Expression of angiogenic growth factors in canine squamous cell cancers.

Skin and subcutaneous tissue tumors are the most common neoplasms in dogs. The most common sites of origin in dogs include digits, skin and the oral cavity including cheek and retromandibular area. We investigated canine squamous cell carcinoma (SCC) samples from 15 dogs and classified them histopathologically according to the degree of differentiation. bFGF, VEGF-C, TGF-ß, PDGF-A, PDGF-C and PDGFR-α expressions were assessed using immunohistochemistry to determine the roles of growth factors during SCC. We found that TGF-ß1 immunolabeling was elevated significantly compared to healthy controls in SCC originating from nailbeds, while expression of other growth factors did not change significantly. Our findings might explain the role of TGF-ß1 in the infiltrative and malignant behavior of SCC originating from nailbeds.

Significance of Angiogenic Growth Factors in Bovine Ocular Squamous Cell Carcinoma.

Bovine ocular squamous cell carcinoma (BOSCC) is the most common and economically significant neoplasm of the eye in cattle. This study investigated the role of angiogenic growth factors in the pathogenesis of BOSCC. Eighteen cases of BOSCC were classified histopathologically according to the degree of differentiation. Normal upper and lower eyelids and third eyelids collected from the right and left eyes of six healthy cattle aged 1-3 years, that had been presented for slaughter to abattoirs, served as controls. Transcription of genes encoding the angiogenic growth factors vascular endothelial growth factor-C (VEGF-C), basic fibroblast growth factor (bFGF), platelet-derived growth factor-C (PDGF-C) and platelet-derived growth factor receptor-α (PDGFR-α) was determined by quantitative real-time polymerase chain reaction (RT-PCR) in tissue obtained from paraffin wax blocks. Immunohistochemistry (IHC) was utilized to detect intensity of expression and tissue distribution of these growth factors. IHC results revealed that bFGF and PDGF-C were elevated significantly (P >0.05) and VEGF-C expression was decreased in BOSCC compared with healthy control tissue. PDGR-α expression was elevated; however, the difference, compared with control tissues, was not significant. RT-PCR results showed an inverse relationship to the results of IHC; where protein levels were elevated their corresponding mRNA levels were decreased or vice-versa. Angiogenic regulators therefore appear to play a role in the pathogenesis of BOSCC.

Periostin alters transcriptional profile in a rat model of isoproterenol-induced cardiotoxicity.

Periostin is an extracellular matrix protein from the fasciclin family that guides cellular trafficking and extracellular matrix organization. Periostin stimulates mature cardiomyocytes to reenter the cell cycle. The molecular mechanism behind such stimulation remains to be explored. A DNA microarray technology constituting 30,429 gene-level probe sets was utilized to investigate effects of recombinant murine periostin peptide on the gene expression pattern in a rat model of isoproterenol (ISO)-induced myocardial injury. The experiment was performed on 84 adult male Sprague-Dawley rats in four groups ( n = 21): (1) control group, (2) only periostin applied group, (3) ISO cardiotoxicity group, and (4) ISO + periostin group. The experiment was continued for 28 days, and rats were killed on days 1, 7, and 28 ( n = 7). Microarray analyses revealed that periostin significantly altered the expression of at least ±2-fold of 2474 genes in the ISO + periostin group compared to the ISO cardiotoxicity group of which 521 genes altered out of 30,429 gene-level probe sets. Ingenuity pathway analysis indicated that multiple pathway networks were affected by periostin, with predominant changes occurring in the expression of genes involved in oxidative phosphorylation, oxidative stress, fatty acid metabolism, and TNF-α NF-κB signaling pathways. These findings indicate that periostin alters gene expression profile in the ISO-induced myocardial injury and modulates local myocardial inflammation, possibly mitigating inflammation through TNF-α NF-κB signaling pathway along with a decreased Casp7 activity and apoptotic cell death.

Periostin normalizes levels of cardiac markers in rats with experimental isoproterenol cardiotoxicity.

