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PubMed1983

Evidence for the origin of Hodgkin and Sternberg-Reed cells from a newly detected small cell population.

To clarify the origin of Hodgkin (H) and Sternberg-Reed (SR) cells, frozen sections of lymph nodes from 25 patients with Hodgkin's disease were immunostained with a large panel of monoclonal antibodies reactive with cells of lymphoid tissue and granulopoiesis. The results showed that (a) H and SR cells are devoid of markers specific to, or characteristic of B cells, macrophages, dendritic reticulum cells, or interdigitating reticulum cells, and (b) the vast majority of H and SR cells contain granulocyte-related antigens detectable with the monoclonal antibodies TU9 and 3C4, but constantly lack other granulocytic cell markers (such as peroxidase and chloroacetate esterase). Monoclonal antibodies raised against a Hodgkin's disease-derived cell line included one, Ki-l, that was found to be selectively reactive with H and SR cells and a minute, but distinct, cell population in normal lymphoid tissue and bone marrow. The latter hitherto unknown cell population appears to be the normal equivalent of H and SR cells.

Antibodies, Monoclonal↗
PubMed1989

The pathology of murine myelogenous leukemias.

Murine myelogenous leukemias can be classified into several distinct subgroups based on morphology, cytochemical staining, and immunoreactivity. The leukemias invariably involve the spleen and the extent of infiltration into other tissues is variable. The myelogenous nature of the leukemia is readily apparent in well-differentiated leukemias on the basis of morphology; with poorly differentiated leukemias, positive staining with chloroacetate esterase, nonspecific esterase, and certain monoclonal antibodies such as Mac-1, is helpful to establish myelogenous differentiation. Subgrouping of myelogenous leukemias depends on the presence or absence of monocytic differentiation, as ascertained by staining with Mac-2, electron microscopy or phagocytosis. Leukemias showing no monocytic differentiation can be classified as myeloblastic, corresponding to the FAB M1 and M2 subtypes in humans. Leukemias exhibiting both monocytic and granulocytic features are myelomonocytic, corresponding to the FAB M4 subtype. Tumors with only monocyte differentiation arise primarily as solid tumors in mice, and a leukemic phase is variable.

Animals↗
PubMed1989

[Release of proteinase inhibitors as a vital reaction in the early post-traumatic interval].

In a pilot study paraffin-embedded sections of open skin wounds (stab and slash wounds, lacerations) were investigated to determine the presence of a vital reaction. Granulocytes were detected by naphthol AS-D chloroacetate esterase, the enzyme "lysozyme", and eight proteinase inhibitors by the indirect immunoperoxidase method. The tissue specimens were taken from consecutive autopsy material. The survival time could be determined in 14 cases (10-165 min) and was unknown in 12 other cases of sudden death due to injury of the major vessels or heart. The controls were cases with injuries inflicted after and cases of sudden death due to massive blunt trauma served death. In vital injuries, accumulations of proteinase inhibitors, particularly alpha-2-macroglobulin and alpha-1-antichymotrypsin, were demonstrable in the corium parallel to the wound surface. In comparison, the reaction of proteinase inhibitors that neutralize only enzymes participating in blood coagulation or complement activation (C1-esterase inhibitor and protein C) was absent or weak. Protein accumulation was observed only sporadically in cases of sudden death and never in cases with wounds inflicted after death. No relationship could be established between semiquantitatively estimated staining and survival time. Granulocytes and lysozyme were first observed in the corium after a survival time of more than 60 min.

Granulocytes↗
PubMed1995

1,25-Dihydroxyvitamin D3 promotes prostaglandin E1-induced differentiation of HL-60 cells.

