Analysis of NOD1, NOD2, TLR1, TLR2, TLR4, TLR5, TLR6 and TLR9 genes in anal furunculosis of German shepherd dogs.

Anal furunculosis (AF) primarily affects German shepherd dogs (GSD) and is characterised by inflammation and ulceration of the perianal tissues with development of cutaneous sinuses or rectocutaneous fistulae. Investigation of pattern recognition receptor (PRR) function has suggested that defective responses might occur in AF-affected GSD. The aim of the current study was to investigate whether canine PRR genes are involved in determining susceptibility to AF in this breed. Chromosomal location and coding sequences for NOD1, NOD2, TLR1, TLR2, TLR4, TLR5, TLR6 and TLR9 were determined and microsatellite markers identified for each gene. Microsatellite genotyping of 100 control GSD and 47 AF-affected GSD showed restricted allelic variation for AHT H91 (associated with TLR5) and REN216 NO5 (associated with both TLR1 and TLR6) compared with non-GSD dogs. Genotyping of single nucleotide polymorphisms identified in canine TLR1, TLR5, TLR6 and NOD2 genes failed to show any significant associations between PRR polymorphisms and AF. The highly restricted PRR genotypes seen in GSD are likely to have resulted from selective breeding and might influence innate immune responses in this breed.

Spontaneous gastroduodenal perforations in dogs: a retrospective study of 15 cases.

The clinical findings and outcomes following surgical management of spontaneous gastroduodenal perforations in 15 dogs were reviewed in a retrospective study to identify related risk factors. Clinical and clinicopathological findings were diverse. Use of multiple diagnostic procedures yielded a strong index of suspicion for gastrointestinal perforation. There was a trend towards improved survival for animals with perforation of the gastric fundus/body compared to pyloric or duodenal perforation, although the difference was not statistically significant. Five dogs were euthanased at surgery; two dogs died within two days after surgery. Seven of the eight surviving dogs had a favourable long-term outcome. Previous administration of NSAIDs was the only identified predisposing factor in ten of 10 of the dogs.

Matrix metalloproteinase mRNA expression in canine anal furunculosis lesions.

Although the aetiology of anal furunculosis (AF) in dogs is poorly understood, there is evidence for an underlying immune dysfunction. This is illustrated by the presence of a T helper type 1 cytokine mRNA profile in AF lesions and the clinical response to ciclosporin therapy. Expression of MMPs 2, 9 and 13 were evaluated in AF lesional biopsies by real-time quantitative RT-PCR. There was significantly increased expression of both MMP-9 and MMP-13 mRNA in AF biopsies compared to controls (p<0.001) but no significant difference in MMP-2 mRNA expression. Since MMP-9 and MMP-13 are primarily produced by macrophages, these data suggest that ulceration could be the result of aberrant activation of this cell type in the tissues. It is feasible that such pathological macrophage activity occurs in response to interferon-gamma secreted by T helper type 1 cells. This could explain why the lesions resolve following treatment with the immunosuppressive drug ciclosporin.

Duplex gall bladder associated with choledocholithiasis, cholecystitis, gall bladder rupture and septic peritonitis in a cat.

A 10-year-old cat was presented with a history of inappetence, pyrexia and weight loss. Clinical investigations showed anaemia, hyperbilirubinaemia, septic peritonitis and a double gall bladder with choleliths in an extrahepatic duct. Initial medical stabilisation was performed. At laparotomy, a duplex gall bladder with two separate cystic ducts was identified. The left gall bladder was thickened and had ruptured at the apex. Multiple choleliths were identified in the left cystic duct. The right gall bladder and cystic duct were grossly normal. The ruptured gall bladder was repaired, the gallstones were removed via a choledochotomy of the left cystic duct and a choledochoduodenostomy was created from the dilated left cystic duct. The cat remained depressed and anorexic, and it was euthanased 72 hours postoperatively at the owners' request. This is the first ante-mortem investigation of extrahepatic biliary disease associated with gall bladder duplication in the cat.

Solitary extramedullary plasmacytoma of the canine larynx.

