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The enzyme telomerase is essential for cell proliferation and tumorigenesis. Telomerase reverse transcriptase (TERT) represents the catalytic subunit of the enzyme. In humans, TERT expression is regulated by several different mechanisms, including alternative splicing. Canis familiaris TERT (dogTERT) has been shown to have a high level of sequence similarity with human TERT, indicating that the dog may represent a suitable animal model for telomerase studies. In the present report we sought to investigate whether dogTERT undergoes alternative splicing. During the analysis of canine mammary tissues (both tumor and paired adjacent to the tumor normal tissues) for dogTERT expression by RT-PCR, we identified eight samples-one tumor and seven adjacent normal-which gave PCR products of unexpected sizes. DNA sequencing revealed two insertions (175 and 28 bp long) and two deletions (17 and 32 bp long), which were encountered in different combinations and gave rise to five different transcripts. The generation of all variants could be explained by the employment of alternative splicing sites within dogTERT genomic sequences. The 175-bp and 28-bp insertions, identified between exons 7 and 8 and between 8 and 9, respectively, constituted unspliced sequences of introns 7 and 8, respectively. Both deletions originated from exon 8 sequence removals due to alternative splicing. All five variants encoded truncated proteins, which lacked essential motifs for reverse transcription and might have thus lost their ability to compose active telomerase enzymes. This is the first identification of alternative splicing events within dogTERT. The results presented here may provide the basis for more thorough studies on the regulation of telomerase activity in canine normal and cancer cells.

A 3.5-year-old intact male American Pit Bull was presented because of urinary incontinence and dysuria. Constipation, followed by diarrhoea, ocular disturbances and finally regurgitation developed over the next 4 years. Autonomic dysfunction was evidenced by clinical presentation, as well as positive ophthalmic pilocarpine test and subnormal Schirmer tear test. Diagnosis, however, was established through histopathological and immunohistochemical examination. Lesions attributable to inflammatory degenerative neuropathy of the autonomic ganglia, which represents one of the various types of human autonomic failure, were detected.

This report describes a case of cardiac right auricle rupture (RAR) in a flock of 11,500 broilers that were 14 days old. The birds were housed at an altitude of 300 m, with an external temperature of -10 degrees C and an internal temperature of 15 degrees C. There was 3.6% mortality, due to sudden deaths, from 10 to 14 days of age. All necropsied birds had haemopericardium due to RAR at the point of the junction with the vena cava, and 85% of them had blood in the oral cavity and external acoustic meatus. The vena cava and its caudal branches, the intestinal vessels, and the sinus durae matris and sinus saggitalis were distended. Histological examination showed haemorrhages into the myocardium, degeneration of the cardiac muscle fibres, as well as oedema of the lungs and hypertrophy of the smooth muscle bundles of the parabronchial walls. Blood in the mouth of the broilers may have been due to haemoptysis, which in humans is caused mainly by mitral stenosis. In broilers, mitral stenosis and/or insufficiency, and left ventricular failure with consequent pulmonary hypertension (PH) were considered as possible triggers for right ventricular failure. The alarm reaction in hypoxaemia, due to secondary factors such as cold, caused tachycardia and tachypnoea, may have induced further elevation of PH, and acute myocardial infarction causing cardiac rupture and haemopericardium in this case. Hypertension and PH, due to possible mitral stenosis/insufficiency in association with acute myocardial ischaemia, were probably the determinant factors causing this acute episode. This opens the possibility that the RAR may be cardiogenic.