BACKGROUND: Although the molecular mechanism of cardiac healing is not fully understood, myocardial infarction is one of the most usual diagnoses in hospitalized patients in industrialized nations while periostin has been recently suggested to have a potential in tissue repairing following myocardial ischemia. OBJECTIVES: To investigate the effects of periostin on the levels of selected cardiac parameters (cardiac troponin I and T, creatine kinase and creatine kinase isoenzyme-MB), antioxidant/lipid peroxidation parameters (superoxide dismutase, catalase, and malondialdehyde), hepatic parameters (alkaline phosphatase, lactate dehydrogenase, aspartate and alanine transaminases) as well as lipids (total cholesterol, triglyceride, high, low and very­low­density lipoproteins) in a rat model of isoproterenol---induced myocardial injury. METHODS: A total of 84 male rats were grouped into saline (Group I), periostin (Group II), isoproterenol (Group III) and isoproterenol+periostin (Group IV) groups (n = 21). Isoproterenol (85 mg/kg/day) and periostin groups were both injected intraperitoneally (1 µg/kg). RESULTS: Our results revealed that periostin has a positive reducing effect on the levels of analysed parameters especially on cardiac troponins and creatine kinases on days 7 and 28 of the recovery period following the induced experimental heart damage in rats. CONCLUSION: It is concluded that periostin could have a potential to increase the rate of myocardial recovery after myocardial infarction (Tab. 5, Ref. 28).

Immunohistochemical detection of autophagy-related microtubule-associated protein 1 light chain 3 (LC3) in the cerebellums of dogs naturally infected with canine distemper virus.

We investigated the expression of microtubule-associated protein 1 light chain 3 (LC3) protein in the cerebellums of dogs infected with canine distemper virus (CDV) using immunohistochemistry to detect autophagy. The cerebellums of 20 dogs infected with CDV were used. Specimens showing demyelination of white matter were considered to have an acute infection, whereas specimens showing signs of severe perivascular cuffing and demyelination of white matter were classified as having chronic CDV. Cerebellar sections were immunostained with CDV and LC3 antibodies. The cytoplasm of Purkinje cells, granular layer cells, motor neurons in large cerebellar ganglia and some neurons in white matter were positive for the LC3 antibody in both the control and CDV-infected dogs. In the infected cerebellums, however, white matter was immunostained more intensely, particularly the neurons and gemistocytic astrocytes in the demyelinated areas, compared to controls. Autophagy also was demonstrated in CDV-positive cells using double immunofluorescence staining. Our findings indicate that increased autophagy in the cerebellum of dogs naturally infected with CDV may play a role in transferring the virus from cell to cell.

Sarcomatoid renal cell carcinoma with scant epithelial components in an Angora cat.

CASE HISTORY: A 6-year-old, neutered, female Angora cat presented with a history of lethargy and anorexia for 2 months and a clinically palpable and gradually enlarging, solid mass in the abdominal cavity extending from the last costal arch to the pelvic cavity. CLINICAL FINDINGS: Examination of the cat revealed jaundice, dehydration and hypothermia. Haematological manifestations included lymphopenia and substantial decrease in haematocrit value. Biochemical analysis of the blood revealed hypoglycaemia, three-fold elevated blood urea nitrogen values, increased level of serum aspartate aminotransferase and increased total bilirubin while the creatinine level was normal. Ultrasonographic examination of the abdomen showed a disrupted and large hypoechoic area around the left kidney. The cat was anaesthetised and the left kidney was removed, but the cat died following surgery. PATHOLOGICAL FINDINGS: On post-mortem examination, the left kidney was markedly enlarged and both the cortical and medullary parenchyma were replaced by confluent, multilobulated, pale tan-white, firm nodular masses protruding above the capsular surface. Metastasis was not observed. Cytological examination revealed a population of spindle-shaped cells of variable size, with abundant coarse chromatin and occasionally prominent nucleoli. Initial sections of the kidney were indicative of undifferentiated sarcoma confirmed by immunohistochemistry revealing vimentin-positive and cytokeratin-negative results in all tumour tissues. Additional sections showed very small amounts of both cytokeratin-positive and vimentin-positive areas. DIAGNOSIS: Sarcomatoid renal cell carcinoma (SRCC) with scant epithelial components originating from left kidney. CLINICAL RELEVANCE: Clinical and pathological features were similar to those of human SRCC, even though there was no evidence of metastases. Immunohistochemistry for vimentin and cytokeratin may be useful for definitive diagnosis of renal cell carcinoma with sarcomatoid differentiation, although staining of sections from several different parts of the tumour may be necessary. When a primary renal tumour is presented, SRCC should be considered as this diagnosis may influence treatment protocols and the clinical outcome.

Immunohistochemical characterization of peroxisome proliferator-activated receptors in canine normal testis and testicular tumours.

Peroxisome proliferator-activated receptors (PPARs) are ligand-activated transcription factors belonging to the nuclear hormone receptor superfamily. Recent studies have demonstrated that PPARs regulate lipid metabolism and are expressed in various cancers. The aim of the present study was to investigate the expression of PPAR-α, -ß and -γ in normal canine testicular tissue and canine testicular tumours (CTTs). Expression of PPAR-α, -ß and -γ was greater (P <0.05) than in normal testicular tissue. PPARs were therefore induced in CTTs and they may play a role in the biology of these tumours.

