ABSTRACT
Phospholipidosis (PLD), an abnormal accumulation of phospholipids within tissues, is observed during the preclinical testing of many pharmaceutical drugs. Diagnosis of PLD is currently based on morphologic criteria. An understanding of the clinical incidence of PLD and its possible relationship to adverse drug reactions has been hampered by the absence of noninvasive biomarkers for PLD. The uncommon phospholipid di-docosahexaenoyl bis(monoacylglycerol) phosphate (di-22:6-BMP) has been proposed as a potential urinary biomarker for PLD, but data on the utility of serum di-22:6-BMP measurements in diagnosing PLD is more limited. In this report, we compared the performance of serum and urinary di-22:6-BMP as biomarkers for PLD in rats treated with the PLD-inducing drugs amiodarone and 4,4'-diethylaminoethoxyhexestrol dihydrochloride or the hepatotoxicant acetaminophen (APAP). Serum levels of di-22:6-BMP showed a higher correlation to a generalized PLD incidence score than did levels in urine, but were unexpectedly elevated in rats with marked levels of APAP-induced liver necrosis. When samples were filtered based on serum ALT or liver histopathology thresholds, the diagnostic accuracy of serum di-22:6-BMP for PLD improved to the high level observed for urinary di-22:6-BMP without sample exclusion. These data help define the potential context-of-use of serum di-22:6-BMP as a non-clinical biomarker of PLD.
Subject(s)
Chemical and Drug Induced Liver Injury/blood , Chemical and Drug Induced Liver Injury/urine , Lipidoses/blood , Lipidoses/urine , Lysophospholipids/blood , Lysophospholipids/urine , Acetaminophen/toxicity , Amiodarone/toxicity , Animals , Biomarkers/blood , Biomarkers/urine , Chemical and Drug Induced Liver Injury/pathology , Histocytochemistry , Male , Microscopy, Electron, Transmission , Phospholipids/blood , Phospholipids/urine , Rats , Rats, Inbred F344ABSTRACT
To provide mechanistic insight in the induction of phospholipidosis and the appearance of the proposed biomarker di-docosahexaenoyl (C22:6)-bis(monoacylglycerol) phosphate (BMP), rats were treated with 150 mg/kg amiodarone for 12 consecutive days and analyzed at three different time points (day 4, 9, and 12). Biochemical analysis of the serum revealed a significant increase in cholesterol and phospholipids at the three time points. Bio-analysis on the serum and urine detected a time-dependent increase in BMP, as high as 10-fold compared to vehicle-treated animals on day 12. Paralleling these increases, micro-array analysis on the liver of treated rats identified cholesterol biosynthesis and glycerophospholipid metabolism as highly modulated pathways. This modulation indicates that during phospholipidosis-induction interactions take place between the cationic amphiphilic drug and phospholipids at the level of BMP-rich internal membranes of endosomes, impeding cholesterol sorting and leading to an accumulation of internal membranes, converting into multilamellar bodies. This process shows analogy to Niemann-Pick disease type C (NPC). Whereas the NPC-induced lipid traffic jam is situated at the cholesterol sorting proteins NPC1 and NPC2, the amiodarone-induced traffic jam is thought to be located at the BMP level, demonstrating its role in the mechanism of phospholipidosis-induction and its significance for use as a biomarker.
Subject(s)
Amiodarone/toxicity , Lipid Metabolism/drug effects , Lipidoses/chemically induced , Lysophospholipids/blood , Lysophospholipids/urine , Animals , Biomarkers/blood , Biomarkers/urine , Cholesterol/blood , Gene Expression Regulation , Glycerophospholipids/blood , Glycerophospholipids/metabolism , Lipidoses/blood , Lipidoses/urine , Liver/pathology , Lung/pathology , Lymphocytes/drug effects , Lymphocytes/pathology , Male , Metabolic Networks and Pathways/drug effects , Oligonucleotide Array Sequence Analysis , Organ Size/drug effects , Phospholipids/blood , Rats , Rats, Sprague-Dawley , Spleen/pathology , ToxicogeneticsABSTRACT
Di-docosahexaenoyl (C22:6)-bis(monoacylglycerol) phosphate (BMP) was identified as a promising phospholipidosis (PL) biomarker in rats treated with either amiodarone, gentamicin, or azithromycin. Sprague-Dawley rats received either amiodarone (150 mg/kg), gentamicin (100 mg/kg) or azithromycin (30 mg/kg) once daily for ten consecutive days. Histopathological examination of tissues by transmission electron microscopy (TEM) indicated different degrees of accumulation of phospholipidosis in liver, lung, mesenteric lymph node, and kidney of drug-treated rats but not controls. Liquid chromatography coupled to mass spectrometry (LC/MS) was used to identify levels of endogenous biochemical profiles in rat urine. Urinary levels of di-docosahexaenoyl (C22:6)-bis(monoacylglycerol) phosphate (BMP) correlated with induction of phospholipidosis for amiodarone, gentamicin and azithromycin. Rats treated with gentamicin also had increased urinary levels of several phosphatidylinositol (PI), phosphatidylcholine (PC), and phosphatidylethanolamine (PE) species.
