小田 民美

Ezetimibe is a cholesterol absorption inhibitor that blocks the intestinal absorption of both biliary and dietary cholesterol, thereby lowering primarily low density lipoprotein-cholesterol (LDL-chol) in human studies. This study aimed to investigate the effects of ezetimibe on dyslipidemia control in nine dogs with hypercholesterolemia. Changes in total cholesterol (T-chol) and each lipoprotein fractions were evaluated at 0, 2, and 4 months following initiation of ezetimibe treatment. A significant decrease in T-chol was observed, and a mean T-chol concentration below 400 mg/dL was achieved at 2 and 4 months. Furthermore, a significant decrease in LDL-chol was observed (-53.3% and -64.3% at 2 and 4 months, respectively). Taken together, treatment of ezetimibe could lower LDL-chol levels in dogs with hypercholesterolemia.

消化酵素剤単独および市販のプロバイオティクス/消化酵素剤投与が11頭の健康な家庭犬の糞便マイクロバイオームに及ぼす影響について検討した.合計9週間の研究を行い、その間すべての犬は同じフードを与えられた。最近の3週間はフードのみ与えられ(試験期間1)、次の3週間において消化酵素剤が添加された(試験期間2)。最後の3週間において、市販のプロバイティクス/消化酵素剤を投与した(試験期間3)。次世代シークエンスの結果、Firmicutes門、Clostridia綱、Clostridiales目、Clostridiaceae、Clostridium属およびBlautia属の割合が試験期間1から試験期間3にかけて有意に上昇した。消化酵素剤単独および市販のプロバイティクス/消化酵素剤は健康な家庭犬の糞便マイクロバイオームに影響を与えることを示した。将来的にはこれらの薬剤が消化器疾患を防ぐことができるかを検討すべきであると考える。(著者抄録)

A 10-year-old castrated male cat showing behavioral (irritation, prowling, and tumbling) and cutaneous abnormalities such as dermal fragility was diagnosed as hyperadrenocorticismwith pituitary macroadenoma, concurrent with insulin dependent diabetes mellitus. Pituitary enlargement (18.0 mm) was observed during magnetic resonance imaging. High endogenous adrenocorticotropic hormone levels (>2,500 pg/ml) were also observed.Although trilostane treatment (5-10 mg/head, daily) was commenced, the clinical signs did not disappear. Insulin and trilostane treatment were discontinued on day 86 after first day of radiation therapy (4 Gy/12 fractions). After radiation therapy, a decreased pituitary tumor size (10.7 mm) was observed on day 301; neurological and dermatological signs exhibited remission. Radiation therapy is the treatment of choice for feline hyperadrenocorticismwith pituitary macroadenoma with neurological signs.

In the present study, we used next-generation sequencing to investigate the impacts of two commercially available prescription diet regimens on the fecal microbiomes of eleven client-owned healthy pet dogs. We tested an anallergenic diet on 6 dogs and a low-fat diet on 5 dogs. Before starting the study, each dog was fed a different commercial diet over 5 weeks. After collecting pre-diet fecal samples, the anallergenic or low-fat diet was administered for 5 weeks. We then collected fecal samples and compared the pre- and post-diet fecal microbiomes. In the dogs on the anallergenic diet, we found significantly decreased proportions of Bacteroides, Ruminococcaceae, and Fusobacteriaceae, belonging to the phyla Bacteroidetes, Firmicutes, and Fusobacteria, respectively. The proportion of the genus Streptococcus belonging to the phylum Firmicutes was significantly increased upon administering the anallergenic diet. In the dogs on the low-fat diet, although the phyla Actinobacteria and Bacteroidetes tended to increase (p=0.116) and decrease (p=0.147) relative to the pre-diet levels, respectively, there were no significant differences in the proportions of any phylum between the pre- and post-diet fecal microbiomes. The anallergenic diet induced a significantly lower diversity index value than that found in the pre-diet period. Principal coordinate analysis based on unweighted UniFrac distance matrices revealed separation between the pre- and post-diet microbiomes in the dogs on the anallergenic diet. These results suggest that, even in pet dogs kept indoors in different living environments, unification of the diet induces apparent changes in the fecal microbiome.

臨床診療の場面で効果的なコミュニケーションを行うためには、獣医療スタッフとクライアントの相互参加が必要である。本研究では、動物病院の診察室におけるコミュニケーションの現状を、Roter Interaction Analysis System(RIAS)により定量的に評価できるか検証を試みた。大学附属動物病院の内分泌科で記録した59例の医療面接から無作為に抽出した12例を対象に、食事管理に関する発話や動物や同伴者に対する発話等の6カテゴリーを追加した上で、RIASに基づいて分析を行った。信頼性の評価として、2名の認定コーダーがそれぞれ独立して分析を行い、両者の結果を確認した。また、各カテゴリーの発話頻度を獣医師とクライアントで比較した。その結果、12例の平均診察時間は10分6秒であることがわかった。コーダー間の信頼性(Spearman's ρ=0.828、P<0.01)は高かった。診察1回あたり平均1回以上の発話があった16のカテゴリーのうち、獣医療スタッフは「あいづち」「理解の確認/明確化のための言い換え」「接続語・移行の合図」が多く、クライアントは「同意・理解」「食事の情報提供」が多かった。今回の研究結果は、対象とした診療科が1つであったため限定的ではあるが、実際の臨床会話の構造と一致しており、妥当なものと考えられる。RIASの有用性が確認されたことで、今後、獣医学的コミュニケーションに関する多様な研究にこの手法を用いることが可能となった。(著者抄録)

