To investigate disparities in hepatic copper concentrations determined by atomic absorption spectroscopy (AAS), inductively coupled plasma mass spectrometry (ICP-MS), and digital image analysis of rhodanine-stained sections.
Medical records of dogs for which hepatic biopsy specimens had been submitted between January 1999 and December 2019 for evaluation of copper content were reviewed. Paired hepatic copper concentrations obtained with digital image analysis and AAS or ICP-MS were compared, and Spearman rank correlation coefficients were calculated to test for correlations between qualitative copper accumulation scores and hepatic copper concentrations. For dogs for which ≥ 4 rhodanine-stained hepatic sections were available, intraindividual variation in copper distribution across hepatic sections was evaluated.
Median hepatic copper concentrations obtained with digital image analysis exceeded concentrations obtained with AAS or ICP-MS. Concentrations were also higher in older dogs (≥ 9 years vs < 9 years), dogs of breeds with a typical body weight ≥ 20 kg (44 lb), and dogs with necroinflammatory changes or uneven copper distribution. Qualitative copper accumulation scores were significantly associated with hepatic copper concentrations; however, the correlation between qualitative score and concentration obtained with digital image analysis (rs = 0.94) was higher than the correlation between qualitative score and concentration obtained with AAS (rs = 0.75) or ICP-MS (rs = 0.57). The coefficient of variation for hepatic copper concentrations obtained with digital image analysis was significantly higher for dogs with higher hepatic copper concentrations.
CONCLUSIONS AND CLINICAL RELEVANCE
Results suggested that spectroscopic-spectrometric analysis of hepatic biopsy specimens commonly underestimated the concentration obtained by digital image analysis of rhodanine-stained sections.
To characterize the frequency and type of bacterial infection by culture- and immunohistochemical (IHC)-based methods and determine the impact of infection on clinical features and survival time in cats with suppurative cholangitis-cholangiohepatitis syndrome (S-CCHS).
168 client-owned cats with S-CCHS (cases).
Clinical features, bacterial culture results, culture-inoculate sources, and survival details were recorded. Cases were subcategorized by comorbidity (extrahepatic bile duct obstruction, cholelithiasis, cholecystitis, ductal plate malformation, biopsy-confirmed inflammatory bowel disease, and biopsy-confirmed pancreatitis) or treatment by cholecystectomy or cholecystoenterostomy. Culture results, bacterial isolates, Gram-stain characteristics, and IHC staining were compared among comorbidities. Lipoteichoic acid IHC staining detected gram-positive bacterial cell wall components, and toll-like receptor expression IHC reflected pathologic endotoxin (gram-negative bacteria) exposure.
Clinical features were similar among cases except for more frequent abdominal pain and lethargy in cats with positive culture results and pyrexia, abdominal pain, and hepatomegaly for cats with polymicrobial infections. Bacteria were cultured in 93 of 135 (69%) cats, with common isolates including Enterococcus spp and Escherichia coli. IHC staining was positive in 142 of 151 (94%) cats (lipoteichoic acid, 107/142 [75%]; toll-like receptor 4, 99/142 [70%]). With in-parallel interpretation of culture and IHC-based bacterial detection, 154 of 166 (93%) cats had bacterial infections (gram-positive, 118/154 [77%]; gram-negative, 111/154 [72%]; polymicrobial, 79/154 [51%]). Greater frequency of bacterial isolation occurred with combined tissue, bile, and crushed cholelith inoculates. Infection and gram-positive bacterial isolates were associated with significantly shorter long-term survival times.
S-CCHS was associated with bacterial infection, pathologic endotoxin exposure, and frequent polymicrobial infection in cats. Combined tissue inoculates improved culture detection of associated bacteria.
Objective—To evaluate the influence of a 1,4-
butanedisulfonate stable salt of S-adenosylmethionine
(SAMe) administered orally on clinicopathologic
and hepatic effects induced by long-term administration
of prednisolone in dogs.
Animals—12 healthy dogs.
Procedure—Following a pilot study (4 dogs), 2 groups
of 4 dogs received prednisolone (2.2 mg/kg) orally
once daily (84-day trial). One group received SAMe
(20 mg/kg/d divided in 2 doses) for 42 days and then
a placebo for 42 days; the other group received treatments
in the reverse order. Before and during the
trial, numerous variables were monitored, including
serum total alkaline phosphatase (ALP) and glucocorticoid-
induced ALP (G-ALP) activities, serum haptoglobin
concentration, and total and oxidized glutathione
(TGSH and GSSG) and thiobarbiturate-reacting
substances (TBARS) concentrations in erythrocytes
and liver tissue (days 0, 42, and 84). Hepatic
specimens also were examined microscopically.
