Shono Saori

Diacron-reactive oxygen metabolite (d-ROM) and biological antioxidant potential (BAP) levels in the serum of horses were measured (ponies, n = 15; thoroughbred, n = 31; other full-sized horses, n = 7). The mean d-ROM levels in horses were significantly higher (p < 0.001) than those in dairy cattle (n = 25) and dogs (n = 31). However, d-ROM levels in horses were lower than the standard levels reported in humans. When d-ROM and BAP levels were plotted graphically, the points for horses with a disease (ringbone in 1 Japanese sports horse, cellulitis in 1 thoroughbred, melanoma in 1 Lipizzaner) fell outside the group of points for other (non-diseased) horses. A similar separation was seen (using data from other authors) for a horse with Rhodococcus equi, a horse following castration surgery, and a mare following delivery. These results, comparing horses, other animals, and humans, are interesting from the standpoint of comparative medicine, and they contribute to the sparse literature available on d-ROM and BAP levels in animals. Because the level of d-ROM and BAP levels were changed depending on the situation of health, those indexes are promising as indices of health in horses.

Insulin degludec (IDeg) is a new insulin formulation that facilitates long-term control of glucose level in humans. In this study, we investigated the effects of IDeg on glycemic control in dogs. Its time-action profiles were monitored in healthy dogs using an artificial pancreas apparatus under euglycemic conditions. At 9.0-13.5 hr post-IDeg injection, an indistinct peak of glucose level was detected. Moreover, the action of IDeg was persistent for >20 hr. Both IDeg and neutral protamine Hagedorn insulin (NPH) lowered blood glucose concentrations in diabetic dogs, but IDeg caused postprandial hyperglycemia and a somewhat lower preprandial glucose level than that caused by NPH. IDeg might be ineffective in concurrently preventing postprandial hyperglycemia and preprandial hypoglycemia in a single-agent administration.

In humans, the rate of acetaminophen（APAP）absorption reflects the rate of gastric emptying. However, few studies have reported the use of the APAP method for evaluating gastric emptying in dogs. The purpose of the current study was to investigate whether a commercial APAP detection kit can measure serum APAP in healthy dogs. First, we evaluated the validity of this kit, measuring canine serum levels after the administration of APAP. Analytical evaluation of serum APAP concentrations indicated acceptable precision,with intra-assay coefficients of variation（CV）not exceeding the commonly accepted 10％limit. However,the inter-assay CV for serum APAP concentrations was 19.4％. A linear relationship was demonstrated with all dilutions tested, up to a 16-fold dilution. Next, to determine the detectable APAP dose using this kit, six healthy dogs were administered the experimental diet alone（0 mg（APAP）/kg（body weight））and the experimental diet with APAP added, at doses of 10 mg/kg and 20 mg/kg. A significant increase was observed in serum APAP concentrations at a dose at 20 mg/kg, compared with doses of 0 mg/kg and 10 mg/kg. In conclusion, an APAP detection kit is able to measure serum APAP concentrations in dogs and may be applicable to determine gastric emptying. Furthermore, the minimum detectable APAP dose in dogs is </tt><tt>approximately 20 mg/kg.</tt></p>

<p><tt>We compared the lipid metabolism in cats by feeding medium-chain triglycerides（MCT）using coconuts oil and long-chain triglycerides（LCT）（using lard and soybean oil）to investigate the effects of dietary MCT on glucose and lipid metabolism. We fed the control diet（low fat diet: 2.8 g of fat content/100 kcal）and control diet with 3 different sources of fat（coconuts oil, lard, and soybean oil）using 6 healthy cats. Each diet was fed for 14 days, and body weight, body fat and physical signs and blood examinations were evaluated. All diet can be fed during study period with no effect on dietary preference detected without any side effects. No significant differences were observed in body weight, body fat and physical signs during the study period. Furthermore, for blood examinations, serum glucose, insulin, triglyceride and non-esterified fatty acid（NEFA）concentrations and plasma glucose-dependent insulinotropic polypeptide（GIP）and glucagon-like peptide 1（GLP-1）concentrations were measured. However, all serum and plasma blood test did not show any significant difference. There is a positive effect of MCT on lipid metabolism in human as an efficient energy source, however, further study with different amount of MCT and log-term study should reveal more information about the kinetics, body fat accumulati</tt><tt>on, and nutrient metabolism in cats. </tt></p>

