Three-dimensional CT angiography of the canine hepatic vasculature

Eight Beagle dogs were anesthetized and were imaged using a single channel helical CT scanner. The contrast medium used in this study was iohexol (300 mg I/ml) and doses were 0.5 ml/kg for a cine scan, 3 ml/kg for an enhanced scan. The flow rate for contrast material administration was 2 ml/sec for all scans. This study was divided into three steps, with unenhanced, cine and enhanced scans. The enhanced scan was subdivided into the arterial phase and the venous phase. All of the enhanced scans were reconstructed in 1 mm intervals and the scans were interpreted by the use of reformatted images, a cross sectional histogram, maximum intensity projection and shaded surface display. For the cine scans, optimal times were a 9-sec delay time post IV injection in the arterial phase, and an 18-sec delay time post IV injection in the venous phase. A nine-sec delay time was acceptable for the imaging of the canine hepatic arteries by CT angiography. After completion of arterial phase scanning, venous structures of the liver were well visualized as seen on the venous phase.     

Microvascularization of the Third Eyelid in Dogs

The third eyelid protects the eye, contribute to the aqueous portion of the pre‐ocular film and its distribution over the corneal surface, as well as removal of foreign material from the anterior surface of the eye. It is rather vulnerable during fights and clinicians often have to perform surgery for third eyelid neoplasia. In dogs, this lid has a triangular shape, being widest at its free margin. It is vascularized by branches of the malar artery. Being of such importance as an adnexa of the eye, the authors concentrated their attention on its vascularization, which is not well defined. Methods: Scanning Electronic Microscopy of vascular corrosion casts and intact tissue. Observations: It was quite interesting to notice the presence of two patterns of vascularization inside the third eyelid. One can say that there are two segmental levels of vessels. In one of these there is a predominance of arteries and veins, while in the other a meshwork of third order arteries, capillaries and post‐capillary venules are present. In fact, the branch of the malar artery that reaches the base of the third eyelid divides into smaller branches that cross almost the complete length of the lid giving only a few collateral vessels, in a monopodical way, and only near the free margin of the lid is possible to observe dicothomical ramifications that became rather anastomotic (being anastomoses of type I and II are present) and contribute to the microvascular meshwork. This microvascular net is drained by post‐capillary venules and veins that empty as a brush into bigger veins that occupy the same segmental level as the third eyelid artery. Remarks: This uncommon angioarchitecture is probably an adaptation to the fact that in the medial canthus of the eye this lid has a smaller space and the way of obtaining an efficient vascularization is by going straight to the top of the lid.     

Contrast harmonic imaging of canine hepatic tumors

Six adult healthy Beagles were used to investigate the hepatic perfusion dynamics of Levovist, a contrast agent used in contrast harmonic imaging (CHI). In addition, 8 dogs with hepatocellular carcinoma (HCC) and 2 dogs with metastatic hepatic hemangiosarcoma (HSA) were used to characterize both the CHI findings with Levovist. In the Beagles, the start of intravenously injected Levovist into the aorta between the cranial mesenteric and renal arteries and the portal vein at the hepatic hilum were 5.47 +/- 1.52 sec and 16.03 +/- 3.39 sec, respectively. As a characteristic CHI finding in the 8 dogs with HCC, the early arterial phase showed a fine network of blood flow enhanced at the surrounding region and within the tumor in all the 8 dogs (100%), and the post vascular phase demonstrated a defect in the whole tumor and an enhancement of the surrounding hepatic tissues in 7 dogs (87.5%). In the 2 dogs with HSA, characteristic finding in which the early arterial and late vascular phases showed a rim contrast enhancement pattern, and the post vascular phase revealed that the whole tumor lacked contrast enhancement and the surrounding hepatic tissues was clearly enhanced. In dogs, the start of the early arterial and late vascular phases, and the characterizations of the CHI findings in HCC and HSA were suggested to be similar to those in humans. Therefore, CHI is thought to be useful for the diagnosis of HCC and metastatic hepatic HSA in dogs as well as in humans.     