Results—The stable salt of SAMe was biologically
available; plasma concentrations of SAMe or prednisolone
were not affected by coadministration.
Compared with baseline values, serum ALP and GALP
activities and haptoglobin concentrations
increased and erythrocyte GSSG and TBARS concentrations
decreased with both treatments. Erythrocyte
TGSH concentration decreased with the prednisolone-
placebo treatment. Administration of SAMe
appeared to conserve erythrocyte TGSH values and
did not inhibit hepatocyte glycogen vacuolation but
increased hepatic TGSH concentration and improved
the hepatic tissue GSSG:TGSH ratio.
Conclusions and Clinical Relevance—In dogs,
administration of 20 mg of SAMe/kg/d may mitigate
the apparent pro-oxidant influences of prednisolone
but did not block development of classic clinicopathologic
or histologic features of vacuolar hepatopathy.
(Am J Vet Res 2005;66:330–341)
Objective—To investigate the influence of dietary supplementation with l-carnitine on metabolic rate, fatty acid oxidation, weight loss, and lean body mass (LBM) in overweight cats undergoing rapid weight reduction.
Procedures—Cats fattened through unrestricted ingestion of an energy-dense diet for 6 months were randomly assigned to 4 groups and fed a weight reduction diet supplemented with 0 (control), 50, 100, or 150 μg of carnitine/g of diet (unrestricted for 1 month, then restricted). Measurements included resting energy expenditure, respiratory quotient, daily energy expenditure, LBM, and fatty acid oxidation. Following weight loss, cats were allowed unrestricted feeding of the energy-dense diet to investigate weight gain after test diet cessation.
Results—Median weekly weight loss in all groups was ≥ 1.3%, with no difference among groups in overall or cumulative percentage weight loss. During restricted feeding, the resting energy expenditure-to-LBM ratio was significantly higher in cats that received l-carnitine than in those that received the control diet. Respiratory quotient was significantly lower in each cat that received l-carnitine on day 42, compared with the value before the diet began, and in all cats that received l-carnitine, compared with the control group throughout restricted feeding. A significant increase in palmitate flux rate in cats fed the diet with 150 μg of carnitine/g relative to the flux rate in the control group on day 42 corresponded to significantly increased stoichiometric fat oxidation in the l-carnitine diet group (> 62% vs 14% for the control group). Weight gain (as high as 28%) was evident within 35 days after unrestricted feeding was reintroduced.
Conclusions and Clinical Relevance—Dietary l-carnitine supplementation appeared to have a metabolic effect in overweight cats undergoing rapid weight loss that facilitated fatty acid oxidation.
To determine whether body weight, age, or sex was associated with ultrasonographically determined adrenal gland thickness (AT) in dogs with non-adrenal gland illness.
Retrospective cross-sectional study.
266 dogs (22 sexually intact and 119 castrated males and 19 sexually intact and 106 spayed females representing 12 breeds) with non-adrenal gland illness.
Thickness of the caudal pole of the left and right adrenal glands was measured on longitudinal ultrasonographic images. Dogs were stratified into age and body weight categories to investigate associations with AT.
AT was significantly lower in dogs that weighed ≤ 12 kg (26.4 lb) than in dogs that weighed > 12 kg and left AT increased with age. Both left and right AT were larger in male than in female dogs that weighed > 12 to ≤ 20 kg, and left AT was larger in male than in female dogs that weighed > 20 to ≤ 30 kg.
CONCLUSIONS AND CLINICAL RELEVANCE
Results suggested that body weight, age, and sex were significantly associated with AT, indicating that these variables should be considered when evaluating AT in dogs with non-adrenal gland illness and when developing reference intervals for AT in dogs. Further, findings indicated that dogs with non-adrenal gland illness that weigh ≤ 12 kg should have an AT no greater than 0.62 cm, whereas dogs that weigh > 12 kg should have an AT no greater than 0.72 cm.
Objective—To determine the diagnostic value of protein C (PC) for detecting hepatobiliary disease and portosystemic shunting (PSS) in dogs.