Dietary therapy with dietary fiber is useful for human obesity and diabetic patients, since it induces delaying glucose absorption and preventing postprandial hyperglycemia. The purpose of this study is to investigate whether two types of dietary fibers（beet pulp; soluble fiber＋insoluble fiber; cellulose; insoluble fiber）affect postprandial blood glucose metabolism parameter in healthy dogs. We used a commercial food as control diet（control diet; low fiber: 0.9g/100kcal）. Furthermore, 10％and 20％of beet pulp and cellulose were added to the control diet, respectively, as supplemented dietary fiber. The total additive dietary fiber amount was 10％（fiber: 3.5g/100kcal）and 20％（fiber: 6.1g/100kcal）respectively. When comparing amount of dietary fiber, 20％supplemented diet induced decreased postprandial blood insulin concentrations,non-esterified free fatty acid（NEFA）concentration and GIP concentration as compared to control diet. Furthermore, 20％supplemented diet induced increased postprandial blood GLP-1 concentration as compared to control diet. Therefore, 20％supplemented diet might help delaying digestion and absorption,thereby preventing obesity and suppressing appetite. However, no significant difference was observed in postprandial blood parameter when the two fiber sources（beet pulp and cellulose）were taken into account. Since beet pulp was mainly composed of insoluble fiber（about 80％）, the characteristics of the soluble fiber might be lacked.

Porphyromonas gulae, Tannerella forsythia and Campylobacter rectus are considered dominant periodontal pathogens in dogs. Recently, quantitative real-time PCR (qRT-PCR) methods have been used for absolute quantitative determination of oral bacterial counts. The purpose of the present study was to establish a standardized qRT-PCR procedure to quantify bacterial counts of the three target periodontal bacteria (P. gulae, T. forsythia and C. rectus). Copy numbers of the three target periodontal bacteria were evaluated in 26 healthy dogs. Then, changes in bacterial counts of the three target periodontal bacteria were evaluated for 24 weeks in 7 healthy dogs after periodontal scaling. Analytical evaluation of each self-designed primer indicated acceptable analytical imprecision. All 26 healthy dogs were found to be positive for P. gulae, T. forsythia and C. rectus. Median total bacterial counts (copies/ng) of each target genes were 385.612 for P. gulae, 25.109 for T. forsythia and 5.771 for C. rectus. Significant differences were observed between the copy numbers of the three target periodontal bacteria. Periodontal scaling reduced median copy numbers of the three target periodontal bacteria in 7 healthy dogs. However, after periodontal scaling, copy numbers of all three periodontal bacteria significantly increased over time (p?0.05, Kruskal-Wallis test) (24 weeks). In conclusion, our results demonstrated that qRT-PCR can accurately measure periodontal bacteria in dogs. Furthermore, the present study has revealed that qRT-PCR method can be considered as a new objective evaluation system for canine periodontal disease.

Reduced expression in immortalized cells (REIC/Dkk-3), a member of the human Dickkopf (Dkk) family, is a growth suppressor in human and canine mammary tumours. Mammary gland tumours are common neoplasms with high malignancy in female cats. The purpose of this study was to clone the feline REIC/Dkk-3 homolog, investigate its expression in cell lines established from feline mammary gland tumours, and test its tumour suppressor function. Western blot analysis revealed that expression of the REIC/Dkk-3 protein was reduced in feline mammary carcinoma cell lines. Forced expression of REIC/Dkk-3 induced apoptosis in feline mammary tumour cell lines. These results demonstrate that REIC/Dkk-3 expression, which is downregulated in feline mammary tumour cell lines, results in the induction of apoptosis in these cells. Our findings suggest that feline REIC/Dkk-3 represents a potential molecular target for the development of therapies against feline mammary cancers.

This study evaluated the accuracy of a newly developed veterinary portable blood glucose meter (PBGM) with hematocrit correction in dogs and cats. Sixty-one dogs and 31 cats were used for the current study. Blood samples were obtained from each dog and cat one to six times. Acceptable results were obtained in error grid analysis between PBGM and reference method values (glucose oxidation methods) in both dogs and cats. Bland-Altman plot analysis revealed a mean difference between the PBGM value and reference method value of -1.975 mg/dl (bias) in dogs and 1.339 mg/dl (bias) in cats. Hematocrit values did not affect the results of the veterinary PBGM. Therefore, this veterinary PBGM is clinically useful in dogs and cats.

