Acute myelomonocytic leukemia negative for alpha-naphthyl acetate esterase stain in a Holstein cow

A 4-year, 7-month-old Holstein cow presented with anorexia. Physical examination revealed masses in the interscapular region and vagina. Blast cells were detected in the masses and peripheral blood by fine needle aspiration cytology and hematological examination. By bone marrow aspiration, blast cells constituted up to 24.2% of all nucleated cells, and 22% and 2% of non-erythroid cells stained positive for myeloperoxidase and alpha-naphthyl acetate esterase (ANAE), respectively. Pathological examination revealed the mass lesions consisted of a proliferation of tumor cells, which were positive for monocytic markers (HLA-DR and Iba-1). The cow was diagnosed with acute myelomonocytic leukemia (AMML). Even when tumor cells are ANAE-negative, AMML cannot be completely ruled out and should be considered when diagnosing cattle with leukemia/lymphoma.     

Characterization of short- and long-term cultured goat peripheral blood monocytes

Goat peripheral blood mononuclear cells were isolated, using Ficoll-sodium diatrozoate. Monocytes were separated by adherence and maintained in culture for up to 50 days. By 24 hours of culture and after removal of nonadherent cells, there were 94.2 +/- 5% of the adherent cells classifiable as monocytes based on nonspecific esterase staining. Greater than 98% of these cells were phagocytic. Approximately 94% had receptors for the Fc portion of bovine immunoglobulin G, and 86% had receptors for equine complement. Cytochemically, goat monocytes were positive for nonspecific esterase, acid phosphatase, glucuronidase, lactate dehydrogenase, succinic dehydrogenase, and glycogen, regardless of culture duration when tested. Results for specific esterase, peroxidase, and Sudan black staining varied from faint to negative. The esterase staining pattern of cultured monocytes was characterized by light and electron microscopies. Ultrastructurally, esterase activity was limited to the cell membrane. Intracytoplasmic esterase activity was not recognizable in normal monocytes or in monocytes containing phagocytized particles.     

Differential cytochemical staining characteristics of channel catfish leukocytes identify cell populations in lymphoid organs

This is one of the first characterizations of channel catfish (Ictalurus punctatus) leukocytes by enzyme cytochemistry. Leukocytes demonstrated cytoplasmic staining patterns very similar to mammalian leukocytes when stained with acid phosphatase, alpha-naphthyl butyrate esterase, beta-glucuronidase, alpha-naphthyl acetate esterase, Sudan Black B and anti-immunoglobulin specific immunohistochemistry. Lymphocytes, monocytes, macrophages, neutrophils, and surface immunoglobulin positive (surface Ig+) cells were present in channel catfish renal hematopoietic tissue and spleen and demonstrated distinctive cytoplasmic foci staining patterns, cytoplasmic blushing or cell membrane staining. Monocytes, macrophages, lymphocytes and surface Ig+ cells were present in the thymus. Thymic and splenic cellular organization appeared very similar to these same mammalian tissues. In the thymus, acid phosphatase positive cells were distributed throughout the parenchyma, while alpha-naphthyl butyrate esterase and beta-glucuronidase positive cells were concentrated in the cortex and the medulla, respectively. Surface immunoglobulin positive cells occurred in the cortex. In the spleen, acid phosphatase positive cells were scattered throughout the parenchyma, while alpha-naphthyl butyrate esterase positive cells were scattered throughout the parenchyma and adjacent to splenic arterioles. Beta-glucuronidase and surface immunoglobulin positive cells were restricted to immediately adjacent to splenic arterioles. Sudan Black B positive cells were scattered throughout the parenchyma, while alpha-naphthyl acetate esterase positive cells occurred adjacent to peri-arteriole lymphoid sheaths and appear very similar to mammalian metallophils.     

