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PubMed1977

[Morphology and monocytopoesis of mycosis fungoides].

Nature and nosology of mycosis fungoides have to be reconsidered because there is now evidence of the t-cell nature of the atypical lymphoid cells within the mycoside infiltrate. Therefore the concept of the reticulum cells in the pathogenesis of mycosis fungoides must be corrected. On the ground of morphological and immunological reasons these cells do not exist in the dermis at all. These new findings are the basis of our morphological study in patients with mycosis fungoides. Concentrating on the polymorphous lymphohistiocytic cells, which cannot be classified further by the method of paraffin-thin sections, light microscopically, the exact differentiation is only possible with semi-thin-sections. With this technique there can be made visible the typical features of nuclei of mycosis fungoides cells. The mycosis fungoides cells are characterized by a large nucleus with hyperconvoluted nuclear membranes, prominent nucleoli, irregular distribution of heterochromatin and sparse cytoplasma. The monocytopoiesis, scheduled by the relative number and the 3H-thymidine labeling indices of promonocytes and the activity of naphthol-AS-D-chloroacetate esterase in blood monocytes was markedly increased. These findings are of special interest, because macrophages are not only able to phagocyte but also play a crucial role in immunology. The elevated monocytopoiesis in all stages of mycosis fungoides points to a stimulation of the immunologic system. This supports the hypothesis of a persistent antigen, which stimulates by means of the functional intact monocyte-macrophage system the lymphocyte system leading to a permanent transformation with augmentation of these immunoblasts (mycosis fungoides cells).

Humans↗
PubMed1996

Testicular extramedullary myeloid cell tumor in a patient with myelodysplastic syndrome.

We report herein a case of extramedullary myeloid tumor arising bilaterally in the testes of a 66-year-old man, who had previously been diagnosed with myelodysplastic syndrome. Light microscopy of the testicular neoplasm demonstrated a tumor composed of large, slightly polygonal cells with pale blue to weakly eosinophilic cytoplasm. The tumor cells were immunoreactive for CD45, myeloperoxidase, lysozyme, CD43, and MB2. Many of the cells also expressed chloroacetate esterase. Peripheral blood and bone marrow findings were consistent with chronic myelomonocytic leukemia (FAB-CMML), particularly in the most recent material, which showed clear cellular dysplasia and an increase in the percentage of blasts in the bone marrow (15% to 20% of all nucleated cells). This case of extramedullary myeloid tumor is unusual in view of the patient's age and the testicular location. It emphasizes the importance of including extramedullary myeloid tumor in the differential diagnosis of histologically undifferentiated large-cell tumors, as well as a need to use a broad panel of immunohistochemical stains in such cases.

Aged↗
PubMed1996

1,25-dihydroxyvitamin D3 differentiates normal neutrophilic promyelocytes to monocytes/macrophages in vitro.

Although it is well established that the addition of 1,25-dihydroxyvitamin D3 (D3) to the culture of normal human granulocyte/macrophage progenitors induces monocyte/macrophage (Mo/M phi) colonies, the target cells of D3 in the Mo/M phi differentiation have not been identified. We examined whether neutrophilic promyelocytes are the target cells. As a source of the promyelocyte fraction, we used colonies after 5 days of culture (5-day colonies) of colony-forming unit-granulocyte. The culture contained granulocyte colony-stimulating factor (G-CSF) as the growth factor and generated only neutrophilic colonies. The promyelocytic nature of the 5-day colonies was confirmed morphologically, cytochemically, and ultrastructurally. After morphological evaluation on part of the individual colonies, they were transferred into new semisolid cultures with or without D3 (10(-7) mol/L) in the presence of G-CSF, then incubated for the subsequent 7 days. With D3, the colonies were loose, and all the constituent cells were morphologically small macrophages, which were positive for alpha-naphthyl butyrate (alpha NB) esterase, strongly positive for CD14 antigen, and plastic-adherent. While without D3, the colonies were rather compact, and all the constituent cells were morphologically mature neutrophils, which were positive for naphthol ASD-chloroacetate esterase and weakly positive for CD14 antigen. Secondary culture of the 8- or 10-day colonies with D3 induced a lower number of alpha NB-positive cells, in proportion to the percentage of promyelocytes at the time of transfer in each colony. Four days of secondary culture with D3 was sufficient to induce alpha NB-positive cells. G-CSF was not an essential factor to induce alpha NB-positive cells. These findings indicate that D3 differentiates normal human neutrophilic promyelocytes into the Mo/M phi lineage in vitro.