An 11-year-old, female, spayed cocker spaniel was presented with dysphonia caused by a solitary laryngeal mass. Excisional biopsy was performed, and a diagnosis of plasmacytoma was made on the basis of histological examination. Further investigations showed no signs of systemic involvement. Coarse fractionated radiation therapy failed to control the tumour. Therapy was successfully instituted with a conventional combination chemotherapy protocol over a period of 14 months. The dog remains disease free 30 months after diagnosis. Most solitary, extramedullary plasmacytomas in dogs arise in the gastrointestinal tract, with fewer reports in other sites. The larynx is an uncommon sight of involvement in any species, and to the authors' knowledge, this is the first report of this tumour type in the canine larynx. In contrast to the therapeutic benefits reported in humans, the combination of surgery and radiation therapy was unsuccessful in this case, although sustained remission was gained following chemotherapy.

Cyclosporine treatment of anal furunculosis in 26 dogs.

OBJECTIVES: To evaluate the effect of cyclosporine on anal furunculosis lesions in 26 dogs. METHODS: Lesions were graded as mild in 11 dogs, moderate in eight and severe in seven. Each dog was treated with approximately 4 mg/kg cyclosporine orally every 12 hours until the lesions resolved or showed no further improvement. Residual lesions were resected surgically. RESULTS: Eighteen dogs (69 per cent) experienced complete resolution, seven (27 per cent) improved but had residual lesions and one (4 per cent) showed no improvement. The mean duration of treatment until resolution or no further improvement was 8.8 weeks (range four to 24 weeks). Nine dogs (35 per cent) experienced recurrence. Six were from the group that had shown complete resolution and three were from the group that had surgery. Fifteen dogs (58 per cent) developed side effects to cyclosporine, although none required treatment to be discontinued. Mean duration of follow-up was 6.8 months (range one to 20 months). CLINICAL SIGNIFICANCE: Cyclosporine was effective at resolving or reducing anal furunculosis lesions in 25 of 26 dogs (96 per cent). However, residual or recurrent lesions remain a potential problem, and surgical resection or long-term cyclosporine treatment may be necessary in some dogs.

The generation of a single nick per plasmid molecule using restriction endonucleases with multiple recognition sites.

Some restriction endonucleases generate a single-stranded nick at their recognition sequences in the presence of ethidium bromide (EtBr). This nick can then be extended to a single-stranded gap in which mutations can be introduced by a variety of techniques. To date, the templates used in these studies have largely contained a single recognition site for a given enzyme. Therefore, we have extended these studies to twelve enzymes for which multiple recognition sites exist in the template and show that, under appropriate conditions, one single-stranded nick is introduced per plasmid molecule.

A possible novel interaction between the 3'-end of 18 S ribosomal RNA and the 5'-leader sequence of many eukaryotic messenger RNAs.

A novel interaction between the 5'-untranslated region of eukaryotic messenger RNAs and non-contiguous sequences in the 18 S ribosomal RNA is proposed. The small ribosomal RNA contains, at its 3'-terminus, a heavily conserved hairpin structure. It is suggested that mRNA 5'-leader sequence stabilises this structure by interacting with other conserved nucleotides which flank it. Sequences closely related to the required sequence (A-U-C-C-A-C-C) occur quite commonly in eukaryotic mRNAs and are often found immediately upstream from the AUG-codon. This interaction may have a role in the events which lead up to the initiation of protein synthesis.

Immunohistochemical demonstration of Wilms tumour gene product WT1 in a canine "neuroepithelioma" providing evidence for its classification as an extrarenal nephroblastoma.

An intradural extramedullary tumour, surgically removed from the spinal canal of a young dog with paraplegia, had the histological appearance of a nephroblastoma. Subsequent necropsy revealed no evidence of a renal primary tumour or of any other tumour. Similar tumours of the spinal canal have been described previously under a variety of names, in particular neuroepithelioma. With an antibody to the human Wilms tumour (nephroblastoma) gene product WT1, labelling of glomeruloid bodies, similar to glomerular podocytes in human fetal kidney, was demonstrated in the tumour. This finding strengthened the suggestion that it was a nephroblastoma.

Comparison of stressed simultaneous urethral pressure profiles between anesthetized continent and incontinent bitches with urethral sphincter mechanism incompetence.

