Endotoxin Concentrations Measured by a Chromogenic Assay in Portal and Peripheral Venous Blood in Ten Dogs With Portosystemic Shunts

A chromogenic Limulus amebocyte lysate assay was used to measure portal and peripheral venous endotoxin concentrations in ten medically managed dogs undergoing surgery for correction of a single extrahepatic portosystemic shunt. In all dogs, both peripheral and portal venous blood samples were obtained at the time of surgical manipulation of the anomalous vessel. In six dogs, peripheral venous samples were obtained an average of 8.0 months after surgery. Five physically normal dogs without biochemical or histologic evidence of liver disease served as controls. Data analysis failed to demonstrate significant differences in peripheral and portal venous endotoxin concentrations between the control and study groups. Postoperatively five of six dogs showed a measurable reduction in peripheral venous endotoxin concentration over intraoperatively obtained values, but the differences were not statistically significant (P = 0.06). Based on results of this study it was concluded that systemic endotoxemia was not present in dogs with a single extrahepatic portosystemic shunt that were medically stable prior to surgery.     

Whole Blood Manganese Concentrations in Dogs with Congenital Portosystemic Shunts

Background: Manganese (Mn) is an essential mineral that is a cofactor for many enzymes required in the synthesis of proteins, carbohydrates, and lipids. Because hepatic clearance is essential in Mn homeostasis, conditions in humans resulting in hepatic insufficiency including cirrhosis and both acquired and congenital portosystemic shunting have been reported to result in increased blood Mn concentrations and increased Mn content in the central nervous system. Because Mn toxicity causes neurologic disturbances, increased Mn concentrations have been implicated in the pathogenesis of hepatic encephalopathy.
Hypotheses: Dogs with congenital portosystemic shunts (cPSS) have significantly higher whole blood Mn concentrations than do healthy dogs or those with nonhepatic illnesses.
Animals: Eighteen dogs with cPSS, 26 dogs with nonhepatic illnesses, and 14 healthy dogs.
Methods: Whole blood Mn was measured by graphite furnace atomic absorption spectrometry. The diagnosis of cPSS was made by ultrasonography or during celiotomy either by visual inspection of a shunting vessel or portovenography.
Results: Dogs with a cPSS had significantly higher whole blood Mn concentrations than did healthy dogs and dogs with nonhepatic illnesses. Whole blood Mn concentrations were not significantly different between healthy dogs and dogs with nonhepatic illnesses.
Conclusion and Clinical Importance: Dogs with a cPSS have significantly increased whole blood Mn concentrations. Additional studies are warranted to investigate the role of Mn in cPSS-associated hepatic encephalopathy.     

Transsplenic Portal Catheterization Surgical Technique and Use in Two Dogs With Portosystemic Shunts

The portal vasculature can be accessed by using a through-the-needle catheter system to pass a catheter through the splenic parenchyma and into a major splenic vein at the hilus. The authors have termed this technique transsplenic portal catheterization (TPC). Transsplenic portal catheterization is indicated for portal angiography, portal pressure measurement, and chronic portal blood sampling. Clinical applications of this technique include use in diagnosis and surgical management of portosystemic shunts and differentiation of prehepatic, hepatic, and posthepatic hypertension. This report describes the technique of transsplenic portal catheterization. Clinical use of this technique in two cases of portosystemic shunts are included.     

Hepatic Organelle Pathology in Dogs with Congenital Portosystemic Shunts

Diversion of portal blood in congenital portosystemic shunts (CPSS) results in liver atrophy and passage of toxins into the systemic circulation causing hepatic encephalopathy. In some dogs, there is indirect evidence for hepatic insufficiency, but histologic findings are equivocal. This study determined whether hepatocyte integrity in PSS is comprised at a subcellular level using analytical subcellular fractionation of liver biopsies. Six dogs with CPSS had hypoproteinemia (6/6), increased serum alkaline phosphatase (6/6) and alanine aminotransferase (4/6) activity, hypocholesterolemia (6/6), and decreased blood urea (2/6). Liver biopsy specimens had increased activities (mU/mg protein) of alkaline phosphatase (17.9 +/- 10.1; controls 5.1 +/- 5.3: P less than 0.01), but not of other plasma membrane enzymes. There were increased activities of endoplasmic reticular (neutral alpha-glucosidase: 1.67 +/- 0.7; controls 0.86 +/- 0.2: P less than 0.01) and lysosomal enzymes (N-acetyl-beta-glucosaminidase: 12.6 +/- 2.3; controls 6.24 +/- 2.7: P less than 0.01; alpha-mannosidase: 0.85 +/- 0.5; controls 0.39 +/- 0.3: P less than 0.05). Subcellular fractionation on reorientating sucrose density gradients showed a high-density peak of alkaline phosphatase suggestive of a specific increase in the biliary canalicular component of enzyme activity. Neutral alpha-glucosidase was shifted to denser fractions, indicative of an increase in the proportion of rough-to-smooth endoplasmic reticulum and consistent with enhanced synthesis of membranous enzymes. There was also evidence for increased fragility of intracellular organelles, particularly lysosomes. In contrast, histology showed either no abnormalities or minor degenerative changes compatible with hepatic underperfusion.(ABSTRACT TRUNCATED AT 250 WORDS)     