Human promyelocytic HL-60 cells can be induced by biochemical agents to differentiate in vitro towards divergent types of myelomonocytic cells. It has been reported that prostaglandin E1 (PGE1) can induce granulocytic differentiation and that 1,25-dihydroxyvitamin D3 (1,25(OH)2D3) can induce monocytic differentiation. We have now examined the effects of these compounds, both alone and in combination, on HL-60 cell differentiation. PGE1 (1 microgram/ml) or 1,25(OH)2D3 (10 nM) each inhibited cell proliferation over 48-96 hours of treatment, but combined treatment with both agents was necessary to produce a strong inhibition. The percentage of HL-60 cells that can reduce nitroblue tetrazolium (NBT) (a characteristic index of early monocytic or granulocytic differentiation) increased 13-fold within 72 hours of PGE1 treatment, and 1,25(OH)2D3 produced a five-fold stimulation. However, combined treatment (PGE1 plus 1,25(OH)2D3) produced a dramatic 35-fold increase. HL-60 cells did not produce significant levels of nitric oxide (NO) before 48 hours in culture, and treatment with PGE1 or 1,25(OH)2D3 did not significantly increase cellular NO elaboration over control levels. However, combined treatment produced a striking 12-fold increase over control levels. Similarly, combined treatment was necessary to obtain the maximal time-dependent stimulation of cellular lactate dehydrogenase (LDH) activity (a marker of granulocytic differentiation) as well as acid phosphatase (ACP) activity. During this same period of time, PGE1, but not 1,25(OH)2D3, markedly stimulated cellular elaboration of interleukin (IL)-1 alpha, IL-6, and tumor necrosis factor (TNF)-alpha, and 1,25(OH)2D3 cotreatment strongly augmented these effects. Thus, combined treatment with 1,25(OH)2D3 plus PGE1 generally augmented the apparent conversion of these cells, producing synergistic (multiplicative) or additive effects. Furthermore, PGE1 induced within 48 hours the more general phenotypic changes classically associated with the differentiation of these cells: increased expression of chloroacetate esterase (ChAE) (a granulocytic marker), decreases in the nuclear/cytoplasmic ratio (characteristic of development beyond the promyelocyte/myelocyte stage), and major alterations in morphology from floating spherical cells to loosely adherent, elliptical polygons. 1,25(OH)2D3 had little effect itself on most of these parameters, but augmented the morphological changes induced by PGE1 treatment. Within 48 hours, the ability of these cells to reduce the tetrazolium salt WST-1, a general measure of cellular metabolic activity, was increased by PGE1, but not by 1,25(OH)2D3; however, the combination of 1,25(OH)2D3 and PGE1 again produced the strongest stimulation. Similarly, only PGE1 significantly reduced intracellular ATP levels, but combined treatments produced a more pronounced decrease. In summary, our findings suggest that PGE1, not 1,25(OH)2D3, is sufficient to promote rapid in vitro differentiation of HL-60 cells along the granulocyte pathway; however, the PGE1-induced conversion of these cells is markedly augmented by cotreatment with 1,25(OH)2D3. In addition, these converted HL-60 cells preferentially utilize the glycolytic pathway, rather than the citric acid cycle, for production of ATP, a metabolic characteristic that resembles that described for mature granulocytes.

Acid Phosphatase↗
PubMed1989

Cooperative effects of gamma-interferon and 1 alpha, 25-dihydroxyvitamin D3 on in vitro differentiation of the blast cells of RAEB and RAEB-T.

We added recombinant human gamma-interferon (gamma-IFN) and 1 alpha, 25-dihydroxyvitamin D3 (1 alpha, 25 (OH)2D3) to the bone marrow cells from six patients with RAEB or RAEB-T in liquid suspension cultures. After cultivation for 7 to 9 days, numerical, morphological and functional changes of the cells were assessed. gamma-IFN and 1 alpha, 25 (OH)2D3 additively suppressed cell growth, especially the number of blast cells decreased. The expression of alpha-naphthylbutyrate esterase (NBE) activity appeared to be promoted but that of naphthol AS-D chloroacetate esterase (NAE) activity was apparently suppressed by the addition of gamma-IFN and/or 1 alpha, 25 (OH)2D3. The percentage of NBT reduction-positive cells and latex-phagocytizing cells was only slightly increased by both agents. These results indicate that gamma-IFN and 1 alpha, 25 (OH)2D3 cooperate to induce monocytoid differentiation of the patients' blast cells. Combination therapy with both agents merits further study.

