Pancreatic surgical biopsy in 24 dogs and 19 cats: postoperative complications and clinical relevance of histological findings.

OBJECTIVE: To assess the immediate postoperative complications associated with pancreatic biopsy in dogs and cats and review the clinical relevance of biopsy findings. METHODS: Retrospective review of clinical records from two referral institutions for cases undergoing pancreatic biopsy between 2000 and 2013. RESULTS: Twenty-four dogs and 19 cats that had surgical pancreatic biopsy had sufficient detail in their clinical records and fulfilled the inclusion criteria. Postoperative complications were seen in 10 cases of which 5 were suggestive of post-surgical pancreatitis. Two patients were euthanased within 10 days of surgery because of the underlying disease; neither suffered postoperative complications. Pancreatic pathology was found in 19 cases, 7 cases showed no change other than benign pancreatic nodular hyperplasia, and no abnormalities were seen in 18 cases. CLINICAL SIGNIFICANCE: Complications may be encountered following surgical pancreatic biopsy, although the risk should be minimal with good surgical technique. Pancreatic biopsy may provide a useful contribution to case management but it is not clear whether a negative pancreatic biopsy should be used to rule out pancreatic disease. Dogs were more likely to have no significant pathology found on pancreatic biopsy than cats, where chronic pancreatitis was the most common finding.

Subtotal epiglottectomy for the management of epiglottic retroversion in a dog.

A six-year-old male neutered Yorkshire terrier was evaluated for severe, acute-onset, inspiratory dyspnoea. Laryngoscopy revealed retroversion of the epiglottis with intermittent occlusion of the rima glottidis during inspiration. The dog underwent both temporary and permanent epiglottopexy procedures that were unsuccessful. Subtotal epiglottectomy was performed and resulted in permanent resolution of dyspnoea without evidence of dysphagia or aspiration. This case highlights potential complications associated with management of this condition as well as describing the successful use of subtotal epiglottectomy in the dog.

Omentalisation as adjunctive treatment of an infected femoral nonunion fracture: a case report.

A three-year-old male working border collie with an infected femoral nonunion fracture was managed in a two-stage procedure involving debridement and omentalisation, followed by stabilisation with a bone plate and an autogenous cancellous bone graft. Osseous union was documented radiographically 16 weeks after surgery. Telephone follow-up one year later revealed the dog had returned to full working function without evidence of lameness. To the authors' knowledge, this is the first clinical case described in the veterinary literature using omentalisation as an adjunct to the management of an infected, biologically inactive nonunion fracture.

Differential effects of prenatal exposure to dexamethasone or cortisol on circulatory control mechanisms mediated by angiotensin II in the central nervous system of adult sheep.

Prenatal exposure to elevated maternal glucocorticoids (dexamethasone (DEX) or cortisol (CORT)) for 2 days early in pregnancy can 'programme' alterations in adult offspring of sheep, including elevated arterial pressure. DEX treatment also results in greater angiotensin II type 1 (AT1) receptor expression in the medulla oblongata in late gestation fetuses than in saline (SAL)- or CORT-exposed animals. We hypothesized that this would result in functional changes in brainstem angiotensinergic control of cardiovascular function in DEX- but not CORT-exposed animals. To test this hypothesis, cardiovascular responses to intracerebroventricular (I.C.V.) angiotensin II were examined in adult male offspring exposed to DEX (0.48 mg h(-1); n = 7), CORT (5 mg h(-1), n = 6) or SAL (n = 9) from 26 to 28 days of gestation. Increases in mean arterial pressure during i.c.v. infusion of angiotensin II (1 or 10 microg h(-1)) were significantly greater in the DEX group (10 +/- 1 mmHg at 1 microg h(-1)) compared with SAL (6 +/- 1 mmHg) or CORT (6 +/- 1 mmHg) animals (P < 0.05). I.C.V. infusion of the AT1 antagonist losartan significantly decreased cardiac output and heart rate in DEX animals, but not in SAL or CORT animals. Thus, increased expression of brainstem AT1 receptor mRNA after prenatal DEX is associated with increased responsiveness of cardiovascular control to activation of brain AT receptors by exogenous and endogenous angiotensin II. The altered role of the brain RAS in sheep exposed prenatally to DEX was not observed in sheep exposed prenatally to cortisol, suggesting these two glucocorticoids have distinct programming actions.

Expression of type II procollagens during development of the human intervertebral disc.