Anion-exchange (AEX)-high-performance liquid chromatography (HPLC) for measurement of cholesterol can be used to separate serum lipoproteins (high-density lipoprotein (HDL); low-density lipoprotein (LDL); intermediate-density lipoprotein (IDL); very-low-density lipoprotein (VLDL)) in humans. However, AEX-HPLC has not been applied in veterinary practice. We had three objectives: (i) the validation of AEX-HPLC methods including the correlation of serum cholesterol concentration in lipoprotein fraction measured by AEX-HPLC and gel permeation-HPLC (GP-HPLC) in healthy dogs and those with hypercholesterolemia was investigated; (ii) the reference intervals of lipoprotein fractions measured by AEX-HPLC from healthy dogs (n=40) was established; (iii) lipoprotein fractions from the serum of healthy dogs (n=12) and dogs with hypercholesterolemia (n=23) were compared. Analytic reproducibility and precision of AEX-HPLC were acceptable. Positive correlation between serum concentrations of total cholesterol (Total-Chol), HDL cholesterol (HDL-Chol), LDL cholesterol (LDL-Chol)+IDL cholesterol (IDL-Chol), and VLDL cholesterol (VLDL-Chol) was noted for AEX-HPLC and GP-HPLC in healthy dogs and dogs with hypercholesterolemia. Reference intervals measured by AEX-HPLC for serum concentrations of Total-Chol, HDL-Chol, and LDL-Chol were determined to be 2.97-9.32, 2.79-6.57, 0.16-3.28mmol/L (2.5-97.5% interval), respectively. Furthermore, there was significant difference in lipoprotein profiles between healthy and dogs with hypercholesterolemia. These results suggest that AEX-HPLC can be used to evaluate lipoprotein profiles in dogs and could be a new useful indicator of hyperlipidemia in dogs.

&ensp;&ensp;Exercise therapy as well as dietary and drug therapy are important therapies for diabetic patients. However, exercise therapy also has the risk of hypoglycemia and any other in diabetic patients. Therefore the purpose of this study was to evaluate influence of dietary fiber amount（fiber diet or non-fiber diet）and starting time of exercise therapy on glucose metabolism in 3 dogs with Type 1 diabetes mellitus. Exercise therapy was performed at postprandial 6 hours（Ex 6hr）and 10 hours（Ex 10hr）. As a result, blood glucose concentrations under Ex 6hr were decreased using in fiber and non-fiber diet groups. However, lowering of blood glucose concentration with fiber diet was stable as compared to that with non-fiber diet. Meanwhile, any changes of blood glucose concentrations was not observed in exercise therapy under Ex 10hr using both diet. In conclusion, post prandial exercise should be started at 10 hours postprandially regardless of dietary fiber content of food.

The concentrations of plasma free amino acids（PFAAs）are reported to change in human patients with cancer before and after chemotherapy. However, it is unclear in cancer dogs. Therefore the purpose of the current study was to investigate the changes in PFAAs before and after chemotherapy in dogs with transitional cell carcinoma（TCC）. We used the 3 TCC dogs in the current study. TCC dogs were administrated mitoxantrone via the intravascular route at the first day of the diagnosis and administration schedule was settled at 0 and 3 weeks. In addition, TCC dogs were orally administrated piroxicam by 6 weeks, once a day. Then, we measured the concentration of different 30 amino acids using liquid chromatography mass spectrometry with pre-prandial plasma samples at 0, 1, 3 and 6 weeks. Significant changes were observed in mean plasma cystathionine concentration between before and after chemotherapy. Cystathionine is gradually decreased at 1 week and 3 weeks as compare to 0 week, thereafter increased at 6 weeks. Plasma cystathionine of TCC dogs might be utilized by the tumor cell. Therefore, these results indicate that PFAAs is affected by administration of anticancer drugs in TCC dogs. Further study is needed to confirm the relationship between TCC and PFAAs profile with more canine patients and longer experimental period.