Changes in lipoprotein profiles occur in dairy cows during the periparturient period and in cows with transition cow disease. Here, the lipoprotein profiles of Holstein-Friesian dairy cows during the periparturient period were obtained by anion-exchange, high-performance liquid chromatography to evaluate the usefulness of lipoprotein profile evaluation during the periparturient period and in cows with fatty liver and milk fever. Lipoprotein levels (including total and high- (HDL-C) and low-density lipoprotein (LDL-C) cholesterol) in 10 healthy cows were low 4 weeks prepartum, with the lowest values at calving or within 1 week of calving; the values increased at 8 weeks postpartum. The lipoprotein levels were measured in 16 cows diagnosed with fatty liver (n=10) or milk fever (n=6) and compared to 10 healthy dairy cows. A significant difference was observed in HDL-C between healthy cows (at calving and 1 week postpartum), and the fatty liver and milk fever cows. Cows with fatty liver and milk fever had a lower mean HDL-C than the 10 healthy dairy cows at calving and 1 week postpartum. HDL-C might be a good indicator of energy balance for differentiating healthy cows from those with transition cow disease.

A 5-year-old castrated male domestic shorthair cat was diagnosed with diabetic ketoacidosis and severe insulin resistance. Although the conventional treatment for diabetic ketoacidosis was provided, the cat required frequent hospitalization because of severe dehydration and repeated diabetic ketoacidosis. We detected anti-insulin antibodies for human in this cat. Serum insulin-binding IgG levels were markedly elevated compared with those in healthy cats and other diabetic cats. We initiated prednisolone to suppress the effects of anti-insulin antibodies. After initiation of prednisolone, the cat was gradually recovered with increasing activity and appetite. Furthermore, satisfactory glycemic control was achieved with combined subcutaneous injection of insulin detemir and insulin degludec.

Insulin degludec (IDeg) is a long-acting basal insulin recently developed for use in humans. This study aimed to investigate the effects of IDeg on glycemic control in diabetic cats. Changes in body weight, IDeg dosage, and glycated albumin (GA) were evaluated at 0, 1, 3, 6, 9, and 12 months following initiation of IDeg. A significant decrease in GA was observed and a mean GA level below 25% was achieved between 3 and 12 months. Furthermore, a significant increase in body weight was observed between 3 and 12 months. The mean IDeg dose was 0.75 ± 0.68 IU/kg/day at 12 months. Taken together, long-term glycemic control was successfully achieved in diabetic cats using IDeg.

ヒトの医学領域および多くの哺乳動物において、食事中の栄養素(特に炭水化物と脂肪)がグルコース依存性インスリン分泌刺激ポリペプチド(GIP)やグルカゴン様ペプチド-1(GLP-1)といったインクレチン分泌を促進することが知られている。さらにヒトでは2つの異なる脂肪源(飽和脂肪酸を多く含むラードと不飽和脂肪酸を多く含む大豆油)によってインクレチンの分泌量が変化することが報告された。本研究では、食事中の異なる脂肪源(ラードおよび大豆油)が健常猫のGIP、GLP-1分泌、そして血糖値、インスリン、中性脂肪、NEFAにどのような影響を及ぼすのかを検討した。低脂肪食であるbasal食と、ラードと大豆油の2種の脂肪をbasal食に加えた高脂肪食を、それぞれ14日間ずつ給与した。結果として高脂肪食給与下ではGIP分泌とNEFA濃度が有意に上昇した。しかし、GLP-1分泌および血糖値やインスリン、TG濃度に有意差は認められなかった。また、異なる脂肪源は猫のGIP分泌に大きな影響は与えない可能性が示唆された。(著者抄録)

The effects of prescription diets on canine intestinal microbiota are unknown. In this study, we used next generation sequencing to investigate the impact of four commercially available prescription diet regimens on the fecal microbiome in six healthy dogs. The diet regimens used were as follows: weight-loss diet, low-fat diet, renal diet, and anallergenic diet. We found a significantly decreased proportion of phylum Actinobacteria with the weight-loss diet compared to the anallergenic diet. There were no significant differences in the proportion of phylum Bacteroidetes between the four diets. The proportion of phylum Firmicutes was significantly decreased with the weight-loss diet compared to the anallergenic diet. The proportion of phylum Fusobacteria was significantly increased with the weight-loss diet compared to the anallergenic diet. There were no significant differences in the proportion of phylum Proteobacteria after consumption of the four diets. We therefore demonstrated that commercial prescription diet influences the fecal microbiome in healthy dogs. These results might be useful when choosing a prescription diet for targeting a disease.

A 12-year-old, castrated male cat with diabetes mellitus was diagnosed with acromegaly and examined with magnetic resonance imaging (enlarged pituitary gland, 8 mm); serum hormone concentrations were measured. After the first course of radiation therapy (4 Gy, 12 fractions), insulin administration was not required from day 420 after diagnosis. Enlarged pituitary tumor (8 mm) recurred, and insulin dosage amount of the cat was increased on day 1,065. The second course of radiation therapy (6 Gy, 4 fractions) was performed on day 1,201 and insulin administration was again discontinued. However, the cat died from lymphoma on day 1,397. Postmortem examination revealed pituitary adenoma. Most tumor cells were positive for chromogranin A, synaptophysin, and growth hormone immunohistochemistry. The pancreatic islet cells revealed diffuse hyperplasia. We achieved long-term successful management of an acromegalic cat with two courses of RT. However, a protocol for a second course of RT for feline recurrent pituitary tumor should be further discussed.