Nonspecific esterase and naphthol-AS-D-chloroacetate esterase in monocytoid and myeloid cells of healthy cattle and cattle suffering from leukosis]

The reaction to non-specific esterase can be used for the identification of the monocytoid cells of the periphery. A negative reaction is exhibited by neutrophile and eosinophile leucocytes and erythrocytes. Varying results are obtained from lymphocytes, rendering it impossible to use this method in the group of lymphoid cells of the periphery or marrow. In the group of large marrow cells (promonocytes), non-specific esterase gave a very strong reaction; this applies both to the marrow of healthy cattle and cattle suffering from leucosis. For the time being, efforts to use this reaction for the solution of the problem of the differentiation of monocytoid cells and cells of similar size in the myeloid series of the bone marrow have not been successful. In neutrophile leucocytes of the periphery, naphtol-AS-D-chloroacetate esterase gives a less intensive reaction than in humans. For this reason, it is less suitable for the differentiation of these cells. Other cell types (eosinophile leucocytes, monocytes, lymphocytes, erythrocytes as well as their bone-marrow stages) give a negative reaction. In the group of large marrow cells of the myeloid series (promyelocytes and neutrophile myelocytes), naphtol-AS-D-chloracetate esterase shows a more intensive reaction in healthy cattle, as compared with cattle suffering from leucosis.     

Establishment of a new canine cell line (CCT) originated from a cutaneous malignant histiocytosis

A new canine cell line, named CCT, was established from the cutaneous malignant histiocytosis in a 4-year-old male Borzoi. CCT proliferated with loose adherence and doubling time was approximately 30 hr. When co-cultured with latex beads, CCT phagocytized beads vigorously. Lysozyme and vimentin were positive by immunostaining, and non-specific esterase and acid phosphatase were positive by cytochemical staining. These features indicated the cells had a histiocytic nature. Furthermore, by subcutaneous injection to nude mice CCT could successfully form tumors with the morphological and immunohistochemical features similar to the original tumor.     

Synthesis of nonspecific esterase in small intestine enterocytes during experimental coccidiosis in suckling pigs]

The activity of nonspecific esterase (EC. 3.1.1.1.) was evaluated in the small intestine mucosa of 21 conventional piglets infected on day 5 after parturition (DAP) with oocysts of the Eimeria debliecki coccidium (infection dose of 200,000 oocysts) for this evaluation a microdensitometric analysis at the level of enterocytes was used. The same examination was also performed in the small intestine mucosa of four control conventional piglets at the age of 2-5 days (Tab. I). The synthesis of nonspecific esterase in the experimentally infected piglets was followed on day 1 to day 10 after infection (DAI). The activity of nonspecific esterase in the small intestine mucosa was found to decrease in a direction from duodenum absorption cells (D mean 34.15) to caudal ones (Fig. 1); ileum enterocytes have the optical density of the enzyme by 8.2% lower (D mean 31.38). The deposition of nonspecific esterase is localized mainly in the supranuclear zone of enterocytes while in the para- and infranuclear zones of absorption cells its concentration is only minute. In the experimentally infected piglets a marked increase in the optical density of nonspecific esterase of enterocytes was observed as soon as on day 1 after infection when the enzyme concentration increased by 19.4% (Tab. II). The maximum increase in the activity of nonspecific esterase of absorption cells was recorded on DAI 9 when the enzyme D mean value was higher by 165% in comparison with the activity of nonspecific esterase demonstrated in the control piglets (Fig. 2, 3, 4). But at the end of experimental infection (DAI 10) the total density of nonspecific esterase of enterocytes decreased by 38.2%.(ABSTRACT TRUNCATED AT 250 WORDS)     

Comparison of Various Staining Procedures in the Identification of Hepatozoon canis Gamonts

The staining characteristics of gamonts of Hepatozoon canis in peripheral blood smears were evaluated. Three stains, Diff-Quik stain, Giemsa stain and a modification of the naphthol-ASD-chloroacetate esterase stain, were compared. The staining characteristics of the infected cell and the parasite were different in each stain used. The modified naphthol-ASD-chloroacetate esterase stain showed the most difference between the infected and noninfected cells, as well as a difference in staining characteristics of the parasite nucleus from those of the host nucleus. Although the parasite was identified with all stains, a definitive diagnosis was more easily obtained with the naphthol-ASD-chloroacetate esterase stain.     