Nesidioblastosis in a simmental calf.

A 2-day-old Simmental calf with arthrogryposis and astasia was subjected to necropsy examination. The calf was normoglycaemic and normoinsulinaemic. Microscopically, pancreatic tissue was hyperplastic with an irregular lobular arrangement of pancreatic islets. Newly-formed islet cells budded from intralobular or intercalated ducts (so-called ductulo-insular complexes) and there were prominent blood vessels with telangiectatic features surrounded by rows of cuboidal-columnar islet cells. The newly-formed islets expressed insulin antigen immunohistochemically. The lesion was diagnosed as nesidioblastosis, an uncommon abnormality previously associated with the double muscling trait in cattle.

Screw fixation is superior to N-butyl-2-cyanoacrylate in onlay grafting procedure: a histomorphologic study.

The aim of this study is the histopathological evaluation of the efficiency of N-2-butyl cyanoacrylate in the fixation of bone block grafts on mandible. Autogenous monocortical block grafts taken from tibial bone were fixed to the outer surface of the angle of the mandible with N-2-butyl cyanoacrylate on the right side and mini screws on the left side. Postoperatively in the first and third months, six rabbits were killed and tissue samples were obtained from the grafted area. No significant difference was found between the cyanoacrylate and screw group with respect to inflammation or foreign body reaction. The level of graft necrosis was found to be significantly higher in the cyanoacrylate group than in the screw group in both the first and third month samples. Bone formation between the graft and recipient bone was evaluated and bone formation was found to be significantly higher in the screw group than in the cyanoacrylate group. No trabecular bone formation was observed between the graft and recipient bone in the cyanoacrylate group. Screw fixation was found to be superior to N-2-butyl cyanoacrylate in all parameters.

Persistent cloaca, fused kidneys, female pseudohermaphroditism and skeletal anomalies in a simmental calf.

A 5-day-old Simmental calf was referred to our department for atresia ani and postural abnormalities caused by skeletal deformities. The calf had a short and deviated tail and a bowed hind limb. The calf appeared like a male because of the prepuce and penis located just near the teats and the absence of female external genitalia. During the necropsy, a horseshoe kidney, single ureter that originated from the kidney, and bilateral uterine horns with one ovary each were detected. The ureter, blind-ended large intestine, and bilateral uterine horns were connected to a dilated cloaca having two sacs, which were filled with a yellowish brown viscous fluid admixed with meconium and urine. Skeletal deformities found included scoliosis, partial synostosis of vertebrae, deviation of rudimentary sacrum and coccygeal vertebrae, and narrowed pelvic cavity. This is the first report of an anomalous combination including urogenital, large intestinal, and skeletal deformities in cattle.

Myocarditis associated with foot-and-mouth disease virus type O in lambs.

The present study describes the pathogenetic mechanisms of myocarditis in 9 lambs that died in a foot-and-mouth disease outbreak in Samsun, Turkey. In all the heart samples tested, ELISA and sequencing for phylogenetic analyses showed that the virus, namely O/TUR/Samsun/05, was associated with the PanAsia pandemic strain of foot-and-mouth disease virus (FMDV) type O. The lambs had myocardial lesions but no typical vesicular lesions. In situ reverse transcription showed that many cardiomyocytes and some interstitial cells were positive for FMDV type O. Inflammatory infiltration, hyaline degeneration, and necrosis of sheets of myocytes were observed. The cellular infiltrates were mononuclear cells, including many lymphocytes, macrophages, a few plasma cells, and neutrophils. Major histocompatibility complex Class II+ dendritic and mononuclear cells, gammadelta T cells, CD172A+ and CD14+ macrophages and monocytes, and IgM+ B cells were detected mainly in the infected hearts. Inducible nitric oxide synthetase (iNOS) was seen mostly in areas of inflammation infiltrated by large numbers of cells. Of the 2 alpha-subunits of integrin known to be used as receptors by FMDV in epithelial tissues, CD49e (integrin alpha5) was detected in the membranes of cardiac myocytes with intercalated discs, but CD51 (integrin alphaV) was not detected in cardiac myocytes from infected or normal lambs. Interstitial and inflammatory cells were positive for both integrin subunits. The terminal deoxynucleotidyl transferase mediated dUTP nick-end labeling (TUNEL)-positive signal was detected in the nuclei of both cardiac myocytes and interstitial cells from infected lambs. These findings suggest that the iNOS expressed by inflammatory cells in lesions may have a deleterious effect on cardiac myocytes in these lesions.