Diagnostic value of serum gamma-glutamyl transferase and alkaline phosphatase activities in hepatobiliary disease in the cat

The diagnostic value of serum gamma-glutamyl transferase (GGT) activity and serum alkaline phosphatase (ALP) activity in the detection of liver disease in the cat (n = 69) was compared. On the basis of histologic examination of the liver, cats were assigned to 8 groups: group 1--complete extrahepatic bile duct obstruction (n = 5), group 2--cholangiohepatitis-cholangitis syndrome (n = 11), group 3--hepatic lipidosis (n = 15), group 4--neoplasia, including lymphosarcoma and myeloproliferative disease (n = 9), group 5--hepatic necrosis (n = 7), group 6--cirrhosis (n = 3), group 7--portosystemic vascular anomaly (n = 4), and group 8--miscellaneous (n = 15). Cats assigned to group 8 lacked substantial histologic abnormalities of the liver. The mean value +/- SD of GGT in 20 clinically normal cats was 0.44 +/- 0.26 IU/L. The highest GGT activity in clinical patients developed in groups 1, 2, and 6. The highest ALP activity developed in groups 1 to 4. Significant correlations between GGT and ALP activities were detected only in groups 2 (P less than 0.001) and 5 (P less than 0.10). Among 54 cats with hepatic disease, only 11% had both the GGT and ALP activities within the normal ranges. Comparatively, 52% had ALP activities within the normal range, and 17% had GGT activities within the normal range.(ABSTRACT TRUNCATED AT 250 WORDS)     

Diagnostic value of alkaline phosphatase isoenzyme separation by affinity electrophoresis in the dog.

Affinity electrophoresis, using wheat germ lectin, was used to separate the alkaline phosphatase isoenzymes in the sera of 150 dogs with alkaline phosphatase values greater than or equal to 150 IU/L. The method provided clearer separation of the liver, bone and steroid-induced alkaline phosphatase isoenzymes commonly observed in canine serum, compared to conventional cellulose acetate electrophoresis. The dogs were divided into four patient groups determined by previous corticosteroid treatment, evidence of elevated endogenous corticosteroid levels, age and alanine aminotransferase values. The isoenzyme pattern of each patient was qualitatively assessed. The isoenzyme pattern most frequently observed was greater than 50% steroid induced alkaline phosphatase, which was present in 76 of 150 dogs. This pattern was observed in 18 of 22 dogs receiving corticosteroid therapy, two of three dogs with hyperadrenocorticism, and in dogs with a variety of other diagnoses. The majority of immature dogs (12 of 20) had an isoenzyme pattern consisting of greater than 50% bone. The majority of dogs with active hepatocellular injury (16 of 27) had greater than 50% liver isoenzyme. The isoenzyme pattern was not specific for certain diseases, therefore the diagnostic usefulness is limited. However the isoenzyme result is useful in some cases to determine which further diagnostic tests are indicated, and to determine the source of alkaline phosphatase elevation.     

Diagnostic value of contrast echocardiography in the horse

M-mode echocardiography is a safe and practical means of using ultrasound to evaluate the dynamic movements of cardiac structures. The technique can be refined by using a simple contrast medium in the form of carbon dioxide mixed with heparinised blood to provide a strong echogenic result. This technique was employed in a series of 15 normal conscious standing horses and in three animals with specific cardiac defects. In the clinical cases it was possible to confirm the diagnosis and differentiate between a congenital septal defect and mitral regurgitation. The method was found to be safe and relatively simple to perform using percutaneous insertion of catheters. The intracardiac catheterisation was trouble free and no clinical side effects to direct injection of the carbon dioxide contrast medium into the heart were demonstrated.     

Diagnostic evaluation of canine serum alkaline phosphatase by immunochemical means and interpretation of results.

Sera of several canine patients contained an isoenzyme of alkaline phosphatase (ALP) that resembled intestinal ALP with respect to heat inactivation, L-phenylalanine inhibition, and sensitivity to anti-canine intestinal ALP antibody, but differed with regard to the electrophoretic migration. The electrophoretic mobility of the isoenzyme was slightly cathodal than that of hepatic ALP, and its migration was reduced, similar to that of hepatic isoenzyme after neuraminidase treatment. This isoenzyme, which could be corticosteroid induced, was in the sera of numerous dogs with hepatobiliary disorders and was different from the hepatic isoenzyme that appeared in the sera of dogs with acute hepatitis, based on anti-canine intestinal ALP antibody interaction, heat inactivation, and electrophoretic migration.     