Morphologic characteristics of a transplantable tumor derived from a spontaneous malignant fibrous histiocytoma in the rat

Malignant fibrous histiocytoma (MFH) developed spontaneously in subcutaneous tissue of the head of a 15-month-old male Fischer 344 rat. The tumor was serially transplanted into syngeneic rats up to the 45th generation and was designated MFH-MT. Light and electron microscopic examinations revealed that the original and serially transplanted tumors were composed of an admixture of fibroblast-like and histiocyte-like cells arranged in a storiform pattern. Neoplastic cells gave positive reactions for acid phosphatase, alkaline phosphatase, nonspecific esterase, alpha-1 antitrypsin and lysozyme. The tumors transplanted into the lungs and cutaneous tissue of the tail had a mixed histologic appearance of storiform, pleomorphic, myxoid and giant cell types. Moreover sclerosing hemangioma-like and osteosarcoma-like structures were also found. MFH-MT grew well in athymic nude mice showing neoplastic proliferation of pleomorphic cells strongly positive for alpha-1 antitrypsin. Development of MFH-MT was significantly retarded by the two antitumor drugs tested. The retarded tumors consisted predominantly of fibroblast-like cells and abundant collagenic fibers, whereas histiocytic cells decreased in number.     

Pulmonary aspergillosis in a horse with myelomonocytic leukemia

Acute myelomonocytic leukemia was diagnosed in a 2-year-old Standardbred mare that had hind limb edema and fever unresponsive to antibiotics. The mare had anemia, thrombocytopenia, and leukocytosis, with circulating myeloblasts and monocytoid cells. A bone marrow specimen was hypercellular, with myeloblasts and monocytoid cells. Peroxidase, chloroacetate esterase, and alpha naphthyl acetate esterase activities were detected in many bone marrow cells. Interstitial pulmonary densities were seen radiographically. The mare was euthanatized and necropsied. Infiltrates of leukemic cells were found microscopically in specimens of spleen, lymph nodes, liver, kidneys, gastrointestinal tract, and lungs. Granulomas containing fungal hyphae were seen microscopically in the lungs, and Aspergillus sp was isolated from the lesions.     

Characterisation of the effector cells in antibody-dependent and spontaneous cell-mediated cytotoxicity in swine against target cells infected with transmissible gastroenteritis virus

When porcine peripheral blood leucocytes were fractionated, lymphocytes were the most active effectors in both antibody-dependent cell-mediated cytotoxicity (ADCC) and spontaneous cell-mediated cytotoxicity (SCMC), although both polymorphs and macrophages showed some activity in ADCC. Adsorption of lymphocytes to antibody-sensitised or unsensitized PK-15-transmissible gastroenteritis (TGE) cells caused similar reductions in ADCC and SCMC effector activities. Over 60 per cent of the target-binding lymphocytes were non-specific esterase positive large lymphocytes, which did not form erythrocyte (E)-rosettes, and about 30 per cent were non-specific esterase positive medium sized lymphocytes, which formed low avidity E-rosettes. The remainder were non-specific esterase negative small lymphocytes, some of which formed high avidity E-rosettes. None of the eluted lymphocytes stained for surface immunoglobulin and all formed low avidity erythrocyte-antibody rosettes. Porcine killer and natural killer cells resembled in many respects those described in humans and rodents.     

A clinical case of acute myelomonocytic leukemia in a Holstein cow

A 2-year, 3-month-old Holstein cow presented with anorexia and enlarged superficial lymph nodes. Fine needle aspiration cytology of the superficial lymph nodes revealed large blast cells. Hematological examination revealed anemia, neutropenia, and blast cells in peripheral blood. Blast cells were the predominant cell type in bone marrow aspirates. Of the non-erythroid cells, 26%, 58%, and 18% were positive for myeloperoxidase, α-naphthyl acetate esterase, and naphthol AS-D chloroacetate esterase, respectively. Pathological examination revealed the proliferation of neoplastic cells, which were positive for monocytic markers, in the affected lymph nodes. The cow was diagnosed with acute myelomonocytic leukemia based on these findings. This report highlights the importance of performing bone marrow aspiration cytology and cytochemical staining when diagnosing bovine myeloid leukemia.     

