Mast cell tumor destruction by deionized water

In a controlled study, malignant murine P815 mastocytoma cells exposed in vitro to distilled and deionized water died as a result of progressive swelling, degranulation, and membrane rupture. A 90% mean cell death occurred when cells obtained directly from culture were exposed to deionized water for 2 minutes. Of 6 cryopreserved malignant murine cell lines, which included Cloudman S91 melanoma, CMT-93 rectum carcinoma, MMT-06052 mammary carcinoma, and S-180 Sarcoma, only P815 mastocytoma and YAC-1 lymphoma were significantly (P less than 0.05) affected by hypotonic shock; Cloudman S91 melanoma cells were the most resistant. Mastocytoma cells were selectively killed by hypotonic solution, and lymphoma cells were also killed by isotonic saline solution. Local mast cell tumor (MCT) recurrence and percentage survival were evaluated in 12 cats (21 MCT) and 54 dogs (85 MCT) subjected to surgery alone or local infiltration of deionized water as an adjunct to surgery. Of all 16 incompletely excised MCT in cats, there was no local recurrence following injection. Four mast cell tumors (2 cats) regressed after being injected in situ. In dogs with clinical stage-I MCT, local recurrence was detected in 50% (5/10), but with injection after incomplete excision, local MCT recurrence was significantly (P less than 0.05) less (6.6%, 1/15). Percentage recurrence was significantly (P less than 0.05) less and survival significantly greater when incompletely excised grade-II MCT were injected. Mean follow-up period after surgery in cats and dogs was 35 and 23.4 months, respectively.     

Mast cell tumour destruction in dogs by hypotonic solution

Distilled water was studied as an hypoosmotic oncolysis agent to prevent mast cell tumour (MCT) recurrence following surgical excision. The results from 118 MCTS excised in 74 dogs were: surgery alone, a recurrence rate of 52·6 per cent (10/19); and surgery followed by distilled water injections into the wound, a 26·2 per cent (26/99) recurrence rate. All of 10 small MCTS (0·03 to 0·25 cm³) injected with distilled water in situ with no surgery, regressed. The present data suggest that distilled water should be considered as an adjunct to surgery for canine MCTS.     

TGF-β and TNF-α stimulate MMP-9 production in murine epidermal keratinocytes

Matrix metalloproteinases 2 and 9 (MMP‐2 and ‐9) are zinc‐dependent metalloenzymes and have gelatin‐degrading activity. Both MMP are known to be secreted by many types of cells and play important roles in several biological changes including tissue remodeling and wound healing. In the present study, a primary culture of murine epidermal keratinocytes was prepared and effects of transforming growth factor‐β (TGF‐β), tumor necrosis factor‐α (TNF‐α) and interferon‐γ (IFN‐γ) on expression of MMP‐2 and MMP‐9 by the keratinocytes was examined. Gelatin zymography revealed that murine epidermal keratinocytes secreted proenzyme forms of MMP‐2 and MMP‐9, but the active forms of both MMP were hardly detectable, indicating that in vitro autoactivation of these proenzymes did not occur. Both TGF‐β and TNF‐α stimulated MMP‐9 production in a dose‐dependent manner, but the MMP‐2 level was not changed. Interferon‐γ hardly affected production of MMP‐2 or MMP‐9. Ribonuclease protection assay demonstrated that TNF‐α increased the level of MMP‐9 mRNA 6‐fold compared to the control, whereas TGF‐β slightly up‐regulated it. These results suggest that expression of MMP‐9 could be regulated by several cytokines in murine epidermal keratinocytes.     

Mast cell tumors in three ferrets

Pruritic open skin lesions (5 mm diameter) on the thorax, neck, or head of 3 ferrets were diagnosed as mast cell tumors. Diagnosis of the tumors was based on histologic examination of biopsy specimens. Histologic diagnosis was necessary because the lesions had been treated as mycotic or bacterial ulcers and did not respond to conventional treatment. Excised mast cell tumors did not recur, and malignant behavior of the tumors was not observed.     

Stimulation and killing of bovine mononuclear leukocytes by bacterial lipopolysaccharide (endotoxin)

Bovine adherent mononuclear leukocytes were incubated with bacterial lipopolysaccharides (LPS) in vitro, and these cells produced a factor that increased the blastogenic reaction of mouse thymocytes to concanavalin A. This factor most resembles interleukin 1. The LPS were also cytotoxic for bovine adherent mononuclear leukocytes in a dose-and time-dependent manner. Cytotoxicosis was determined by the release of cytoplasmic lactic dehydrogenase. This cytotoxicosis was blocked by treating the cells with corticosteroids. Variation in the reaction to LPS occurred in cells collected from the same cow on different days and from cells collected from different cows.     

Mast cell tumors in the dog.

The most common skin tumor in dogs is the mast cell tumor (MCT), with an incidence of close to 20% in the canine population. MCTs range from relatively benign to extremely aggressive, leading to metastasis and eventual death from systemic disease. Although surgical removal with or without radiation therapy may cure most patients with low-grade MCTs, there are no effective treatments for dogs with aggressive high-grade MCTs. This article reviews the current understanding of MCT biology with regard to diagnosis, staging, identification of prognostic indicators, and appropriate treatment planning.     