<p>Incretins such as glucose-dependent insulinotropic peptide（GIP）and glucagon like peptide-1（GLP-1）were shown to exert their insulinotropic effects through a variety of mechanisms. In the presence of matched glucose concentrations, insulin secretion is greater following ingestion of glucose than following infusion of glucose. This was referred to as "the incretin effect"and GIP and GLP-1 are released by gut endocrine cells in response to a meal and regulate blood glucose levels by stimulating glucose-dependent insulin secretion. The purpose of this study was to investigate the effect of incretin secretion for glucose and insulin metabolism in healthy cats. In order to determine changes in incretin secretion with different nutritional composition （control, high-carbohydrate, high-fat, high-fiber）, different diets were fed in 5 healthy cats. We measured postprandial glucose, insulin, GIP and GLP-1 concentrations. No significant changes in mean glucose and insulin concentrations. Furthermore, GLP-1 concentration was not significantly difference in 4 type diets. Meanwhile, GIP secretion was increased in high-fat diet. Furthermore, GIP was increased in direct proportion to fat content of these diets. As the result of GIP secretion was increased in high- fat diet, similarly as with humans and rodents. However, GLP-1 was not consistent with human objects, and we suggest that cats have been different digestive tissue and feeding habit.</p>

<p>Managing cat diabetes mellitus recommended that diets with low carbohydrate content and protein content may be advantageous, the carbohydrate type of the diets in those studies were not well characterized. The purpose of this study was to the effects of carbohydrate sources（glucose, Glc; maltose, Mal; trehalose,Tre; cornstarch, Corn）on glucose metabolism in healthy cats. As a result, Glc and Mal diets were significant increase was observed in Glucose-AUC than Tre and Corn diet. Meanwhile, no significant changes in Insulin AUC of all diets. Furthermore, there was a significant increase in NEFA-AUC of Tre diet compared to Glc and Mal diets. In conclusion, these results indicate that carbohydrate might be useful as an energy source in healthy cats. Especially, the slow absorption carbohydrate as cornstarch was considered as an available to correct hyperglycemia in obese and diabetes cats.</p>

This study investigated the changes in lymphocyte subsets during the trilostane medication of Pituitary-dependent hyperadrenocorticism (PDH) dogs. The cortisol level and lymphocyte subsets of eight dogs with PDH were monitored 0, 1, 3, 6, 9 and 12 months after the initiation of trilostane treatment. White blood cells (WBC), lymphocytes, CD3(+) (T lymphocyte), CD4(+) (helper T lymphocyte), CD8(+) (cytotoxic T lymphocyte) and CD21(+) (B lymphocyte) cells were measured. Although the post-ACTH stimulation test cortisol level was significantly lower during trilostane treatment, changes in the CD3(+), CD4(+), CD8(+) and CD21(+) counts were not observed. Meanwhile, significant decrease was observed in WBC counts during trilostane treatment. These indicate that long-term trilostane treatment has little effect on the lymphocyte subsets in PDH dogs.

&emsp;Changes in blood glucose concentration due to exercise were reported to induce various side effects such as hypoglycemia and delayed digestion and absorption depending on timing of mealtime. Postprandial hypoglycemia has been reported in the dog during hunting, however, its mechanism is unknown. Therefore the purpose of this study was to evaluate blood biochemical parameters before and after oral glucose tolerance test（OGTT）in healthy 4 dogs. Exercise was performed after 30 minutes from the beginning of OGTT. The results suggested that exercise induce decreasing blood glucose and insulin concentrations after OGTT. As such, conditions with both hyperglycemia and hyperinsulinemia might have higher risk of lowering blood glucose concentratsons. Moreover, exercise in fasting state induced lipolysis. <br> &emsp;In the future, we should evaluate relationship between exercise and diet（include various nutrient component）, since OGTT was only performed in the present study.

Carbohydrates have a large effect on postprandial glycemic control compared to nutrient content. Therefore, if slower digestion of glucose was achieved, which would be inhibition of postprandial hyperglycemia. Recently, the mulberry leaf includes 1-deoxynojirimycin（DNJ）, which inhibit postprandial glucose absorption. In human patients, it was reported that the postprandial hyperglycemia was significantly improved after administration of mulberry leaf. However, the hypoglycemic effects for dogs are unknown. Therefore, the objective of this study was to investigate the changes in glucose and lipid metabolism after administration of mulberry leaf in healthy dogs. In 6 healthy dogs, mulberry leaf powder（0.5 g/head, DNJ: 0.97 mg/head）was administered with a high carbohydrate diet for 4 days. After 4 days, blood examination results significant reduction in blood glucose level of postprandial 30 minutes was observed in the administered group as compared with the no administered group. In addition, slightly lower trend in blood insulin level of postprandial 30 and 180 minutes was observed in the administered group as compared to the no administered group. However, no significant difference in blood triglyceride level was observed between the administered group and the no administered group. From the results, administration of mulberry leaf might inhibit postprandial elevation of blood glucose and insul in secretion in healthy dogs.