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.

The purpose of this study was to determine the therapeutic and/or adverse effects of radiation therapy (RT) against pituitary tumors in dogs with pituitary-dependent hypercortisolism, as monitored by frequent post-RT detailed MRI examinations, clinical signs, and changes in hormone concentrations. Nine dogs with an adrenocorticotropic hormone (ACTH)-secreting pituitary mass diagnosed by magnetic resonance imaging (MRI) underwent RT for 4 weeks (total of 48 Gy in 4-Gy fractions). Pituitary height/brain area (P/B) value, clinical signs, basal plasma ACTH concentrations, serum cortisol concentrations (pre- and post-ACTH stimulation test) and adverse effects of RT were evaluated before and post-RT. The P/B value was significantly lower in all nine dogs post-RT. One dog lacking any neurological signs demonstrated no change in clinical signs pre and post-RT. Out of 8 dogs which exhibited neurological signs pre-RT, half of them demonstrated complete resolution of their signs, whereas the other half showed transient resolution. In all animals with recurrence of neurological signs, pituitary tumor regrowth was not observed however, MRI revealed moderate to severe pituitary hemorrhage. Late adverse effect (bilateral otitis media) was observed in three of nine dogs post-RT. RT did not induce any significant changes in the dogs’ basal plasma ACTH concentration and pre- and post-ACTH serum cortisol concentrations. In conclusion, RT is effective to reduce pituitary size and the mass effect, but does not appear to affect blood hormone concentrations, necessitating additional medical treatment against hypercortisolism. Periodic MRI imaging post-RT enables early detection of adverse effects of RT.

<tt>本研究では、中鎖トリグリセリド（MCT）が健常猫にどのような影響を与えるか検討するため、MCTを豊富に含むココナッツ油を添加した食事、および長鎖脂肪酸脂肪（LCT）を含むラード、大豆油を添加した食事を給与した場合の糖、脂質代謝の変化を比較した。健常猫6頭を用いて、3種の異なる脂肪を添加した食事を給与した。3種の脂肪添加食をそれぞれ14日間ずつ給与し、体重および体脂肪率の測定、臨床症状の有無の評価、血液検査を実施した。全ての食事において試験期間中、全頭で嗜好性に問題はなく副作用も認められなかったため、脂肪添加食は安全に給与できた。さらに、血液検査項目の血糖値、インスリン濃度、GIP濃度、GLP-1濃度、中性脂肪（TG）、遊離脂肪酸（NEFA）濃度を測定した所、全ての検査項目において3種の食事間で有意な違いは認められなかった。ヒトにおいてMCTは代謝が速く効率の良いエネルギー源とされ、また脂肪蓄積抑制効果なども認められているが、猫においては今後さらに検討</tt><tt>が必要である。 </tt>

ヒトの肥満者や糖尿病患者において、食物の消化吸収を遅延させ、食後高血糖を緩やかにする食物繊維を積極的に摂取する食事療法が行われている。本研究では健常犬に対し、ビートパルプ（可溶性＋不溶性繊維）およびセルロース（不溶性繊維）を添加したフードにおける食後の血中糖代謝パラメーター（グルコース、インスリン、NEFA）およびインクレチン分泌（GIP、GLP-1）の比較を行った。コントロール食として、繊維含有量の少ないイヌ用ドライフード（fiber:0.9g/100kcal）を用いた。繊維添加量はフード量（g）の10％（fiber:3.5 g/100kcal）および20％（fiber:6.1g/100kcal）とし、コントロール食に添加した。結果として、コントロール食よりも20％添加食の方がインスリン、NEFAおよびGIP分泌が抑制され、GLP-1分泌が促進されたことから、20％添加食は消化吸収遅延、肥満防止および食欲抑制に繋がると考えられる。しかし、種類間では大きな差異がみられなかった。これは、ビートパルプの可溶性繊維：不溶性繊維の割合が2：8であり、可溶性繊維の特徴である食物移行遅延に乏しいためであることが示唆された。

<tt>ヒトにおいてアセトアミノフェン（APAP）の吸収速度は、胃内容排出速度を反映することが報告されている。しかし、胃内容排出時間を評価するためのAPAP法は、健常犬での報告は少ない。本研究では、ヒトに使用されている市販のAPAP検出キットがイヌにおいて血清APAP濃度を測定可能かを検討することを目的とした。まず、APAP投与後のイヌ血清を用いたAPAP検出キットの再現性および希釈直線性を評価した。同時再現性では、変動係数（CV）は10％以下と良好な結果を示した。しかし、日差再現性のCVは、19.4％であった。次に、本キットを用いてイヌにおいて検出可能なAPAP投与量を決定するため、6頭の健常犬にAPAPを食事のみ（0 mg/kg（体重））、食事と10 mg/kgおよび食事と20 mg/kgをそれぞれ投与した。0 mg/kgおよび10 mg/kgと比較し、20 mg/kgで血清APAP濃度の有意な増加が認められた。結論として、APAP検出キットはイヌの血清APAP濃度を測定することが可能であり、胃内容排出時間を評価するために応用することができると考えられた。さらに、イヌにおいてAPAP検出キットを使用</tt><tt>した場合、鋭敏に検出可能なAPAPの用量は20 mg/kgであることが分かった。 </tt>