Collagen fiber arrangement in canine hepatic venules

Cell-maceration/scanning electron microscopy, serial sections and scanning electron microscopy of vascular resin casts were employed to demonstrate the arrangement of collagen fibers in the terminal hepatic venules, involving the central, intercalated and collecting veins in dog liver. In cell-maceration specimens, each collagen fiber was observed to run in various directions, forming a sheath with a compact meshwork of collagen fibers. The collagenous meshwork in the hepatic venules was looser than those of the terminal portal venules and hepatic arterioles. Some collagen fibers formed bundles with an elongated spiral arrangement encircling the wall of the terminal hepatic venules. In resin casts, these venules were observed as a twisted configuration caused by spiral collagen bundles. A helical modification of such connective tissue bundles might provide a mechanically stable vascular structure and permit reversible changes in linear and circumferential vascular dimensions at the terminal tributaries of veins. Round or oval pores with diameters of approximately 9 microns were also observed in the sheath of collagen fibers. These pores, together with the relatively loose collagenous meshwork in the hepatic venules, might play a role in lymphocyte migration from these venules into the surrounding tissue and provide high permeability to the venule walls. No such helical configuration and pores were observed in either the portal venules or the hepatic arterioles.     

Binding affinity of anti-xylitol antibodies to canine hepatic vessels

Xylitol is used as a sugar substitute in food products. Dogs have been reported to experience lethal liver injury after accidental ingestion of xylitol. Because liver injury may be a serious consequence of canine immune-mediated reactions, antibodies produced against xylitol may attack the liver. Therefore, in the present study, we evaluated whether binding sites for xylitol antibodies are located at the liver or not. Anti-xylitol antibodies were generated by immunization of rabbits with a xylose-bovine serum albumin conjugate. Immunohistological examination showed that binding sites for the anti-xylitol antibodies were located in the hepatic arteries and the portal veins. Western blotting analyses by using a canine liver homogenate showed 4 protein bands with different molecular weights which reacted with anti-xylitol antibodies. Therefore, binding of anti-xylitol antibodies to the vessels may be the first step in an immune-mediated pathogenic response in xylitol toxicity. Further studies are necessary to determine the effects of anti-xylitol antibodies on the liver in the pathogenesis of xylitol toxicity.     

1例犬肝腺瘤的诊断与治疗

为了诊断8岁萨摩耶患犬腹腔不明肿块,采用临床检查、实验室检查、影像学检查、病理诊断等方法,检测其一般体征、血液指标、影像表现及病理组织形态。结果显示:超声显示有一巨大异常不均质回声团块,后壁回声增强,团块边界不清,形态不规整,与肝脏邻接且血流异常丰富;CT增强扫描显示肿块略低密度或等密度占位性病变,边界清楚;病理组织学诊断可见肿瘤呈扩张性生长,边缘没有浸润性,正常肝小叶结构丧失,缺乏门静脉束,小梁取向异常。结论:该犬所患为肝腺瘤,需手术切除并后续跟踪治疗。期望该病例能丰富肝腺瘤病例资料,以供类似疾病诊疗时作为参考。     

Correlation of ultrasonographic and patho-morphological findings in canine hepatic diseases

Findings of hepatic ultrasonography were analysed in 22 dogs with liver disease and compared with the results of final morphological diagnoses. Ultrasonographic appearance of the liver demonstrated focal alterations in 11 dogs (50 per cent): multifocal lesions in hepatic neoplasia (six), hepatic cirrhosis (one), generalised mycosis (one) and unifocal lesions in haemangiosarcoma (one), nodular hyperplasia (one) and misdiagnosed intestinal invagination (one), Diffuse ultrasonographic alterations were found in 11 dogs (50 per cent): hyperechoic liver of normal/enlarged size in lymphosarcoma (four) and hepatic lipidosis (two); hyperechoic ‘bright’ but small liver in atrophic cirrhosis (two); hypoechoic to normal intensity liver of normal size in liver dystrophy (two) and hepatic venous distension (one). Gallbladder abnormalities were detected in 14 of 20 dogs (70 per cent). Correct ultrasonographic diagnoses were made in 11 dogs (50 per cent). The best results were achieved by combining the clinicolaboratory and ultrasonographic findings, providing a correct diagnosis in 17 dogs (77-3 per cent).     

Accelerated neutrophil apoptosis in 2 canine cases of hepatic disorder

Accelerated neutrophil apoptosis was confirmed by TUNEL assay in two canine cases of hepatic disorder. One dog was diagnosed as having lymphocytic hepatitis and the other lymphocytic cholangitis by histopathology of liver biopsy specimen.     

