Nobuko Mori

Predisposition for primary hyperlipidemia in Miniature Schnauzers and Shetland sheepdogs as compared to other canine breeds.

Miniature Schnauzers are the first canine breed, in the United States, reported to suffer from primary hyperlipidemia, but this has yet to be documented in other regions. Using over 900 canine plasma samples collected from over seven different veterinary clinics across Japan, the aim of this study was to compare plasma triglyceride (TG) and cholesterol concentrations between Miniature Schnauzers and other purebreeds in Japan. In addition, we investigated the influence of aging and sex on changes to hyperlipidemia incidence in purebred dogs. Our results indicated that both Miniature Schnauzers and Shetland sheepdogs in Japan exhibited remarkably high concentrations of plasma TG and total cholesterol, which are considered to be signs of hyperlipidemia, as compared to other purebred and mixed (Mongrel) canine breeds. Interestingly, the cause and conditions of primary hyperlipidemia in Miniature Schnauzers and Shetland sheepdogs might be different, with hypertriglyceridemia predominantly occurring with Miniature Schnauzers and hypercholesterolemia occurring in Shetland sheepdogs. However, with the influence of aging, the hyperlipidemia evolves into both hypercholesterolemia and hypertriglyceridemia in both groups indicating that the severity of hyperlipidemia positively correlates with aging. Gender differences were also observed with regards to severity. In fact, a higher severity was prevalent with female Miniature Schnauzers than their male counterparts whereas it was more balanced between genders for Shetland sheepdogs.

Obesity induced changes to plasma adiponectin concentration and cholesterol lipoprotein composition profile in cats.

Feline obesity generally results in aberrations to plasma metabolite levels, such as lipid concentrations and lipoprotein composition. This study sought to investigate the resultant effect of obesity on cholesterol lipoprotein composition and circulating adiponectin concentrations in cats. Plasma glucose, lipids (triglyceride, cholesterol and free fatty acid), insulin and adiponectin concentrations, and cholesterol lipoprotein composition were measured and compared between body condition score (BCS) determined normal healthy control and obese cats. Although the obese group demonstrated higher levels of plasma cholesterol, glucose, and triglycerides, as compared to healthy controls, the difference was insignificant thus indicating that the BCS determined obese cats may have been overweight and not morbidly obese. Plasma insulin levels were significantly higher (25-30%) versus healthy control animals thereby possibly hinting at the ensuing emergence of obesity induced insulin resistance. However, the BCS determined obese cat demonstrated a significant reduction (p<0.05) in plasma adiponectin concentration and a significant increase (p<0.05) in LDL-cholesterol % as compared to age matched healthy control animals. This would indicate that changes in plasma adiponectin concentration and cholesterol lipoprotein composition may be good early indicators of obesity in cats.

Potential predictive biomarkers of obesity in Burmese cats.

Australian Burmese cats are predisposed to diabetes mellitus and, compared to other breeds, have delayed triglyceride clearance that may result in subtle changes within cells and tissues that trigger specific alterations in gene expression within peripheral blood leucocytes (PBLs). Expression of genes involved in energy metabolism (glucose-6-phosphate dehydrogenase and malate dehydrogenase), lipogenesis (ATP citrate lyase [ACL], fatty acid synthase [FAS] and sterol regulatory binding protein-1c [SREBP-1c]), and insulin signalling (insulin receptor substrates 1 and 2, and phosphatidylinositol-3 kinase), as well as cholesterol lipoprotein subfraction profiling were carried out on PBLs from lean Burmese cats and compared with similar profiles of age and gender matched lean and obese Australian domestic shorthaired cats (DSHs) in an attempt to identify possible biomarkers for assessing obesity. For the majority of the genes examined, the lean Burmese cats demonstrated similar PBL gene expression patterns as age and gender matched obese Australian DSH cats. Lean Burmese had increased expression of ACL and FAS, but not SREBP-1c, a main upstream regulator of lipid synthesis, suggesting possible aberrations in lipogenesis. Moreover, lean Burmese displayed a 3- to 4-fold increase in the very low density cholesterol fraction percentage, which was double that for obese DSH cats, indicating an increased degree of lipid dysregulation especially in relation to triglycerides. The findings suggest that Burmese cats may have a particular propensity for dysregulation in lipid metabolism.

Hypertriglyceridemia with increased plasma insulin concentrations in cats.