A 12-year-old female Shih-Tzu with hyperadrenocorticism and hypothyroidism developed concurrent refractory generalized demodicosis that did not respond to doramectin treatment. Although amitraz treatment was effective, the dog developed severe diabetes, which resulted in the cessation of amitraz and trilostane. Attempts to control the diabetes were unsuccessful, and its hyperadrenocorticism was left untreated, leading to the recurrence of demodicosis. However, demodicosis went into complete remission with a single dose of fluralaner. Transient erythematous papules appeared on the trunk three days after the administration of fluralaner, but no other adverse reactions were noted. We demonstrated that fluralaner is a potent treatment for demodicosis, and skin eruptions are possible after the first dose of the drug.

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.

&lt;p&gt;糖尿病猫はしばしば感染症に罹患しやすいといわれており、またストレス性の高血糖を生じやすく、血糖値を降下させるのも不得手な動物種である。このような特性をもつ猫の糖尿病管理においては、血糖コントロールを悪化させる原因が数々存在するため、糖尿病猫の入院管理は時に困難をともなう。そこで本学に来院した、慢性感染症を併発した猫、入院によるストレスで血糖コントロールが困難となった猫、食事変更が必要となった猫について、その問題と対策について検討した。症例によってストレスを感じる事象は様々あり、その対処も大きく異なるものであった。同時に、院内で起こる特有の問題からの学びもあり、退院後の自宅でのケアに生かすことができた。病院という制限ある集団管理においても、可能な限りその個別性を大切に、それぞれに合った生活環境を整える工夫ができるかが動物看護師の重要な仕事であると考える。&lt;/p&gt;

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, 0.16-3.28 mmol/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.

糖尿病患者において、運動療法は食事療法や薬物療法と併用し実施する治療法の一つである。しかし、1型糖尿病患者においては、運動時間帯や食事の栄養組成によっては、低血糖など二次的弊害を引き起こす危険性がある。そこで本研究では、糖尿病犬に食物繊維量の異なるフードを給与し、運動開始時間の違いおよびフード内の食物繊維量が運動時の血糖変動に及ぼす影響を明らかとすることを目的とした。結果より、食事給与6時間後の運動実施では、血糖値が急激に低下した。しかし、食物繊維の多い食事を与えると血糖値の急激な低下がわずかに抑制された。食事給与10時間後の運動実施においては、運動後の血糖値の低下はほぼ認められず、またフード内の食物繊維量が増加しても、運動時の血糖値に大きな変動を認めないことが明らかとなった。本研究より、食物繊維の量に関係なく血糖値の急激な低下を伴わない運動開始時間は、食事給与10時間後であることがわかった。

<p>糖尿病猫の食事には血糖値を緩やかに上昇させる炭水化物の使用が好ましいが、炭水化物の違いが血糖変動に与える影響を検討した報告は少ない。そこで本研究では、異なる炭水化物（グルコース：Glc、マルトース：Mal、トレハロース：Tre、コーンデンプン：Corn）を健常猫4頭に給与し、血中グルコース、インスリンおよびNEFA濃度に与える影響を検討した。グルコース曲線下面積（Glucose-AUC0-10h）は、Tre食、Corn食と比較してGlc食、Mal食で有意に高値を示した。Insulin-AUC0-10hは全ての群間で有意差は認められなかった。また、NEFA-AUC0-10hは、Glc食、Mal食と比較してTre食で有意に高値を示したが、Corn食では有意差は認められなかった。コーンデンプンは血糖上昇およびインスリン分泌が緩やかであり、また、血中NEFA濃度の上昇が認められなかったことから、炭水化物代謝の不得手な猫にとっても利用率が高い有用な炭水化物源となることが示唆された。</p>

<p>GLP-1およびGIPは共に血糖依存的にインスリンを分泌させる他、様々な臓器で多様な作用を持ち糖代謝に深く関与している消化管ホルモンである。本研究では栄養組成の違いがネコのインクレチン分泌におよぼす影響について検討することを目的とした。日本獣医生命科学大学で飼育管理されている健常猫5頭に、コントロール食、高炭水化物食、高脂肪食、高繊維食の4種を給与し、血糖値、インスリン濃度およびインクレチン濃度を測定した。GLP-1濃度変動は4種の食事で差はなかった。この結果は、ヒトでは炭水化物と脂質がGLP-1分泌に大きく影響をおよぼしているという報告とは異なるものであった。これはネコの食性や消化管構造の違いによる影響と考えられた。一方、GIP濃度変動はコントロール食と比較し高脂肪食で有意に高値を示した。また、食事中の脂肪含量が多いほどGIP濃度も高値を示した。本研究により、ネコにおいて栄養組成の違いがGIP分泌におよぼす影響について明らかとなったが、GLP-1分泌については今後さらなる検討が必要である。</p>

