Methods for measuring proteasome activity: current limitations and future developments.

The proteasome has been validated as a therapeutic target, with proteasome inhibitors showing particular efficacy in the treatment of Multiple Myeloma. A wide range of methods have been developed to profile proteasome activity. These include the current method of choice fluorogenic peptide substrates, as well as bioluminescent imaging, immunological methods, and more recently, site-specific fluorescent probes. The aim of this review is to evaluate the currently available methods for profiling proteasome activity and their suitability for use in translational studies. Ongoing development of techniques for profiling proteasome activity will facilitate future research into proteasome-related pathologies, thus accelerating the development of more specific drug regimes.

Gastrointestinal cryptosporidiosis in a puppy.

An 8-week-old female Yorkshire terrier with a history of weakness, diarrhea and intestinal isosporiasis was euthanized and a post mortem examination performed. Histologically, there was severe gastrointestinal cryptosporidiosis, severe intestinal isosporiasis and thymic lymphoid depletion. PCR revealed visible bands for the actin and 18S rRNA genes but not for the acetyl CoA synthetase gene for Cryptosporidium spp. The PCR product for the actin gene was sequenced and found to have a 97.6-99.8% similarity to that of Cryptosporidium canis. To our knowledge, this is the first report of gastric cryptosporidiosis in a canine.

Forensic web watch--medicolegal aspects of paediatric pathology.

The trials over the deaths of Matthew Eappen and Victoria Climbie have highlighted the importance of forensic evidence in cases of suspected child abuse. The debate as to whether bruises, fractures or head injuries have been sustained as a result of previous trauma or non-accidental injury is central to these, and other, cases. A variety of subjects are encountered in forensic paediatric pathology, including Shaken Baby Syndrome, non-accidental injury, retinal haemorrhage, skeletal injury, Sudden Infant Death, sexual abuse and Munchausen Syndrome by Proxy. The coverage of these areas on the internet was assessed using two search engines (Google and the meta-search engine Mamma) and revealed patchy coverage. The majority of sites uncovered were, unsurprisingly, aimed at the layperson concerned by such issues; however, several sites containing useful information for the professional are available.

Angiolipomatous tumors in dogs and a cat.

Variants of lipoma are uncommon, although fibrolipoma and infiltrative lipoma have been well documented. This report describes two cases of rare angiolipoma in dogs and the first documentation of angiofibrolipoma and infiltrating angiolipoma in a cat and a dog, respectively. Tumors were solitary, and most were located on the thorax of middle-aged patients. Angiolipomas were composed of mature adipose tissue mixed with variable numbers of blood vessels. In addition to the adipose and vascular components, the angiofibrolipoma contained bundles of collagenous connective tissue. The infiltrative angiolipoma had a primary mass external to the muscle and was histologically similar to a mixed intramuscular hemangioma that was confined to the muscle. Both disrupted bundles of striated muscle and were associated with segmental degeneration and loss of myofibers.

Endometritis in postparturient cattle associated with bovine herpesvirus-4 infection: 15 cases.

Suppurative, ulcerative endometritis associated with bovine herpesvirus-4 (BHV-4) infection was identified in 15 postparturient dairy cows from 5 separate dairies. Characteristic eosinophilic to amphophilic intranuclear viral inclusion bodies were identified within degenerate endometrial lining epithelium and endothelial cells. Bovine herpesvirus-4 was confirmed as the etiology by a combination of fluorescent antibody assays, viral isolation, heminested PCR, ultrastructural examination of the uterus and inoculated tissue culture cells, and negative-stain electron microscopy of tissue culture supernatant. Viral particles measuring 70-95 nm were demonstrated in uterine epithelial and endothelial cells by electron microscopy. Bacteria including Arcanobacterium pyogenes, Escherichia coli, and an alpha-Streptococcus isolate were isolated from all uteri. Bovine herpesvirus-4-associated endometritis has been previously reported in sporadic cases in Europe but has not been previously reported in the United States. Endometritis associated with BHV-4 appears to be an emerging syndrome in Georgia dairy herds.