Adult↗
PubMed1989

Cytochemical studies of normal feline blood and bone marrow cells.

Blood and bone marrow cells of ten clinically healthy cats were stained for alkaline phosphatase (ALP), peroxidase (PO), chloroacetate esterase (CAE), alpha-naphthyl butyrate esterase (NBE), sudanophilia, and periodic acid-Schiff (PAS) reaction. Mature neutrophils in blood and bone marrow were devoid of ALP and NBE, but exhibited modest to strong PO, CAE, sudanophilia, and PAS reaction. In bone marrow, sudanophilia, PO, and CAE were prominent at the promyelocyte stage and diminished with cellular differentiation and maturation, while PAS reactivity increased with cell maturation usually from the myelocyte stage onwards. Myeloblasts were negative for all cytochemical reactions, but some large unidentifiable cells reacted strongly for ALP. Eosinophils were slightly reactive for ALP, CAE, and PAS, but not for PO, sudanophilia, and NBE. Basophil granules stained strongly for CAE, revealed PAS positivity, and stained negatively for PO, NBE, ALP, and sudanophilia. Slight ALP activity was detected in the intergranular cytoplasm of basophils. Lymphocytes and monocytes, with few exceptions, stained negatively. An occasional lymphocyte revealed slight globular NBE activity (NaF-resistant) and diffuse PAS reaction, while an occasional monocyte contained a few PO-positive and sudanophilic granules. Monocytes reacted modestly, whereas bone marrow macrophages reacted strongly for NBE (NaF-sensitive). Cells of the erythroid series stained negatively for all cytochemical reactions, megakaryocytes were PAS-positive, and platelets gave positive reactions for PAS and CAE.

Alkaline Phosphatase↗
PubMed1988

TdT expression in acute myeloid leukaemia. Haemopoietic immaturity or maturational asynchrony?

A total of 412 cases of acute leukaemia were examined for the presence of nuclear terminal deoxynucleotidyl transferase (TdT) by indirect immunofluorescence. Of the 129 cases of acute myeloblastic leukaemia (AML FAB groups M1/M2) examined, 18% (n = 23) had significant proportions (greater than 10%) of TdT-positive blasts. Although most of these AML cases (n = 18) were of poorly differentiated (M1) type; 5 cases of AML showing features of granulocytic differentiation (M2) were also found to be TdT-positive. Even though TdT was generally more strongly expressed in the M1 group and associated with other markers of myeloid immaturity (Ia positive and lack of chloroacetate esterase), there was no inverse relationship with Sudan black or myeloperoxidase activity. In addition, although the proportion of AML-M1 cases with increased TdT-positive cells was slightly higher (18/95, 19%) than for the AML-M2 group (5/34, 15%) the results suggest that the presence of nuclear TdT in leukaemic myeloblasts may not only reflect cellular immaturity but may also be due to maturational asynchrony in otherwise well-differentiated blasts.

Bone Marrow↗
PubMed1982

Increased acetone exhalation induced by metabolites of halogenated C1 and C2 compounds.

Rats were exposed, in a closed desiccator jar chamber, to concentrations of various halogenated C1 and C2 compounds at which the metabolizing capacities were saturated (Vmax conditions). Within the exposure period of 50 h concentrations of the xenobiotic and of exhaled acetone were monitored in the gas phase of the system. The quantitative extent of acetone exhalation was dependent on the individual compound examined. Acetone exhalation was stimulated in presence of vinyl chloride, vinyl bromide, vinyl fluoride, vinylidene fluoride, cis- and trans-1,2-dichloroethylene, trichloroethylene, perchloroethylene, methylene chloride, chloroform, carbon tetrachloride and 1,1,2-trichloroethane. No stimulation of acetone exhalation occurred with 1,1,1-trichloroethane and with the reference hydrocarbon n-hexane. Also, acetone exhalation was evoked by infusions of either fluoroacetate or chloroacetate, two anticipated or proven metabolites of some haloethylenes; the infusion rates of which were based on calculations of the metabolic rates of vinylidene fluoride and of vinyl chloride, respectively.