Animals—238 clinically ill dogs with (n = 207) and without (31) hepatobiliary disease, including 105 with and 102 without PSS.
Procedures—Enrollment required routine hematologic, serum biochemical, and urine tests; measurement of PC activity; and a definitive diagnosis. Total serum bile acids (TSBA) concentration and coagulation status, including antithrombin activity, were determined in most dogs. Dogs were grouped into hepatobiliary and PSS categories. Specificity and sensitivity were calculated by use of a PC cutoff value of 70% activity.
Results—Specificity for PC activity and TSBA concentrations was similar (76% and 78%, respectively). Best overall sensitivity was detected with TSBA, but PC activity had high sensitivity for detecting PSS and hepatic failure. Protein C activity in microvascular dysplasia (MVD; PC ≥ 70% in 95% of dogs) helped differentiate MVD from portosystemic vascular anomalies (PSVA; PC < 70% in 88% of dogs). A receiver operating characteristic curve (PSVA vs MVD) validated a useful cutoff value of < 70% activity for PC.
Conclusions and Clinical Relevance—Combining PC with routine tests improved recognition of PSS, hepatic failure, and severe hepatobiliary disease and signified a grave prognosis when coupled with hyperbilirubinemia and low antithrombin activity in hepatic failure. Protein C activity can help prioritize tests used to distinguish PSVA from MVD and sensitively reflects improved hepatic-portal perfusion after PSVA ligation.
Objective—To determine disorders associated with vacuolar hepatopathy (VH), morphologic hepatic and clinicopathologic abnormalities, and affiliation with steroidogenic hormone excess in dogs.
Design—Retrospective case series.
Animals—336 dogs with histologically confirmed moderate or severe VH.
Procedures—Information on signalment, results of diagnostic testing, definitive diagnoses, and exposure to glucocorticoids (ie, exogenous glucocorticoid administration or high endogenous concentrations of steroidogenic hormones) was obtained from medical records. Dogs were grouped by underlying disorder, glucocorticoid exposure, acinar zonal distribution of lesions, and histologic severity.
Results—12 disease groups (neoplastic, acquired hepatobiliary, neurologic, immune-mediated, gastrointestinal tract, renal, infectious, cardiac disease, diabetes mellitus, portosystemic vascular anomaly, adrenal gland dysfunction, and miscellaneous disorders) were identified. There were 186 (55%) dogs with and 150 (45%) dogs without evidence of glucocorticoid exposure. Acinar zonal distribution of hepatic vacuolation and clinicopathologic values did not differ between dogs with and without evidence of glucocorticoid exposure. However, a 3-fold increased likelihood of severe VH was associated with steroidogenic hormone exposure. Of 226 dogs with high serum alkaline phosphatase activity, 102 (45%) had no evidence of glucocorticoid exposure.
Conclusions and Clinical Relevance—Results suggest that neoplasia and congenital or acquired hepatobiliary disease are common in dogs with VH and provide support for the suggestion that VH, high alkaline phosphatase activity, and illness-invoked physiologic stress may be associated. Histologic confirmation of VH should initiate a diagnostic search for a primary disease if glucocorticoid treatment and hyperadrenocorticism are ruled out.
To determine hepatic copper concentrations and zonal distribution in ferrets with and without hepatobiliary disease, validate rhodanine-based qualitative copper scoring and digital copper quantification in ferret hepatic samples, and ascertain whether clinical features predicted copper accumulation.
34 ferrets, including 7 with necroinflammatory disease, 5 with hepatocellular carcinoma, 13 with non-necroinflammatory disease, and 9 with no hepatobiliary disease.
Rhodanine-based digital copper quantification was validated by use of liver dually measured by atomic absorption spectroscopy and digital scanning (R2 = 0.98). Clinical features and hepatic copper scores and concentrations (dry weight liver) were compared between groups. Zonal copper distribution was determined.
Hepatic copper concentration was strongly correlated with copper scores (ρ = 0.88). Ferrets with hepatobiliary disease were significantly older and had significantly higher serum alkaline phosphatase and γ-glutamyltransferase activities and creatinine concentrations. Centrilobular copper accumulated in 23 of 34 (64%) ferrets with (n = 15) and without (8) hepatobiliary disease. Median copper concentrations were not significantly different between ferrets with and without hepatobiliary disease but were significantly higher within neoplastic hepatic tissue in ferrets with hepatocellular carcinoma. Hepatic copper concentrations exceeded feline (> 180 µg/g) and canine (> 400 µg/g) reference limits in 19 and 9 ferrets, respectively. Hepatic copper > 1,000 µg/g occurred in 5 ferrets with and 2 without hepatobiliary disease. Clinical features did not predict copper accumulation.
