rhodanine and Liver-Diseases

Canine hepatic copper content has been increasing. Recognition of canine copper-associated hepatopathies is becoming more common.. The purpose of the study was to assess the diagnostic performance of Wright-Giemsa (WG) and rhodanine staining for detection of increased canine hepatic copper following a proposed cytologic protocol for semi-quantitative evaluation of liver aspirates and the effect of previous WG staining.. Retrospectively, 40 canine hepatic WG-stained cytology cases were rhodanine stained. Diagnostic performance of WG staining for increased hepatic copper was evaluated. A rhodanine-stained cytologic copper grading system was developed. Prospectively, 67 canine liver samples with quantitative copper measurement, a WG-then rhodanine-stained slide, and a non-WG rhodanine-stained slide were used to assess the performance of the grading system and the effect of previous WG staining.. Copper was not described in 40 retrospective cases on initial cytologic evaluation; 8/40 cases had increased copper content after rhodanine staining or quantitative copper assessment. Prior WG staining and destaining significantly affected the cytologic copper grade but not the diagnostic performance as measured by receiver-operating characteristic curve analysis. Quantitative copper concentration and previously WG-stained copper grade were moderately correlated (n = 67, ρ = .79 [.68-.87]). For detection of ≥ 600 ppm, dry weight (dw) copper, sensitivity was .75 and specificity was .97. For detection of ≥ 1500 ppm, dw copper, sensitivity was 1.0 and specificity was .97.. Wright-Giemsa staining alone does not reliably detect hepatic copper. Grading of rhodanine-stained canine hepatic cytologic samples demonstrates acceptable diagnostic performance for detection of copper content.

Topics: Animals; Azure Stains; Biopsy, Needle; Coloring Agents; Copper; Cytodiagnosis; Dog Diseases; Dogs; Female; Liver; Liver Diseases; Male; Reproducibility of Results; Retrospective Studies; Rhodanine; Sensitivity and Specificity; Staining and Labeling

To evaluate the accuracy of digitally scanned rhodanine-stained liver biopsy specimens for determination of hepatic copper concentration and compare results with qualitatively assigned histologic copper scores in dogs.. 353 liver biopsy specimens from dogs.. Specimens (n = 139) with quantified copper concentration ranging from 93 to 6,900 μg/g were allocated to group 1 (< 400 μg/g [37]), group 2 (401 to 1,000 μg/g [27]), group 3 (1,001 to 2,000 μg/g [34]), and group 4 (> 2,001 μg/g [41]); stained with rhodanine; and digitally scanned and analyzed with a proprietary positive pixel algorithm. Measured versus calculated copper concentrations were compared, and limits of agreement determined. Influence of nodular remodeling, fibrosis, or parenchymal loss on copper concentration was determined by digitally analyzing selected regions in 17 specimens. After method validation, 214 additional liver specimens underwent digital scanning for copper concentration determination. All sections (n = 353) were then independently scored by 2 naive evaluators with a qualitative scoring schema. Agreement between assigned scores and between assigned scores and tissue copper concentrations was determined.. Linear regression was used to develop a formula for calculating hepatic copper concentration ≥ 400 μg/g from scanned sections. Copper concentrations in unremodeled specimens were significantly higher than in remodeled specimens. Qualitative scores widely overlapped among quantitative copper concentration groups.. Calculated copper concentrations determined by means of digital scanning of rhodanine-stained liver sections were highly correlated with measured values and more accurate than qualitative copper scores, which should improve diagnostic usefulness of hepatic copper concentrations and assessments in sequential biopsy specimens.

Topics: Animals; Biopsy; Copper; Dog Diseases; Dogs; Image Processing, Computer-Assisted; Linear Models; Liver; Liver Diseases; Rhodanine

We surveyed histochemically detectable copper in various liver diseases with emphasis on chronic biliary disease (CBD) and venous outflow impairment. Using rhodanine, we graded copper accumulation in 298 liver specimens: venous outflow impairment (n = 64), CBD (n = 123), Wilson disease (WD) (n = 12), chronic hepatitis C (n = 32), steatohepatitis (n = 28), sarcoidosis (n = 15), cholestatic hepatitis (n = 12), and acute large bile duct obstruction (n = 12). Copper was detected in 39% of specimens; all had chronic liver disease. Copper increased with increasing fibrosis. CBD accumulated copper more frequently than other chronic diseases (except WD), both in early (61% vs 3%) and late (94% vs 59%) stages and in larger amounts. Rhodanine was positive in 73% of livers with CBD, 20% with sarcoidosis, 9% with chronic hepatitis C, and 7% with steatohepatitis. Copper was detected in 14% of chronic venous outflow impairment specimens; with 1 exception, stainable copper was absent in early stages but detected in 38% of cirrhotic livers. In conclusion, rhodanine helps differentiate CBD from other conditions, including venous outflow impairment; in the absence of advanced fibrosis, rhodanine positivity strongly favors CBD. In contrast, rhodanine positivity is nonspecific in cirrhosis, but the absence of copper in that setting excludes CBD.

Topics: Acute Disease; Adult; Aged; Biliary Tract Diseases; Chronic Disease; Copper; Diagnosis, Differential; Female; Hepatic Veins; Humans; Immunohistochemistry; Liver Diseases; Male; Middle Aged; Peripheral Vascular Diseases; Regional Blood Flow; Rhodanine; Staining and Labeling; Young Adult

Three different histochemical methods for copper detection were compared. Atomic absorption analysis was used to substantiate the tissue stains. There was good correlation between rhodanine staining and rubeanic acid-stained tissue sections. The orcein reaction for copper-associated protein did not consistently correlate with the methods demonstrating copper. Prolonged staining (72 hours) with rubeanic acid more consistently and clearly detected increased copper in canine livers than did staining with rhodanine. Seventy-two hour staining with rubeanic acid is the method of choice for histochemical detection of copper in canine liver.

Topics: Animals; Copper; Dog Diseases; Dogs; Female; Liver; Liver Diseases; Metallothionein; Oxazines; Rhodanine; Spectrophotometry, Atomic; Staining and Labeling; Thioamides

The incidental finding of orcein positive granules, indicating copper associated protein, in alpha-1-antitrypsin (AAT) positive liver biopsies stimulated a histochemical search for evidence of copper and copper-binding protein in a series of 46 liver biopsies with histological evidence of AAT accumulation. Hepatic accumulation copper and copper-binding protein occurred in all 19 cirrhotics (100%) and in 14 out of 27 non-cirrhotic livers (51.85%). The overall percentage was 71.73%. AAT and copper deposits coexisted in the same periportal hepatocytes. AAT globules showed positive reactivity both to rhodanine and orcein stains. The severity of chronic liver damage correlated with increasing amounts of copper deposition. It is suggested that in AAT storage, not only is the metabolism of this substance disturbed, but also that of proteins involved in copper metabolism and excretion, resulting in copper accumulation within hepatocytes.

Topics: Adolescent; Adult; Aged; alpha 1-Antitrypsin; Carrier Proteins; Cholestasis, Intrahepatic; Copper; Cytoplasmic Granules; Female; Humans; Liver; Liver Cirrhosis; Liver Diseases; Male; Middle Aged; Rhodanine