Calcitriol↗
PubMed1996

Interleukin-8 in Hodgkin's disease. Preferential expression by reactive cells and association with neutrophil density.

Hodgkin's disease (HD) shows rare neoplastic Hodgkin and Reed-Sternberg cells embedded in an abundant reactive infiltrate containing, among other cell types, neutrophilic granulocytes. Interleukin (IL)-8 is chemotactic for neutrophils. The expression of IL-8 was tested by in situ hybridization with 35S-labeled IL-8-specific RNA probes on 38 cases of HD. Reactive lesions, non-Hodgkin's lymphomas of B and T phenotype, and Langerhans cell histiocytosis served as controls. IL-8 expression was observed in Hodgkin and Reed-Sternberg cells in 3 of 33 cases of classical HD and in reactive cells in 20 of 33 HD cases as evidenced by combined isotopic in situ hybridization and immunohistology for the demonstration of cell-type-characteristic antigens or enzyme histochemistry for chloroacetate esterase. IL-8-positive cells were more numerous in cases of nodular sclerosing HD as compared with the mixed cellularity histotype (P = 0.01). The number of IL-8-positive cells and the density of neutrophils were positively correlated (P < 0. 01). In 5 cases of lymphocyte-predominant HD, IL-8 expression was not displayed. Non-Hodgkin's lymphoma cases contained IL-8 transcripts only in 1 of 23 cases in sparse reactive cells. In 4 of 7 cases of Langerhans cell histiocytosis, IL-8-specific signals were displayed in S100-negative cells. In conclusion, IL-8 expression in HD is largely confined to reactive cells and associated with infiltration by neutrophils. Elaboration of other cytokines by Hodgkin and Reed-Sternberg cells and reactive cells may explain the frequent expression of this cytokine in HD, particularly in the nodular sclerosing type.

Cell Count↗
PubMed1996

Role of immunophenotyping in characterisation of blast crisis of chronic myeloid leukemia--a study of 25 cases.

The blast cell populations of 25 patients of chronic myeloid leukemia in blast crisis (CML-BC) were studied for morphological, cytochemical and immunophenotypic features. The patients were divided into 6 broad groups based upon the pattern of surface marker positivity-myeloblastic, mixed myeloblastic, megakaryoblastic, mixed lineage, lymphoid and undifferentiated blast crisis. Myeloperoxidase (MPO), Sudan Black B (SBB) and Chloroacetate esterase (CAE) stains showed 100% specificity for the myelomonocytic lineage but the sensitivity was low. Periodic acid Schiff (PAS) stain was neither specific nor sensitive for the lymphoid lineage. Immunophenotyping as compared to morphologic and cytochemical assessment, was seen to be most useful for assigning a lineage to leukemic cells in CML-BC.

Adult↗
PubMed1979

Acute myelomonocytic leukemia: coexistent cytochemical markers for monocytes and granulocytes in the leukemic cells.

A 22-year-old man presented with mediastinal lymphoma and lymphoblastic leukemia. The leukemic cells in his blood and marrow did not stain positively with Sudan black B, peroxidase, chloroacetate esterase, or nonspecific esterase, and were considered as either lymphoblasts or stem cells. One year after the initial presentation, acute myelomonocytic leukemia developed and he died. The leukemic cells possessed the morphologic and cytochemical characteristics of both monocytes and granulocytes. This case illustrates the close ontogenetic relationship between the monocytes and the granulocytes. It also demonstrates that our present concept of acute myelomonocytic leukemia should be broadened to include cases in which the leukemic cells possess the morphologic and cytochemical characteristics of both the monocytes and the granulocytes.

Adult↗
PubMed1997

Renal involvement in myeloproliferative and lymphoproliferative disorders. A study of autopsy cases.