Transplantable mononuclear cell leukemia in F344 rat with particular reference to nodular tumor developing at the transplant site.

A transplantable mononuclear cell leukemia (MCL) was established from spontaneous MCL in an F344 rat. In this work, we paid special attention to a nodular tumor, named MCL-YSK, developed at the subcutaneous transplant site. MCL-YSK was serially passaged in subcutaneous tissue of syngeneic rats up to the 19th generation. Transplants from MCL-YSK grew into nodules 3 cm in diameter and 11.3 g in weight 9 weeks after subcutaneous implantation. Neoplastic cells forming the nodules had azurophilic cytoplasmic granules, which ultrastructurally appeared to be lysosomes. The cells reacted positively for acid phosphatase and nonspecific esterase, but not for alkaline phosphatase, alpha-1 antitrypsin and lysozyme, nor reacted with anti-rat monocyte/macrophage monoclonal antibody and anti-rat CD-8 monoclonal antibody. They possessed Fc-receptor. Leukemic cells first appeared in the peripheral blood 6 weeks after transplantation when subcutaneous nodules reached an average diameter of one cm. Subsequently, leukemic changes progressed in recipients as MCL-YSK grew larger. The recipients died during the period from 8 to 12 weeks after transplantation, showing anemia, depression, splenohepatomegaly and lymph node enlargement. Diffuse or focal proliferation of sprinkled tumor cells was present in many organs. Hematologic changes suggestive of hemolytic anemia, elevated plasma enzymes and decreased drug-metabolizing enzymes, indicative of hepatic malfunction, were seen in transplant recipients. MCL-YSK was easily transplanted into athymic nude mice. The transplanted mice showed leukemic changes similar to those observed in rats with transplanted MCL-YSK.     

家蚕蛾溶茧酶分泌器官及蛾吐出液的研究

家蚕蛾溶茧酶可能来源于下颚、嗉囊和中肠,嗉囊是否具有分泌溶茧酶的能力至今缺乏直接的证据。分别用光学显微镜和透射电镜观察了家蚕蛾下颚、嗉囊和中肠的组织结构,发现嗉囊不具备分泌器官的形态特征,是由薄的膜围成的囊状结构,没有观察到分泌细胞及其分泌物;而下颚和中肠具有分泌细胞,存在大量的分泌物质。对蛾吐出液进行酶活性检测,发现蛾吐出液除具有酯酶活力外,还具有纤溶酶活力。分别对下颚、嗉囊和中肠的组织蛋白作酯酶活力和纤溶酶活力测定,发现嗉囊组织蛋白不具有这两种酶活力,而下颚和中肠的组织蛋白具有较强的酯酶和纤溶酶活力。以上实验结果表明:家蚕蛾溶茧酶的来源不是嗉囊,溶茧酶可能来源于下颚和中肠。     

Morphologic and cytochemical characteristics of blood cells from the desert tortoise (Gopherus agassizii).

Morphologic and cytochemical staining characteristics of erythrocytes, leukocytes, and thrombocytes of the desert tortoise (Gopherus agassizii) were evaluated, using blood smears prepared from 23 healthy tortoises of Kern County, Calif. Special emphasis was placed on differentiating features of the various leukocytes and thrombocytes. A variety of cytochemical stains, including benzidine peroxidase, Sudan black B, chloroacetate esterase, alpha-naphthyl butyrate esterase, acid phosphatase, leukocyte alkaline phosphatase, periodic acid-Schiff, and toluidine blue were used. Heterophils had a characteristic, large, focal area of positive staining with chloroacetate esterase, alpha-naphthyl butyrate esterase, and acid phosphatase. Eosinophils stained diffusely positive with benzidine peroxidase, allowing differentiation of this leukocyte from heterophils. Thrombocytes stained focally positive with periodic acid-Schiff, allowing differentiation of these cells from lymphocytes, which stained uniformly negative. An intracytoplasmic body, commonly observed within erythrocytes, was considered ultrastructurally to represent a degenerate organelle.     

