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PubMed1993

Candidate reference methods for determining target values for cholesterol, creatinine, uric acid, and glucose in external quality assessment and internal accuracy control. II. Method transfer.

We describe the testing of transferability of candidate Reference Methods developed by INSTAND for cholesterol, creatinine, uric acid, and glucose. The methods are based on isotope dilution-gas chromatography--mass spectrometry. The study consisted of two parts: setup of the methods and self-evaluation for readiness in the collaborating laboratory, followed by independent measurements in parallel with INSTAND. Criteria used for judging the transferability and general reliability of the candidate Reference Methods were: the accuracy and precision of the collaborating laboratory and the agreement between the two laboratories. The accuracy was judged from the results on the Standard Reference Material 909 from the National Institute of Standards and Technology. For all analytes except glucose the bias from the certified value was < 0.7%. The mean intralaboratory imprecision ranged from 0.66% to 1.24%. The agreement between the results was tested by an advanced linear-regression analysis and Student's t-test. In general, the results demonstrate that the candidate Reference Methods developed by INSTAND can be successfully transferred without loss of their inherent precision and accuracy.

Blood Glucose↗
PubMed1994

Localization of uric acid oxidase activity in core and matrix of peroxisomes as detected in unfixed cryostat sections of rat liver.

Because of controversial data in the literature, we studied the localization of uric acid oxidase (UAOX) activity in rat liver by light microscopy (LM) and electron microscopy (EM). UAOX is partially inactivated by aldehyde fixation and therefore we developed a technique that permits the use of unfixed cryostat sections for both LM and EM studies. Sections of rat liver were mounted on a semipermeable membrane stretched over a gelled incubation medium containing urate as specific substrate for UAOX and cerium ions to capture H2O2 produced by oxidase activity. The specificity of the reaction was checked by comparing incubations in the presence of substrate with incubations either in the absence of substrate or in the presence of substrate and 2,6,8-trichloropurine, a competitive inhibitor of UAOX. After incubation the sections were either fixed immediately for EM or visualized for LM with a second-step incubation. At the LM level, final reaction product was found in a granular form, homogeneously distributed throughout the hepatocytes. EM revealed excellent subcellular morphology and electron-dense reaction product in both the core and the matrix of peroxisomes, but not in other organelles or the cytoplasmic matrix. After incubations without substrate or with substrate and inhibitor, hardly any reaction product was found. We conclude that, because of the use of unfixed tissue, UAOX is not inactivated, which results in localization of UAOX activity not only in the core of peroxisomes but also in the peroxisomal matrix.

Animals↗
PubMed1998

[Simultaneous determination of creatinine, pseudouridine and uric acid in serum and urine by high performance liquid chromatography].

In this paper a sensitive, rapid and simple HPLC method for the simultaneous determination of creatinine (Cr), pseudouridine (Pu) and uric acid(UA) has been established. We have evaluated clinical value of the method in the early diagnosis of diabetic nephropathy (DN). Separation was obtained using Shim-pack CLC-ODS 15 x 0.6 cm column and mobile phase of buffer solution of phosphate (pH 3.0, 0.02 mol/L) with flow rate of 1 mL/min. Detection was performed with UV detector at an automatic adjustment of wavelength. The recoveries of Cr, Pu and UA were 101.4%, 104.9% and 105.0% respectively. The calibration curves were linear within the concentration range of 8.6-274.6 mumol/L for Cr, 0.72-22.93 mumol/L for Pu and 19.1-612.7 mumol/L for UA (n = 6, r > 0.999, p < 0.01). The CV for within day and between day measurements were < 2.5% and < 5.0% respectively. In addition, Pu, Cr and UA were simultaneously determined in serum and urine of 39 patients with diabetic mellitus (DM). 15 patients with nephrotic syndrome (NS) and 53 normal subjects by the method. Results include: 1. the levels of serum Pu in all DM patients (100%) with microalbuminuria (MiAU) and macroalbuminuria (MaAU) were greater than maximum of normal subjects (X + 2S). In addition, in 9 DM patients (41%) with normal albuminuria (NAU) the levels were also greater than the maximum. 2. In patients with DM, there was no correlation between the serum Pu and the urinary albumin excretion (UAE), whereas the serum Pu correlated closely with the serum Cr. However, the incidence for serum Pu increase was significantly greater in patients. 3. The levels of serum Pu in patients with NS were greater than those in control subjects and patients with DM. Conclusion is that the determination of serum Pu could be used as sensitivity index for the early diagnosis of DN.