The popular urodynamic technique of stressed urethral pressure profilometry used for investigation of genuine stress incontinence in women was adapted and applied to bitches. The aim was to assess the suitability and reproducibility of the technique in the canine species, and to determine whether differences seen in continent and incontinent women were found in bitches. Resting and stressed simultaneous urethral pressure profiles were obtained for 25 continent and 25 incontinent bitches, the latter diagnosed as having urethral sphincter mechanism incompetence. The stressed urethral pressure profiles were produced by ballottement of the abdomen during catheter withdrawal. The degree of stress induced was consistent and had got short-term reproducibility. Highly significant (P < 0.001) differences in the percentage of negative spikes extending below the resting intravesical pressure were found between continent and incontinent bitches. Significant differences were not observed in the pressure transmission profiles between continent and incontinent bitches; both groups had a gradual decrease in pressure transmission from the bladder neck to the external urethral orifice. The distance from the start of the urethral pressure profile to the first negative peak (attributable to respiration or ballottement) on the subtracted profile was compared with the radiographic distance that the bladder neck was positioned with respect to the cranial pubic brim, taking body weight and continence status into account. Body weight and continence status did not have significant effect on the relation in either instance.(ABSTRACT TRUNCATED AT 250 WORDS)

Comparison of two catheter withdrawal speeds during simultaneous urethral pressure profilometry in anesthetized bitches.

Maximal urethral closure pressure, functional profile length, and number of respiratory peaks on the resting urethral pressure profile, expressed as a percentage of those occurring on the bladder pressure recording, were compared at catheter withdrawal speeds of 1 and 3 mm/s in 30 anesthetized bitches. Significant (P less than 0.001) differences were found in maximal urethral closure pressure and percentage of transmission of respiratory peaks between the 2 speeds. Significant difference was not detected in functional profile length.

Suitability of the cranial portion of the vagina as a site for measurement of intra-abdominal pressure variations in dogs.

OBJECTIVE: To determine whether the cranial portion of the vagina of dogs is a suitable site for measuring intra-abdominal pressure during cystometry. ANIMALS: 16 bitches (8 sexually intact and 8 spayed). PROCEDURE: 2 types of vaginal catheters were used to measure intra-abdominal pressure changes in anesthetized dogs. Catheters were inserted in the rectum and cranial portion of the vagina. RESULTS: Intra-abdominal pressure variations were detected with greater magnitude in the rectum than the cranial portion of the vagina, regardless of type of catheter used and reproductive status (sexually intact vs spayed) of the dogs. CONCLUSIONS AND CLINICAL RELEVANCE: The rectum was the better site for measuring intra-abdominal pressure changes in female dogs. Measurement of intra-abdominal pressure with concomitant measurement of intravesical pressure to determine detrusor pressure during cystometry in female dogs is more reliably detected from the rectum than the cranial portion of the vagina.

Effect of body position on intravesical pressure in the anaesthetised bitch.

The position adopted by 50 recumbent dogs was recorded while they were sleeping or resting; the commonest orientation of their hindquarters was right lateral recumbency. The intravesical pressure in the urinary bladder of 24 anaesthetised bitches lying in right lateral recumbency was compared with the intravesical pressure recorded when the same bitches were supported in a standing position. The intravesical pressure was significantly higher when the bitches were in right lateral recumbency (P less than 0.001) and the difference was unaffected by whether they were continent or not, and had no significant correlation with bodyweight. These findings help to explain why bitches with an incompetent urethral sphincter mechanism tend to leak predominantly when they are recumbent.

Ultrasonography of the ureterovesicular junction in the dog: a preliminary report.

Contrast radiography followed by ultrasonography of the urinary bladder was performed on 10 incontinent dogs. The ureterovesicular junctions and ureteral jets were identified by ultrasonography in all the dogs except one that had a caudally displaced bladder. An ectopic ureter in one dog was diagnosed by ultrasonography on the basis of the absence of a ureteral jet on the affected side, and the direct identification of the ureter passing caudal to the bladder neck. Ultrasonography appears to be a simple, repeatable test for identifying the distal ureters.

Expression of cytokine mRNA in canine anal furunculosis lesions.

The pattern of expression of cytokine mRNA in the lesions of anal furunculosis was evaluated in tissue biopsies from 15 dogs, and compared with the pattern in control skin samples from 24 dogs, by reverse-transcriptase PCR using canine cytokine-specific primers and a semi-quantitative multiplex PCR assay. Interleukin-2 (IL-2) was detected in 11 of the 15 affected dogs but in only one of the controls, and interferon-gamma was detected in 14 of the affected dogs but none of the controls. In contrast, IL-4 was detected only in one of the affected dogs. Increased expression of mRNA for IL-1beta, IL-6, tumour necrosis factor alpha, IL-8, IL-10 and transforming growth factor beta1 was detected in the biopsies from the lesions of anal furunculosis relative to the control tissues (P < 0.05).