Microvascularization and angiogenic activity of equine corpora lutea throughout the estrous cycle

Corpus luteum growth and endocrine function are closely dependent on the formation of new capillaries. The objectives of this study were to evaluate (i) tissue growth and microvascular development in the equine cyclic luteal structures; (ii) in vitro angiogenic activity of luteal tissues in response to luteotrophic (LH, PGE₂) and luteolytic (PGF₂) hormones and (iii) to relate data to luteal endocrinological function. Our results show that microvascular density was increased in the early and mid luteal phase, followed by a fall in the late luteal phase and a further decrease in the corpus albicans. Hyperplasia of luteal tissue increased until the mid luteal phase and it was followed by tissue regression. Luteal explants were cultured with no hormone added, or with PGF₂, LH, PGE₂, LH + PGE₂ or LH + PGF₂. Media conditioned by equine luteal tissue from different stages of the luteal phase were able to stimulate mitogenesis of bovine aortic endothelial cells (BAEC), suggesting the presence of angiogenic activity. No difference was observed among luteal structures on their mitogenic capacity, for any treatment used. Nevertheless, Late-CL conditioned-media with PGF₂ showed a significant decrease in BAEC proliferation (p < 0.05) and LH + PGF₂ a tendency to reduce mitogenesis. Thus, prostaglandin F₂ may play a role on vascular regression of the CL during the late luteal phase in the mare. These data suggest that luteal angiogenesis and vascular regression in the mare are coordinated with the development of non-vascular tissue and might be regulated by many different factors.     

Novel markers of inflammatory response and hepatic dysfunction in canine leishmaniasis

Dogs are the main host of Leishmania infantum, and the clinical presentation may range from asymptomatic to systemic manifestations. The immune mechanisms in infected, but clinically healthy dogs, prevails Th1 response mediated by cytokines. In this sense, adenosine deaminase (ADA) and butyrylcholinesterase (BChE) are considered as key enzymes in several physiological processes, including the modulation of inflammatory process. Considering the variable immune response against Leishmania and the known participation of ADA and BChE, the aim of this study was to assess the relation between these two enzymes with the inflammatory response as well as hepatic function in dogs naturally infected with L. infantum. For this purpose, the activity of ADA and BChE was assessed in sera of 24 dogs naturally infected with L. infantum, plus 17 healthy dogs. The naturally infected dogs had clinical signs compatible with leishmaniasis and sera activities of ADA (P < 0.01) and BChE (P < 0.05) decreased, when compared to the healthy group. The reduction of ADA activity probably represented an effect on inflammatory response, especially due to the decreased hydrolysis of extracellular adenosine, might in order to protect against tissue damage and, also, setting a down-regulation on pro-inflammatory cytokines. BChE enzyme had no effect on modulating the immune response in leishmaniasis, but it decreased, a fact may related to deficiency of synthesis in the liver. Therefore, ADA and BChE activities reduced probably in order to protect against extra tissue damage and due failure in synthesis, respectively.     

The plasma superoxide scavenging activity in canine cancer and hepatic disease

To clarify the relationship between plasma antioxidant activity and diseases in dogs, plasma samples were collected from 6 healthy dogs and 16 diseased dogs (6 dogs with cancer, 5 dogs with hepatic disease, and 5 dogs with inflammation ), and measured superoxide anion scavenging activities. Antioxidant activities of canine plasma were evaluated by measuring their superoxide anion (O(2)(-.)) scavenging activities with electron spin response spectroscopy combined with spin trapping reagent, 5,5-dimethyl-1-pyrroline-N-oxide (DMPO). Total O(2)(-.) scavenging activities in the presence of plasma of diseased dogs tended to be higher than those in healthy controls, especially significant higher activities in the presence of canine plasma of hepatic disease and inflammation were observed. In diseased dogs, KCN-insensitive activities, suggesting the activity of manganese-containing superoxide dismutase (Mn-SOD), were significantly higher than those in healthy controls. Therefore, it seems that there is a possibility of utilizing of plasma O(2)(-.) scavenging activity as one of clinical indicators for oxidative-related diseases such as cancer, hepatic disease and inflammation in dogs.     

Expression of blood hepatocyte-derived microRNA-122 in canine multicentric lymphoma with hepatic involvement

Veterinary Research Communications - This study was performed to evaluate the hepatocyte-derived microRNA (miR)-122 as novel diagnostic biomarker in canine lymphoma. Fifteen dogs were enrolled in...     