Subject(s)
Anti-Arrhythmia Agents/adverse effects , Anti-Bacterial Agents/adverse effects , Lipidoses/urine , Lysophospholipids/urine , Phospholipids/urine , Amiodarone/adverse effects , Animals , Azithromycin/adverse effects , Biomarkers/urine , Chromatography, Liquid , Dose-Response Relationship, Drug , Gentamicins/adverse effects , Kidney/drug effects , Kidney/pathology , Lipidoses/chemically induced , Lipidoses/pathology , Liver/drug effects , Liver/pathology , Lung/drug effects , Lung/pathology , Lymph Nodes/drug effects , Lymph Nodes/pathology , Magnetic Resonance Imaging , Male , Microscopy, Electron , Phosphatidylcholines/urine , Phosphatidylethanolamines/urine , Phosphatidylinositols/urine , Rats , Rats, Sprague-Dawley , Spectrometry, Mass, Electrospray Ionization , Time FactorsABSTRACT
3 male and 2 female infants with Canavan disease proven in some by brain biopsy, whose symptoms appeared within the first 4 months of life, are presented. Urinary organic acids were analyzed by gas chromatography/mass spectrometry. All excreted large amounts of N-acetylaspartic acid, probably secondary to decreased activity of its hydrolase. The pathogenetic mechanism is not well understood. Analysis of urinary organic acids can replace brain biopsy in the diagnosis of this condition, and the diagnosis can now be made prenatally.
Subject(s)
Acids/urine , Demyelinating Diseases/urine , Lipidoses/urine , Aspartic Acid/analogs & derivatives , Aspartic Acid/urine , Female , Humans , Infant , MaleABSTRACT
Pathognomonic ultrastructural inclusions were found in the lymphocytes of 16 patients with neuronal ceroid lipofuscinosis (Batten's disease) and in the urine of three of four of such patients. These inclusions remained type specific for each patient and were identical with deposits in nerve cells and other body tissues. They persisted in lymphocytes in short-term tissue culture, but disappeared with stimulation by phytohemagglutinin. Siblings had identical inclusions. None were present in heterozygotes. Examination of lymphocytes in the peripheral blood cells is an easy method to use in the diagnosis of ceroid lipofuscinosis, but it cannot be used to detect carriers.
Subject(s)
Lipidoses/blood , Lymphocytes/ultrastructure , Adolescent , Child , Female , Humans , Inclusion Bodies/ultrastructure , Lipidoses/urine , Male , ParentsABSTRACT
Urinary sediment from a patient with the juvenile form of Batten disease was examined by electron microscopy. Membrane bound fingerprint profiles were found which were similar to those previously described in a biopsy from this patient's thyroid gland. These findings might represent a useful diagnostic tool.
Subject(s)
Lipidoses/urine , Humans , Lipidoses/pathology , Microscopy, ElectronSubject(s)
Lipidoses/diagnosis , Urine/cytology , Centrifugation , Child , Child, Preschool , Female , Humans , Inclusion Bodies , Lipidoses/urine , Male , Microscopy, Electron , Pigments, BiologicalABSTRACT
We compared the sphingolipid content of urine from a patient with Farber's disease with that of control urine. The ceramides were measured by high-performance liquid chromatography. The patient's urine contained 1.2 mug of ceramides per milligram of creatinine, more than 200-fold the normal amount. The urinary ceramides were isolated by high-performance liquid chromatography for further identification. They contained mainly nonhydroxy fatty acids and only a small quantity of those with 2-hydroxy fatty acids. This contrasts with the previously described composition of the patient's renal and cerebellar tissue. The fatty acid and long-chain base compositions of the urinary ceramides containing nonhydroxy fatty acids were nearly identical to those of the patient's kidney.
