Assessment of proliferative activity of canine dermal mast cells by bromodeoxyuridine and proliferating cell nuclear antigen labelling

Cutaneous mast cells from skin biopsies of three healthy dogs and three dogs with atopic dermatitis were assessed for their proliferative potential using bromodeoxyuridine and proliferating cell nuclear antigen labelling. Mast cells isolated from the skin of two healthy dogs were also studied using bromodeoxyuridine labelling. Mast cells in skin biopsy specimens and mast cells isolated from the skin of healthy dogs did not incorporate bromodeoxyuridine. Two mast cells expressing proliferating cell nuclear antigen were seen around two superficial vessels in the dermis of one atopic dog. Epidermal cells, glandular epithelial cells, fibroblasts and endothelial cells incorporated bromodeoxyuridine and showed positive staining for proliferating cell nuclear antigen. These results suggest that canine mature mast cells do not proliferate in the dermis.     

Distribution and ultrastructure of mast cells in the equine respiratory tract

Mast cells in the equine respiratory mucosa were studied at both light--and transmission electron--microscope levels. Mast cells were identified at all levels of the tract, with the greatest cell density in the nasopharynx. The majority (57 to 94 per cent) of this cell population were located within the connective tissue of the lamina propria. Up to 20 per cent of these cells were associated with the mucosal glandular tissue, whilst small numbers were present within the surface epithelium and in association with nodular lymphoid tissue. In the peripheral lung tissue 20 per cent of the mast cell population was associated with the small airways, 25 per cent with the pleura, and 32 per cent with blood vessels. The fine and ultrastructural features of these mast cells resemble those described in other species, and their granules consist of two types which resemble human mast cells.     

Acute myeloid leukemia with mastocytic and megakaryocytic differentiation in a calf

A diagnosis of acute myeloid leukemia was made in a 10-month-old Holstein female calf. The leukemia was macroscopically characterized by great enlargement of the spleen and moderate enlargement of some lymph nodes. Histochemical and immunohistochemical examination disclosed the presence of neoplastic cells either containing metachromatic and tryptase-positive granules or expressing factor VIII-related antigen. The granules, which were positive for naphthol AS-D chloroacetate esterase and did not have particulate contents, were distinct from those of basophilic leukemia cells. This leukemia was thought to be derived from a common myeloid progenitor capable of giving rise to megakaryocyte-erythrocyte progenitors and granulocyte-monocyte progenitors with the ability to differentiate into mast cells.     

Mast cells and canine mast cell tumours. A review

Summary

This article reviews the literature on mast cells and tumours derived from mast cells in the dog. Mast cells play a central role in inflammatory and inunune reactions. Mast cells, normal and neoplastic, contain and release important biologically active substances: heparin, histamine, eosinophilic chemotactic factor and proteolytic enzymes.

Mast cell tumours occur in the dog, particularly in the boxer and related breeds, in the skin and less frequently in the intestines. Cytology usually provides an accurate diagnosis, but histological examination adds further information concerning the histologic grade and the completeness of surgical therapy. Cutaneous mast cell tumours should be regarded as potentially malignant and therefore be removed widely (3 cm. margin). Local recurrence, regional and distant metastases together with paraneoplastic disorders may cause the death of the pet. Histologic grading (2 or 3 grades) and clinical staging together with kinetic parameters and breed (boxers have relatively benign tumours) are important prognostic parameters. Based on prognostic criteria, surgical treatment should be completed with adjuvant radiotherapy, corticosteroids and eventually with combined chemotherapy. A novel, promising therapy is the application of the receptor kinase inhibitor. The study of the pathogenesis of mast cell tumours received new impetus by the finding of mutations, deletions and duplications, in exons 11 and 12 of the C‐kit oncogene. Further study of physiological and oncological aspects of mast cells are favoured by the availability of mast cells isolated from spontaneous mast cell tumours and of cultured cell lines.     

Canine Transmissible Venereal Tumour: Asessment of Mast Cell Numbers as Indicators of the Growth Phase