Quantification of bone alkaline phosphatase in canine serum

An assay was developed and proven accurate and precise for the quantification of canine serum alkaline phosphatase of bone origin (BAP). The assay uses wheat germ lectin (WGL) which selectively precipitates SAP but not liver alkaline phosphatase (LAP) in serum preincubated for 1 hour at 37 degrees C before conducting the assay. Although a large percentage of corticosteroid-induced alkaline phos- phatase (CAP) is also precipitated by WGL, the activity of this isoenzyme can be determined by utilizing the automated levamisole inhibition assay and BAP determined by subtraction except in those cases in which CAP is very markedly increased. Use of these two assay techniques in combination allows the quantification of LAP, BAP, and CAP activity in canine serum. In sera from adult dogs of various ages, BAP activity represents a mean of 21.27 -/+ 11.4 U/L; however, there was a statistical decrease in BAP activity with age. This allowed the determination of 95% confidence interval for a reference range dependent on age of the dog. Bone AP activity in puppies drops dramatically within the first 3 months, reaching a magnitude of activity consistent with that of the adult dog by approximately 15 months. BAP was increased over adult reference range in five of five dogs tested with osteosarcoma. This assay will now allow conducting a clinical study of the diagnostic significance of bone AP activity in neoplastic and metabolic diseases of bone.     

Genetic dominant serum alkaline phosphatase activity in goats

Serum alkaline phosphatase activity was determined in 290 Norwegian goats, from 8 months to 10 years of age. The investigations revealed a wide range of individual enzyme activities which constituted two distinct peaks with mean values of 2.37 ± 1.42 and 37.73 ± 14.78 “Sigma” units per ml respectively. Forty-one of the 290 goats were examined by 15 successive samplings for 18 months without obtaining lapping-over values between the two groups. The segregation of the high enzyme level indicated a genetic dominant inheritance.     

The estimation of serum alkaline phosphatase in the dog

Abstract— Four readily available methods for the estimation of serum alkaline phosphatase have been compared using dog sera with varying levels of the enzyme's activity. It has been demonstrated that it is possible to adapt two of the newer methods which are available commercially to permit simple visual assessments to be made without using an absorbtiometer. Résumé— Quatre méthodes aisément disponibles pour l'évaluation de la phosphatase de sérum alcalin ont été comparées en utilisant des sérums de chien à des niveaux différents d'activité de l'enzyme. Il a été démontré qu'il est possible d'adapter deux des méthodes plus nouvelles disponibles dans le commerce pour permettre de faire des évaluations visuelles sans utiliser un absorptiomètre. Zusammenfassung— Vier leicht zugängliche Methoden zur Schätzung von Serum-Alkaliphos-phatase wurden unter Verwendung von Hundeserum mit unterschiedlichen Spiegeln in der Enzym-aktivität vergliechen. Es wurde die Möglichkeit erwiesen, zwei der neuesten, verfügbaren Methoden durch kommerzielle Anpassung zu verwenden, urn einfache, visuelle Bewertungen ohne Benutzung eines Absorptiometers zu gestatten.     

Origin of serum alkaline phosphatase in the dog.

The origin of canine serum alkaline phosphatase (ALP) was investigated by various means. On the basis of electrophoretic migration, neuraminidase treatment, thermal denaturation, and chromatographic fractionation, canine serum was found to contain ALP principally of hepatic origin. There was evidence of only a minor portion of ALP being of osseous origin. Intestinal ALP was not detected in canine serum when monitored by immunochemical technique, L-phenylalanine inhibition, and thermal denaturation.     

The isoelectric focusing properties of serum alkaline phosphatase in disease and following prednisolone and phenylbutazone administration in the horse.