Microcytosis and Iron Status in Dogs With Surgically Induced Portosystemic Shunts

Microcytosis is common in dogs with congenital portosystemic shunts (PSS) and acquired liver disease. The objective of this study was to determine if microcytosis could be induced in normal dogs by surgical creation of PSS, and to characterize the changes in hematology and iron status. Hematocrit, mean cell volume, mean cell hemoglobin, and mean cell hemoglobin concentration decreased linearly from 45.5%. 69.1 fL, 22.8 g/dL and 33.1% to 39.5%. 55.9 fL, 17.8 g/dL and 31.9%. respectively, 18 weeks after creation of PSS. The erythrocyte count did not change, but red cell distribution widths indicated a shift to a heterogenous population with decreased volume. Mean cell volume and mean cell hemoglobin decreased rapidly after induction of PSS and were significantly ( P < .05) different from presurgery values within 2 weeks. Serum iron and copper concentrations and total iron binding capacity were decreased in dogs with PSS. Liver iron concentration doubled after creation of PSS, with the majority of stainable iron located in Kupffer cells. The changes in erythrocyte indices and measures of iron status in dogs with surgically induced PSS were similar to those in dogs with congenital PSS. Microcytosis developed rapidly in dogs after induction of PSS. These results indicate that iron deficiency was not the cause of microcytosis in these dogs.     

Long-Term Results of Complete and Partial Ligation of Congenital Portosystemic Shunts in Dogs

The medical records of 65 dogs that underwent complete or partial ligation of a single congenital portosystemic shunt (CPSS) were reviewed to determine the long-term clinical results. Information retrieved from the records included age at surgery, preligation (baseline) portal pressure, postligation portal pressure, change in portal pressure from baseline, complete or partial occlusion of the shunting vessel and fasting, and 2-hour postprandial bile acids from the preoperative, early postoperative (PO), and greater than 1 year PO time periods. A clinical rating score derived from a follow-up examination greater than 1 year PO was assigned to each dog. Of the 56 dogs that survived the perioperative period, 29 (52%) had complete and 27 (48%) had partial ligations. Age at surgery, pre- and postligation portal pressure, change in portal pressure from baseline and serum bile acid concentrations were not related to long-term clinical outcome. Clinical rating scores were significantly greater for dogs with partial CPSS ligations compared with dogs with complete ligations, indicating a less favorable clinical outcome for partial ligations. Fasting and 2-hour postprandial bile acid values at both PO time intervals were significantly greater in partial versus complete ligation groups. Follow-up information for more than 1 year was available on 18 of 29 dogs (62%) with complete ligations. All were clinically normal. Of 27 dogs with partial ligations, 11 dogs (41%) developed recurrence of clinical signs resulting in presentation to the university or referring veterinarian for additional surgery, medical management, or euthanasia. Only three dogs with partial CPSS ligation (11%) were clinically normal. Another nine dogs (33%) were operated on again before the possible development of clinical signs and four dogs (15%) were unavailable for follow-up. It was concluded that partial ligation of CPSS is associated with a greater recurrance of clinical signs and patient morbidity than complete ligation.     

Markers of Angiogenesis Associated with Surgical Attenuation of Congenital Portosystemic Shunts in Dogs

Background

Dogs with congenital portosystemic shunts (CPSS) have hypoplasia of the intrahepatic portal veins. Surgical CPSS attenuation results in the development of the intrahepatic portal vasculature, the precise mechanism for which is unknown, although new vessel formation by angiogenesis is suspected.