Mast cells are immune cells that are involved mainly in type 1 hypersensitivity reactions, and they have been implicated in tumour angiogenesis. In this study we assessed the presence of mast cell numbers and microvessel density during the progression and regression stages of natural spontaneous canine transmissible venereal tumours (CTVT). Mast cells were demonstrated by histochemical staining with toluidine blue, alcian blue and safranin O. Microvessel counts were demonstrated by immunohistochemical labelling with an antibody against the endothelial cell marker factor VIII. Mitotic cells, apoptotic cells and tumour infiltrating lymphocytes were counted from haematoxylin–eosin-stained sections. Tumour fibrosis was evaluated on Masson's trichome-stained sections. The results showed that progressing tumours had significantly higher mast cell counts and microvessel counts at the invasive edges of the tumours than did regressing tumours. In both the progressing and regressing tumours, microvessel counts were significantly positively correlated with mast cell counts. Regressing tumours had significantly higher mast cell counts of the whole tumour than progressing tumours. The results also showed that progressing tumours had significantly higher mitotic rate than regressing tumours, and fibrosis and apoptosis were significantly higher in regressing tumours than progressing tumours. There were no significant differences between the biochemical and haematological values of dogs with progressing and regressing tumours. These results suggests that mast cells play a role in CTVT progression probably by promoting vascularization at the invasion front during the progression phase, and that mast cell count could be used as one of the histological factors to indicate growth stage of CTVT.     

Mast cells and canine mast cell tumours. A review

This article reviews the literature on mast cells and tumours derived from mast cells in the dog. Mast cells play a central role in inflammatory and immune reactions. Mast cells, normal and neoplastic, contain and release important biologically active substances: heparin, histamine, eosinophilic chemotactic factor and proteolytic enzymes. Mast cell tumours occur in the dog, particularly in the boxer and related breeds, in the skin and less frequently in the intestines. Cytology usually provides an accurate diagnosis, but histological examination adds further information concerning the histologic grade and the completeness of surgical therapy. Cutaneous mast cell tumours should be regarded as potentially malignant and therefore be removed widely (3 cm. margin). Local recurrence, regional and distant metastases together with paraneoplastic disorders may cause the death of the pet. Histologic grading (2 or 3 grades) and clinical staging together with kinetic parameters and breed (boxers have relatively benign tumours) are important prognostic parameters. Based on prognostic criteria, surgical treatment should be completed with adjuvant radiotherapy, corticosteroids and eventually with combined chemotherapy. A novel, promising therapy is the application of the receptor kinase inhibitor. The study of the pathogenesis of mast cell tumours received new impetus by the finding of mutations, deletions and duplications, in exons 11 and 12 of the C-kit oncogene. Further study of physiological and oncological aspects of mast cells are favoured by the availability of mast cells isolated from spontaneous mast cell tumours and of cultured cell lines.     

Comparison of tracheal aspirates and bronchoalveolar lavage in racehorses. 1. Evaluation of cytological stains and the percentage of mast cells and eosinophils

OBJECTIVE: To compare a fast Romanowsky cytological stain (Diff-Quik) and Leishman's stain for the detection of mast cells in samples from the lower airways of racehorses, and to compare the proportion of mast cells and eosinophils in the total inflammatory cells in tracheal aspirate (TA) with those in paired bronchoalveolar lavage (BAL) samples. DESIGN: Retrospective case series of 48 young Thoroughbred and Standardbred racehorses. PROCEDURE: Fifty-one paired TA and BAL samples were collected after treadmill exercise from 48 horses with poor racing performance. Two slides were prepared from each sample; one was stained with Diff-Quik stain and the other with Leishman's stain. Differential cell counts of eosinophils and mast cells were recorded from each slide. Comparison of the suitability of the stains for the detection of mast cells, and comparisons of eosinophil and mast cell percentages in TA and BAL samples were analysed using the non-parametric Wilcoxon matched pairs test. RESULTS: Percentages of mast cells were significantly higher in Leishman than in Diff-Quik stained slides in both TA (P = 0.03) and BAL samples (P < 0.0001). Mast cell percentages were significantly higher in BAL than in TA samples using Leishman's stain (P < 0.0001). There was no significant difference in eosinophil percentages between TA and BAL samples (P = 0.07). CONCLUSIONS: Fast Romanowsky type stains (for example Diff-Quik) are not appropriate for the detection of mast cells in samples from the equine lower respiratory tract. Therefore, a metachromatic stain that reliably identifies mast cells (for example Leishman's) should be used if evaluation of mast cells in lower respiratory tract is undertaken. Mast cells are predominantly found in the distal small airways and alveoli sampled with a BAL. In contrast, eosinophils appear to be evenly distributed in the lower respiratory tract. However, high percentages of eosinophils are occasionally found only in TA samples. We recommend that both a TA and BAL be used for the evaluation of eosinophils and mast cells within the equine lower respiratory tract.     