Mice lacking type II collagen fail to develop intervertebral discs. The present study describes the distribution of the developmentally expressed type IIA procollagen molecule, as well as types I and III collagens, in human IV disc specimens ranging from 42 to 101 days gestation. Type IIA procollagen contains the alternatively spliced exon 2 which encodes a 69-amino-acid cysteine-rich domain. By radioactive in situ hybridization and fluorescence immunohistochemistry, we identified changes in the localization patterns of type IIA procollagen, particularly between days 54 and 101. At day 54, the developing disc was divided into the outer annulus containing types I and III collagens, the inner annulus containing type IIA procollagen and the notochord consisting of all three fibrillar collagens. Specifically, the IIA N-terminal propeptide was localized in the extracellular matrix at day 54 but, by day 101, was only observed in the cytoplasm of the inner annulus cells. A functional role for the IIA N-terminal propeptide during this specific stage of disc development seems apparent. This function may involve regulation of growth factors since the exon 2-encoded domain of type IIA procollagen has previously been shown to bind to bone morphogenetic protein-2 and transforming growth factor-beta. We aim to explore this mechanism further.

Type IIA procollagen in development of the human intervertebral disc: regulated expression of the NH(2)-propeptide by enzymic processing reveals a unique developmental pathway.

Type II collagen can be synthesized in two forms generated by alternative splicing of the precursor mRNA. Type IIA procollagen, which contains a cysteine-rich domain in the NH(2)-propeptide (exon 2), is produced by precartilage and noncartilage epithelial and mesenchymal cells, and type IIB procollagen, without the cysteine-rich domain, is characteristic of chondrocytes. Mice lacking type II collagen fail to develop intervertebral discs. We have previously shown that the human intervertebral disc and notochord synthesize primarily the type IIA form of procollagen. Therefore, we investigated the distribution of type IIA procollagen during early disc development in humans. By processes of radioactive in situ hybridization and fluorescence immunohistochemistry, we localized mRNA and protein of type IIA procollagen, type I collagen, and type III collagen in fetal intervertebral disc specimens ranging from day 42 (embryonic stage 17) to day 101 (week 14.5) of gestation. Antibodies to the three distinct domains of type IIA procollagen: the NH(2)-propeptide, the fibrillar domain, and the COOH-propeptide were used. The earliest stage of developing intervertebral disc (42 days, stage 17) was characterized by diffuse synthesis of types I and III collagens in the dense zone (intervertebral area) and synthesis of type IIA procollagen by the chondrocyte progenitor cells surrounding the disc. The notochord cells synthesized and deposited into the notochordal sheath all three fibrillar collagens. By 54 days (stage 22), the developing disc was clearly divided into three regions: 1.) the outer annulus, characterized by synthesis and deposition of types I and III collagens; 2.) the inner annulus, characterized by synthesis and deposition of type IIA collagen containing the NH(2)-propeptide but devoid of the COOH-propeptide (pN-procollagen); and 3.) the notochord, the cells of which synthesized and deposited of all three fibrillar collagens. In later stages of fetal development (72-101 days), a change in type IIA procollagen processing was observed in the cells of the inner annulus: even though these cells continued to synthesize type IIA procollagen, they deposited into the extracellular matrix (ECM) only the processed fibrillar domain, with the NH(2)-propeptide removed. This finding indicates that there is a developmentally regulated change in the processing of type IIA procollagen NH(2)-propeptide in the cells of the inner annulus. This mechanism is in contrast to previously shown developmental regulation of the cysteine-rich domain of the NH(2)-propeptide by alternative splicing of the precursor mRNA. Although the cells of the inner annulus have been identified as chondrocytes, based on their shape and synthesis of characteristic ECM components, they appear to represent a distinct developmental pathway characterized by their synthesis and differential processing of type IIA procollagen. This developmental pattern may prove important for disc regeneration.

The amino-terminal heptad repeats of the coiled-coil neck domain of pulmonary surfactant protein d are necessary for the assembly of trimeric subunits and dodecamers.

