Cloning and characterization of the cDNA and gene for human epitheliasin.

Previously, we reported cloning and characterization of the mouse gene, epitheliasin. In the present work we cloned the cDNA of the full-length human orthologue and characterized its gene including 2 kb of 5' flanking sequence. Analysis of epitheliasin gene expression in adult tissues shows that it is expressed as 3.4 kb and 2 kb transcripts. The major 3.4 kb transcript is observed in the following order: prostate > colon > small intestine > pancreas > kidney > lung > liver. Epitheliasin transcripts in fetal tissues are observed only in kidney and lung. In situ hybridization analysis of tissues revealed that epitheliasin was preferentially expressed in epithelial cells. The gene consists of 14 exons and 13 introns based on comparison with its cDNA sequence. In the 5' flanking region, we identified two transcription start sites and three CpG islands encompassing a number of potential regulatory elements including SP1, SREBP, GRE/PRE and ERE. The region upstream of the transcription sites lacks a TATA box but contains an initiator-like element as well as a downstream promoter-like element. In vitro experiments with lymph node carcinoma of prostate (LNCaP) cells revealed that the epitheliasin gene was induced by androgens and the induction was not blocked by cycloheximide indicating that the induction required no intermediate protein factors. Immunoprecipitation analysis showed that androgens strongly increased epitheliasin protein levels.

Cloning, genomic organization, chromosomal assignment and expression of a novel mosaic serine proteinase: epitheliasin.

We report the isolation of a cDNA encoding a novel murine serine proteinase, epitheliasin. The cDNA spans 1753 bp and encodes a mosaic protein with a calculated molecular mass of 53529 Da. Its domains include a cytoplasmic tail, a type II transmembrane domain, a low-density lipoprotein receptor class A domain, a cysteine rich scavenger receptor-like domain and a serine proteinase domain. The proteinase portion domain shows 46-53% identity with mouse neurotrypsin, acrosin, hepsin and enteropeptidase. The gene, located in the telomeric region in the long arm of mouse chromosome 16, consists of 14 exons and 13 introns and spans approximately 18 kb. Epitheliasin is expressed primarily in the apical surfaces of renal tubular and airway epithelial cells.

Cell-mediated immune response in calves to single-dose, trickle, and challenge infections with Fasciola hepatica.

A peripheral blood mononuclear cell (PBMC) proliferation assay was used to study the cell-mediated immune response in eight calves experimentally infected with Fasciola hepatica. Hypersensitivity-related eosinophil and mast-cell responses were also assessed. The primary infection of 500 metacercariae was administered either as a single-dose or as a trickle infection over a 4-week period. Calves were challenge-infected 4 months later with 100 metacercariae and slaughtered 24 weeks postprimary infection. Skin eosinophil counts (SEC) were determined prior to infection on the basis of the intradermal reaction (IDR) to phytohaemagglutinin (PHA). These counts correlated negatively with the mean fluke length but not with the fluke burden found at necropsy. At the end of the experiment, non-specific (PHA) and specific (excretory-secretory parasite, products, FhESAg, and whole-worm extract, FhSomAg) immediate type hypersensitivity IDR were elicited in contrast to delayed type hypersensitivity (DTH) responses. The SEC correlated with blood eosinophilia but not with parasite parameters. These findings suggest that the eosinophil response does not correlate clearly with the development of resistance to F. hepatica infection in cattle. A specific mononuclear cell response to FhSomAg was detectable as early as 7 days after infection in both infected groups, being significantly higher during the very early migratory phase of the juveniles in the single-dose infected calves than in the trickle infected calves. This response remained significantly higher in infected groups than in the control group throughout the experiment. Challenge elicited a significant proliferative response, less pronounced than after primary infection. No production of gamma-interferon (INF-gamma) was recorded 3 weeks after challenge. At necropsy, the mean number of flukes recovered was similar in both infected groups, suggesting that the rate at which the infection is administrated has no effect on protective immunity. Hepatic lesions, similar in both infected groups, were characterised by marked eosinophil and mast-cell infiltration. Liver biopsies were performed and their diagnostic value is discussed. All results suggest that F. hepatica infection predominantly induces a Type-2 response in cattle, and that this response has little protective effect.

Nonresponsive generalized bacterial infection associated with systemic lupus erythematosus in a Beauceron.

A case of concurrent canine systemic lupus erythematosus (SLE) and generalized bacterial infection in a six-year-old female Beauceron is reported. The dog presented with purulent nasal and ocular discharges, skin lesions (including seborrhea, hyperkeratotic areas, and papules as well as ecchymoses around the eyes, on both sides of the pinnae, and on the vulva), generalized lymph node enlargement, a mitral murmur, and lameness. Later, facial swelling, a retrobulbar abscess, and a cough also developed. Occurrence of a generalized bacterial infection was established by culture of group-C, beta-hemolytic Streptococcus from the throat, the mouth, a biopsy site (popliteal lymph node area), the retrobulbar abscess, and the lung. The diagnosis of SLE was based on the clinical signs and particularly on the occurrence of antinuclear antibody (ANA) and antidoublestranded-desoxyribonucleic acid (ds-DNA) antibody. Interestingly, the latter type of antibodies were also detected in two young female puppies whelped by this dog. Salient histological findings included an extreme cell depletion of the lymph nodes and spleen and severe pneumonitis and peribronchiolitis. The results of this case indicate that a definite diagnosis of canine SLE can, at times, be made on the basis of the presence of serum ANA and ds-DNA antibodies.

Postmortem investigations on winter stranded sperm whales from the coasts of Belgium and The Netherlands.

During winter 1994-95, four and three sperm whales (Physeter macrocephalus) were stranded along the Belgian and the Dutch coasts, respectively. Necropsies and tissue samplings were collected 24 hrs post mortem. Lesions on several whales included round and linear skin scars, ventral skin abrasions, acute skin ulcers, acute ulcerative stomatitides, acute to chronic external otitides, and passive visceral congestion. In addition, these sperm whales appeared to be debilitated with severe weight deficit, had blubber thickness reduction, the absence of abdominal fat, and the intestinal tracts were almost empty. Three categories of lesions and their possible relation with the stranding were evaluated. Cutaneous scars observed on the seven whales appeared to have no relation with the stranding. The poor body condition and acute integument ulcerative lesions were present before the stranding. Ventral skin abrasions and visceral passive congestion were caused by the strandings. Absence of food in the alimentary tracts, evidence of weight loss and blubber thickness reduction were compatible with an extended presence of the sperm whales in the North Sea, where adequate food is not available. This might lead to progressive weakness, predisposing the animals to secondary pathogens such as viral diseases. Finally, the coastal configuration of the southern North Sea makes it a trap for sperm whales which have entered the area during their wanderings.

Bronchopulmonary and disseminated granulomatous disease associated with Aspergillus fumigatus and Candida species infection in a golden retriever.

A seven-year-old, female golden retriever was referred for a paroxysmal, chronic cough and dyspnea, dysphagia, facial pruritus, anterior uveitis, and deteriorating general condition. A severe, mixed interstitial and alveolar pattern, with poorly defined amorphous lesions, was seen on thoracic radiographs. Multiple, whitish nodules disseminated on the hyperemic respiratory mucosa were noted on bronchoscopy. Escherichia coli and Aspergillus fumigatus were cultured from the bronchoalveolar lavage. Granulomatous lesions in numerous organs were identified during necropsy, and Aspergillus fumigatus and Candida spp. were cultured from lung and kidney tissues. Microscopic granulomatous lesions were compatible with mycotic infection; however fungal organisms were not observed.