The distribution and heterogeneity of mast cells in tongue from five different avian species

This study was conducted with the aim of determining the morphology, distribution and heterogeneity of mast cells in the tongues of seagull (Larus fuscus), common buzzard (Buteo buteo), goose (Anser anser), white stork (Ciconia ciconia) and Gerze rooster. The study used five samples of tongue material from each of the healthy adult avian species. The samples were fixed in 10% neutral‐buffered formalin (NBF) solution, then, after routine tissue follow‐up, the samples blocked with paraplast. Cross‐sections with 5–6 μm of thickness were stained with the 0.5% toluidine blue and alcian blue/safranin O (AB/SO). In all five avian species, it was found that the mast cells were in different sizes and round, oval or spindle‐shaped based on their place of distribution. Mast cell numbers were determined in stained with toluidine blue, examined ×40 objectives in a 1 mm² area. It was observed that mast cell density in subepithelial lamina propria and microscopic papilla was higher in the tongues of all species. Mast cell distribution and heterogeneity varied through the tongue, and there were more mast cells in the dorsal side of the tongue than the ventral side. The highest amount of mast cells was found in the tongue of the Gerze rooster among all five species. In the tongue cross‐sections stained with the combined method of alcian blue/safranin O (AB/SO), the mast cells were stained as AB (+), SO (+) and AB/SO (+) (mixed).     

Distribution and ultrastructure of mast cells in the duck

The distribution and ultrastructure of mast cells in the duck were studied. Carnoy's fluid was found to be the best fixative, and Padawer's method was found to be useful in demonstrating the metachromasia. Mast cells were seen in many of the organs and were most numerous in the proventriculus. Perivascular location was frequently seen. The ultrastructure resembled that of the chicken mast cell. The cell contained different types of membrane-bound granules, some of which were electron-dense and mottled.     

Mast cells in the ovary of Gallus gallus domesticus

1. Histamine or mast cells are involved in mammalian ovary function. Their role in the avian ovary is not known. In the present study mast cell distribution in the ovary of the domestic fowl was studied. 2. Mast cells were distributed throughout the ovary, both in the stroma of medullary and cortical regions as well as in the thecae of normal and atretic follicles. In the stroma, mast cells were especially abundant in the area just below the germinal epithelium (GE) and followed the contours of the GE. 3. In the follicles, mast cells were more obvious in the thecae of small non-yolky follicles, whereas they were compressed and scattered in the larger yolky follicles. They were more frequently seen in the theca externa than in the theca interna and in their ultrastructure showed characteristic mast cell granules. 4. Some of the atretic follicles showed increased mast cells in their thecae. Postovulatory follicles had very few mast cells. 5. The possible role of the mast cells in the ovarian activity of domestic fowl is discussed.     

Culture and initial characterization of the secretory response of neoplastic cat mast cells.

Mast cells isolated from feline splenic mastocytomas were cultured to study their structural and functional properties. Isolated cells from various cats were grown as monolayer cultures for a mean of 56 days (range, 30 to 76 days). Cat mast cells released allergic mediators in response to compound 48/80, anti-cat serum antibodies, and concanavalin A. On the basis of the finding that secretion from cat mast cells was stimulated by anti-cat serum antibodies and concanavalin A, these cells contain surface-bound immunoglobulins. The presence of mast cell-sensitizing antibodies has been suspected in cats, but never before directly demonstrated. Cultured cat mast cells have cytochemical and functional characteristics common to connective tissue-type mast cells and provide one of the few non-rodent models of cultured cells for the study of this type of mast cell.     

On the Ultrastructure of Mast Cells and Globule Leucocytes in the Common Bile Duct of Cattle Chronically Infected With Fasciola Hepatica

Mast cells and globule leucocytes in the walls of the common bile ducts of seven uninfected slaughtered cows and of five cows chronically infected with Fasciola hepatica were studied by electron microscopy. The mast cells displayed numerous small intracytoplasmic granules, the matrixes of which were either strongly electron-opaque and homogeneous or less dense with a fine-granular ultrastructure. Unattached ribosomes were abundant in the cytoplasm of the cells. Many mast cells in the walls of chronically infected bile ducts were normal-looking while the cells in the subepithelial tissues often showed striking irregularities in the ultrastructure of the granules. Globule leucocytes were most numerous in the surface epithelium and were not connected by desmosomes to the epithelial cells. The nuclei of the globule leucocytes were similar to those in the mast cells except that indentations caused by the globules were more usually seen. The globules were considerably larger than the mast cell granules but bounded by similar agranular capsules. The matrix of most globules was dense and homogeneous while the other globules showed dense reticulate or lamellate structures. The relationship between the mast cell and globule leucocyte in the liver is briefly discussed with reference to recent reports on these cells in the intestinal wall.     

