High level of phosphatidylcholines/lysophosphatidylcholine ratio in urine is associated with prostate cancer.

The altered levels of phospholipids (PLs) and lysophospholipids (LPLs) in prostate cancer (CaP) and benign tissues in our previous findings prompted us to explore PLs and LPLs as potential biomarkers for CaP. Urinary lipidomics has attracted increasing attention in clinical diagnostics and prognostics for CaP. In this study, 31 prostate tissues obtained from radical prostatectomy were assessed using high-resolution matrix-assisted laser desorption/ionization imaging mass spectrometry (HR-MALDI-IMS). Urine samples were collected after digital rectal examination (DRE), and urinary lipids were extracted using the acidified Bligh-Dyer method. The discovery set comprised 75 patients with CaP and 44 with benign prostatic hyperplasia (BPH) at Kyoto University Hospital; the validation set comprised 74 patients with CaP and 59 with BPH at Osaka University Hospital. Urinary lipidomic screening was performed using MALDI time-of-flight MS (MALDI-TOF/MS). The levels of urinary lysophosphatidylcholine (LPC) and phosphatidylcholines (PCs) were compared between the CaP and BPH groups. The (PC [34:2] + PC [34:1])/LPC (16:0) ratio was significantly higher (P < .001) in CaP tissues than in benign epithelial tissues. The urinary PCs/LPC ratio was significantly higher (P < .001) in the CaP group than in the BPH group in the discovery and validation sets.

Acute kidney disease and acute kidney injury biomarkers in coronary care unit patients.

BACKGROUND: Acute kidney disease (AKD) describes acute or subacute damage and/or loss of kidney function for a duration of between 7 and 90 days after exposure to an acute kidney injury (AKI) initiating event. This study investigated the predictive ability of AKI biomarkers in predicting AKD in coronary care unit (CCU) patients. METHODS: A total of 269 (mean age: 64 years; 202 (75%) men and 67 (25%) women) patients admitted to the CCU of a tertiary care teaching hospital from November 2009 to September 2014 were enrolled. Information considered necessary to evaluate 31 demographic, clinical and laboratory variables (including AKI biomarkers) was prospectively recorded on the first day of CCU admission for post hoc analysis as predictors of AKD. Blood and urinary samples of the enrolled patients were tested for neutrophil gelatinase-associated lipocalin (NGAL), cystatin C (CysC) and interleukin-18 (IL-18). RESULTS: The overall hospital mortality rate was 4.8%. Of the 269 patients, 128 (47.6%) had AKD. Multivariate logistic regression analysis revealed that age, hemoglobin, ejection fraction and serum IL-18 were independent predictors of AKD. Cumulative survival rates at 5 years of follow-up after hospital discharge differed significantly (p < 0.001) between subgroups of patients diagnosed with AKD (stage 0A, 0C, 1, 2 and 3). The overall 5-year survival rate was 81.8% (220/269). Multivariate Cox proportional hazard analysis revealed that urine NGAL, body weight and hemoglobin level were independent risk factors for 5-year mortality. CONCLUSIONS: This investigation confirmed that AKI biomarkers can predict AKD in CCU patients. Age, hemoglobin, ejection fraction and serum IL-18 were independently associated with developing AKD in the CCU patients, and urine NGAL, body weight and hemoglobin level could predict 5-year survival in these patients.

Urinary Lipidomics: evidence for multiple sources and sexual dimorphism in healthy individuals.