イオン交換高速クロマトグラフィー（AEX-HPLC）法は近年、ヒト領域で開発され、血清リポ蛋白質を高密度リポ蛋白質（HDL）、低密度リポ蛋白質（LDL）、中間密度リポ蛋白質、および超低密度リポ蛋白質に分けて測定することが可能となった。しかしながら、今までウシでは測定されていない。本試験の目的として、ウシの血清を用いてAEX-HPLC法による測定が可能か、AEX-HPLC法と超遠心法およびゲルろ過高速液体クロマトグラフィー（GP-HPLC）法との相関性を調査し、さらに1酪農家で飼養された泌乳ステージの異なったウシ24頭を用いてリポ蛋白質分画を評価した。AEX-HPLC法の再現性は良好な結果が得られ、超遠心法およびGP-HPLC法との測定値比較において、総コレステロール、HDL、およびLDLで有意な正の相関が得られた。また、泌乳ステージの異なる成乳牛のリポ蛋白質分画は、HDLおよびLDLともに泌乳初期から泌乳中期まで上昇し、その後乾乳期にかけて低下した。以上の結果から、AEX-HPLC法は乳牛のリポ蛋白質分画を評価するのに優れた方法であることが示唆された。

ヒトのがん患者の研究において、化学療法前後の血漿遊離アミノ酸濃度（PFAAs）が変化することが知られているが、イヌにおいては明らかではない。そこで本研究では、移行上皮癌（TCC）罹患犬における化学療法前後のPFAAsの変化を検討することを目的とした。TCC罹患犬3頭を用いた。TCC罹患犬にミトキサントロンを診断時および診断から3週間後に静脈内投与した。また、化学療法中はピロキシカムも6週間目まで毎日投与した。そして、0（抗がん剤投与前）、1、3、6週間目に採取した絶食時の血漿を高速液体クロマトグラフィー質量分析法にて異なる30種類のアミノ酸濃度を測定した。結果として化学療法前後で血漿中のシスタチオニン濃度に有意な変化が認められた。シスタチオニンは0週目と比較し、1および3週間後で減少、6週間後に増加した。TCC罹患犬の血漿中のシスタチオニンは、腫瘍細胞で利用された可能性が考えられた。以上より、TCC罹患犬におけるPFAAsは抗がん剤投与に影響されることが示された。今後は症例数を増やし、さらに長期間にわたり調べることで、TCCとPFAAsの関係をさらに検討していく必要がある。

Cat's AB blood group system (blood types A, B, and AB) is of major importance in feline transfusion medicine. Type A and type B antigens are Neu5Gc and Neu5Ac, respectively, and the enzyme CMAH participating in the synthesis of Neu5Gc from Neu5Ac is associated with this cat blood group system. Rare type AB erythrocytes express both Neu5Gc and Neu5Ac. Cat serum contains naturally occurring antibodies against antigens occurring in the other blood types. To understand the molecular genetic basis of this blood group system, we investigated the distribution of AB blood group antigens, CMAH gene structure, mutation, diplotypes, and haplotypes of the cat CMAH genes. Blood-typing revealed that 734 of the cats analyzed type A (95.1%), 38 cats were type B (4.9%), and none were type AB. A family of three Ragdoll cats including two type AB cats and one type A was also used in this study. CMAH sequence analyses showed that the CMAH protein was generated from two mRNA isoforms differing in exon 1. Analyses of the nucleotide sequences of the 16 exons including the coding region of CMAH examined in the 34 type B cats and in the family of type AB cats carried the CMAH variants, and revealed multiple novel diplotypes comprising several polymorphisms. Haplotype inference, which was focused on non-synonymous SNPs revealed that eight haplotypes carried one to four mutations in CMAH, and all cats with type B (n = 34) and AB (n = 2) blood carried two alleles derived from the mutated CMAH gene. These results suggested that double haploids selected from multiple recessive alleles in the cat CMAH loci were highly associated with the expression of the Neu5Ac on erythrocyte membrane in types B and AB of the feline AB blood group system.

Acarbose (AC) and Sitagliptin (STGP) are oral hypoglycemic agents currently used either alone or in conjunction with human diabetic (Type 2) patients. AC has been used with diabetic cats, but not STGP thus far. Therefore, the objective of this study was to determine the potential use of AC or STGP alone and in combination for diabetic cats, by observing their effect on short-term post-prandial serum glucose, insulin, and incretin hormone (active glucagon-like peptide-1 (GLP-1) and total glucose dependent insulinotropic polypeptide (GIP)) concentrations in five healthy cats, following ingestion of a meal with maltose. All treatments tended (p &lt; 0.10; 5-7.5% reduction) to reduce postprandial glucose area under the curve (AUC), with an accompanying significant reduction (p &lt; 0.05, 35-45%) in postprandial insulin AUC as compared to no treatment. Meanwhile, a significant increase (p &lt; 0.05) in postprandial active GLP-1 AUC was observed with STGP (100% higher) and combined treatment (130% greater), as compared to either AC or no treatment. Lastly, a significant reduction (p &lt; 0.05) in postprandial total GIP AUC was observed with STGP (21% reduction) and combined treatment (7% reduction) as compared to control. Overall, AC, STGP, or combined treatment can significantly induce positive post-prandial changes to insulin and incretin hormone levels of healthy cats. Increasing active GLP-1 and reducing postprandial hyperglycemia appear to be the principal mechanisms of combined treatment. Considering the different, but complementary mechanisms of action by which AC and STGP induce lower glucose and insulin levels, combination therapy with both these agents offers great potential for treating diabetic cats in the future. (C) 2016 Elsevier Ltd. All rights reserved.

