Search

OBJECTIVE To investigate effects of changes in analytic variables and contrast medium osmolality on glomerular filtration rate estimated by CT (CT-GFR) in dogs. ANIMALS 4 healthy anesthetized Beagles. PROCEDURES GFR was estimated by inulin clearance, and dogs underwent CT-GFR with iodinated contrast medium (iohexol or iodixanol) in a crossover-design study. Dynamic renal CT scanning was performed. Patlak plot analysis was used to calculate GFR with the renal cortex or whole kidney selected as the region of interest. The renal cortex was analyzed just prior to time of the second cortical attenuation peak. The whole kidney was analyzed 60, 80, 100, and 120 seconds after the appearance of contrast medium. Automated GFR calculations were performed with preinstalled perfusion software including 2 noise reduction levels (medium and strong). The CT-GFRs were compared with GFR estimated by inulin clearance. RESULTS There was no significant difference in CT-GFR with iohexol versus iodixanol in any analyses. The CT-GFR at the renal cortex, CT-GFR for the whole kidney 60 seconds after appearance of contrast medium, and CT-GFR calculated by perfusion software with medium noise reduction did not differ significantly from GFR estimated by inulin clearance. The CT-GFR was underestimated at ≥ 80 seconds after contrast medium appearance (whole kidney) and when strong noise reduction was used with perfusion CT software. CONCLUSIONS AND CLINICAL RELEVANCE Selection of the renal cortex as region of interest or use of the 60-second time point for whole-kidney evaluation yielded the best CT-GFR results. The perfusion software used produced good results with appropriate noise reduction. IMPACT FOR HUMAN MEDICINE The finding that excessive noise reduction caused underestimation of CT-GFR suggests that this factor should also be considered in CT-GFR examination of human patients.

Canine progressive rod-cone degeneration (PRCD) is a middle- to late-onset, autosomal recessive, inherited retinal disorder caused by a substitution (c.5G>A) in the canine PRCD gene that has been identified in 29 or more purebred dogs. In the present study, a TaqMan probe-based real-time PCR assay was developed and evaluated for rapid genotyping and large-scale screening of the mutation. Furthermore, a genotyping survey was carried out in a population of the three most popular breeds in Japan (Toy Poodles, Chihuahuas and Miniature Dachshunds) to determine the current mutant allele frequency. The assay separated all the genotypes of canine PRCD rapidly, indicating its suitability for large-scale surveys. The results of the survey showed that the mutant allele frequency in Toy Poodles was high enough (approximately 0.09) to allow the establishment of measures for the prevention and control of this disorder in breeding kennels. The mutant allele was detected in Chihuahuas for the first time, but the frequency was lower (approximately 0.02) than that in Toy Poodles. The mutant allele was not detected in Miniature Dachshunds. This assay will allow the selective breeding of dogs from the two most popular breeds (Toy Poodle and Chihuahua) in Japan and effective prevention or control of the disorder.

Canine degenerative myelopathy (DM) is characterized by progressive degeneration of the spinal cord. Although atrophic changes in the intercostal muscles were previously reported in the late stage of DM in Pembroke Welsh Corgis (PWCs), changes in respiratory function have not yet been examined. In the present study, we performed an arterial blood gas analysis and measured respiratory movements over progressive disease stages to document changes in respiratory function in DM-affected PWCs. We found that respiratory dysfunction progressed during the later stages of DM and correlated with a change in respiratory movement to the abdominal breathing pattern. These results suggested that hypoventilation occurred due to dysfunctional changes in the intercostal muscles and resulted in hypoxemia in the later stages of DM.

OBJECTIVE To measure cerebral blood flow (CBF) and cerebral blood volume (CBV) by means of perfusion CT in clinically normal Holstein calves. ANIMALS 9 Holstein calves. PROCEDURES Each of the 9 calves (mean age, 20.2 days) was anesthetized and received an injection of iodinated contrast medium into the right jugular vein at a rate of 4.0 mL/s. Dynamic CT scanning of the head at a level that included the mandibular condyle was initiated at the time of the contrast medium injection and continued for 100 seconds. A deconvolution method was used as an analytic algorithm. RESULTS Among the 9 calves, the mean ± SD CBF in the cerebral cortex, white matter, and thalamus was 44.3 ± 10.3 mL/100 g/min, 36.1 ± 7.5 mL/100 g/min, and 40.3 ± 7.5 mL/100 g/min, respectively. The CBF in white matter was significantly lower than that in the cerebral cortex or thalamus. The mean CBV in the cerebral cortex, white matter, and thalamus was 6.8 ± 1.0 mL/100 g, 5.2 ± 1.0 mL/100 g, and 5.7 ± 0.7 mL/100 g, respectively. The CBV in the cerebral cortex was significantly higher than that in the white matter or thalamus. CONCLUSIONS AND CLINICAL RELEVANCE Measurement of CBF and CBV in clinically normal calves by means of perfusion CT was feasible. The data obtained may be useful as baseline values for use in future research or for comparison with findings from calves with CNS diseases. Investigations to determine the lower limit of blood flow at which brain function can still be restored are warranted.