Urinary lipidomics may add new valuable biomarkers to the diagnostic armamentarium for early detection of metabolic and kidney diseases. Sources and composition of urinary lipids in healthy individuals, however, have not been investigated in detail. Shotgun lipidomics was used to quantify lipidomic profiles in native urine samples from 16 individuals (eight men, eight women) collected in five fractions over 24 h. All probands were comprehensively characterized by urinary and clinical indices. The mean total urinary lipid concentration per sample was 0.84 µM in men and 1.03 µM in women. We observed significant intra- and interindividual variations of lipid concentrations over time, but failed to detect a clear circadian pattern. Based on quantity and subclass composition it seems very unlikely that plasma serves as major source for the urinary lipidome. Considering lipid metabolites occurring in at least 20% of all samples 38 lipid species from 7 lipid classes were identified. Four phosphatidylserine and one phosphatidylethanolamine ether species (PE-O 36:5) were detectable in almost all urine samples. Sexual dimorphism has been found mainly for phosphatidylcholines and phosphatidylethanolamines. In men and in women urinary lipid species were highly correlated with urinary creatinine and albumin excretion, reflecting glomerular filtration and tubular transport processes. In women, however, lipid species deriving from urinary cells and cellular constituents of the lower genitourinary tract considerably contributed to the urinary lipidome. In conclusion, our study revealed the potential of urinary lipidomics but also the complexity of methodological challenges which have to be overcome for its implementation as a routine diagnostic tool for renal, urological and metabolic diseases.

Distinct urinary lipid profile in children with focal segmental glomerulosclerosis.

BACKGROUND: Focal segmental glomerulosclerosis (FSGS) accounts for the majority of new-onset end-stage renal disease (ESRD) during adolescence. FSGS treatment is a great challenge for pediatric nephrologists due to intertwined molecular pathways underlining its complex pathophysiology. There is emerging evidence showing that perturbed lipid metabolism plays a role in the pathophysiology of FSGS. METHODS: We postulate that the nephrotic milieu in FSGS differs from minimal change disease (MCD) and that urinary lipidomics can be used as a tool for early diagnosis of FSGS. We explored the urinary lipid profile of patients with FSGS and MCD using an unbiased metabolomics approach. RESULTS: We discovered a unique lipid signature characterized by increased concentration of fatty acid (FA) and lysophosphatidylcholines (LPC) and a decrease in urinary concentration of phosphatidylcholine (PC) in patients with FSGS. These findings indicate increased metabolism of membrane phospholipid PC by phospholipase A2 (PLA2), resulting in higher urinary concentrations of LPC and FA. CONCLUSIONS: We propose that increased PC by-products can be used as a biomarker to diagnose FSGS and shed light on the mechanism of tubular and podocyte damage. Validation of identified urinary lipids as a biomarker in predicting the diagnosis and progression of FSGS in a larger patient population is warranted.

Quantitative analysis of phosphatidylcholines and phosphatidylethanolamines in urine of patients with breast cancer by nanoflow liquid chromatography/tandem mass spectrometry.

Phosphatidylcholines (PCs) and phosphatidylethanolamines (PEs) from urine of patients with breast cancer were qualitatively and quantitatively analyzed by nanoflow liquid chromatography-electrospray ionization tandem mass spectrometry (nLC-ESI-MS-MS). Urinary phospholipids (PLs) were extracted from three different categories of patients (non-cancer controls and breast cancer patients before and after surgery) by first lyophilizing only 1 mL of urine sample to enrich PLs. Next, nLC-ESI-MS-MS analysis of intact urinary phospholipids was performed, resulting in structural identification of 21 PCs and 12 PEs, followed by quantitative analysis using a multiple standard addition method. This study demonstrated that nLC-ESI-MS-MS can be powerfully utilized for the study of relative changes in the contents and concentration of urinary PCs and PEs from breast cancer patients: total concentration of PCs and PEs of patient sample increased to (144 +/- 9)% and (171 +/- 11)%, respectively, compared to control sample but they decreased significantly following surgery.

Occurrence of novel branched-chain fatty acids in Refsum's disease.

Two novel branched-chain fatty acids, which appear to be unsaturated analogs of phytanic acid, have been observed in sera and urine of patients with Refsum's disease. They occur in both phospholipids and neutral lipids, and have been isolated and characterized.

A case of prune-belly syndrome: prenatal diagnostic problems.

The authors discuss a case of "Prune-belly syndrome" which could be diagnosed prenatally thanks to echography; they stress the importance of echographic controls since the first gestational weeks to better recognize pathological findings, and describe their attempts to drain the megavesica. However these attempts could not reverse the ominous prognosis.