This study evaluated the accuracy and reproducibility of a human portable blood glucose meter (PBGM) for canine and feline whole blood. Reference plasma glucose values (RPGV) were concurrently measured using glucose oxidation methods. Fifteen healthy dogs and 6 healthy cats were used for blood sampling. Blood glucose concentrations and hematocrits were adjusted using pooled blood samples for our targeted values. A positive correlation between the PBGM and RPGV was found for both dogs (y = 0.877, x = -24.38, r = 0.9982, n = 73) and cats (y = 1.048, x = -27.06, r = 0.9984, n = 69). Acceptable results were obtained in error grid analysis between PBGM and RPGV in both dogs and cats; 100% of these results were within zones A and B. Following ISO recommendations, a PBGM is considered accurate if 95% of the measurements are within +/- 15 mg/dl of the RPGV when the glucose concentration is &lt;100 mg/dl and within +/- 15% when it is &gt;= 100 mg/dl; however, small numbers of samples were observed inside the acceptable limits for both dogs (11%, 8 of 73 dogs) and cats (39%, 27 of 69 cats). Blood samples with high hematocrits induced lower whole blood glucose values measured by the PBGM than RPGV under hypoglycemic, normoglycemic, and hyperglycemic conditions in both dogs and cats. Therefore, this device is not clinically useful in dogs and cats. New PBGMs which automatically compensate for the hematocrit should be developed in veterinary practice.

Dietary therapy for human diabetic patients is mainly focusing on dietary carbohydrate level. In the present study, five healthy dogs were fed different sources of carbohydrate diets to investigate their effect on postprandial glucose metabolism parameter. We used three commercial diets which include three different major carbohydrate sources（rice, corn and wheat）. As a result, no significant difference was observed in postprandial blood concentrations between three diets. Furthermore, from the results of insulin secretion, a tendency for digestion and absorption speeds were tend to increase wheat, corn, in the order of rice in three diets. In addition, GIP secretion is stimulated by the amount of dietary fat, rather than carbohydrate sources. No significant differences were observed in postprandial GLP-1 concentrations between three diets. In conclusion, different sources of dietary carbohydrate did not influence for glucose and GLP-1 concentrations in healthy dogs, however, they might relate to insulin secretion. Lastly dietary fat contents might relate to GIP secretion.

Hyperadrenocorticism (HAC) is a common endocrine disorder in dogs, in which excess glucocorticoid causes insulin resistance. Disturbance of insulin action may be caused by multiple factors, including transcriptional modulation of insulin signal molecules which lie downstream of insulin binding to insulin receptors. In this study, gene expressions of insulin signal molecules were examined using neutrophils of the HAC dogs (the untreated dogs and the dogs which had been treated with trilostane). Insulin receptor substrate (IRS)-1, IRS-2, phosphatidylinositol 3-kinase (PI3-K), protein kinase B/Akt kinase (Akt)-2 and protein kinase C (PKC)-lambda were analyzed in the HAC dogs and compared with those from normal dogs. The IRS-1 gene expressions decreased by 37% and 35% of the control dogs in the untreated and treated groups, respectively. The IRS-2 gene expressions decreased by 61% and 72%, the PI3-K gene expressions decreased by 47% and 55%, and the Akt-2 gene expressions decreased by 45% and 56% of the control dogs, similarly. Collectively, gene expressions of insulin signal molecules are suppressed in the HAC dogs, which may partially contribute to the induction of insulin resistance.

Therapeutic exercise is a beneficial treatment for diabetes mellitus that ameliorates insulin sensitivity and increases glucose uptake into skeletal muscle. In this study, we investigated whether therapeutic exercise affects blood biochemical parameters and muscular mRNA expression in diabetic dogs. No significant difference was observed between the fasting blood glucose concentrations before and after exercise. However, the levels of glycated albumin（GA）and non-esterified fatty acids were significantly decreased after exercise. No significant differences were found between before and after exercise in the mRNA expression of insulin signaling and glucose metabolism genes including insulin receptor substrate （IRS）-1, IRS-2, phosphatidylinositol 3-kinase, Akt kinase 2, glucose transporter 4, AMP-activated protein kinase, uncoupling protein 3, and acetyl-CoA carboxylase. In summary, therapeutic exercise decreased GA levels and thus ameliorated glycemic control in diabetic dogs.