To clarify the immunohistochemical features of amyloid deposits and cerebral amyloid angiopathy (CAA), the distribution of the amyloid beta-protein subtypes Abeta40, Abeta42, Abeta43 and Abeta precursor protein (APP) were examined in the brains of fourteen aged cats (7.5-21 year-old). Two types of plaques were detected. The first type was characterized by Ass positive antigenic material and detected in the cortical layers of the frontal and parietal lobes of all examined cats. The second type was characterized by diffuse positive immune staining representing diffuse plaques, which were detected only in the very aged cats (17-21 years old) and distributed throughout the cortical layers of the parietal lobes. Vascular amyloid and the amyloid deposits were strongly positive-stained with the antibody Abeta42. APP was exhibited in neurons and axons while the staining was stronger in the very aged cats (17-21 years old). Our findings suggest that the feline forms a spontaneous model for understanding the early changes of normal brain aging and the early stage of amyloid beta-protein deposition.

A 3-year-old spayed female dog was admitted with a history of episodes of dyspnoea and coughing. Severe tracheal stenosis was found on radiography. The dog died during an episode of refractory dyspnoea. Necropsy revealed an obstruction of the thoracic part of the trachea because of a chronic granulomatous inflammation protruding into the tracheal lumen. Histological examination revealed nematodes, which were identified as Onchocerca sp. according to their morphological characteristics. In contrast to the common ocular manifestation in dogs, obstructive tracheitis caused by Onchocerca infection has not been reported before.

A total of 216 local crossbred sheep from 16 scrapie-affected Greek flocks and 210 purebred sheep of the milk breeds Chios and Karagouniko from healthy flocks were analysed for scrapie-linked polymorphisms in the prion protein (PrP) gene. Of the 216 sheep in this case-control study, 96 sheep were clinical cases, 25 subclinical cases (asymptomatic at the moment of euthanasia but positive by histopathology and/or ELISA detecting proteinase-resistant PrP) and 95 healthy controls (negative by all evaluations). Polymorphisms at codons 136, 154 and 171 were determined by denaturing gradient gel electrophoresis, followed by RFLP and sequencing. Scrapie, both clinical and subclinical, was associated with the genotypes ARQ/ARQ (88 of 110 sheep of that genotype), ARQ/TRQ (9 of 13), ARQ/AHQ (15 of 38) and VRQ/VRQ (9 of 17). Histopathological lesions were more severe in the clinical cases. Genotypes ARQ/ARR (26 sheep), ARQ/ARK (seven sheep), AHQ/ARR (one sheep), ARH/ARH (one sheep) and ARR/ARH (three sheep) were detected exclusively in healthy control sheep. In the purebred survey, four genotypes were present in the Chios sheep (ARQ/ARQ, ARQ/TRQ, ARQ/AHQ and ARQ/ARR) and four in the Karagouniko sheep (ARQ/ARQ, ARQ/AHQ, ARQ/ARR and ARQ/ARH).

In this study we described four cases of bilateral ectropion in pigeons that were investigated in Greece. Anemia, leukocytosis, and increased levels of enzymes lactate dehydrogenase, aspartate aminotransferase, creatine phosphokinase, and total serum proteins were found. Chlamydial elementary bodies were observed by modified Ziehl-Neelsen stain in direct smears of conjunctiva, liver, and spleen as well as in yolk sac samples after egg inoculation with eyelid material. Histologically, significant hyperplasia of the conjunctival epithelium was observed. Using immunohistochemical methods, chlamydial antigen was revealed in eyelid, liver, and spleen paraffin sections. This study suggests that Chlamydia spp. was the causative agent that induced ectropion.

The ultra-structural lesions of lead poisoning and the deposition of lead in liver, kidney, peritoneum, cerebellum and retina of dogs were studied. Specimens were obtained from 5 3-4 mo-old crossbreed dogs each injected i.p. with a total dose of 120 mg lead acetate, divided in 10 equal doses of 12 mg, administered every other day. Two dogs were controls. Histopathological examination revealed degeneration of the epithelial cells of the urinary tubules, the endothelial cells of the renal capillaries and the hepatocytes. Characteristic lead inclusion bodies were observed intracytoplasmically and intranuclearly in mesothelial and giant cells of the peritoneum. Lead needle-like inclusions were intracytoplasmically in the interstitial connective tissue cells of the kidney, and substantial quantities of lead were in collagen fibers of the interstitial kidney tissue.