Hypothesis

That the degree of portal vascular development and the increase in portal vascularization after CPSS attenuation is significantly associated with hepatic vascular endothelial growth factor (VEGF) and VEGF receptor 2 (VEGFR2) gene expression and serum VEGF concentration.

Animals

Client‐owned dogs with CPSS undergoing surgical treatment. Forty‐nine dogs were included in the gene expression data and 35 in the serum VEGF data.

Materials and Methods

Dogs surgically treated by partial or complete CPSS attenuation were prospectively recruited. Relative gene expression of VEGF and VEGFR2 was measured in liver biopsy samples taken at initial and follow‐up surgery using quantitative polymerase chain reaction. Serum VEGF concentration was measured before and after CPSS attenuation using a canine specific ELISA. Statistical significance was set at the 5% level (P ≤ .05).

Results

There was a significant increase in the mRNA expression of VEGFR2 after partial attenuation (P = .006). Dogs that could tolerate complete attenuation had significantly greater VEGFR2 mRNA expression than those that only tolerated partial attenuation (P = .037). Serum VEGF concentration was significantly increased at 24 (P < .001) and 48 (P = .003) hours after attenuation.

Conclusions and Clinical Importance

These findings suggest that intrahepatic angiogenesis is likely to occur after the surgical attenuation of CPSS in dogs, and contributes to the development of the intrahepatic vasculature postoperatively.     

Splanchnic Surface Oximetry during Experimental Portal Hypertension and Surgical Manipulation of Portosystemic Shunts in Dogs

Surface oxygen tension (PSO2) was measured in dogs during experimental manipulation of the portal vein and hepatic artery, and during surgery to correct portosystemic shunting. There was no alteration in PSO2 of liver, pancreas, duodenum, or jejunum during partial (50%) or complete reduction of hepatic artery flow. After 100% reduction in portal vein blood flow, PSO2 was lower in jejunum, duodenum, and liver. With 50% reduction in portal flow, PSO2 was significantly decreased only in jejunum. In six dogs with single extrahepatic shunts, there was a significant correlation between portal pressure and jejunal PSO2. It was concluded that measurement of visceral organ PSO2 represents an accurate noninvasive means of obtaining objective data on the effect of reduction in hepatic blood flow on perfusion of other select splanchnic organs.     

Soy Protein Isolate versus Meat-Based Low-Protein Diet for Dogs with Congenital Portosystemic Shunts

Background: Both presurgical preparation and long-term support of nonoperable dogs with congenital portosystemic shunts (CPSS) require optimal dietary management. Studies suggested that protein source may play an important role, with vegetable and dairy protein sources having better effects on hepatic encephalopathy (HE) than meat proteins.
Objectives: Determine whether a low-protein test diet with soy as its main protein source results in better scores than a control diet with the same composition but with poultry as its main protein source in dogs with CPSS.
Methods: In a double-blind cross-over study, 16 dogs received each diet for 4 weeks. Dogs in group T first received the test diet and then the control diet, whereas dogs in group C were fed the diets in the opposite order. Different variables (body weight, body condition score, HE score, fecal score, CBC, plasma tests of liver function including NH₃, and coagulation tests) were measured at the start of the study and after completion of each diet.
Results: One-way repeated measures ANOVA was performed. Plasma NH₃ was significantly lower after the test diet than after the control diet. The test diet also resulted in significantly higher fibrinogen concentrations and lower prothrombin times. The HE score improved with both diets, with no significant difference between the 2 diets.
Conclusions: Both diets achieved a significant improvement in HE score. The influence of the soy-based diet on plasma NH₃ concentration and coagulation parameters suggests that such a diet decreases the risk for HE and gives better support of liver function.     

Hepatic Volume Measurements in Dogs with Extrahepatic Congenital Portosystemic Shunts before and after Surgical Attenuation

Background: In dogs with congenital portosystemic shunts (CPSS), the ability of the hypoplastic liver to grow is considered important for recovery after surgical shunt attenuation.
Objectives: This study investigated hepatic growth after extrahepatic shunt attenuation in dogs using magnetic resonance imaging (MRI) and computed tomography (CT).
Animals: Ten client-owned dogs with single extrahepatic CPSS.
Methods: Abdominal MRI, CT, or both were performed before and 8 days, 1, and 2 months after shunt attenuation. Liver volumes were calculated from the areas of the MRI or CT images.
Results: Before surgery, median liver volume was 18.2 cm³/kg body weight. Liver volume increased significantly after surgery. Growth was highest between days 0 and 8 and decreased afterward. Median liver volume was 28.8 cm³/kg at 2 months after attenuation. No significant differences in growth were found between dogs with complete or partial shunt closure or between dogs with complete or incomplete metabolic recovery. Volumes measured from consecutively performed MRI and CT images correlated well ( r = 0.980), but volumes from MRI images were significantly larger than volumes from CT images (6.8%; P = .008).
Conclusion and Clinical Importance: After shunt attenuation, rapid normalization of liver size was observed. Hepatic growth was not decreased in dogs after partial closure of CPSS or in dogs with subclinical, persistent shunting 2 months after surgery. CT is the preferred imaging method for volumetric estimation because of speed.     