Rhodanine-based digital copper quantification and qualitative copper scoring discerned liver copper accumulation in ferrets. Ferrets with and without hepatobiliary disease displayed a propensity for centrilobular hepatic copper accumulation of uncertain clinical importance. Clinical and clinicopathologic features could not exclusively implicate pathologic copper accumulation.
Objective—To evaluate prognostic factors, survival,
and treatment protocols for immune-mediated
hemolytic anemia (IMHA) in dogs.
Animals—151 dogs with IMHA not associated with
underlying infectious or neoplastic disease.
Procedure—Information recorded from review of medical
records included signalment at the time of initial
evaluation; vaccination history; 30-, 60-, and 365-day follow-up outcomes; laboratory data; results of imaging
studies; and necropsy findings. Dogs were grouped
according to the presence of spherocytes, autoagglutination,
a regenerative erythrocyte response, and treatments
received (azathioprine, azathioprine plus ultralowdose
aspirin, azathioprine plus mixed–molecular-weight
heparin [mHEP], or azathioprine plus ultralow-dose
aspirin plus mHEP) for comparisons. All dogs received
Results—Cocker Spaniels, Miniature Schnauzers,
neutered dogs, and female dogs were overrepresented.
Alterations in certain clinicopathologic variables were
associated with increased mortality rate. Rates of survival
following treatment with azathioprine, azathioprine
plus ultralow-dose aspirin, azathioprine plus mHEP, and
azathioprine plus ultralow-dose aspirin plus mHEP were
74%, 88%, 23%, and 70%, respectively, at hospital discharge;
57%, 82%, 17%, and 67%, respectively, at 30
days; and 45%, 69%, 17%, and 64%, respectively, at 1
year. In comparison, mean survival rates at discharge
and at 30 days and 1 year after evaluation collated from
7 published reviews of canine IMHA were 57%, 58%,
and 34%, respectively.
Conclusions and Clinical Relevance—Treatment
with a combination of glucocorticoids, azathioprine,
and ultralow-dose aspirin significantly improved short-and
long-term survival in dogs with IMHA. (J Am Vet
Med Assoc 2005;226:1869–1880)
Objective—To compare morphologic diagnoses determined
from needle biopsy specimens obtained from
the livers of dogs and cats with morphologic diagnoses
determined from wedge biopsy specimens.
Animals—124 dogs and cats.
Procedure—2 needle biopsy specimens were
obtained from each animal; wedge biopsy specimens
were obtained from the same liver lobe during laparotomy
or postmortem examination. Histologic features
were scored independently by 3 individuals; a morphologic
diagnosis was rendered after histologic features
were scored. Cases were included only if at least 2 of
the 3 examiners agreed on the morphologic diagnosis;
the definitive diagnosis was considered to be the morphologic
diagnosis rendered for the wedge biopsy
specimen. Physical characteristics (length, width, surface
area, degree of fragmentation, and number of portal
triads for needle biopsy specimens and surface area
for wedge biopsy specimens) were determined.
Results—Definitive diagnoses included hepatic necrosis
(n = 10), cholangitis-cholangiohepatitis (13), chronic
hepatitis-cirrhosis (12), canine vacuolar hepatopathy
(11), portosystemic vascular anomaly-microvascular
dysplasia (17), neoplasia (10), miscellaneous hepatic
disorders (18), and no hepatic disease (33). For individual
examiners, the morphologic diagnosis assigned
to needle biopsy specimens agreed with the morphologic
diagnosis assigned to wedge biopsy specimens
for 56 and 67% of the specimens. All 3 examiners
agreed on the morphologic diagnosis assigned to needle
and wedge biopsy specimens for 44 and 65% of
the specimens, respectively. Morphologic diagnoses
assigned to needle biopsy specimens concurred with
the definitive diagnosis for 59 of 124 (48%) animals.
Conclusions and Clinical Relevance—Results
suggest that needle biopsy specimens of the liver
from dogs and cats must be interpreted with caution.
(J Am Vet Med Assoc 2002;220:1483–1490)