A considerable proportion of cases of myeloproliferative and lymphoproliferative disorders exhibit renal involvement. However, it is unclear whether the cytologic features, immunophenotype or grade of malignancy of the cells infiltrating the kidney differ from those of the primary tumor. This study was performed on 120 autopsy cases with the following diagnoses: acute myelogenous leukemia (AML, n = 22; subtypes M1 + M2, n = 12, subtype M4, n = 10), chronic myelogenous leukemia (CML, n = 7), agnogenic myeloid metaplasia/myelofibrosis (AMM/MF, n = 6), acute lymphocytic leukemia (ALL, n = 6), chronic lymphocytic leukemia (CLL, n = 9), other low-grade non-Hodgkin's lymphomas (low-grade NHL, n = 24), high-grade NHL (n = 21) and multiple myeloma (MM, n = 25). Renal involvement was investigated by light microscopy and immunohistochemistry. It was found in 34% of the cases, and was most common in ALL (83%) and low-grade NHL (50%) and least common in high-grade NHL (10%) and MM (12%). Dense infiltration of almost the entire kidney was most commonly seen in AML, low-grade NHL and ALL. Infiltration was bilateral and involved both the cortex and medulla in the majority of cases. When involvement of other organs was compared with that of the kidney, the lung was found to be involved in approximately the same number of cases, but liver involvement was more common and heart involvement less common. Reactive lymphocytic infiltration of the kidney was found in 18 of the 120 cases (15%), and was distinguished from scanty tumorous infiltration by immunohistochemical staining. No major phenotypical differences were found between the tumor cells infiltrating the kidney and those of the primary tumors in the bone marrow or lymph nodes. However, in one case of CML, the cells infiltrating the kidney were negative for KP1 and chloroacetate esterase, but could be identified by reactivity for CD34. The grade of malignancy in NHL was similar in both the nodal and renal manifestations.

Adult↗
PubMed1997

Granulocytic sarcoma of the testis: a report of two cases of a neoplasm prone to misinterpretation.

We report the cases of two men, aged 48 and 71 years, with granulocytic sarcoma of the testis. Both presented with left testicular swelling and underwent orchiectomy, which revealed cream-colored to yellow-tan, rubbery-to-firm, testicular tumors with extensive paratesticular spread. The tumor in the younger patient was composed of a uniform population of primitive cells with scant cytoplasm and was initially misinterpreted as malignant lymphoma. Staging revealed no extrascrotal spread. The patient was treated with radiation and chemotherapy and remained free of disease for 12 years, at which time he died of unrelated causes. The older patient had a history of a myelodysplastic syndrome. His tumor contained cells with bright eosinophilic, occasionally granular cytoplasm, consistent with myeloid lineage. Because of a prominent component of myelocytes, with round, eccentric nuclei and moderately abundant cytoplasm, and because of an associated chronic inflammatory cell infiltrate that contained mature plasma cells, the tumor was initially misinterpreted as a plasmacytoma, although it was reinterpreted as a granulocytic sarcoma before initiation of therapy. Tumor cells in both cases were positive with a chloroacetate esterase stain. Immunohistochemical staining revealed expression of myeloperoxidase, lysozyme, leukocyte common antigen, and CD43, but not of B-cell-specific or T-cell-specific antigens in both cases. Granulocytic sarcomas are apt to be misinterpreted as other hematolymphoid tumors, which may result in a significant error in management. The diagnosis should at least be thought of any time the diagnosis of malignant lymphoma or plasmacytoma of the testis is being considered.

Aged↗
PubMed1977

Histochemistry of blood and bone marrow smears of the straw-coloured fruit-eating bat, Eidolon helvum.

Blood and bone marrow cells of the straw-coloured fruit bat, Eidolon helvum were examined for Sudan black B and periodic acid-Schiff (PAS)-positive material, peroxidase, naphthol ASD chloroacetate esterase and beta-glucuronidase. A strong peroxidase reaction was given by developing granulocytes and monocytes in bone marrow smears, but the mature cells in the peripheral blood smears showed no reactivity. Further, developing and mature erythrocytes in both blood and bone marrow smears showed intense peroxidase reactivity.

Animals↗
PubMed1997

Synthesis and antimicrobial screening of some novel thiazoles, dithiazoles and thiazolylpyridines.