Comparison of urethral pressure profilometry and contrast radiography in the diagnosis of incompetence of the urethral sphincter mechanism in bitches.

Three diagnostic indicators of urethral sphincter mechanism incompetence were compared in 25 continent and 25 incontinent anaesthetised bitches: the resting urethral pressure profile, the stressed urethral pressure profile and the radiographic position of the bladder neck. Logistic regression indicated that the best predictor of continence status was the stressed urethral pressure profile as assessed by the percentage of negative peaks extending below the resting intravesical pressure; it classified 43 of the 50 dogs correctly. The radiographic position of the bladder neck was a better predictor of continence than either the measurement of functional profile length or the maximum urethral closure pressure from the resting urethral pressure profile, whether alone or in combination. By combining the percentage of negative peaks on the stressed profile with the position of the bladder neck, 46 of the 50 dogs were classified correctly. Cut-off values for the percentage of negative peaks on the stressed urethral pressure profile, and for the radiographic position of the bladder neck are suggested for use in evaluating incontinent bitches in clinical practice.

Urethral conformation and position in relation to urinary incontinence in the bitch.

The conformation of the urethra, the position of the bladder neck and the distance between the external urethral orifice and the cranial pubic brim were compared in 30 continent and 30 incontinent bitches with incompetence of the urethral sphincter mechanism, using the measurements made from vagino-urethrograms. The bladder neck was significantly (P less than 0.001) further caudal in incontinent dogs than in continent dogs. Its position was not affected by neutering and could not be explained by the degree of urethral curvature. The distance from the external urethral orifice to the cranial pubic brim was correlated (P less than 0.001) with bodyweight but was not significantly different in the continent and incontinent bitches. In neutered animals, the distance between the cranial pubic brim and the external urethral orifice was significantly (P less than 0.05) shorter than in entire animals.

Evaluation of the canine tympanic membrane by positive contrast ear canalography.

Positive contrast ear canalography was described briefly in 1973 as a method for detecting rupture of the tympanic membrane in dogs with otitis media. The purpose of this study was to evaluate the sensitivity and usefulness of the technique. The ears of 10 normal canine cadavers and 31 dogs with clinical signs of ear disease were examined using otoscopy, radiography and contrast radiography after infusing 2 to 5 ml of positive contrast medium into the ear canals. These examinations were repeated in the cadavers after the tympanic membrane had been punctured with a Spreull needle. In the cadavers 14 of 19 (74 per cent) of the tympanic membranes were visible otoscopically; contrast medium did not enter the tympanic bulla of any of the ears before the tympanic membrane was ruptured, but was visible in the bulla in every ear after rupture. In the clinical study, 40 of 61 (66 per cent) of the tympanic membranes were visible otoscopically, and 12 appeared to be ruptured. Radiographic signs of otitis media (increased opacity and/or thickening of the tympanic bulla) were identified in seven ears. Canalography was positive for rupture of the tympanic membrane in 13 ears, including four in which it appeared to be intact otoscopically. In normal canine ears, canalography was a more accurate method for detecting iatrogenic tympanic membrane rupture than otoscopy. In dogs with ear disease, canalography may be more sensitive for otitis media than either otoscopy or survey radiography.

Middle ear disease associated with congenital palatine defects in seven dogs and one cat.

Medical records of eight dogs and one cat with congenital palatine defects were reviewed retrospectively. Five of the dogs had nasal discharge and seven had radiographic signs of middle ear disease, but no clinical signs of ear disease were identified in any of the dogs, nor were any reported by their owners during a one- to five-year follow-up period. One dog had an ipsilateral impairment of hearing detected by brainstem auditory evoked responses. The cat had clinical and radiographic signs of middle ear disease. These findings suggest that, as in humans, congenital palatine defects in dogs and cats may predispose to middle ear disease. Any associated deafness could cause problems for working dogs.

Parotid duct dilation in two dogs.

Two cases of parotid duct dilation are described; the clinical signs on initial examination were a tubular swelling over the lateral aspect of the face with an associated intraoral submucosal distension. Surgical exploration confirmed that the distended structure was the dilated parotid duct and intraoral marsupialisation resulted in no recurrence postsurgery.