Acetone↗
PubMed1982

Toxicokinetics of chloroethanol in the rat after single oral administration.

The excretion and tissue distribution of 14C-labelled chloroethanol were studied in rats following single oral administration of 5 and 50 mg/kg body weight. At both dose levels, the radioactivity was rapidly eliminated, mainly in the urine. On the first day after application of 5 mg/kg body weight, 77.2% of the dose were found in the urine, 1.7% in the faeces, and 1.0% as carbon dioxide in the expired air. Only 2.8% were excreted by these routes during the following 3 days. The residual radioactivity remaining in the tissues after 4 days was almost equally distributed and amounted to about 0.4% of the dose in the liver and 3% in the whole organism. At the higher dose level, excretion rates and tissue concentrations were similar. Examination of the urine by anion exchange chromatography on DEAE-Sephadex revealed two metabolites which were identified by GC/MS analysis as thiodiacetic acid and thionyldiacetic acid. These metabolites represented almost the whole urinary radioactivity. They were excreted in approximately equal amounts at the low dose whereas the thiodiacetic acid predominated with about 70% of the urinary radioactivity at the high dose. Unchanged chloroethanol, chloroacetic acid, S-carboxymethylcysteine and sulphonyldiacetic acid were not found in the urine.

Animals↗
PubMed1986

Electron microscopic and immunohistochemical observations of differentiation of human myeloid leukemia line, PL-21.

A new human myeloid leukemia cell line, PL-21, consisting of promyelocytes, was microscopically and immunohistochemically studied for their maturation induced by 12-O-tetradecanoyl-phorbol-13-acetate (TPA) and retinoic acid (RA). More than 80% of PL-21 cells cultured for 4 days with 10 ng/ml of TPA became macrophage-like cells with functional and histochemical properties consistent with monocytes/macrophages. The cells adhered to the plastic flask, developed phagocytic activity and macrophage-specific intracytoplasmic alpha subunit of S-100 protein, and had reduced myeloid-specific cytochemical markers. In contrast, RA-treated PL-21 cells displayed mature neutrophil-like morphology after 7 days of exposure. Most of the cells had reduced nitro blue tetrazolium and acquired phagocytic activity with persistence of myeloid-specific cytochemical markers such as peroxidase, naphthol-AS-D chloroacetate esterase, and Sudan black B. These results indicate that the PL-21 cell line ca be induced to mature into two directions of macrophages and neutrophils by chemical inducers, and will provide a useful tool for studying the differentiation of leukemic cells and searching for other differentiation inducers.

Cell Differentiation↗
PubMed1979

Large basophilic cells in the bone marrow in iron deficiency anemia.

Many large cells with a strongly basophilic cytoplasm and large nuclei with a delicate chromatin pattern and large nucleoli were studied in 1 mu sections of core biopsies in 8 patients with iron deficiency anemia and in 5 patients with pernicious anemia. In 2 normal controls these cells were rare. Possible features of granulocytic differentiation were assessed with the Giemsa stain and with the naphtol-ASD-chloroacetate esterase reaction which is indicative of early granule formation: the large cells were constantly negative. In thin sections the morphological appearances of the large basophilic cells are rather similar in such different haematological disorders as pernicious anemia or iron deficiency anemia. A clear difference can be demonstrated by karyometry, which shows that the large basophilic cells in pernicious anemia are significantly larger than those in iron deficiency anemia or in the normal control.

Anemia, Hypochromic↗
PubMed1989

Enzyme cytochemistry of unfixed leukocytes and bone marrow cells using polyvinyl alcohol for the diagnosis of leukemia.