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.

Nesfatin-1 is an anorexic peptide derived from a predursor, nucleobindin-2 (NUCB2), which is distributed in various organs, coexists with ghrelin in the gastric X/A-like cells and closely relates to an appetite control in rodents and humans. Nesfatin-1 may be a significant factor addressing the satiety also in veterinary medicine, however, there are few reports about nesfatin-1 in dogs. In the present study, we detected canine NUCB2/nesfatin-1 mRNA in various tissues, especially abundant in pancreas, gastrointestinal tracts, testis and cerebellum. We examined circulating nesfatin-1 concentrations and NUCB2/nesfatin-1 mRNA expressions in upper gastrointestinal tracts (gastric corpus, pyloric antrum and duodenum) in dogs fed on different types of diets. Plasma nesfatin-1 concentrations in the dogs were approximately 4 ng/ml and they did not change after feeding through the study, however, NUCB2/nesfatin-1 mRNA expressions in pyloric antrum were 1.84-fold higher in the dogs fed on a High fiber/High protein diet (P&lt;0.001), 1.48-fold higher in the dogs fed on a High fat/Low protein diet (P&lt;0.05) and 1.02-fold higher in the dogs fed on a Low fat/High carbohydrate diet (not significant) comparing to those on a control diet. It was concluded that High fiber/High protein and High fat/Low protein diets increased NUCB2/nesfatin-1 production in canine gastrointestinal tracts. These results may set the stage for further investigations of canine NUCB2/nesfatin-1, which may relate to satiety effects in dogs.

血糖変動に最も影響を与える栄養素は炭水化物であり、グルコースの消化吸収速度を緩やかにできれば、食後の血糖上昇を抑制できる。桑の葉にはグルコースと構造が類似したDNJが含まれ、グルコース吸収を抑制すると考えられている。ヒトでは桑の葉投与により食後の血糖上昇が抑制されたが、イヌでの効果は明らかにされていない。そこで、本研究では、桑の葉が健常犬の糖・脂質代謝に与える影響を検討した。6頭の健常犬に高炭水化物食とともに桑の葉粉末を0.5 g/頭で4日間給与した結果、桑の葉給与群では無給与群と比較して食後30分の血糖値が有意に低値を示し、血糖上昇が抑制された。さらに、食後30、180分の血中インスリン濃度に低下傾向が見られ、インスリン分泌もやや抑制された。しかし、血中中性脂肪濃度は給与群と無給与群で有意な差は認められなかった。本研究により桑の葉給与はイヌにおいても血糖上昇を抑制することが分かった。

運動による血糖変動は、食事内容および食事時間によって低血糖や消化吸収遅延など、様々な弊害を引き起こすことが報告されている。犬でも、食後に猟を行った猟犬において低血糖と思われる意識障害が報告されているが、その詳細は不明である。そこで本実験では、健常犬（n＝4）に対する運動実施前の糖摂取が糖脂質代謝に及ぼす影響を検討した。運動は経口糖負荷試験開始30分後の血中グルコースおよびインスリン濃度が高値を示す時間帯で行った。結果より、血中グルコースおよびインスリン濃度共に高値を示す時間帯での運動実施は、筋への糖取り込み作用とインスリン作用とが合わさり、血中グルコース濃度の急激な低下を引き起こすことが示唆された。また、絶食時の運動は脂肪分解を亢進することがわかった。しかし、これらの結果はあくまで糖質のみを摂取した場合の変動であり、今後実際に様々な栄養素を含む食事を摂取した場合の変動を検討していく必要がある。

Carbohydrate is an important source of energy, which can significantly affect postprandial blood glucose and insulin levels in cats. In healthy animals, this is not a big concern; however, in obese and diabetic animals, this is an important detail. In the present study, the impact of four different carbohydrate sources (glucose, maltose, corn starch, and trehalose) on short-term post-prandial serum glucose, insulin, and non-esterified fatty acid (NEFA) concentrations was investigated with four obese cats. Each of the carbohydrate sources was added to a commercial wet food diet for feeding the animals. A significant difference was observed in postprandial glucose, insulin, and NEFA area under the curve (AUC) values between each carbohydrate source in obese cats. Furthermore, glucose and maltose induced the highest postprandial glucose and insulin AUC values, whereas trehalose induced the lowest postprandial glucose and insulin AUC value amongst all carbohydrate sources, respectively, in obese cats. However, trehalose has a higher risk of inducing side effects, such as diarrhea, as compared to other carbohydrate sources. As such, different carbohydrate sources appear to have a very significant impact on post-prandial glycemia and subsequent insulin requirement levels in obese cats. These results might be useful when selecting a prescription diet for obese or diabetic cats. In addition, maltose appears to be capable of inducing experimentally evoked postprandial hyperglycemia in obese cats, which may serve as a good tool for use to check the impact and effectiveness of newly developed oral hypoglycemic drugs or supplements for cats in future experiments.

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.