Chromatography, High Pressure Liquid↗
PubMed2002

Square wave voltammetric sensing of uric acid using the self-assembly of mercaptobenzimidazole.

The self-assembled monolayer of a heterocyclic thiol, mercaptobenzimidazole (MBI) on gold (Au) electrode is successfully utilized for the voltammetric sensing of uric acid (UA). The self-assembly of MBI separates the voltammetric signal of UA from the interfering ascorbate (AA). Selective detection of UA in the presence of a large excess of AA or the simultaneous detection of UA and AA is achieved at the MBI monolayer-modified electrode. This electrode can detect as low as 1 microM of UA in the presence of 100-fold excess of AA with excellent reproducibility. The practical utility of the electrode is demonstrated by measuring the concentration of UA in human serum.

Benzimidazoles↗
PubMed2005

[Multifactorial disorder: molecular and evolutionary insights of uric acid nephrolithiasis].

Nephrolithiasis is a common multifactorial disorder affecting about 10% of the Western populations and it is characterized by the presence of small crystals and stones in the urinary tract. Uric acid nephrolithiasis (UAN) accounts for 20% of all stones but its prevalence varies between countries. Nephrolithiasis is likely caused by several factors but a genetic component has clearly been demonstrated. While studying an ancient founder population in Sardinia, we recently identified a susceptibility locus for UAN on chromosome 10. In this region we identified a missense mutation in a specific isoform of a novel gene is strongly associated with UAN. Through a comparative genomic approach, we did not found a mouse homolog even if we were able to identify the corresponding genomic region, while in Old World monkey we found a canonical gene structure with several stop codons preventing protein production. We detected expression in New World monkeys while in humans we observe a functional protein. It seems, therefore, that, to avoid human disease, a fierce selection worked to develop a renal-haematic urate homeostasis system against excessive hyperuricaemia. ZNF365 emerged during primate evolution and assumed its role in parallel with the disappearance of uricase, probably against a disadvantageous excessive hyperuricaemia.

Animals↗
PubMed2003

Determination of uric acid in human saliva by high-performance liquid chromatography with amperometric electrochemical detection.

The aim of the present study is to establish a highly sensitive method for the determination of uric acid (UA) in human saliva. The monitoring of UA levels in less invasive biological samples such as saliva is suggested for the diagnosis and therapy of gout, hyperuricemia, and the Lesch-Nyhan syndrome, and for detecting such conditions as alcohol dependence, obesity, diabetes, high cholesterol, high blood pressure, kidney disease, and heart disease. Reversed-phase high-performance liquid chromatography with electrochemical detection (HPLC-ED) was employed for the determination of UA obtained by solid-phase extraction from saliva. To quantify UA, we compared the ED efficiencies of an amperometric ED (Ampero-ED) with a single electrode and a coulometric ED (Coulo-ED) with a multiple electrode array. The results showed that the detection limits (S/N=3) were 3 nM for Ampero-ED and 6 nM for Coulo-ED, and the linearity of the calibration curves of 60-6000 nM had correlation coefficients exceeding 0.999. In addition, the total analytical time was 10 min. In the sample preparation of UA in saliva, an Oasis MAX solid-phase cartridge was used. The recoveries of UA spiked at 0.6 and 3 microM in saliva were above 95% with a relative standard deviation (RSD) of less than 15%. Therefore, the present method may be used in the routine and diagnostic determination of UA in human saliva.