Transportal Approach for Attenuating Intrahepatic Portosystemic Shunts in Dogs

A novel surgical approach, using portal venotomy during total hepatic vascular occlusion, was used to locate and attenuate congenital intrahepatic portosystemic shunts in nine dogs. Shunt location was consistent with a persistent ductus venosus in only two dogs. In the remaining seven dogs the shunts were window-like orifices arising from either the left (two dogs) or right portal vein branch (five dogs) and communicating with the ipsilateral hepatic vein or caudal vena cava. The transportal approach using total hepatic vascular occlusion consistently provided good access to the portosystemic shunts, including those with window-like communications. A 7 to 16 minute period of total vascular occlusion was well-tolerated hemodynamically, with few intraoperative complications. Intrahepatic shunts were successfully attenuated in eight dogs, while one dog with portal atresia was euthanatized. The postoperative course was complicated by high protein pulmonary edema (one dog), an encapsulated biliary pseudocyst (one dog) and uncontrollable hemorrhage caused by an uncharacterized coagulopathy (one dog). Three dogs required a second operation to further attenuate their shunts. The clinical condition of all seven surviving dogs was improved after surgery.     

Intravascular Occlusion for the Correction of Extrahepatic Portosystemic Shunts in Dogs

Background: Congential extrahepatic portosystemic shunts (EHPSS) are common in dogs. An effective minimally invasive technique for correction of EHPSS could result in reduced morbidity, reduced costs, and reduced hospitalization times. Hypothesis: Use of an intravascular occlusion device can effectively and safely result in acute complete occlusion of EHPSS in dogs. Animals: Seven dogs with naturally occurring EHPSS that presented to the Purdue University Veterinary Teaching Hospital. Methods: Prospective, clinical trial. The 7 dogs were consecutively enrolled over a 2‐year period. Results of serum biochemistry, total serum bile acids, fasting plasma ammonia, abdominal radiography, and ultrasonography suggested the diagnosis of portosystemic shunts in all dogs. Definitive diagnosis of EHPSS was achieved with cranial mesenteric arterial portography and acute occlusion was attempted by the deployment of the Amplatzer vascular plug (AVP). Results: EHPSS were identified in all dogs consisting of 5 portocaval and 2 portoazygous variants; 1/7 dogs (14%) were intolerant to temporary complete occlusion of the EHPSS. Of the remaining 6 dogs, 5 (83%) had complete occlusion of the EHPSS by the AVP. There were no complications and resolution of abnormal clinical signs and laboratory values was achieved in 4/5 (80%) dogs with complete occlusion. Conclusions and Clinical Importance: Intravascular correction of EHPSS by the AVP is a viable option to surgical correction while larger studies will be required to determine the clinical applicability of this procedure in the broader portosystemic shunt population.     

Iron Status and Erythrocyte Volume in Dogs With Congenital Portosystemic Vascular Anomalies

Microcytosis, hypochromasia, and low mean corpuscular hemoglobin are frequent hematologic abnormalities in dogs with portosystemic vascular anomalies (PSVA). The relationship of iron status to these abnormalities is unclear. We evaluated iron status and hematologic and biochemical parameters in dogs with congenital PSVA before (25 dogs) and after (11 dogs) partial ligation of the vascular anomaly. Serum iron concentration and total iron binding capacity were subnormal in 56% and 20% of dogs with PSVA, respectively. Transferrin saturation was normal in 68%, decreased in 20%, and increased in 12% of the dogs. Plasma ferritin concentration was either normal (56%) or high (44%), and was not associated with increases in ceruloplasmin concentration. Hepatic stainable iron was increased in 10 of 16 dogs. Mean corpuscular volume (MCV), mean corpuscular hemoglobin, and mean corpuscular hemoglobin concentration were decreased in more than 60% of dogs with PSVA. Serum biochemical abnormalities included high bile acid concentration and alanine transaminase (ALT) and alkaline phosphatase (ALP) activities; and low urea, creatinine, cholesterol, and total protein concentrations. Serum iron concentration and clinical status (normal or PSVA) significantly influenced MCV ( P = .003 and P < .001, respectively), whereas age, ceruloplasmin, ferritin, cholesterol, bile acids, and total iron binding capacity did not. Partial ligation of PSVA was associated with resolution of clinical signs and the return to normal of iron status and all clinicopathologic abnormalities, except total fasting bile acid concentrations. These findings indicate that iron status is frequently abnormal in dogs with PSVA and that low serum iron concentration appears to be related to the development of microcytosis. The normalization of iron status and clinicopathologic abnormalities after treatment suggests that they are direct consequences of PSVA.     