Phosphatidylarsenocholine, one of the major arsenolipids in marine organisms: synthesis and metabolism in mice.

Marine organisms contain arsenic at high levels, both in water-soluble and lipid-soluble forms. In contrast to the accumulated knowledge on water-soluble arsenic compounds, toxicological properties of lipid-soluble arsenic compounds (arsenolipids) have been little understood. Therefore, this study was aimed to clarify the metabolism of phosphatidylarsenocholine, one of the major arsenolipids so far identified in marine organisms. Phosphatidylarsenocholine (dipalmitoyl) was synthesized from phosphatidylcholine (dipalmitoyl) and arsenocholine by the transphosphatidylation reaction with phospholipase D and its synthesis was confirmed by LC/ESI-MS analysis. When phosphatidylarsenocholine was orally administered to mice at 45 µg As/mouse, arsenic was excreted mainly in urine almost in parallel with the time elapsed. The excretion rate was considerably slow compared to the case of water-soluble arsenic compounds but more than 90% of the administered arsenic was excreted within 144 h after administration. Analysis by LC/ESI-MS revealed that the major urinary metabolite was arsenobetaine, although small amounts of arsenocholine were detected in urine up to 72 h. These results allowed us to conclude that phosphatidylarsenocholine is mostly absorbed from the gastrointestinal tract in mice, metabolized to arsenobetaine and slowly excreted mainly in urine.

Biomarkers to monitor drug-induced phospholipidosis.

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.

Population pharmacokinetics of amphotericin B lipid complex in neonates.

The pharmacokinetics of amphotericin B lipid complex (ABLC) were investigated in neonates with invasive candidiasis enrolled in a phase II multicenter trial. Sparse blood (153 samples; 1 to 9 per patient, 1 to 254 h after the dose) and random urine and cerebrospinal fluid (CSF) samples of 28 neonates (median weight [WT], 1.06 kg; range, 0.48 to 4.9 kg; median gestational age, 27 weeks; range, 24 to 41 weeks) were analyzed. Patients received intravenous ABLC at 2.5 (n = 15) or 5 (n = 13) mg/kg of body weight once a day over 1 or 2 h, respectively, for a median of 21 days (range, 4 to 47 days). Concentrations of amphotericin B were quantified as total drug by high-performance liquid chromatography. Blood data for time after dose (TAD) of <24 h fitted best to a one-compartment model with an additive-error model for residual variability, WT0.75 (where 0.75 is an exponent) as a covariate of clearance (CL), and WT as a covariate of volume of distribution (V). Prior amphotericin B, postnatal age, and gestational age did not further improve the model. The final model equations were CL (liters/h) = 0.399 x WT(0.75) (interindividual variability, 35%) and V (liters) = 10.5 x WT (interindividual variability, 43%). Noncompartmental analysis of pooled data with a TAD of >24 h revealed a terminal half-life of 395 h. Mean concentrations in the urine after 1, 2, and 3 weeks ranged from 0.082 to 0.430 microg/ml, and those in CSF ranged from undetectable to 0.074 microg/ml. The disposition of ABLC in neonates was similar to that observed in other age groups: weight was the only factor that influenced clearance. Based on these results and previously published safety and efficacy data, we recommend a daily dosage between 2.5 and 5.0 mg/kg for treatment of invasive Candida infections in neonates.

Determination of alkylphosphocholines by high-performance liquid chromatography with light-scattering mass detection.

A rapid and sensitive method for the determination of five different alkylphosphocholines including the antineoplastic phospholipid analogues hexadecylphosphocholine and octadecylphosphocholine is presented. The method is based on the separation of the lipids by high-performance liquid chromatography and quantitation by light-scattering mass detection. The lower limit of detection is approximately 50 pmol for each alkyphosphocholine tested. Quantitation is linear over the range 0.05-75 nmol. Hexane-isopropanol extracts of cultured cells can be applied to the column without further cleanup. The high resolution of separation and the sensitivity of detection render this method useful for pharmacokinetic investigations dealing with the uptake of alkylphosphocholines into different types of cells.