Metabolite, insulin and adiponectin concentrations and LDH, AST and ALT activities were measured in plasma of 142 client-owned cats (1-13years old, 16 breeds) to set up a new criterion of hypertriglyceridemia (hyper-TG) with increased plasma insulin concentrations for early diagnosis of lipid metabolism abnormality including obesity. 25 cats with over 165mg/dl of plasma triglyceride (TG) concentrations were decided as hyper-TG with increased plasma insulin concentrations, and prevalence of hyper-TG was 16.7% in young (1-6years old) and 18.3% in old (>7years old) cats examined. In the hyper-TG cats, their plasma TG concentrations increased to 6.6-7.4-fold of the values of control cats with 35-50mg/dl of plasma TG and their plasma cholesterol, FFA and insulin concentrations and LDH and ALT activities increased significantly, whereas their plasma adiponectin concentrations decreased significantly compared to those in the control cats. Hyper-TG cats with significantly increased body weights and plasma insulin and decreased plasma adiponectin seemed to be in early stage of obesity accompanying increased plasma insulin concentrations. Increased TG, insulin, LDH and ALT and decreased adiponectin values in plasma seemed to be key factors for diagnosis of lipid metabolism abnormality at early stage in cats.

Supplementing five-point body condition score with body fat percentage increases the sensitivity for assessing overweight status of small to medium sized dogs

Background and methods Currently, five-point body condition scoring (BCS) is widely used by veterinarians and clinicians to assess adiposity in dogs in Japan. However, BCS score assignment is subjective in nature, and most clinicians do not score with half points, instead preferring to round off values, thereby rendering less accurate assessments. Therefore, we sought to determine whether assessing body fat percentage using simple morphometric measurements and supplementing this with five-point BCS can have increased sensitivity for detecting increasing adiposity in overweight small-medium sized dog breeds via plasma metabolite validation. Results Overall, lean body fat percentage was determined to be 15%-22% for male (non-neutered/neutered) dogs and 15%-25% for female (nonspayed/spayed). Dogs categorized as overweight by BCS had significantly higher levels of nonesterified fatty acids (P = 0.005), whereas animals categorized as overweight by BCS + body fat percentage were observed to have significantly higher levels of nonesterified fatty acids (P = 0.006), total cholesterol (P = 0.029), and triglycerides (P = 0.001) than lean animals. The increased sensitivity due to body fat percentage for gauging alterations in plasma metabolite levels may be due to increased correlation strength. Body fat percentage correlated positively with plasma insulin (r = 0.627, P = 0.002), nonesterified fatty acids (r = 0.674, P < 0.001), total cholesterol (r = 0.825, P < 0.0001), triglycerides (r = 0.5823, P < 0.005), blood urea nitrogen (r = 0.429, P < 0.05), creatinine (r = 0.490, P = 0.021), and total protein (r = 0.737, P < 0.0001) levels, which all tend to increase as a result of increasing adiposity. Conclusion Supplementing body fat percentage with five-point BCS appears to increase the likelihood of validating overweight status in small-medium sized dog breeds by detecting changes in plasma metabolite levels, especially lipids, induced as a result of increasing adiposity.

Impact of feline AIM on the susceptibility of cats to renal disease.

Renal failure is one of the most important social problems for its incurability and high costs for patients’ health care. Through clarification of the underlying mechanism for the high susceptibility of cats to renal disease, we here demonstrates that the effective dissociation of serum AIM protein from IgM is necessary for the recovery from acute kidney injury (AKI). In cats, the AIM-IgM binding affinity is 1000-fold higher than that in mice, which is caused by the unique positively-charged amino-acid cluster present in feline AIM. Hence, feline AIM does not dissociate from IgM during AKI, abolishing its translocation into urine. This results in inefficient clearance of lumen-obstructing necrotic cell debris at proximal tubules, thereby impairing AKI recovery. Accordingly, mice whose AIM is replaced by feline AIM exhibit higher mortality by AKI than in wild-type mice. Recombinant AIM administration into the mice improves their renal function and survival. As insufficient recovery from AKI predisposes patients to chronic, end-stage renal disease, feline AIM may be involved crucially in the high mortality of cats due to renal disease. Our findings could be the basis of the development of novel AKI therapies targeting AIM-IgM dissociation, and may support renal function in cats and prolong their lives.