Comparison of four staining methods for detection of mast cells in equine bronchoalveolar lavage fluid

Mast cells normally are present in equine bronchoalveolar lavage fluid (BALF), but usually represent <2% of all cells in healthy horses. An increased percentage of mast cells has been associated with airway hyperactivity and inflammatory airway diseases, but marked differences are reported between studies in normal and diseased horses. Because an abnormal mast cell count may be of clinical relevance, we compared the ability of a fast Romanowsky method to stain mast cell granules with that of 3 metachromatic stains: automated Romanowsky, May-Grünwald Giemsa, and toluidine blue stains. The BALF cells from 24 horses were studied. A differential cell count was performed blindly on 400 cells. The percentages of mast cells obtained were analyzed by means of repeated-measures analysis of variance and Fischer's PLSD test. The Bland and Altman method was used to assess agreement among stains. The mean percentage of mast cells in BALF was significantly lower with the fast Romanowsky than with the automated Romanowsky, May-Grünwald Giemsa, and toluidine blue stains. With the fast Romanowsky stain, the metachromatic granules of mast cells were not stained, and their identification was based on morphologic criteria. Toluidine blue staining allowed detection of the highest mean percentage of mast cells, but was inadequate for performing a differential cell count on other cell types. In conclusion, fast Romanosky stain may be inadequate for detection of mast cells in equine BALF, whereas automated Romanowsky, May-Grünwald Giemsa, and toluidine blue stains provide metachromatic staining of mast cell granules.     

PUFA-dependent alteration of oxidative parameters of a canine mastocytoma cell line

Mast cells play a key role in the immune response. Thereby, the balance of oxidative metabolism is of importance in mast cell mediator synthesis and release. Fatty acids may modify mast cell function in several ways. In this study, we investigated the influence of polyunsaturated fatty acids (PUFAs) on oxidative parameters of a canine mastocytoma cell line. C2 cells were cultured in media supplemented with linoleic acid, arachidonic acid, alpha-linolenic acid and eicosapentaenoic acid, respectively. Production of reactive oxygen species (ROS) as well as lipid peroxides was tested. Furthermore, stressor-induced DNA damage was measured. Exposure of the cells to PUFAs resulted in a significant increase in the synthesis of both ROS and lipid peroxides. Distinct differences between the PUFAs tested underline the impact of the unsaturation degree of fatty acids as well as the position of double bonds on mast cells.     

小鼠胃、小肠和精囊黏膜肥大细胞的组织学观察

取健康小鼠胃、肠管和精囊放入生理盐水中冲洗干净,放入10%中性甲醛固定液中固定、透明、浸蜡、包埋,对胃、肠管和精囊进行连续切片,并用苏木精-伊红和甲苯胺蓝染色。然后,用光学显微镜观察小鼠胃、肠管和精囊的肥大细胞并计数,再用生物统计学方法分析、比较了胃、肠管和精囊肥大细胞的分布规律。结果发现,胃内的肥大细胞最多,小肠较少,而精囊中几乎看不到。胃和小肠内肥大细胞主要分布在血管和腺管周围。     

Mast cell tumour and cutaneous histiocytoma excision wound healing in general practice

Objective : To assess whether wounds from incomplete mast cell tumour excisions are at greater risk of healing complications than wounds from complete excisions, or cutaneous histiocytomas. Methods : Mast cell tumours and cutaneous histiocytomas submitted to Nationwide Laboratories between November 1, 2007 and April 30, 2008 were selected. Questionnaires were sent to submitting veterinarians requesting details of tumour characteristics, clinical approach to the tumour and wound healing. Results : Three hundred and eighty‐six mast cell tumours and 524 cutaneous histiocytomas were identified. One hundred and eighty‐five mast cell tumours and 244 cutaneous histiocytomas questionnaires were returned (47% response). Wound complications arose in 20% of mast cell tumours and 21% of cutaneous histiocytomas. Multivariable analysis confirmed that larger tumours, tumours on the feet and a soft/“baggy” appearance, were significantly associated with a greater frequency of problems, leading to delayed wound healing and dehiscence. Clinical Significance : Incomplete mast cell tumour excision does not lead to greater risk of wound complications. Mast cell tumour surgical wounds have a similar rate of wound complications as cutaneous histiocytoma wounds.     