[Urinary excretion of emulsified lipids in urolithiasis]. / Ekskretsiia s mochoi ému'lgirovannykh lipidov pri moche- kamennoi bolezni.

Examinations of the urine emulsified lipid phospholipid composition carried out by thin-layer chromatography have revealed that phospholipid excretion is the minimal in normal subjects and is represented only by sphingomyelin and phosphatidylcholine fractions. In subjects with a history of cholelithiasis (CL) phospholipid excretion is essentially elevated and the total phospholipid spectrum, including the minor fractions, is present in the urine. In CL patients phospholipiduria+ is still more marked, phosphatidylethanolamine appears in the urine, that is absent in the other groups of examinees. This method is recommended for the early diagnosis of CL.

Evaluation of human fetal urine as a source of amniotic fluid phospholipids.

The accumulation of surface active material in amniotic fluid during gestation is assumed to result from lung fluid secretion through the trachea. Some animal studies, however, have indicated that virtually all of the tracheal fluid is swallowed, whereas little if any enters the amniotic cavity. Following these observations fetal urine has been considered by some authors as an alternative source of amniotic fluid phospholipids. However, the phospholipid content of human fetal urine has not yet been determined. We have determined the L/S ratio and lamellar body concentration in human amniotic fluid and fetal urine obtained during 3 term cesarean deliveries. While both the L/S ratio and lamellar body particle concentration in the amniotic fluid samples were equivalent to values reported in term pregnancies, no measurable lecithin or sphingomyelin peaks were demonstrated in the urine samples and very few, if any, particles were counted. The lack of similarity between determinants of surface activity in human amniotic fluid and fetal urine does not support a major contribution of fetal urine to the phospholipid content of amniotic fluid.

Characterization of urinary sulfatides in metachromatic leukodystrophy using electrospray ionization-tandem mass spectrometry.

Metachromatic leukodystrophy is an inherited disorder characterized by a deficiency of the lysosomal enzyme arylsulfatase A and the subsequent accumulation of sulfatide in neural and visceral tissues. Clinical diagnosis is usually confirmed by in vitro analysis of arylsulfatase A activity, but may be complicated in cases of arylsulfatase A pseudodeficiency and sphingolipid activator protein deficiency. Large quantities of sulfatide can be detected in the urinary sediment of affected individuals and its measurement can aid in diagnosis. A number of complex methods have been described for the measurement of urinary sulfatide excretion. We have developed a rapid, sensitive, and specific mass spectrometric method for determining urinary sulfatide concentration of metachromatic leukodystrophy patients. Sulfatides are extracted from urine and then directly analyzed using electrospray ionization-tandem mass spectrometry. A sulfatide internal standard has been employed for quantification. The assay has demonstrated significant elevations in the concentrations of several hydroxy and nonhydroxy molecular species of sulfatide in the urine of metachromatic leukodystrophy patients compared to age-matched controls. Analysis of urinary sulfatides in arylsulfatase A pseudodeficiency patients showed a mild elevation in some individuals when related to urinary phosphatidylcholine.

Persistent skin ulcers, mutilations, and acro-osteolysis in hereditary sensory and autonomic neuropathy with phospholipid excretion. Report of a family.

We observed three children in a Turkish family who from early childhood had deformations of the feet and torpid ulcers with subfocal osteomyelitis and osteolysis, which subsequently led to amputations. The fingers showed ainhumlike constriction bands and spontaneous amputations. Neurologic studies revealed an almost complete sensory and autonomic loss affecting all modalities and a marked involvement of motor fibers. The clinical symptoms fulfill many of the hallmarks of hereditary sensory and autonomic neuropathy type II, including autosomal recessive inheritance, onset of symptoms in childhood, and mutilating acropathy. A high urinary excretion of sphingomyelin and lecithin suggests that the pathogenic mechanism may be a disorder of phospholipid metabolism.