This study was undertaken to ascertain if the isoelectric focusing pattern of serum alkaline phosphatase (AP) from sick horses with high activity is useful for determining its tissue origin. The effect of oral prednisolone and phenylbutazone therapy on this enzyme in healthy horses was also investigated. The sick horses were divided into three groups: hepatic, intestinal and miscellaneous. All sera had approximately thirteen bands of AP activity when focused on agarose gels with a pH gradient of 3.5 to 9.5. All the horses in the liver disease group had greater than 65% of enzyme activity in bands 3 to 7 (counted from the anode) whereas the other two groups had at least 30% and up to 80% of activity in bands 8 to 13. This was true even in the several cases of primary intestinal disease that had additional biochemical evidence of liver damage. All bands were heat sensitive indicating that little if any AP was of small intestinal or renal origin. Oral prednisolone and phenylbutazone for 20 and 12 days respectively had no affect on serum AP activity or isoelectric pattern. We concluded that the AP in bands 3 to 7 is of liver origin but the origin of bands 8 to 13 remains undetermined although small intestinal or renal origin is unlikely. Isoelectric focusing of serum AP shows promise in differentiating cases of primary from secondary liver disease but further studies are required correlating serum patterns and tissue patterns in animals with diseases.     

Phenylalanine inhibited p-nitrophenyl phosphatase activity in the serum as an indication of intestinal cellular disruption in the horse.

Examination of tissues obtained from thoroughbred horses showed that the 'intestinal' phosphatase activity could be differentiated from other phosphatases by analysis at a pH of 9-5 and inhibition with 15 mM L-phenylalanine. A simple method for the measurement of 'intestinal' phosphatase in heparinised plasma or serum is described. Application of the technique to serum or plasma from normal and diseased horses indicates that the increase in the activity of 'intestinal' phosphatase is associated with cases showing clinical, biochemical and haematological evidence of intestinal damage.     

Immunological investigation of intestinal, liver, kidney, bone, placental and serum alkaline phosphatase in cattle

The purpose of this study is to investigate the immunological difference between intestinal, liver, kidney, bone, placental and serum alkaline phosphatase (ALPs) in cattle. Kidney, bone and placental ALPs were purified from each tissue by gel filtration and ion-exchange chromatographies, and intestinal, liver, and placental ALPs were obtained from a commercial source. The antibody to each tissue ALP was prepared by immunizing rabbits and tested by double immunodiffusion analysis in cross reactivities. The results indicated that the bovine tissue ALPs are divided into four groups in antigenicity, that is, intestinal, bone, liver and kidney-placental ALPs. The fetus, calf and cow sera contained bone ALP, and calf and cow sera also contained kidney or placental ALP, but all of the sera did not contain intestinal and liver ALP.     

Isoenzymes of alkaline phosphatase in serum of lambs and ewes.

Electrophoresis in polyacrylamide gels was used to identify isoenzymes of serum alkaline phosphatase in sheep. Skeletal isoenzyme predominated in the serum of lambs and liver, skeletal and intestinal isoenzymes were found in the serum of adult ewes. In r ewes (R-r-i blood group system) intestinal isoenzyme activity was 67 per cent of total serum activity; in R ewes intestinal activity was only 36 per cent of total activity. Relative activities of the three isoenzymes differed greatly within individual ewes and between individual ewes during pregnancy and lactation.     

Changes of serum alkaline phosphatase activity in dry and lactational cows

Serum alkaline phosphatase (ALP) activities were measured during dry and lactational periods to investigate the influence of lactation on serum ALP activity in cows. Higher levels of serum ALP activity were seen in lactational periods than in dry periods. The serum activities of bone-specific ALP (BALP), liver ALP (LALP), tartrate resistant acid phosphatase (TRAP) and aspartate aminotransferase also increased in lactational periods. ALP activities in the bone extract and in whey were decreased at similar rates by the addition of lectin. Moreover, since the ALP band in whey was observed to have the same migration in polyacrylamide gel (PAG) disk electrophoresis as that of the bone extract, analysis of ALP isoenzymes by lectin affinity or PAG disk electrophoresis could not distinguish ALP originating from the mammary gland from that of bone. In this study, it was clear that the increased level of serum ALP activity was due to increases of BALP and LALP in lactational periods. However, the extent of the influence of ALP originating from the mammary glands on serum ALP activity was unknown. Judging from changes of BALP and TRAP activities in the serum and the correlation between the both, it was guessed that ALP originating from the mammary glands influenced serum ALP activity.     

Comparison of techniques for quantifying alkaline phosphatase isoenzymes in canine serum

Three methods for quantifying the steroid-induced and liver isoenzymes of alkaline phosphatase in canine serum were compared on a group of 29 canine serum samples with increased total alkaline phosphatase activity. Levamisole inhibition, heat inactivation, and affinity electrophoresis with densitometry yielded results that correlated strongly. The relationship between levamisole inhibition and heat inactivation test values was a simple linear one, whereas the relationship between their values and those of electrophoresis was better fitted to an exponential model. The levamisole inhibition and heat inactivation tests provided essentially the same information regarding the relative proportions of steroid-induced and hepatic isoenzymes in canine serum; either test was judged practical for routine clinical application.     