Mushroom toxicity in a horse with meningioangiomatosis.

We describe a fatal case of mushroom intoxication in an 18-y-o horse presumably due to Amanita verna. Horses are normally regarded as too fastidious to eat the ill-flavored toadstools. In this case, the horse had a rare benign brain tumor, meningioangiomatosis, which may have altered the horse's normal eating behavior resulting in consumption of the mushrooms.

162 cases of aldicarb intoxication in Georgia domestic animals from 1988-1998.

A 10-y retrospective study of aldicarb intoxication in domestic animals from 1988 to 1998 is provided from animals submitted to the University of Georgia Veterinary Diagnostic and Investigational Laboratory. Within the period examined, 162 separate cases were confirmed and 18 additional cases were suspected based on bioassay but could not be confirmed. Some cases involving as many as 15 affected animals. Dogs were most often involved, but cats, horses, cattle and goats were also poisoned. Most intoxications appeared intentional or malicious. "Baits" including frankfurters, ham or ground beef were consistently found in stomach contents. The restriction of aldicarb to an agricultural soil application limits accidental exposure to domestic animals. Its use is regulated through license in Georgia as a systemic insecticide for pecan trees, peanuts and cotton. Unfortunately, aldicarb seems favored as a malicious poison, possibly due to its quick lethality.

Multiple persistent vitelline duct cysts in a dog.

Persistent vitelline duct remnants, with the exception of Meckel's diverticulum in pigs and horses, are rare in animals. During an ovariohysterectomy of an 8-month-old Labrador Retriever, multiple fibrous nodules with cystic centers were found attached to the ileal serosa and in a mesodiverticular band attached to the abdominal wall. Histologic and ultrastructural evaluation revealed that the cysts were composed of well-differentiated intestine with mucosa, submucosa, and muscularis layers surrounded by a thick layer of fibrous connective tissue. The morphology and arrangement of lesions were consistent with multiple persistent vitelline duct cysts, a distinct condition related to Meckel's diverticulum. This case in a dog represents a unique presentation of this congenital anomaly in domestic animals.

Protection of pigs by vaccination of pregnant sows against eastern equine encephalomyelitis virus.

Serum-virus neutralizing antibodies were detected in serum and colostrum of sows vaccinated during pregnancy with commercially available vaccines against eastern equine encephalomyelitis virus (EEEV), and antibodies were detected in serum from nearly all pigs from vaccinated sows following colostrum uptake. Serum-virus neutralizing antibody (SVN) test titers were measured in colostrum and pigs at the next farrowing, and additional vaccination of sows prior to the third farrowing led to elevated SVN titers in serum, colostrum and all pigs. Six pigs from vaccinated sows challenged at 8 to 9 days of age with 1 x 10(6) TCID50 EEEV did not develop the high temperatures or signs of central nervous system disease that 6 pigs from non-vaccinated sows developed. Virus was isolate from blood and oropharyngeal swabs from all pigs from non-vaccinated sows with blood virus titers as high as 9.3 x 10(4) TCID50, while only low levels of virus were detected in blood and oropharyngeal swabs from pigs from vaccinated sows. Virus was also isolated from tonsils collected at necropsy from 3 pigs from non-vaccinated and 1 pig from vaccinated sows. Vaccination of pregnant sows leads to development of maternal antibodies that are transmitted via colostrum to pigs and are protective against clinical EEEV related disease after experimental challenge with EEEV. In addition, vaccination prevents amplification of virus in infected pigs and could result in protection of animals and farm labor in the environment of infected pigs.

Eastern equine encephalomyelitis virus infection in swine.

In 1 of 2 outbreaks of eastern equine encephalomyelitis virus (EEEV) infection in swine, 280 of 350 pigs died. Histopathologic findings in the brain were typical of EEEV infection. In addition, multifocal necrosis and inflammation were seen in the myocardium. Follow-up investigation in 1 of the herds revealed antibody response to EEEV in surviving pigs and in adult swine. Growth retardation was noticed in surviving pigs. Histologic examination revealed mild lesions in the brain of the survivors.