Liver Size,Bodyweight, and Tolerance to Acute Complete Occlusion of Congenital Extrahepatic Portosystemic Shunts in Dogs

Objective— To investigate the relationship between preoperative liver size, bodyweight, and tolerance to shunt occlusion in dogs with congenital extrahepatic portosystemic shunt(s) (CPSS). Study Design— Longitudinal cohort study. Animals— Dogs with CPSS (n=35). Methods— Ultrasonography was used to measure preoperative maximum transverse dimension of the liver (TS) of each dog. Intraoperative portal pressures were measured, before and after CPSS occlusion, via a jejunal vein catheter. Tolerance to shunt occlusion was judged on gross visceral observations, and on changes in portal pressure, central venous and mean arterial pressures. Results— TS was significantly related to bodyweight (P<.05). Mean ratios for TS/bodyweight were calculated for dogs tolerant and intolerant of acute complete shunt occlusion. Dogs tolerant to occlusion had significantly higher TS/bodyweight ratios than dogs intolerant to occlusion (P=.025). Dogs with a TS/bodyweight ratio of >7 were more likely to tolerate CPSS occlusion than dogs with a TS/bodyweight ratio of <5 (P=.036). A model was generated to predict portal pressure rise after shunt occlusion, based on liver dimensions and bodyweight (R=0.668). Intestinal oxygenation did not correlate significantly with tolerance to CPSS occlusion (P=.29). Conclusion— In dogs with CPSS, liver size (relative to bodyweight) is significantly greater (P=.025) in dogs that are tolerant of full ligation than intolerant of occlusion. Clinical Relevance— Preoperative measurement of bodyweight and liver size help indicate the likelihood of tolerance to acute complete occlusion of CPSS in dogs.     

Hematologic Changes Associated With Serum and Hepatic Iron Alterations in Dogs With Congenital Portosystemic Vascular Anomalies

Serum and hepatic iron determinations and hematologic parameters were measured in 10 dogs with congenital portosystemic vascular anomalies. Anemia, hypoferremia, and microcytosis were present in 70%, 70%, and 60% of the dogs, respectively. An increase in hepatic iron content was observed in all dogs. These results suggest a relationship between altered hepatic blood flow and abnormal iron metabolism in dogs with congenital portosystemic vascular anomalies.     

Gradual Occlusion of Extrahepatic Portosystemic Shunts in Dogs and Cats Using the Ameroid Constrictor

Gradual occlusion of the splenic vein, using a specialized device (ameroid constrictor), was evaluated experimentally in three normal beagle dogs. Splenoportograms were used to verify that total occlusion of the splenic vein had occurred in all dogs within 4 to 5 weeks after application of the device. The ameroid constrictor (AC) was also evaluated as a method of gradual vascular occlusion in 12 dogs and two cats with single, extrahepatic, portosystemic shunts (PSS). Serum bile acid (SBA) concentrations were measured and portal scintigraphy (PS) was performed on all 14 animals preoperatively and 10, 20, 30, and 60 days postoperatively. Two dogs (14%) died from portal hypertension in the early postoperative period. One dog and one cat developed multiple acquired PSS, confirmed by mesenteric portography 90 days after the operation. Portal scintigraphy confirmed total occlusion of the primary shunt in the other 10 animals. Shunt fractions (SF), as measured by PS on postoperative days 30 and 60, declined significantly from preoperative values. Significant decreases were noted between preoperative and postoperative values for preprandial SBA on postoperative day 60 and for postprandial SBA on postoperative day 30. SBA concentrations did not correlate with SF. Based on this study, gradual vascular occlusion using the AC is recommended as a method for treatment of single, extrahepatic, PSS.