Isoenzymes of alkaline phosphatase in serum of newly born lambs.

In the serum of lambs at birth most of the circulating alkaline phosphatase, identified by electrophoretic analysis, was of skeletal origin. Suckling was associated with a rapid increase in activity of intestinal alkaline phosphatase isoenzymes in serum. There was a coincidental increase in level of circulating gamma globulin. Activity of the intestinal isoenzymes in serum began to fall between 15 and 21 h after birth and this fall preceded a fall in serum gamma globulin concentration. The electrophoretic characteristics of the intestinal isoenzymes in serum changed as activity of these isoenzymes increased and then decreased.     

Prognostic significance of serum alkaline phosphatase activity in canine appendicular osteosarcoma

Sixty-one dogs with appendicular osteosarcoma were treated with amputation and chemotherapy of cisplatin and doxorubicin. Serum samples were obtained before and after treatment for determination of total alkaline phosphatase (TALP) activity as well as the activities of the constituent bone (BALP), liver (LALP), and corticosteroid-induced (CALP) isoenzymes. The relationship between alkaline phosphatase activities and survival was examined by Cox proportional hazards regression analysis and Kaplan-Meier log rank analysis. Mean activity of TALP, BALP, and LALP decreased significantly after treatment (P < .001). TALP and LALP activities before treatment were significantly correlated with survival (P = .006 and .001, respectively). The correlation between BALP activity before treatment and survival approached significance (P = .054). CALP activity and TALP, BALP, and LALP activities after treatment were not significantly correlated with survival. Dogs with normal pretreatment TALP and BALP activities survived significantly longer than dogs with increased pretreatment activities (P = .001 and .003, respectively). Median survival times for dogs with normal or increased TALP activities before treatment were 12.5 and 5.5 months, respectively; and median survival times for dogs with normal or increased BALP activities before treatment were 16.6 and 9.5 months, respectively. In the design of future clinical trials involving dogs with osteosarcoma, consideration should be given to stratifying the randomization according to alkaline phosphatase activity. In addition, alkaline phosphatase activity should be a factor considered by clinicians attempting to tailor the aggressiveness of adjuvant chemotherapy to the needs of individual patients or owners.     

The diagnostic and prognostic value of alkaline phosphatase activity in serum and peritoneal fluid from horses with acute colic

Alkaline phosphatase (ALP) is an enzyme present in intestinal mucosa, bile, bone, and renal tubule cells. We sought to assess the diagnostic and prognostic relationships of total ALP (ALPt) activity and that of intestine-derived ALP (ALPi) in serum and peritoneal fluid of 126 horses with colic. ALPt and ALPi activities were measured in both serum and peritoneal fluid by using both standard and L-phenylalanine-based buffers, respectively. Neither ALPt nor ALPi activity were useful in classifying type or severity of intestinal damage. ALPt and ALPi activities in peritoneal fluid were lowest in horses suffering from simple medical colic (39 international units [U]/L [19-60 U/L]; versus 31 U/L [16-44 U/L], median [interquartile range], P < .001) and nonstrangulated surgical lesions (45 U/L [30-62 U/L] versus 36 U/L [23-54 U/L], P < .001), and highest in surgical cases with suspected ulceration (109 U/L [60-1,113 U/L] versus 83 U/L [52-970 U/L], P < .001), strangulation (114 U/L [69-240 U/L] versus 94 U/L [56-191 U/L], P < .001), peritonitis (313 U/L [110-2,227 U/L] versus 283 U/L [91-1,800 U/L], P < .001) or intestinal rupture (687 U/L [205-852 U/L] versus 564 U/L [166-732 U/L], P < .001). Higher ALPt and ALPi activities in peritoneal fluid were associated with greater intestinal damage, increased probability of surgery, and a worse prognosis. The use of L-phenylalanine buffer in both serum and peritoneal fluid did not improve the sensitivity of the test. Based on these results, total ALP activity in peritoneal fluid may help in identifying ischemic or inflammatory bowel lesions in horses with acute colic.