子宫内膜基质金属蛋白酶-2、-9 mRNA表达与奶牛子宫内膜炎关系的研究

为探讨基质金属蛋白酶(MMPs)-2、MMP-9在奶牛子宫内膜炎中的作用机制,选用产后6～10d健康及患急性化脓性子宫内膜炎的中国荷斯坦奶牛各10头,分别为对照组和试验组,通过ELISA检测PGF2a、孕酮浓度;免疫组化检测子宫内膜Ⅰ型胶原、Ⅳ型胶原的表达;SYBR Green I荧光定量PCR检测子宫内膜MMP-2和MMP-9mRNA表达。结果表明:试验组PGF2a浓度极显著低于对照组(P<0.01),孕酮含量显著高于对照组(P<0.05),PGF2a与孕酮浓度呈显著负相关(r=0.893);试验组子宫内膜中Ⅰ型胶原、Ⅳ型胶原的表达量均高于对照组,差异极显著(P<0.01);MMP-2和MMP-9mRNA水平则显著低于对照组(P<0.01)。结果提示:产后子宫内膜炎病牛子宫内膜MMP-2、MMP-9低表达,导致Ⅰ型和Ⅳ型胶原降解受阻,使细胞外基质为子宫内膜细胞提供的刺激信号不足,子宫分泌PGF2a量少,黄体消退障碍,血清孕酮含量高;说明子宫内膜MMP-2、MMP-9mRNA表达对产后子宫内膜炎的发生和发展起促进作用。     

Matrix-metalloproteinases-2 and -9 production in bovine endometrial cell culture

In vitro cell culture is a convenient tool for studying cellular mechanisms. In the present study, production of matrix-metalloproteinases (MMPs) in bovine endometrial (containing both epithelial and stromal cells) monolayer cells was examined. Blastocysts attached to the endometrial cells in a monolayer culture were examined for their effects on MMP-2 production. Initial attachment of blastocysts to the monolayer inhibited MMP-2 production by endometrial cells. But once trophoblast cells began to migrate into the endometrial cell layer, MMP-2 production increased, and at the same time MMP-9 production also became evident in the medium. In order to understand how blastocysts affected MMP-2 production, we examined the effect of progesterone, estradiol, insulin-like growth factors (IGFs), tumor necrosis factors (TNFs), and interferon-tau (IFN-tau) supplementation. It was IFN-tau that inhibited the production of MMP-2. In addition, progesterone at a lower dose appeared to inhibit MMP-2 production. Both TNF-alpha and TNF-beta strongly stimulated the production of MMP-2 and MMP-9, whereas IGFs had no effect. Based on these findings, it appears that conceptus has the capacity to inhibit MMP activity.     

Profiles of type-II pneumocytes in rats inoculated intratracheally with bacterial lipopolysaccharide

Ultrastructural and morphometric profiles of type-II pneumocytes (P-II) were investigated in rats killed 18 or 24 hours after a single intratracheal inoculation of bacterial (Escherichia coli) lipopolysaccharide (LPS). Inoculation with LPS induced pulmonary injury and inflammation, as measured by increased lactate dehydrogenase and alkaline phosphatase activities and increased numbers of polymorphonuclear neutrophils in fluid collected by bronchoalveolar lavage. Marked ultrastructural changes and desquamation of a few P-II developed at the time of high activity of lactate dehydrogenase and alkaline phosphatase in bronchoalveolar lavage fluid. Ultrastructural changes included swollen mitochondria and localized cisternal dilatation of the endoplasmic reticulum in which was contained membrane-bound homogenous material of medium electron density. Twenty-four hours after LPS inoculation, point-count stereologic analysis and digitizing morphometry revealed greater than 50% increase in P-II size. Changes in cell size corresponded with ultrastructural finding of swollen cells. Results obtained by point-count stereologic analysis and digitizing morphometry were highly correlated (r = 0.95). Lamellar bodies (LB) comprised 12 to 15% of P-II volume. Volume density and number of LB remained unaltered in LPS-injured P-II, and evidence of accelerated release of LB was not detected after LPS inoculation. Exudated polymorphonuclear neutrophils and pulmonary alveolar macrophages were involved actively in the phagocytosis of LB originating from necrotic and desquamated P-II. On the basis of measurement of enzyme activity (enzymes released into the bronchoalveolar space), considerable ultrastructural alterations developed in P-II when maximal LPS-induced pulmonary cell injury took place.     

Mast cell numbers in normal and glaucomatous canine eyes

Numbers of mast cells in the cornea, sclera, choroid, ciliary body, iris, and retina of sections of globes from 35 clinically normal dogs and 34 dogs with secondary glaucoma was determined. Fixed globes were trimmed along a vertical midsagittal plane and embedded in paraffin. Tissue sections, approximately 6 microns thick, were stained with toluidine blue for identification of mast cells. In normal globes, most of the mast cells were observed in the anterior portion of the uvea, and fewer mast cells were seen in the choroid and sclera. Mast cells were not observed in the retina and were seldom observed in the cornea of dogs with or without glaucoma. In sections of glaucomatous globes, mast cells were distributed evenly in the uvea and sclera, and fewer mast cells were present than in normal globes, regardless of the cause of glaucoma.     

Mast cell tumor invading the cornea in a horse

A 3‐year‐old Marwari mare was presented for evaluation of an irregular, reddish mass protruding from behind the right third eyelid. The mass appeared to arise at the ventral limbal area, involved the perilimbal bulbar conjunctiva and widely extended into corneal tissue. No other ocular or systemic abnormalities were detected at the time of presentation. The mass was surgically removed by lamellar keratectomy, with defocused CO₂ laser used as adjunctive therapy to treat the surgical exposed area and its surroundings. Histopathologic evaluation showed sheets of densely packed, well‐differentiated neoplastic mast cells separated by fibrovascular connective tissue. Nuclear staining for Ki‐67 was performed, and an average of 370 cells were positive per 1000 counted cells. Two months postoperatively, the surgical site was filled with flat fibrovascular and pigmented tissue, while the surrounding cornea was transparent with no superficial vascularization around the fibrotic scar. Thirty‐two months after treatment, no recurrence of the neoplasia was reported.