Sequential study of lesion development in experimental haemophilus pleuropneumonia.

Recently weaned pigs were infected aerogenically with Haemophilus (Actinobacillus) pleuropneumoniae, serotype 5. At three, six, 12, and 18 hours and one, two, four and seven days after exposure to haemophili a pair of animals were killed and necropsied. Pulmonary oedema with multifocal petechial haemorrhages and a diffuse neutrophilic bronchiolitis and alveolitis were observed at three and six hours after infection. Focal areas of coagulative necrosis developed in areas of intense suppuration at 12 and 18 hours after infection. At one and two days after infection, necrotic areas were surrounded by dense bands of degenerating leucocytes and contained unidentifiable round and elongated cells characteristic of this disease. In subacute lesions a thick layer of granulation tissue formed around the outer margin of developing abscesses. Most of the round and elongated cells in alveolar exudates could not be identified by enzyme histochemistry or electron microscopic examination. Neutrophils in various stages of degeneration and deterioration provided strong evidence that some of the cells were of this type. These findings suggest that neutrophils may play an early and significant role in development of lesions.

Myospherulosis in the subcutis of a pony.

Myospherulosis was diagnosed in a mature pony. Several parent bodies containing many spherules were observed microscopically in biopsy material from an area of cellulitis. The spherules are altered red blood cells that form as the result of prolonged contact with necrotic fat or petrolatum-based ointments. These structures must be differentiated from fungal elements. The recommended treatment is surgical excision.

Acute inflammatory effects of intratracheally instilled Escherichia coli endotoxin and sonicated suspension of Haemophilus pleuropneumoniae in swine.

A single bolus of either Escherichia coli endotoxin, sonicated suspension of Haemophilus pleuropneumoniae, or pyrogen-free normal saline was intratracheally instilled in six week old specific-pathogen-free pigs. Pigs exposed to E. coli endotoxin developed fever, leukopenia followed by leukocytosis, and endotoxemia. Leukocytosis was the only clinical abnormality noted in pigs receiving the sonicated suspension of H. pleuropneumoniae. At one day postexposure, focal areas of atelectasis and consolidation were observed in the caudal lung lobes of animals receiving either E. coli endotoxin or the sonicated suspension of H. pleuropneumoniae. Lesions were characterized by a neutrophilic bronchitis and bronchiolitis with alveolitis in the surrounding tissue. Increased numbers of alveolar macrophages and evidence of phagocytosis were observed by light and electron microscopy. No clinical abnormalities or lesions were observed in animals receiving normal saline. Lesions typical of acute porcine Haemophilus pleuropneumonia were not produced by either E. coli endotoxin or sonicated suspension of H. pleuropneumoniae, indicating that multiple virulence factors are probably involved in lesion development.

Canine aflatoxicosis: a continuing problem.

Aflatoxins are hepatotoxic in many species including dogs. In two separate outbreaks, the primary signalment was high morbidity and mortality in hunting dogs presenting with clinical signs of icterus, anorexia and listlessness. Preliminary laboratory examinations revealed toxic hepatitis, bilirubinuria and anemia. In the first case, a feed sample was not available and the diagnosis was established by confirming the presence of significant levels of aflatoxin B1 in tissues. In the second case, cornmeal utilized in formulating the ration contained 511 ng aflatoxin B1 and B2/g. These cases illustrate that aflatoxicosis is a continuing problem despite widespread awareness and testing for aflatoxin.

Canine laryngopharyngeal rhabdomyoma resembling an oncocytoma: light microscopic, ultrastructural and comparative studies.

A laryngopharyngeal rhabdomyoma was diagnosed in a four-year-old dog with clinical signs of upper respiratory obstruction. In people, rhabdomyomas have a predilection for the head and neck area. Features of this tumor were compared with an oncocytoma with which it may be confused.