Elevated Plasma Adiponectin Level and Peripheral Blood Leukocyte Adiponectin Receptor Expression in Dogs Suffering from Insulin Deficiency

The aim of this study is to determine and compare the effect of obesity or insulin deficiency on plasma adiponectin level and peripheral blood leukocyte (PBL) adiponectin receptor 1 and 2 (ADIPOR1 and -R2) expression in dogs. Obese and insulin deficient (ID) dogs demonstrated a significant reduction (60% lower) and significant increase (2x higher) in circulating adiponectin level, respectively, as compared to normal controls. PBL ADIPOR1 and -R2 mRNA expression was also significantly higher in obese (R1=333 mean fold higher, R2=64 mean fold higher) and ID dogs pre- insulin treatment (R1 =48 mean fold higher, R2=16 mean fold higher) as compared to normal PBL. Insulin treatment reduced ADIPOR1 (3 mean fold higher) and -R2 (1.5 mean fold higher) expression back down closer to control PBL levels in ID animals. The upregulation of adiponectin receptor expression might reflect an increased need for adiponectin signaling; however, the increase may have a different implication between obese and ID dogs. This may be due to adiponectins contradictory paradoxical dual role, having both anti-inflammatory and pro-inflammatory properties on peripheral leukocytes, especially on monocytes depending on the biological context and adiponectin isoform.

Preliminary analysis of modified low-density lipoproteins in the serum of healthy and obese dogs and cats

Oxidized low-density lipoprotein (LDL) is thought to play an important role in the inflammatory response associated with human obesity. The purpose of this preliminary study was to determine oxidized LDL concentrations in healthy dogs and cats, and to evaluate whether obesity affects oxidized LDL concentration, using 39 cats and 19 dogs that had visited two different veterinary clinics in Japan. We hypothesized that oxidized LDL concentrations measured against body condition score (BCS) may have a potential value in evaluating the qualities of accumulated or circulating lipids in obese dogs and cats that do not show signs of metabolic diseases. The mean oxidized LDL value in BCS3 dogs (2.4 ± 0.9 μg/dl) was very similar to that of BCS5 dogs (2.2 ± 0.3 μg/dl). The mean oxidized LDL value of BCS4 dogs was 7.2 ± 10.3 μg/dl and the highest among three groups. BCS4 dogs included two dogs whose oxidized LDL values were higher than the mean oxidized LDL value of healthy humans (11.2 ± 0.3 μg/dl). On the other hand, the mean oxidized LDL value of BCS3 cats was 2.5 ± 0.9 μg/dl, and those of BCS4 and 5 cats were higher than that of BCS3, but there was no significant difference. The BCS4 cat group included one cat with a higher oxidized LDL value, and the BCS5 group also included two cats with oxidized LDL values higher than the mean oxidized LDL value of healthy humans. Interestingly, the oxidized LDL values in two obese dogs and three obese cats were indeed higher than the mean oxidized LDL value of humans with coronary artery disease (20.1 ± 1.1 μg/dl). In conclusion, this preliminary study showed reference ranges of oxidized dogs and cats against BCS. Obesity alone does not appear to have any direct effect on serum oxidized LDL values in healthy dogs and cats.

Short communication: molecular characterization of dog and cat p65 subunits of NF-kappaB.

Nuclear factor kappa B (NF-κB) plays an important role in the immune system. The p65 subunit is an important part of NF-κB unit, and studies of dog and cat p65 subunits of NF-κB (dp65 and cp65) are important in understanding their immune function. In this study, we described the molecular characterization of dp65 and cp65. The dp65 and cp65 complementary DNA encoded 542 and 555 amino acids, respectively, showing a high sequence homology with the mammalian p65 subunit (>87.5%). Quantitative polymerase chain reaction revealed that the p65 messenger RNA is highly expressed in the dog stomach and cat heart and adipose tissue. Functional NF-κB promoter-luciferase reporter vectors revealed that our isolated dp65 and cp65 cDNA encodes a functionally active protein. Transiently expressed dp65 and cp65 up-regulated pro-inflammatory cytokine expression levels in dog and cat, respectively. These findings suggest that dp65 and cp65 play important roles in regulating immune function.

Sirtuin 1 suppresses nuclear factor κB induced transactivation and pro-inflammatory cytokine expression in cat fibroblast cells

Nuclear factor κB (NF-κB) is a key factor in the development of chronic inflammation and is deeply involved in age-related and metabolic diseases development. These diseases have become a serious problem in cats. Sirtuin 1 (SIRT1) is associated with aging and metabolism through maintaining inflammation via NF-κB. In addition, fibroblasts are considered an important factor in the development of chronic inflammation. Therefore, we aimed to examine the effect of cat SIRT1 (cSIRT1) on NF-κB in cat fibroblast cells. The up-regulation of NF-κB transcriptional activity and pro-inflammatory cytokine mRNA expression by p65 subunit of NF-κB and lipopolysaccharide was suppressed by cSIRT1 in cat fibroblast cells. Our findings show that cSIRT1 is involved in the suppression of inflammation in cat fibroblast cells.