Pulmonary surfactant protein D (SP-D), a lung host defense protein, is assembled as multimers of trimeric subunits. Trimerization of SP-D monomers is required for high affinity saccharide binding, and the oligomerization of trimers is required for many of its functions. A peptide containing the alpha-helical neck region can spontaneously trimerize in vitro. However, it is not known whether this sequence is necessary for the complete cellular assembly of disulfide-cross-linked, trimeric subunits and dodecamers. For the present studies, we synthesized mutant cDNAs with deletions or site-directed substitutions in the neck domain of rat SP-D, and examined the assembly of the newly synthesized proteins after transfection of CHO-K1 cells. The neck domain contains three "classical" heptad repeat motifs with leucine residues at the "d position," and a distinctive C-terminal repeat previously suggested to drive trimeric chain association. Deletion of the highly conserved core of the latter repeat (FSRYLKK) did not interfere with the secretion of dodecamers with lectin activity. By contrast, deletion of the entire neck domain or deletion of one or two amino-terminal repeats resulted in defective molecular assembly. The secreted proteins eluted in the position of monomers by gel filtration under nondenaturing conditions. In addition, the neck + carbohydrate recognition domain of SP-D was necessary and sufficient for the trimerization of a heterologous collagen sequence located amino-terminal to the trimeric coiled-coil. These studies provide strong evidence that the amino-terminal heptad repeats of the neck domain are necessary for the intracellular, trimeric association of SP-D monomers and for the assembly and secretion of functional dodecamers.

Age-related changes in the synthesis and mRNA expression of decorin and aggrecan in human meniscus and articular cartilage.

OBJECTIVE: To determine if the biosynthesis of aggrecan and decorin in the human meniscus and the potential of the cells to express these macromolecules (mRNA), is affected by the age of the individual and that if any changes are observed are they different to those measured in articular cartilage obtained from the same joint. DESIGN: Radiolabelling of tissue explants, anion-exchange chromatography and agarose-polyacrylamide gel electrophoresis were used to analyze newly synthesized proteoglycans. A quantitative, competitive reverse-transcriptase polymerase chain reaction was developed and applied to the tissue to measure the expression of decorin and aggrecan mRNA. RESULTS: Proteoglycan synthesis in the meniscus was higher in young donors (1-5 mmoles sulfate incorporated/h/mgDNA, under 20 years of age) than in adult tissues (0.5-1 mmoles incorporated/h/mgDNA, 20-62 years of age) and decorin was the major proteoglycan synthesized at this time. An age-related increase in the proportion of aggrecan synthesis in the meniscus was also observed using agarose-polyacrylamide gel electrophoresis. Both decorin (five-fold) and aggrecan (eight-fold) mRNA expression increased with age in meniscus whereas levels were relatively constant in articular cartilage. In addition, the synthesis of decorin and aggrecan and the expression of their mRNA was different in meniscus and articular cartilage from the same knee joint. CONCLUSION: The synthesis and turnover of aggrecan and decorin in the human meniscus is influenced by the age of the individual and is not the same as that observed for articular cartilage.

Persistent depletion of I kappa B alpha and interleukin-8 expression in human pulmonary epithelial cells exposed to quartz particles.

Chronic inflammation and fibrosis following quartz inhalation has been associated with persistent up-regulation of several "pro-inflammatory" genes, which are commonly regulated by nuclear factor kappa-B (NF-kappaB). Transcription of the NF-kappaB-inhibitor IkappaBalpha is also under NF-kappaB control, and its de novo synthesis is considered to comprise a negative feedback loop in transient inflammation. To investigate this mechanism in particle inflammation, we have studied IkappaBalpha degradation in A549 cells exposed to DQ12-quartz or TiO(2), in relation to the expression of IL-8. Although both quartz and TiO(2) were found to cause IkappaBalpha degradation, only quartz elicited a mild IkappaBalpha depletion, first appearing at 4 h. TiO(2) was found to cause a higher short-term increase in IkappaBalpha mRNA-expression compared to quartz, whereas the early enhancement of IL-8 expression and release was similar for both particles. Up-regulation of IL-8 expression was found to persist with quartz only. Cotreatment with PDTC and curcumin reduced particle-elicited IL-8 response, whereas cycloheximide caused enhancement of IL-8 mRNA expression in both the quartz- and TiO(2)-treated cells. Our results demonstrate that mineral dusts cause IkappaBalpha degradation, a transient increase in de novo synthesis of IkappaBalpha, and enhanced IL-8 expression in human pulmonary epithelial cells. While IkappaBalpha degradation and early IL-8 expression seem to be general particle phenomena, particle-specific characteristics impact on activation of IkappaBalpha gene transcription, apparently accounting for the different proinflammatory IL-8 responses seen with quartz and TiO(2) in the longer term. These observations may provide an explanation for the transient versus the persistent pulmonary inflammatory status and subsequent differences in pathogenic potency of TiO(2) and quartz.