The purpose of this study was to evaluate the in vivo biocompatibility of Apexit (a calcium hydroxide) and Pulp Canal Sealer (a classic zinc oxide-eugenol), root canal sealers, after implantation in rat connective tissue. Forty-four white female Wistar-Furth rats were used. Each sealer was placed in Teflon tubes and implanted into specific dorsal subdermal tissues sites. Implants were removed after 5, 15, 60, and 120 days; fixed; and histologically prepared for microscopical evaluation. Severe inflammatory reactions with differing extensions of necrosis were observed with Apexit on the 5th and 15th days. The intensity of the reaction had diminished by the 60th day, and this reduction continued progressively through the 120th day. It was characterized by the presence of connective tissue with a few macrophages. Moderate to severe inflammation with confined areas of necrosis was observed in the Pulp Canal Sealer specimens on the 5th day. The intensity of the reaction diminished by the 15th, 60th, and 120th days, but remained slightly greater than Apexit through longterm observation periods.

Ketac-Endo, a new glass-ionomer cement for endodontic use as root canal sealer, and Tubli-Seal were tested for tissue biocompatibility in rat connective tissue. Forty-four white female Wistar-Furth rats were used. Each sealer was placed in Teflon tubes and implanted subcutaneously. The implants were removed after 5, 15, 60, and 120 days; fixed; and histologically prepared for microscopical evaluation. Mild inflammatory reaction was observed with Ketac-Endo on the 5th day. The connective tissue was infiltrated with plasma cells. Lymphocytes and macrophages were observed. The intensity of the reaction diminished by the 15th day, and this reduction continued progressively through the 60th and 120th days. Severe inflammation with differing extensions of necrosis was observed with Tubli-Seal on the 5th and 15th days, and the material remained irritating even after long-term implantation periods (60 and 120 days).

Infections caused by the nematode Trichinella spiralis is characterized in the host by an inflammatory response with cytokine production. In these studies we have detected TNF alpha, IL-6, IFN gamma, IL-4 and IL-10 in the serum of 10 mice infected with T. spiralis. Moreover, we detected, for the first time, these cytokines in the serum of mice treated with 4-DPD, a potent antagonist of vitamin B6 coenzyme which has anti-inflammatory properties. 4-DPD was used at 100, 400, 800 micrograms/bolus for 20 days, starting one day before the infection. After 15 days of T. spiralis infection, TNF alpha reached a maximum level, while IL-6 was maximal after 7 days, IFN gamma at 20 days and IL-4 at 14 days. IL-10 was not affected by the T. spiralis infection. When the animals were treated with 4-DPD at the reported dosages and infected with T. spiralis the inhibition of TNF alpha and IL-6, were dose-dependent in the first 7 days while IL-4 was reduced only at 400-800 micrograms/bolus. 4-DPD-treated mice did not statistically (P > 0.05) affect the generation of IFN gamma. In healthy animals the production of cytokines were not measurable, just as it was in non-infected animals treated with 4-DPD. The increase of cytokines such as, TNF alpha and IL-6 may be related to the severity of the disease, boosting the host's resistance to the pathogen and inhibiting parasite survival. In addition, the augmentation of IL-4 production enhances T and B cells and macrophage responses and may stimulate T-cell antibody-mediated response to the pathogen. 4-DPD, an inhibitor of IL-1 and inflammatory reactions, proved to be most effective on TNF alpha and IL-6, which are mainly produced by macrophages.

Formalin-fixed tissue samples obtained from puppies experimentally infected with canine parvovirus type 2 (CPV-2) were investigated immunohistologically with the direct immunoperoxidase method. Besides cardiac muscle and lymphatic tissues, bone marrow, intestine, liver, kidney, pancreas and to a lesser degree lung stained positive for viral antigen and are considered as sites of viral replication. The distribution of viral antigen reveals a tropism of CPV-2 to numerous organs in puppies infected during the first week of life.