Cytochemical methods for the demonstration of enzyme activities in blood and bone marrow cells were systematically improved by the addition of an inert polymer, polyvinyl alcohol (PVA), to the incubation medium and by using optimized reaction media. The methods investigated were tetrazolium salt methods for lactate, glucose-6-phosphate, succinate and glutamate dehydrogenase, the indoxyl-tetrazolium salt method for alkaline phosphatase, the diaminobenzidine method for peroxidase, and diazonium salt methods for chloroacetate esterase, beta-glucosaminidase, beta-glucuronidase, acid phosphatase, and dipeptidylpeptidase II and IV. PVA in the media preserved the morphology of cells very well and prevented leakage of large molecules such as enzymes from the cells. Therefore, fixation or long periods of air-drying prior to incubation leading to substantial loss of enzyme activity could be avoided. A brief period of drying (2 min at 37 degrees C) of the cell preparations just before the incubation was sufficient for making the cells permeable. Localization of enzyme activities was very precise and precipitation of the final reaction product was confined to sites which are known to contain the enzyme under study (granules, mitochondria, lysosomes). These advantages advocate the use of PVA in haematological enzyme cytochemistry and especially for diagnosis of leukemia.

Bone Marrow↗
PubMed1981

The thymus as haematopoietic tissue of non-lymphoid cells.

Non-lymphoid haematopoiesis in the thymus was studied in 33 cases ranging from 14 weeks gestation up to 15 months postnatally. All thymuses displayed focally granulopoiesis in the connective tissue septa and in the lymphoid tissue, where it was localized in the peripheral cortex and at the cortico-medullary junction. Within the lymphoid tissue the granulocytic series showed many precursor cells with large rounded nuclei. Their origin could only be identified by the naphtol AS D chloroacetate esterase stain. In cases with thymic atrophy these precursor cells were present in relatively large numbers. Within the connective tissue septa many granulocytes with nuclear segmentation were intermingled with precursor cells. The erythroid series were detected with an immunohistochemical method for demonstration of haemoglobin. The positively stained nucleated cells occurred mainly in nests mostly localized in the peripheral cortex. These precursor cells were usually found in different places from the granulopoietic cells. No megakaryocytes were found. With appropriate techniques non-lymphoid haematopoiesis can be demonstrated in the developing thymus and must be considered as an expression of normal growth.

Connective Tissue Cells↗
PubMed1986

Contraluminal transport of small aliphatic carboxylates in the proximal tubule of the rat kidney in situ.

In order to study the characteristic of contraluminal transport of hydrophylic small fatty acids the in situ stopped flow microperfusion technique [12] has been applied. By measuring with 4 s contact time the decrease in the contraluminal concentration of the respective radiolabelled substances the concentration dependence of the influx into the cortical cells was tested. The 4 s decrease in contraluminal concentration of chloroacetate, L-lactate, D-lactate, 3-hydroxybutyrate and acetoacetate was between 26% and 31%. For each substance the percent decrease was the same, no matter whether it was offered in a concentration of 0.1 or 10 mmol/l. Contraluminal disappearance of 0.1 mmol/l L-lactate was not influenced by 5 mmol/l H2DIDS, probenecid, phloretin, mersalyl or cyanocinnamate, but it was significantly (37%) inhibited by 5-nitro-2-(phenyl-propyl-amino) benzoate, a blocker of the nonspecific anion channel. The percent decrease in propionate uptake was somewhat larger - between 36% and 39% - but again not different at 0.01, 0.1, 1.0 and 10 mmol/l. With pyruvate the contraluminal decrease was 20% at 0.1 mmol/l and 31% at 10 mmol/l. The percent disappearance of the aromatic pyrazinoate was 38% and 34% at 0.1 and 10 mmol/l and for nicotinate 42% and 22%, respectively. The disappearance of nicotinate (0.1 mmol/l) was significantly inhibited by 10 mmol/l pyrazinoate and paraaminohippurate (PAH). The data are in agreement with the hypothesis that the hydrophilic small fatty acids traverse the contraluminal cell side by simple diffusion, possibly via the unspecific anion channel [14], pyruvate via the dicarboxylic acid pathway in a cooperative manner and pyrazinoate, as well as nicotinate, via the PAH pathway.

Acetates↗
PubMed1989

Myeloblastoma of the brain.