Several thiazolines 2a-c, and 4-thiazolidinones 3a-c were synthesized through the reaction of the cyanoacetic acid hydrazide derivatives 1a-c with phenacyl bromides or chloroacetic acid, respectively. The dithiazoles 4a, b were obtained from 2a, b through their reaction with sulphur and phenyl isothiocyanate. The dithiazoles 6a, b were prepared from 4a through its reaction with dimethyl sulphate followed by reaction of the produced 2-methylthiothiazolium salt 5 with either malononitrile or ethyl cyanoacetate. The thiazolyl-pyridines 7a, b and 8a, b were obtained from 2a, b through their reaction with arylidene malonitrile and arylidene ethyl cyanoacetate, respectively. The prepared compounds were screened for their antibacterial and antifungal activities in vitro. Some of them showed weak effects.

Anti-Bacterial Agents↗
PubMed1997

Expression of constitutively activated human c-Kit in Myb transformed early myeloid cells leads to factor independence, histiocytic differentiation, and tumorigenicity.

The cDNAs encoding wild type (WT) human receptor tyrosine kinase c-Kit and a constitutively activated mutant, V816Kit, were introduced into granulocyte-macrophage colony-stimulating factor (GM-CSF )-dependent early murine hemopoietic cells, which had been transformed with activated Myb. WTKit cells were able to grow in the presence of the human ligand for Kit, stem cell factor (SCF ), but displayed reduced growth and clonogenic potential in either SCF or GM-CSF compared with the parental cells in GM-CSF. In contrast, V816Kit cells grew without factor at a higher rate than the parental cells in GM-CSF and displayed increased clonogenicity. Dissection of the growth characteristics in liquid culture showed that in the presence of appropriate factors, the different populations had similar proliferation rates, but that V816Kit profoundly increased cell survival compared with WTKit or parental cells. This suggests that the signals transduced by WTKit activated with SCF, and by V816Kit, were not identical. Also, WTKit and V816Kit-expressing cells both varied from the early myeloid progenitor phenotype of the parental cells and gave rise to a small number of large to giant adherent cells that expressed macrophage (alpha-naphthyl acetate) esterase and neutrophil (naphtol-AS-D-chloroacetate) esterase, were highly phagocytic and phenotypically resembled histiocytes. Thus, WTKit activated by SCF and V816Kit were able to induce differentiation in a proportion of Myb-transformed myeloid cells. The factor independent V816Kit cells, unlike the parental and WTKit expressing cells, were shown to produce tumors of highly mitotic, invasive cells at various stages of differentiation in syngeneic mice. These results imply that constitutively activated Kit can promote the development of differentiated myeloid tumors and that its oncogenic effects are not restricted to lineages (mast cell and B-cell acute lymphoblastic leukemia), which have been reported previously. Furthermore, the mixed populations of cells in culture and in the tumors phenotypically resembled the leukemic cells from patients with monocytic leukemia with histiocytic differentiation (acute myeloid leukemia-M5c), a newly proposed subtype of myeloid leukemia.

3T3 Cells↗
PubMed1998

[Synthesis and properties of 5(4,6-diphenyl-2-pyrimidin-2-yl)-1,2,4-triazolin-3-thione and derivatives ].

By treatment of 4,6-diphenyl-pyrimidin-2-carboxyl chloride (1) with thiosemicarbazide the acylthiosemicarbazide 3 has been synthesized. Compound 3 could also be obtained by action of hydrazide on potassium thiocyanate. 5-(4,6-Diphenyl-pyrimidin-2-yl)-1,2,4-triazolin-3-thione (5) could be prepared either by basecatalyzed cyclization of acylthiosemicarbazide or by the reaction of methyl 4,6-diphenyl-pyrimidin-2-carboxylate with thiosemicarbazide in the presence of sodium methoxide. Alkylation of the 1,2,4-triazolin-3-thione 5 with chloroacetic acid resulted in formation of the S-carboxymethyl derivative 6. On heating, with a mixture of glacial acetic acid/acetic anhydride, compound 6 afforded 6-(4,6-diphenyl-pyrimidin-2-yl)thiazolo[3,2-b]-1,2,4-triazol-3(2H) -one (7). Condensation of 7 with aromatic aldehydes yielded 2-arylidene-6-(4,6-diphenyl-pyrimidin-2-yl)thiazolo[3,2-b]-1,2,4-+ ++triazol-3(2H)-ones, which could also be prepared by treatment of the S-carboxymethyl derivative 6 with an appropriate aldehyde in the presence of acetic acid and acetic anhydride. Some of the synthesized compounds possess a weak antiinflammatory activity.

