Ahmet Akkoç

Cardiac metastasising rhabdomyosarcoma in a great Dane

perineal area and skin of dogs (Kim and others 1996, Kuwamura and others 1998, Lascelles and others 1998, Barnhart and Lewis 2000, Ueno and others 2002, Machida and others 2003, Brockus and Myers 2004). Dissemination to the lymph nodes, lungs, heart, liver, spleen, adrenal glands and kidneys has been reported (Ware 2001). In the present case there were metastases in various organs. A combination of haematoxylin and eosin, immunohistochemistry, PTAH and PAS staining is used to differentiate Cardiac metastasising rhabdomyosarcoma in a great Dane

Suspected congenital generalised tuberculosis in a newborn calf

BOVINE tuberculosis remains an important disease in many countries (Morris and others 1994, Szewzyk and others 1995, Chalmers and others 1996, Phillips and others 2003). It is mainly a chronic disease of middleto old-aged animals, although congenital forms of the disease can occur when the disease involves the dam’s genital tract or placenta. In this context, tuberculosis bacilli are introduced into the fetus haematogenously via the umbilical vein, or via infected amniotic fluid ingested or aspirated in utero or at birth (Dungworth 1993, Stahelin-Massik and others 2002). This short communication describes a case of tuberculosis in a 15-day-old female calf. A 15-day-old Holstein calf, weighing 30 kg, was referred to a large animal clinic because of chronic coughing, variable appetite and gradual weight loss during the previous 10 days. The calf had not responded to previous antibacterial (amoxicillin and clavulanic acid) and non-steroidal anti-inflammatory drug (flunixin meglumine) treatments. Clinically, the calf had a soft, moist, chronic cough, pale mucous membranes, a slight mucopurulent nasal discharge, generalised muscular wasting and depression. Body temperature (39·6°C) and respiratory rate (44 breaths/minute) were slightly raised. Lung auscultation revealed crackles and wheezes in the anterio ventral parts of the chest. Routine haematology indicated leucocytosis (13,500 leucocytes/ ml) with neutrophilia (75 per cent). However, the haematocrit (21 per cent), erythrocyte (3·7 x 1012 erythrocytes/l) and haemoglobin levels (67 g/l) were low. The calf was diagnosed with chronic pneumonia and was treated with 10 mg/kg tylosin (Tylan; Lilly) and 10 mg/kg oxytetracycline (Primamycin; Pfizer) for five days. There was no clinical improvement, and the calf died seven days after the initiation of treatment. At postmortem examination, the lungs were found to be severely consolidated by numerous pale, coalescent necrotic areas, ranging from 0·5 to 2 cm diameter in size in all lobes (Fig 1). The colour of the cut surface of the lesions was greyish and calcified areas were noted. No lesions were found macroscopically in any other organs. Tissue samples of lung, liver, kidney, heart and bronchial lymph nodes were fixed in 10 per cent neutral-buffered formalin, paraffin-embedded and cut in 5 μm thick sections. All slides were stained with haematoxylin and eosin and by the Ziehl-Neelsen (ZN) method. Selected lung sections were stained immunohistochemically using polyclonal rabbit anti-Mycobacterium bovis serum (Dako). After blocking endogenous peroxidase activity with 3 per cent hydrogen peroxide in methanol for 20 minutes, the slides were treated with trypsin solution at 40°C for 10 minutes; they were then transferred into citrate buffer solution and incubated in a microwave oven to effect antigen retrieval. A blocking solution (normal goat serum) was then applied for 20 minutes at 40°C. Slides were incubated with rabbit anti-M bovis serum (1:500 dilution) at 40°C for one hour, and then with biotinylated goat anti-rabbit immunoglobulin G for 10 minutes. After incubation with streptavidin-peroxidase for 20 minutes, aminoethyl carbazole was applied for five minutes for colour development. The slides were counterstained with Mayer’s haematoxylin for 15 minutes. Negative controls were achieved by replacing the primary antibody with normal rabbit serum. Previously confirmed cases were used as positive controls. Histopathologically, the lung lesions were characterised by central areas of caseous necrosis and calcification surrounded by epithelioid cells, multinucleate giant cells, lympho cytes and an outer fibrous capsule (Fig 2). A few acid-fast bacilli were visualised by ZN staining in the necrotic areas or in the cytoplasm of multinucleate giant or epithelioid cells. Large numbers of tubercles were seen in both the cortical and medullary regions of the bronchial lymph nodes; the centre of these tubercles was slightly necrotic and was surrounded by epithelioid cells, lymphocytes and giant cells. No calcification or prominent connective tissue was present. Similar granulomatous lesions without caseous necrosis or calcification were found in the liver and in both kidneys, although no agent could be detected after ZN staining. The tubercles