食後の血糖上昇に最も影響を与えるのは炭水化物であり、摂取した炭水化物量によって追加インスリン療法を行い、食後の血糖値をコントロールするカーボカウントという方法が近年注目されている。そこで本研究では、炭水化物摂取量の違いが糖尿病犬の血糖値およびインスリン必要量に与える影響について明らかにすることを目的とし、速効型インスリン1.0 Uを注射することによって摂取できる炭水化物量、つまり犬におけるカーボカウントの算出について検討した。血糖値をリアルタイムで確認できる人工膵臓装置を糖尿病犬に接続し、コントロール食と高炭水化物食（コントロール食＋コーンスターチ粉）を給与して、速効型インスリンによる連続的静脈内投与を行い、正常血糖範囲（血糖値80-120 mg/dl）を維持した。結果は、高炭水化物食のインスリン必要量はコントロール食と比較して増加し、速効型インスリン1.0 Uあたり炭水化物をおよそ12.7 g摂取できることがわかった。この炭水化物摂取量は糖尿病犬のインスリン効果値によって左右されるため、あらかじめ個体ごとのインスリン効果値を算出する必要があると考えられた。本研究より、食事中の炭水化物量は糖尿病犬におけるインスリン必要量の決定に関する重要な因子であることが示唆された。

G protein-coupled receptor (GPR) 120 is an unsaturated fatty acid receptor, which is associated with various physiological functions. It is reported that the genetic variant of GPR120, p.Arg270His, is detected more in obese people, and this genetic variation functionally relates to obesity in humans. Obesity is a common nutritional disorder also in dogs, but the genetic factors have not ever been identified in dogs. In this study, we investigated the molecular structure of canine GPR120 and searched for candidate genetic variants which may relate to obesity in dogs. Canine GPR120 was highly homologous to those of other species, and seven transmembrane domains and two N-glycosylation sites were conserved. GPR120 mRNA was expressed in lung, jejunum, ileum, colon, hypothalamus, hippocampus, spinal cord, bone marrow, dermis and white adipose tissues in dogs, as those in mice and humans. Genetic variants of GPR120 were explored in client-owned 141 dogs, resulting in that 5 synonymous and 4 non-synonymous variants were found. The variant c.595C&gt;A (p.Pro199Thr) was found in 40 dogs, and the gene frequency was significantly higher in dogs with higher body condition scores, i.e. 0.320 in BCS4-5 dogs, 0.175 in BCS3 dogs and 0.000 in BCS2 dogs. We conclude that c.595C&gt;A (p.Pro199Thr) is a candidate variant relating to obesity, which may be helpful for nutritional management of dogs.

ヒト糖尿病患者において、血糖値に影響を与えやすいとされる炭水化物に着目した食事療法が多く用いられている。本研究では健常犬に対し、市販食で炭水化物源の異なる食事を与え、食後の糖代謝パラメーターの比較検討を行った。主要な炭水化物源が米・トウモロコシ・小麦である3種の市販食を用いた。その結果、食後の血糖変動は3種のフードの間に差は認められなかった。食後のインスリン分泌の結果から、小麦、トウモロコシ、米の順で消化・吸収速度が速くなる傾向がみられた。また、インクレチンであるGIPは炭水化物源の違いよりも食事中の脂質によって分泌刺激されることが分かった。また、もう一つのインクレチンであるGLP-1は炭水化物源の違いにより影響を受けることはなかった。これらのことから、健常犬において市販食での炭水化物源の違いは血糖値およびGLP-1分泌に影響を与えないが、フード中の炭水化物源がインスリン分泌に関与し、</tt><tt>脂質含有量がGIP分泌量に関与していることが示唆された。

Sitagliptin is a dipeptidyl peptidase-4 inhibitor aimed at treating Type 2 diabetes mellitus (T2DM) and T1DM, by increasing blood levels of Glucagon-like peptide 1 (GLP-1) and insulin. The objective of this preliminary study is to characterize Sitagliptin's ability for glycemic control, in healthy dogs under an oral glucose tolerance test (OGTT) environment. Overall, Sitagliptin did not result in any significant changes to temporal glucose and insulin concentrations. However, a similar to 55% increase in median total GLP-1 AUC(0-120min) was observed, as compared to baseline control in healthy dogs (n=5), thus indicating a similar mode of action of Sitagliptin between healthy dogs and humans. Future studies to validate the use of Sitagliptin with dogs suffering from insulin independent diabetes are warranted.

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 (P13-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 P13-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.

運動療法は、インスリン感受性を改善し、骨格筋へのブドウ糖取り込みを増加させることから、糖尿病患者において有用な治療法である。本研究では、糖尿病犬に対する運動療法が血液生化学パラメーターおよび骨格筋遺伝子発現にどのような影響を及ぼすかを検討した。空腹時血糖値は運動前後で有意な変動は認められなかった。しかし、糖化アルブミン（GA）および遊離脂肪酸は運動後有意に低下した。インスリンシグナリングおよび糖代謝に関連する遺伝子（インスリンレセプター基質1および2、ホスファチジルイノシトール3キナーゼ、aktキナーゼ2、グルコーストランスポーター4、AMP活性化プロテインキナーゼ、脱共益蛋白3およびアセチルCoAカルボキシラーゼ）の発現には運動前後で有意な変化は認められなかった。本実験結果より、運動療法を行うことでGAの低下をもたらし、糖尿病犬において血糖コントロールが改善された。