Effects of long-term primidone and phenytoin administration on canine hepatic function and morphology

Primidone, phenytoin, or phenytoin and primidone in combination were given to healthy Beagle dogs for 6 months. Serum biochemical changes in dogs given primidone alone or phenytoin and primidone in combination for the entire 6-month test period included increased activities of alanine aminotransferase, alkaline phosphatase (AP), and gamma-glutamyltransferase, and decreased concentrations of albumin and cholesterol. Changes in dogs given phenytoin alone were limited to increased AP activity and decreased albumin concentration. Sulfobromophthalein excretion and conjugated bile acid concentration were within normal limits. All dogs given primidone alone or phenytoin alone remained clinically healthy throughout the treatment period. Three of 8 dogs given both drugs in combination became clinically ill after 9, 14, and 15 weeks of treatment, and were euthanatized. Two of the dogs developed clinical jaundice. In addition to the serum biochemical abnormalities observed in clinically healthy dogs, these dogs developed hyperbilirubinemia, delayed sulfobromophthalein excretion, and increased conjugated bile acid concentrations. Histologic examination of the liver showed intracanalicular casts of bile pigment typical of intrahepatic cholestasis in all 3 dogs. Histologic findings characteristic of treated dogs included hepatocellular hypertrophy attributable to hyperplasia of the smooth endoplasmic reticulum. Single-cell necrosis and multifocal lipidosis were observed in individuals of all treatment groups. Electron microscopy of the liver showed dilated bile canaliculi and damaged sinusoidal epithelium in dogs given both drugs. The elevated serum AP activity, associated with anticonvulsant drug therapy, was found to be exclusively the liver isoenzyme by cellulose acetate electrophoresis. The hepatic AP was localized to primarily the canalicular membranes by enzyme histochemistry. There was a statistically significant positive correlation between the AP activities of liver and serum. The results of this study indicate that long-term administration of anticonvulsant drugs to dogs is associated with clinical, serum biochemical, and histologic evidence of hepatic dysfunction. High drug dosage contributed most to abnormal serum biochemical test results, and combining phenytoin with primidone was responsible for more severe electron microscopic lesions of the liver of surviving dogs and for the death of 3 dogs.     

Immunochemical study of canine intestinal, hepatic, and osseous alkaline phosphatases.

Partially purified alkaline phosphatase (ALP) from canine intestine, liver, and bone were injected into rabbits to elicit anti-canine intestinal, hepatic, and osseous ALP antibodies, respectively. The antibody formed a soluble enzyme-antienzyme complex when directly interacted with the ALP antigen. In order to form an insoluble complex, it was then necessary to interact the initial soluble complex with the goat anti-rabbit gamma-globulin antibody in the 2nd step. Antiintestinal ALP antibody was highly specific and did not cross react with canine hepatic, osseous, splenic, and renal ALP. Antiliver and antibone ALP antibodies, on the other hand, did cross react with hepatic, osseous, splenic, and renal ALP, but not with the intestinal ALP.     

Dimethylnitrosamine-induced hepatotoxicosis in dogs as a model of progressive canine hepatic disease.

A model of toxin-induced progressive hepatitis is described in Beagles. The toxin, dimethylnitrosamine, was administered orally to 18 Beagles; 6 dogs comprised a control group. Clinical signs and laboratory test results were monitored as disease progressed and were used to determine the end point of disease. Following euthanasia, histologic lesions were scored and used to derive a total severity score for each dog. Severity scores were then used to allot the 18 dogs to 3 groups of hepatic disease, defined as mild, moderate, or severe. Changes in clinical laboratory test results, including tests of hepatic function, and clinical signs indicative of liver disease were described chronologically for all dogs. Group means of clinical laboratory test results and quantifiable clinical signs (eg, weight loss and ascitic fluid accumulation) were compared. This model offers several advantages, compared with other experimental models of canine hepatic disease. These include hepatospecificity, similarity to natural disease (eg, the development of multiple extrahepatic portosystemic shunts), and the ability to titrate the disease to a desired end point. The major disadvantages of this model were the toxic nature of the drug to human beings and the variation in individual animal response to the toxin, which precludes preassignment of animals into groups.     

Identification of hepatic stem/progenitor cells in canine hepatocellular and cholangiocellular carcinoma

Hepatic progenitor cells (HPCs) are bipotential stem cells residing in human and animal livers that are able to differentiate towards the hepatocytic or cholangiocytic lineages. HPCs are present in both hepatocellular (HCC) and cholangiocellular carcinoma (CC) in humans; and a small percentage of HCC can originate from cancer stem cells. However, its distribution in canine liver tumour has not been studied. Herein, we searched for stem/progenitor cells in 13 HCC and 7 CC archived samples by immunohistochemical analysis. We found that both liver tumours presented a higher amount of K19‐positive HPCs. Besides, 61.6% of HCC cases presented immature CD44‐positive hepatocytes. Nevertheless, only two cases presented CD133‐positive cells. As observed in humans, hepatic canine tumours presented activated HPCs, with important differentiation onto hepatocytes‐like cells and minimal role of cancer stem cells on HCC. These findings reiterate the applicability of canine model in the search for new therapies before application in humans.