Erythrocyte pyruvate kinase (PK) deficiency is an inherited glycolytic erythroenzymopathy caused by mutations of the PKLR gene. A causative mutation of the feline PKLR gene was originally identified in Abyssinian and Somali cats in the U.S.A. In the present study, a TaqMan probe-based real-time PCR genotyping assay was developed and evaluated for rapid genotyping and large-scale screening for this mutation. Furthermore, a genotyping survey was carried out in a population of four popular purebred cats in Japan to determine the current mutant allele frequency. The assay clearly displayed all genotypes of feline PK deficiency, indicating its suitability for large-scale survey as well as diagnosis. The survey demonstrated that the mutant allele frequency in Abyssinian and Somali cats was high enough to warrant measures to control and prevent the disease. The mutant allele frequency was relatively low in Bengal and American Shorthair cats; however, the testing should still be carried out to prevent the spread of the disease. In addition, PK deficiency should always be considered in the differential diagnosis of anemia in purebred cats in Japan as well as worldwide.

GM2 gangliosidosis variant 0 (Sandhoff disease, SD) is a fatal, progressive neurodegenerative lysosomal storage disease caused by mutations of the HEXB gene. In canine SD, a pathogenic mutation (c.283delG) of the canine HEXB gene has been identified in toy poodles. In the present study, a TaqMan probe-based real-time PCR genotyping assay was developed and evaluated for rapid and large-scale genotyping and screening for this mutation. Furthermore, a genotyping survey was carried out in a population of toy poodles in Japan to determine the current mutant allele frequency. The real-time PCR assay clearly showed all genotypes of canine SD. The assay was suitable for large-scale survey as well as diagnosis, because of its high throughput and rapidity. The genotyping survey demonstrated a carrier frequency of 0.2%, suggesting that the current mutant allele frequency is low in Japan. However, there may be population stratification in different places, because of the founder effect by some carriers. Therefore, this new assay will be useful for the prevention and control of SD in toy poodles.

Whole exome sequencing (WES) has become a common tool for identifying genetic causes of human inherited disorders, and it has also recently been applied to canine genome research. We conducted WES analysis of neuroaxonal dystrophy (NAD), a neurodegenerative disease that sporadically occurs worldwide in Papillon dogs. The disease is considered an autosomal recessive monogenic disease, which is histopathologically characterized by severe axonal swelling, known as "spheroids," throughout the nervous system. By sequencing all eleven DNA samples from one NAD-affected Papillon dog and her parents, two unrelated NAD-affected Papillon dogs, and six unaffected control Papillon dogs, we identified 10 candidate mutations. Among them, three candidates were determined to be "deleterious" by in silico pathogenesis evaluation. By subsequent massive screening by TaqMan genotyping analysis, only the PLA2G6 c.1579G>A mutation had an association with the presence or absence of the disease, suggesting that it may be a causal mutation of canine NAD. As a human homologue of this gene is a causative gene for infantile neuroaxonal dystrophy, this canine phenotype may serve as a good animal model for human disease. The results of this study also indicate that WES analysis is a powerful tool for exploring canine hereditary diseases, especially in rare monogenic hereditary diseases.

A single nucleotide substitution (c.-6-180T>G) associated with resistance to phenobarbital therapy has been found in the canineMDR1/ABCB1gene in Border Collies with idiopathic epilepsy. In the present study, a PCR-restriction fragment length polymorphism assay was developed for genotyping this mutation, and a genotyping survey was carried out in a population of 472 Border Collies in Japan to determine the current allele frequency. The survey demonstrated the frequencies of the T/T wild type, T/G heterozygote, and G/G mutant homozygote to be 60.0%, 30.3%, and 9.8%, respectively, indicating that the frequency of the mutant G allele is extremely high (24.9%) in Border Collies. The results suggest that this high mutation frequency of the mutation is likely to cause a high prevalence of phenobarbital-resistant epilepsy in Border Collies.

Collie eye anomaly (CEA) is an inherited canine ocular disorder causing regional choroidal hypoplasia and coloboma of the optic disk or adjacent areas, and is associated with a 7.8-kilo base pair (bp) deletion in the canine NHEJ1 gene (Parker and others 2007). A 4-bp deletion in the canine MDR1 / ABCB1 gene ( MDR1 gene defect) causes neurotoxicosis after administration of P-glycoprotein substrates such as ivermectin (Gramer and others 2011). These two mutations are common in collie-related and sighthound breeds, suggesting a possible parallel selection of the two defects, although these canine genes are located on different chromosomes: NHEJ1 on chromosome 37 and MDR1 on chromosome 14 (Parker and others 2007, Gramer and ­others 2011). Recently, a very high frequency of the CEA mutation has been reported in the Hokkaido inu, a traditional Japanese breed; this is the first identification of a CEA-predisposed breed not classified as collies or sighthounds (Mizukami and others 2012a). A native Japanese breed, the shiba inu, has been previously localised near collies in multibreed cluster analyses (Parker and others 2007), suggesting that collies and such native Asian breeds share more genetic traits than one may realise. However, prevalence of the MDR1 gene defect in the Hokkaido inu is unknown. Furthermore, epidemiologic surveys targeting potential breeds likely to possess these two mutations have not been performed. This study investigated allele frequencies of these multiple dog breeds and checked whether a parallel and simultaneous selection for these two mutations has occurred. In this study, 1331 DNA samples were collected from nine dog breeds including two collie-related breeds (collie and border collie), one sighthound (Saluki), two non-collie European …