Animals↗
PubMed1998

Granulocytic sarcoma presenting as a giant breast tumor in a pregnant woman: a case report.

We report a case of granulocytic sarcoma (chloroma) presenting as a giant breast tumor in a pregnant woman with no history of leukemia. The case was initially diagnosed as medullary carcinoma on a biopsy specimen and a modified radical mastectomy was performed. The diagnosis of granulocytic sarcoma requires the pathologist's high index of suspicion. The presence of immature eosinophils was an important clue. Leder's chloroacetate esterase stain; immunostaining for myeloperoxidase, CD34, CD43, CD68, and lysozyme; and ultrastructural finding of cytoplasmic lysosomal granules and Auer bodies all aided in confirming the diagnosis. It is imperative to recognize granulocytic sarcoma to avoid unnecessary surgery. Granulocytic sarcoma should be included in the differential diagnosis of breast tumors, especially in tumors with diffuse proliferation of small tumor cells.

Adult↗
PubMed1998

Specific signals generated by the cytoplasmic domain of the granulocyte colony-stimulating factor (G-CSF) receptor are not required for G-CSF-dependent granulocytic differentiation.

Granulocyte colony-stimulating factor (G-CSF) is the principal growth factor regulating the production of neutrophils, yet its role in lineage commitment and terminal differentiation of hematopoietic progenitor cells is controversial. In this study, we describe a system to study the role of G-CSF receptor (G-CSFR) signals in granulocytic differentiation using retroviral transduction of G-CSFR-deficient, primary hematopoietic progenitor cells. We show that ectopic expression of wild-type G-CSFR in hematopoietic progenitor cells supports G-CSF-dependent differentiation of these cells into mature granulocytes, macrophages, megakaryocytes, and erythroid cells. Furthermore, we show that two mutant G-CSFR proteins, a truncation mutant that deletes the carboxy-terminal 96 amino acids and a chimeric receptor containing the extracellular and transmembrane domains of the G-CSFR fused to the cytoplasmic domain of the erythropoietin receptor, are able to support the production of morphologically mature, chloroacetate esterase-positive, Gr-1/Mac-1-positive neutrophils in response to G-CSF. These results demonstrate that ectopic expression of the G-CSFR in hematopoietic progenitor cells allows for multilineage differentiation and suggest that unique signals generated by the cytoplasmic domain of the G-CSFR are not required for G-CSF-dependent granulocytic differentiation.

Animals↗
PubMed1998

Species- and sex-related differences in metabolism of trichloroethylene to yield chloral and trichloroethanol in mouse, rat, and human liver microsomes.

Trichloroethylene (TRI) has been shown to cause a variety of tumors, particularly in mouse liver and lung and rat kidney. However, a clear association between exposure to TRI and cancer development in humans has not been established. Because TRI metabolism by cytochrome P450s has been implicated in the mechanisms of TRI-induced carcinogenicity in mice, the purpose of the present study was to characterize the kinetics of TRI oxidation in male and female mouse, rat, and human liver microsomes to possibly allow for a better assessment of human risk. Methods were developed to detect and quantitate chloral, trichloroethanol, trichloroacetic acid, dichloroacetic acid, chloroacetic acid, glyoxylic acid, and oxalic acid, known TRI metabolites in rodents or humans. However, only chloral and its further metabolite, trichloroethanol, were consistently detected in the various liver microsomes in the presence of NADPH. Chloral was the major metabolite detected, and its levels were species- and sex-dependent; the amounts of trichloroethanol detected were also species- and sex-dependent but never exceeded 15% of total metabolites. Double-reciprocal plots of metabolite formation with male and female rat and human liver microsomes indicated biphasic kinetics, but this trend was not observed with microsomes from male or female mouse liver. The Vmax data are consistent, with male and female mice being more susceptible to TRI-induced liver carcinogenicity than male rats. However, the Vmax/Km ratios in male and female rat liver microsomes, in comparison with the male mouse liver microsomes, did not correlate with tumor incidences in these tissues. Furthermore, as only two out of six human liver samples examined exhibited Vmax/Km ratios similar or higher than the ratio obtained with male mouse liver, humans may vary in their toxic response after TRI exposure.