Reduced mucosal injury of SPF chickens by mast cell stabilization after infection with very virulent infectious bursal disease virus

Recent studies here have demonstrated that increased mast cell populations and tryptase activity contribute to lesion formation in regions of immune organs in special-pathogen-free chickens after infection with very virulent infectious bursal disease virus (vvIBDV). Mast cells and their mediators have been implicated in acute inflammatory injury after vvIBDV infection, but their precise role in this process remains elusive. In this study, the role of mast cells in the vvIBDV infection process was examined using ketotifen, a mast cell membrane stabilizer. On days 1, 2, and 3 postinfection, the bursa of Fabricius (BFs) were collected to quantify mast cells, tryptase and histamine contents by cytochemistry, immunohistochemistry and fluorospectrophotometry analyses, respectively. The results showed that the mast cell populations, tryptase expression, and histamine released increased significantly in the BFs (p < 0.01) of infected birds compared to controls, and acute inflammatory responses were observed in the former. In contrast, in infected chickens pretreated with ketotifen, mast cells, tryptase, and histamine were markedly decreased (p < 0.01) and probably as a result, the BFs remitted significantly. The overall results suggest that mast cells are positively involved in BF injury induced by vvIBDV infection. Inhibition of mast cell degranulation and concurrent mediator release may represent a novel strategy to modulate this process. This study, thus, advances the understanding of the acute inflammatory injury mechanisms triggered by vvIBDV infection and the contribution of mast cell activity in this process.     

Morphological and immunocytochemical investigations on mast cells in porcine ureter

Morphological, morphometric, histochemical and immunocytochemical investigations on mast cells, located in the wall of ureter of 8 months aged pigs were performed. Mast cells were found in all three layers of ureteral wall, but their distribution was irregular and the number unequal. It was established that alcian blue (AB)-positive mast cells were significantly more than toluidine blue (TB)-positive mast cells. A statistically significant smaller number of both AB and TB-stained mast cells were observed in the tunica mucosa. The largest number of mast cells was found in the tunica muscularis. In the adventitia, mast cells were higher in number in the main connective tissue than in the connective tissue near the blood vessels. Mast cells stained with TB showed variably expressed gamma-metachromasia, which was best visible in those situated in the lamina propria of the mucosa. The prevailing parts of mast cells, however, were AB-positive after AB-safranin staining. This was mostly found in mast cells of the tunica muscularis and in mast cells of perivascular location in the tunica adventitia. Immunocytochemically, mast cells were found to be positive for histamine and vasoactive intestinal polypeptide in the muscle coat, and to histamine in the adventitia, as well. On the basis of obtained results it was presumed that the mast cells in porcine ureter most probably took part not only in keeping of local homeostasis, but played also an important role of mobility of smooth muscle cells in the middle layer of ureter on one hand, and, on the other, in the adventitial blood vessels.     

Ultrasonographic findings in abdominal mast cell disease: a retrospective study of 19 patients

A retrospective survey from January 1989 to January 1999 of Tufts University Foster Hospital for Small Animals radiology records of 12 dogs and seven cats with cytologically or histopathologically confirmed abdominal mast cell disease was performed. Ultrasound changes in hepatic mast cell infiltration in dogs included a subjective increase in size, a diffuse increase in echogenicity, and one or more hypoechoic nodules. Ultrasound findings in the affected canine spleen included one or more hypoechoic nodules and a subjective increase in size. Two ultrasonographically unremarkable canine livers and one unremarkable spleen were found to be infiltrated by mast cells. The mast cell-infiltrated feline spleen was subjectively increased in size, mottled, irregular, or contained nodules. The affected lymph nodes in both dogs and cats were hypoechoic or inhomogeneous, subjectively increased in size, and rounded. Gastrointestinal involvement in cats was characterized by a thickened ileocecocolic junction or colon with loss of wall layering. Mast cells were not found in the gastrointestinal tract in any dog. One dog with mast cell infiltrate of the kidneys had multiple hypoechoic nodules in the cortex that distorted the outer contour of the kidney. Although these findings are not specific to the disease in either species, abdominal ultrasound is considered a useful tool for determining the extent of disease in small-animal patients with mast cell tumor if used in conjunction with histopathology or cytology.     

Globule leukocyte and mast cell in bile ducts of cattle naturally infected with liver flukes

In n previous report dealing with the pathology of bovine fascio-liasis the author described an unknown cell type in the epithelium of bile ducts. The histological and histochemical investigations published in this paper suggest that the cell may be considered a globule leukocyte. Globule leukocytes are rare in uninfected livers but are occurring in abundance in main bile ducts of cattle with spontaneous fascioliasis and also in small perilobular ducts in dicrocoeliasis. Liver fluke infection causes an increase in the population of subepithelial mast cells. Mast cell and globule leukocyte present similarities In their cytochemical properties. However, at low pH toluldinc blue shows a stronger but Alcian blue a weaker affinity for mast cells than for globule leukocytes.