Subject(s)
Ceramides/urine , Lipidoses/urine , Ceramides/metabolism , Cerebrosides/urine , Cholesterol/urine , Chromatography , Fatty Alcohols/urine , Humans , Intellectual Disability/urine , Kidney/metabolism , Leukodystrophy, Metachromatic/urine , Lipids/isolation & purification , Lipids/urine , Male , Pigmentation Disorders/urine , Spectrophotometry, Infrared , Sphingomyelins/urine , Sulfoglycosphingolipids/urineSubject(s)
Galactosidases/analysis , Gangliosides , Hexosaminidases/analysis , Lipidoses/enzymology , Liver/enzymology , Sphingolipidoses/enzymology , Carbon Radioisotopes , Catalysis , Child, Preschool , Dehydroepiandrosterone , Glucosamine/analogs & derivatives , Glucosamine/urine , Humans , Infant , Kinetics , Lipidoses/urine , Sphingolipidoses/urine , TestosteroneSubject(s)
Glucosidases/metabolism , Glucuronidase/metabolism , Glycoside Hydrolases/metabolism , Hexosaminidases/metabolism , Isoenzymes/metabolism , Kidney/enzymology , Chromatography, Gel , Glucosidases/urine , Glucuronidase/urine , Glycoside Hydrolases/urine , Hexosaminidases/urine , Humans , Isoelectric Focusing , Isoenzymes/urine , Lipidoses/enzymology , Lipidoses/urine , MannoseSubject(s)
Glycopeptides/urine , Mucopolysaccharidoses/urine , Sphingolipids/urine , Child , Child, Preschool , Chromatography, Paper , Female , Gangliosides , Gaucher Disease/urine , Glycosaminoglycans/urine , Hexoses/urine , Humans , Infant , Lipidoses/urine , Male , Mucopolysaccharidosis IV/urine , Neuraminic Acids/urine , Sulfates/urine , Uronic Acids/urineSubject(s)
Hexosaminidases/urine , Isoenzymes/urine , Acetamides , Chromatography, DEAE-Cellulose , Electrophoresis, Polyacrylamide Gel , Female , Heterozygote , Hexosaminidases/metabolism , Humans , Hydrogen-Ion Concentration , Kinetics , Lipidoses/enzymology , Lipidoses/urine , Male , Sex Factors , Spectrophotometry , Time FactorsSubject(s)
Galactosidases/metabolism , Glycopeptides/urine , Lipid Metabolism, Inborn Errors/metabolism , Lipidoses/metabolism , Adolescent , Chromatography , Galactosidases/analysis , Galactosidases/blood , Glucosidases/analysis , Glucosidases/blood , Glycosaminoglycans/urine , Hexosaminidases/analysis , Hexosaminidases/blood , Humans , Kidney/pathology , Lipid Metabolism, Inborn Errors/pathology , Lipid Metabolism, Inborn Errors/urine , Lipidoses/pathology , Lipidoses/urine , Liver/enzymology , Liver/pathology , Male , Rectum/enzymologySubject(s)
Brain/metabolism , Gangliosides/metabolism , Glycolipids/metabolism , Lipidoses/metabolism , Sphingolipids/metabolism , Brain/pathology , Glycolipids/blood , Glycolipids/urine , Humans , Infant , Lipidoses/blood , Lipidoses/pathology , Lipidoses/urine , Liver/pathology , Male , Myocardium/pathology , Sphingolipids/blood , Sphingolipids/urineSubject(s)
Amino Acids/urine , Lipidoses/urine , Adolescent , Adult , Autoanalysis , Child , Female , Histidine/urine , Humans , Imidazoles/urine , MaleSubject(s)
Carbohydrate Metabolism, Inborn Errors/urine , Glycoside Hydrolases , Lipid Metabolism, Inborn Errors/urine , Oligosaccharides/urine , Acetates/urine , Chemical Precipitation , Chromatography , Chromatography, Ion Exchange , Chromatography, Paper , Colorimetry , Ethanol , Fucose/urine , Galactose/urine , Gangliosides/metabolism , Glucosamine/urine , Glucose/analysis , Glycoproteins/metabolism , Glycoproteins/urine , Hexosamines/urine , Hexosaminidases , Hexoses/urine , Humans , Infant , Lipidoses/urine , Mannose/urine , Monosaccharides/urine , Neuraminic Acids/urine , Uronic Acids/urineABSTRACT
In an ever larger number of neurologic conditions, the diagnosis depends on biochemical procedures. These range in complexity from simple screening tests on urine or blood, to analyses of nervous tissue lipids or metabolic studies or explants of brain tissue. Examples of each these are provided.