Morphology and cytochemistry of bovine bone marrow mononuclear phagocytes.

Bone marrow samples were collected from five normal calves and mononuclear cells were separated using Ficoll-Hypaque. Mononuclear cells were cultured on coverslips in Leighton tubes for six hours. The adherent cells were differentiated using Wright's and nonspecific esterase stains. Monoblasts, promonocytes and monocytes were present in the proportion of 1:2.31:4.96.     

二倍体和四倍体野生鸭茅遗传特性比较研究

采用细胞遗传学，生化遗传学的方法，对我国亚热带地区几个二倍体和四倍体野生鸭茅的核型，花粉母细胞减数分裂染色体配对行为，不同生育期的酯酶和过氧化物酶同功酶进行比较研究。结果表明，供试野生二倍体鸭茅之间，各四倍体之间的同源性较高，遗传差异较小；二倍体与四倍体之间，在核型，花粉数量，种，子千粒重，酯酶和过氧化物酶等方面都有很大差异；二倍体和四倍体野生鸭茅的花粉母细胞减数分裂虽然正常，但花粉育性和结实率不     

Morphologic and cytochemical characteristics of blood cells of juvenile loggerhead sea turtles (Caretta caretta)

A morphologic classification based on the cytochemical characteristics of blood cells of 35 juvenile loggerhead sea turtles (Caretta caretta) is described. Cytochemical stains included benzidine peroxidase, chloroacetate esterase, alpha-naphthyl butyrate esterase (with and without sodium fluoride), acid phosphatase (with and without tartaric acid), Sudan black B, periodic acid-Schiff, and toluidine blue. The morphologic characteristics of erythrocytes were similar to those reported in green turtles. Six types of white blood cells were identified: heterophils, eosinophils, basophils, lymphocytes, monocytes and thrombocytes. Except for the basophils, the rest of the white blood cells from loggerhead turtles had different cytochemical characteristics compared to blood cells from other sea turtle species. The leukocyte differential count was different from that reported for other sea turtle species. Heterophils were the most numerous leukocytes from these loggerhead turtles, followed by lymphocytes, eosinophils, monocytes and basophils. This paper provides a morphologic classification of blood cells of loggerhead sea turtles that is useful for veterinary surgeons involved in sea turtle conservation.     

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.     

The effect of insulin on primary cultures of rat preadipocytes grown in fetal or postnatal pig serum

Stromal vascular cell cultures, prepared from the inguinal pads of 50-g Sprague Dawley rats, were exposed to media with 10% fetal pig serum which is inherently low in insulin, for the first 3 to 5 d of culture. Insulin was supplemented to media for periods of 2 to 6 d. In cultures treated (2 to 4 d) with 10(-9), to 10(-10) or 10(-11) M insulin, differentiated cells (lipid and esterase staining) appeared 1.5 to 2 times wider than differentiated cells in control cultures. At 10(-9) M insulin (4 to 5 d), in cultures grown in the presence of fetal pig serum the number of esterase reactive cells was increased twofold to threefold. The percentage of total cells that were esterase reactive was elevated 50 to 300% relative to control cultures. Insulin-treated preadipocytes were more reactive for lipoprotein lipase activity (histochemical assay) compared with reactivity of control cells. Quantitative analysis of percentage of light transmittance (Zeiss photometer) through stained cells indicated an increase (P less than .001) in lipoprotein lipase staining at 10(-9), 10(-11) and 10(-13) M insulin (2 d). The specific activity of glycerol phosphate dehydrogenase was elevated twofold to threefold (P less than .05) and soluble protein elevated 50 to 100% (P less than .05) in cultures treated (3 to 6 d) with 10(-9) M insulin. Decreasing the cell plating density (50%) in cultures grown in the presence of pig serum reduced the elevation in enzyme activity induced by insulin in preadipocyte cultures. Physiological levels of insulin enhanced lipogenic enzyme activity in preadipocytes and may enhance the conversion of stromal cells to preadipocytes.