HER-2/neu (c-erbB-2) oncoprotein in hyperplastic endometrial polyps detected in two cats

The presence of HER-2/neu (c-erbB-2) oncoprotein, oestrogen-α receptor (ER), and progesterone receptor (PR) in hyperplastic endometrial polyps (EPs) of two cats with cystic endometrial hyperplasia-pyometra (CEH-P) complex was investigated. Immunohistochemistry assay for ER, PR and c-erbB-2 oncoprotein in the glandular and stromal tissue of the EPs was performed. ER and c-erbB-2 immunoreactivity was observed in the glandular epithelium of the EPs whereas PR immunoreactivity was detected only in the stromal fibroblasts. The c-erbB-2 oncoprotein may play a role with the ER in the pathogenesis of the hyperplastic EPs, although the role of this oncoprotein in the pathogenesis of EPs has yet to be determined.

Expression of matrix metalloproteinases in Marek's disease tumours

Mareks disease is an important lymphoproliferative neoplasm of chickens, caused by oncogenic strains of herpes virus. Visceral lymphomas and invasion of both central nervous tissues and peripheral nerves by lymphoid cells are common findings. Matrix metalloproteinases (MMPs) are a group of enzymes responsible for extracellular matrix degradation and tumour metastases. The localisations of MMPs in tumour cells were evaluated immunohistochemically. As a result, positive reactions for MMP-9 and MMP-2 enzymes were found in the lymphoid tumour cells. These enzymes may take part in the pathogenesis of Mareks disease causing extracellular matrix degradation and metastasis.

Severe granulomatous hepatitis caused by Capillaria hepatica in a puppy

* Correspondence: aalasonya@uludag.edu.tr

Hypoxia enhances secretion of matrix metalloproteinases in bovine dermal fibroblasts: an in vitro approach to bovine digital dermatitis

ABSTRACT Dermal fibroblasts are of great importance in skin homeostasis. They are responsible for the synthesis and remodelling of extracellular proteins by the way of synthesis of some specific kind of Matrix Metalloproteinases (MMPs). Imbalance between MMPs and their tissue inhibitors (TIMPs) are known to involve ulcerative wounds in many tissues, including skin. In veterinary medicine, bovine digital dermatitis (BDD) is an ulcerative skin disorder of dairy cattle. To date, Koch’s postulate has not yet been fulfilled for the pathogenesis. In present study, we tested in vitro role of hypoxia if it evokes a response in MMP-1, MMP-2 and MMP-13 enzymes and their inhibitors in bovine dermal fibroblasts. Dermal fibroblasts were exposed to hypoxia in various time periods (6, 12 and 24 h). Immunoblotting revealed 6-, 12- and 24-h hypoxia, which resulted in 43.5%, 75% and 65% increase in MMP-1, 76%, 61% and 55% increase in MMP-2 and 80%, 78% and 55% increase in MMP-13 enzymes levels, respectively. We observed slight increases in TIMP-1 and TIMP-2 enzymes, but differences between treatment and control groups did not show any statistical significance. Our findings indicate that hypoxia induces MMP-1, MMP-2 and MMP-13 enzymes and may contribute to pathogenesis of BDD through in the development of in vivo lesions.

Effects of hypoxia and hyperosmosis on the expression of matrix metalloproteinases in broiler lung fibroblasts

This study investigated the effects of hypoxic and hyperosmotic stresses on the matrix metalloproteinases 2 and 9 (MMP-2, MMP-9) and their tissue inhibitors 2 and 1 (TIMP-2, TIMP-1) respectively in cultured lung fibroblasts from broiler chickens. Isolated and sub-cultured primary lung fibroblasts from 1-day-old broilers were exposed to hypoxia and hyperosmosis. The presence of the enzymes and the effects of hypoxia and hyperosmosis on the enzymes in cultured lung fibroblasts, and in the culture media, were evaluated with the streptavidin-biotin-peroxidase method and western blotting using commercially available primary monoclonal antibodies. Short (6 hours) and long term (12 hours) hypoxia, hypoxia + hyperosmosis (6 hours) and hyperosmosis (6 hours) triggered the release of both MMP-2 and TIMP-1 in experimental groups when compared with their controls. Neither MMP-9 nor TIMP-2 antibodies stained the fibroblast cells kept in normoxic, hypoxic or hyperosmotic conditions.