Acetates↗
PubMed1998

Neutrophils deficient in PU.1 do not terminally differentiate or become functionally competent.

PU.1 is an ets family transcription factor that is expressed specifically in hematopoietic lineages. Through gene disruption studies in mice we have previously shown that the expression of PU.1 is not essential for early myeloid lineage or neutrophil commitment, but is essential for monocyte/macrophage development. We have also shown that PU.1-null (deficient) neutrophils have neutrophil morphology and express neutrophil-specific markers such as Gr-1 and chloroacetate esterase both in vivo and in vitro. We now demonstrate that although PU.1-null mice develop neutrophils, these cells fail to terminally differentiate as shown by the absence of messages for neutrophil secondary granule components and the absence or deficiency of cellular responses to stimuli that normally invoke neutrophil function. Specifically, PU.1-deficient neutrophils fail to respond to selected chemokines, do not generate superoxide ions, and are ineffective at bacterial uptake and killing. The failure to produce superoxide could, in part, be explained by the absence of the gp91 subunit of nicotinamide adenine dinucleotide phosphate oxidase, as shown by our inability to detect messages for the gp91(phox) gene. Incomplete maturation of PU.1-deficient neutrophils is cell autonomous and persists in cultured PU.1-deficient cells. Our results indicate that PU.1 is not necessary for neutrophil lineage commitment but is essential for normal development, maturation, and function of neutrophils.

Animals↗
PubMed1998

Morphologic and cytochemical characteristics of blood cells from Hawaiian green turtles.

OBJECTIVE: To identify and characterize blood cells from free-ranging Hawaiian green turtles, Chelonia mydas. SAMPLE POPULATION: 26 green turtles from Puako on the island of Hawaii and Kaneohe Bay on the island of Oahu. PROCEDURE: Blood was examined, using light and electron microscopy and cytochemical stains that included benzidine peroxidase, chloroacetate esterase, alpha naphthyl butyrate esterase, acid phosphatase, Sudan black B, periodic acid-Schiff, and toluidine blue. RESULTS: 6 types of WBC were identified: lymphocytes, monocytes, thrombocytes, heterophils, basophils, and eosinophils (small and large). Morphologic characteristics of mononuclear cells and most granulocytes were similar to those of cells from other reptiles except that green turtles have both large and small eosinophils. CONCLUSIONS: Our classification of green turtle blood cells clarifies improper nomenclature reported previously and provides a reference for future hematologic studies in this species.

Animals↗
PubMed1998

Identification of basophilic cells that express mast cell granule proteases in the peripheral blood of asthma, allergy, and drug-reactive patients.

Metachromatic cells in the peripheral blood of patients with asthma, allergy, or an allergic drug reaction were evaluated for their nuclear morphology, surface expression of the mast cell (MC) marker c-kit, surface expression of the basophil marker Bsp-1, and granule expression of MC proteases. Consistent with previous findings by others, Bsp-1+/metachromatic cells represented <1% of the cells in the peripheral blood of normal individuals. These cells generally contained segmented nuclei. Very little, if any, tryptase (Try), chymase (Chy), or carboxypeptidase A (CPA) was found in their granules, and very little, if any, c-kit was observed on their surfaces. The number of metachromatic cells increased in the peripheral blood of the three groups of patients. Like the basophils in normal individuals, most of these metachromatic cells contained segmented nuclei and expressed Bsp-1. However, in contrast to the basophils in normal individuals, many of the metachromatic cells in the three patient groups expressed c-kit, Try, Chy, and/or CPA. That the metachromatic cells in the blood of our patients have some features of MCs and some features of basophils suggests that human basophils and MCs are derived from a common progenitor. As assessed by the chloroacetate esterase cytochemical assay, the immunoreactive Chy in the peripheral blood of these patients is enzymatically active. Because MC proteases regulate numerous immunologic and other biologic systems, the expression of Try, Chy, and/or CPA in a peripheral blood-localized cell in an individual having asthma, allergy, or an allergic drug reaction has important clinical implications.

Acute Disease↗