Glucagon-like peptide 1 (GLP-1) is a glucose-lowering, intestinal-derived factor with multiple physiological effects, making it attractive for diabetes therapy. However, the therapeutic potential of endogenous GLP-1 is limited, because of rapid inactivation by dipeptidyl peptidase-4. Recently, enhanced incretin preparations, such as liraglutide, have emerged, which are more resistant to degradation and longer lasting. Liraglutide is a long-acting acylated human GLP-1 receptor agonist, with a 97% amino acid sequence identity to endogenous human GLP-1, and 100% amino acid sequence homology with canine GLP-1.Since liraglutide has yet to be examined for use in dogs, and the incretin effect has been reported to exist in dogs, we sought to initially characterize liraglutide's ability for glycemic control in healthy dogs, under an oral glucose tolerance test (OGTT) environment initially. This was followed up a more realistic scenario involving food with insulin injection +/- liraglutide injection resulting in a glucose curve based study involving dogs suffering from Type 1 diabetes mellitus (T1DM). Overall, liraglutide had a stabilizing effect on glucose levels, maintaining circulating levels between 77.0 and 137.0mg/ml throughout the OGTT test period, resulting in a significant reduction of 13.8% in glucose AUC0-120min (total area under the curve for 0-120min) as compared to baseline control in healthy dogs (n=5). Interestingly, the liraglutide associated reduction in circulating glucose was not accompanied by any significant increase in insulin. Moreover, T1DM dogs (n=4) responded favorably to liraglutide treatment, which lead to a significant reduction of 46.0% in glucose AUC0-12h (total area under the curve for 0-12h), and a significant reduction of 66.5% in serum glucose as compared to baseline controls (insulin treatment only). Therefore, liraglutide's prandial glucagon suppressive ability appears to play a key role in its glucose-lowering capability, and offers great potential for use with dogs suffering from T1DM. © 2013 Elsevier Ltd.

Diet therapy is an important treatment component available for obese cats. In this study, the impact of four commercially available prescription diet regimens (1 for general use and 3 aimed at treating obesity and diabetes mellitus (DM)) on short-term postprandial serum glucose, insulin, triglyceride and nonesterified fatty acid (NEFA) concentrations was investigated with five obese cats. The diet regimens used were as follows: C/D dry (general use: moderate protein, moderate fat, high carbohydrate and low fiber), M/D dry (DM: high protein, high fat, low carbohydrate and high fiber), W/D dry (DM: high protein, low fat, high carbohydrate and high fiber) and Diabetic dry (DM: high protein, low fat, low carbohydrate and high fiber). A significant reduction (10-13%) in postprandial glucose (area under the curve; AUC) was observed with the MID and Diabetic diets, which both contained lower concentrations of carbohydrates than the C/D diet. An accompanying significant reduction (30-36%) in postprandial insulin AUC was also observed with the three DM diets, which all had higher amounts of fiber, as compared with the C/D diet. Lastly, a significant increase (32-65%) in postprandial NEFAAUC was observed with the M/D and Diabetic diets as compared with the C/D diet. Therefore, dietary amounts of carbohydrates and fiber, as opposed to protein content or dietary fat, appear to have a very significant impact on postprandial glycemia and subsequent insulin requirement levels in obese cats. In addition, dietary amounts of carbohydrates may also impact lipid metabolism in obese cats.

Monitoring of blood glucose concentration is important to evaluate the diabetic status of dogs. Continuous glucose monitoring systems (CGMS) have been applied in veterinary medicine for glucose monitoring in diabetic dogs. The purpose of the study was to evaluate the daily glycemic profiles obtained with CGMS and compare glucose fluctuations between day- and night-time in diabetic dogs. Five diabetic dogs were used in this study and were treated with either NPH insulin or insulin detemir. For data analyses, day-time was defined as 9:00 am-9:00 pm and night-time as 9:00 pm-9:00 am. Using glucose profiles, we determined the mean glucose concentrations (1-and 12-hr intervals), and times spent in hyperglycemia &gt;200 mg/dl or hypoglycemia &lt;60 mg/dl. None of the parameters differed significantly between day-time and night-time in dogs treated with NPH insulin or insulin detemir. In conclusion, this study confirmed, using CGMS, that there are no differences in glucose fluctuations between day-and night-time, in diabetic dogs on a similar feeding regimen and insulin administration.

Euglycemic-hyperinsulinemic glucose clamp (EHGC) method is a gold standard for assessing insulin resistance in humans. However, this method has yet to be commonly used with dogs, due to the requirement of frequent blood sampling for glucose measurement and adjusting glucose infusion rate (GIR). The purpose of this study was to evaluate insulin resistance, induced either by Cushing Syndrome (CS) or diestrus in dogs, as determined by GIR by EHGC, using an artificial pancreas apparatus. Twenty animals were used in this study with ten (7 females and 3 males) serving as healthy controls, four (3 females, 1 male) diagnosed with CS, and six (all females) undergoing diestrus. A higher GIR value indicates increased insulin sensitivity and lower insulin resistance. GIR of healthy control animals was determined to be within a reference range of [10.6-21.3] with a median of 15.2 mg/kg/min. In comparison, the CS group had a median of 5.4 mg/kg/min; whereas the diestrus group exhibited a median of 8.9 mg/kg/min. Insulin resistant animals suffering from CS and undergoing diestrus demonstrated reductions of 65 and 40% in GIR, respectively; thus indicating differences in degree of insulin insensitivity can be discerned using the EHGC method.