antigen and the carcinoma antigen-125 (CA-125). In addition, the FbEM-1 cells were found to be moderately positive for periodic acid Schiff's (PAS) solution but were negative for alpha-naphthyl acetate esterase, peroxidase and chloroacetate esterase activities. Using specific antibodies against the progesterone, androgen and oestrogen receptors, approximately 40% and 5-10% of the cells immunostained for progesterone and androgen receptors respectively, while oestrogen receptors were not detected. On cytogenetic analysis using R-banding, these cells showed numerous chromosomal aberrations, including loss of one chromosome X, 4q+, 5q+, trisomy 7,8 and 10 and tetrasomy of chromosomes 17, 18, 19 and 20. These data show that the continuously growing FbEM-1 cell line established from endometriotic implants may be useful in achieving better understanding of the histogenesis of endometriosis.

Animals↗
PubMedDate not supplied

Determination of sodium monofluoroacetate (compound 1080) in tissues and baits as its benzyl ester by reaction-capillary gas chromatography.

A reaction-capillary gas-chromatographic procedure using photo-ionization (PID) or flame-ionization (FID) detection was developed for the determination of sodium monofluoroacetate (compound 1080), a pesticide, in tissues and baits. Fluoroacetic acid from tissue (1 g) and bait (10 g) extracts was first partitioned into ethyl acetate and then into 0.5 M benzyldimethylphenylammonium hydroxide. Benzylation was achieved by pyrolysis of the quaternary ammonium salt in the injection port. Chloroacetic acid was used as the internal standard. A linear relationship (r = 0.999) was observed between the peak area ratio of the substrate/internal standard and the fluoroacetic acid concentration. The detection limit for compound 1080 using the described analytical procedure, was 15 micrograms/kg with PID and 100 micrograms/kg with FID.

Animals↗