Two patients developed intraparenchymal myeloblastomas (granulocytic sarcomas) of the brain. One patient with acute myelogenous leukemia in hematological remission had multiple myeloblastomas in the cerebrum and thalamus. The second patient, who had Philadelphia-chromosome-positive chronic myelogenous leukemia, developed a solitary lesion of the cerebrum without systemic evidence of blastic transformation of leukemia. By light microscopy, each biopsy specimen revealed a tendency for the malignant cells to appear in perivascular locations, and the presence of cytoplasmic naphthol AS-D chloroacetate esterase activity in some cells. Ultrastructural analysis demonstrated the presence of primary granule-like lysosomal structures in these cells, confirming their granulocytic origin. The lack of continuity of the myeloblastomas with the dura mater and the perivascular clusters of blast cells suggest that the pathogenesis of these lesions in the present cases could best be accounted for by the direct, hematogenous spread of mitotically competent leukemic cells to brain parenchyma.

Adult↗
PubMed1994

Genetics and biochemistry of 1,2-dichloroethane degradation.

Dichloroethane (1,2-DCE) is a synthetic compound that is not known to be formed naturally. Nevertheless, several pure microbial cultures are able to use it as a sole carbon source for growth. Degradation of 1,2-DCE proceeds via 2-chloroethanol, chloroacetaldehyde and chloroacetate to glycolate. The genes encoding the enzymes responsible for the conversion of 1,2-DCE to glycolic acid have been isolated. The haloalkane dehalogenase and an aldehyde dehydrogenase are plasmid encoded. Two other enzymes, the alcohol dehydrogenase and the haloacid dehalogenase, are chromosomally encoded. Sequence analysis indicates that the haloacid dehalogenase belongs to the L-specific 2-chloroproprionic acid dehalogenases. From the three-dimensional structure and sequence similarities, the haloalkane dehalogenase appears to be a member of the alpha/beta hydrolase fold hydrolytic enzymes, of which several are involved in the degradation of aromatic and aliphatic xenobiotic compounds.

Alcohol Dehydrogenase↗
PubMed1986

Immunohistological study of histiocytic necrotizing lymphadenitis.

Immunohistological study of 18 cases of histiocytic necrotizing lymphadenitis (HNL) demonstrated numerous helper/inducer cells (OKT-4) and suppressor/cytotoxic cells (OKT-8) with activation (Tac) and proliferation (OKT-9) markers, and histiocytes (lysozyme, alpha-1 anti-chymotrypsin, OK-M1) in the affected areas. However, B cells (B-1), NK cells (Leu-7 and Leu-11), complement proteins and receptor (C4 and C3d receptor), and neutrophils (chloroacetate esterase) were scanty or absent in these foci. Activity of NK cells was also decreased in the peripheral blood of 2 cases examined. The results suggest that HNL might be induced by the abnormal T cell-histiocyte response against some causative agents which induce a similar reaction of delayed hypersensitivity type.

Adolescent↗
PubMed1989

Heterogeneous malignant non Hodgkin's lymphomas as a causative disorder in lethal midline granuloma.

The present report describes the results of a combined morphological, enzyme- and immunohistochemical analysis of nine cases of malignant non Hodgkin's lymphomas (NHL) clinically presenting as lethal midline granuloma. In a previous report written before antibodies directed against B and T lymphocytes were available, a histiocytic origin of such neoplasms had been suggested. A panel of antibodies reactive with most B cells (L26, MB1, KiB3) and a majority of T cells (MT1, UCHL1) was applied on paraffin sections of formalin fixed tissues as well as antibodies directed against leukocyte common antigen (LCA), myeloid/histiocyte antigen (MAC 387), lysozyme, alpha-1-antitrypsin, alpha-1-antichymotrypsin, S-100 protein, prekeratin and immunoglobulin light chains. Enzyme histochemistry included tests for non-specific acid esterase, acid phosphatase, beta-glucuronidase and chloroacetate esterase. As a result, five T, two B and two unclassified (malignant histiocytosis probable) NHL were identified, indicating distinct heterogeneity of NHL as causative disorders in lethal midline granuloma.

Adolescent↗