Chromatography, High Pressure Liquid↗
PubMed2005

[Serum uric acid and C-reactive protein levels in patients with chronic kidney disease].

Either inflammation or hyperuricemia has been related with increased cardiovascular risk and mortality. A hypothetical relationship between serum uric acid levels (SUA) and inflammatory markers has never been tested in chronic kidney disease (CKD) patients. The purpose of this study was to determine the prevalence of increased C-reactive protein (CRP) levels in CKD patients, and to test the hypothesis of a relationship between SUA and CRP levels. The study group consisted of 337 patients (174 males, mean age 63 +/- 16 years) with advanced chronic renal failure not yet on dialysis. None of them had overt inflammatory or infectious diseases. High sensitivity CRP levels were analyzed as a binary (above or below median value), or continuous variable (log-transformed CRP), by multiple logistic or linear regression analysis, respectively. Demographics, clinical, and biochemical characteristics, including SUA levels, were the variables tested in these analysis. In a subset of 169 patients without diabetes, the same analysis were carried out, with the inclusion of fasting insulin levels and HOMA-IR as independent variables. Median CRP level was 3.25 mg/L, and mean SUA level was 7.59 +/- 1.94 mg/dl. Patients with CRP levels above the median had significantly higher mean SUA level than that of the rest of study patients (7.93 +/- 1.79 vs 7.24 +/- 2.03 mg/dl, p = 0.001). SUA levels correlated significantly with log-transformed CRP levels (r = 0. 16, p = 0.0022). The relationship between SUA and CRP levels remained statistically significant after adjustment for age, sex, comorbid index, obesity, residual renal function, diuretic and allopurinol treatment, in the multivariate logistic and linear regression models (OR: 1.296, p = 0.0003; and beta: 0.204, p = 0.0002). The significant association between SUA and CRP levels did not change when HOMA-IR and fasting insulin levels were included as independent variables in the subset of 169 patients without diabetes. In conclusion, SUA levels are related with CRP levels in CKD patients.

C-Reactive Protein↗
PubMed1980

Anaerobic degradation of uric Acid by gut bacteria of termites.

A study was done of anaerobic degradation of uric acid (UA) by representative strains of uricolytic bacteria isolated from guts of Reticulitermes flavipes termites. Streptococcus strain UAD-1 degraded UA incompletely, secreting a fluorescent compound into the medium, unless formate (or a formicogenic compound) was present as a cosubstrate. Formate functioned as a reductant, and its oxidation to CO(2) by formate dehydrogenase provided 2H + 2e needed to drive uricolysis to completion. Uricolysis by Streptococcus UAD-1 thus corresponded to the following equation: 1UA + 1formate --> 4CO(2) + 1acetate + 4NH(3). Urea did not appear to be an intermediate in CO(2) and NH(3) formation during uricolysis by strain UAD-1. Formate dehydrogenase and uricolytic activities of strain UAD-1 were inducible by growth of cells on UA. Bacteroides termitidis strain UAD-50 degraded UA as follows: 1UA --> 3.5 CO(2) + 0.75acetate + 4NH(3). Exogenous formate was neither required for nor stimulatory to uricolysis by strain UAD-50. Studies of UA catabolism by Citrobacter strains were limited, because only small amounts of UA were metabolized by cells in liquid medium. Uricolytic activity of such bacteria in situ could be important to the carbon, nitrogen, and energy economy of R. flavipes.

Journal Article↗
PubMed1984

Chemolysis of uric acid stones.

This paper illustrates one of the more satisfying aspects of the treatment of urinary calculous disease. A small series of 8 patients is presented to emphasize the possibility of dissolving uric acid stones by conservative measures thereby saving the patients extensive and often destructive surgical procedures. Patient selection is important for successful therapy. The stone must not be calcified and there must be sufficient function in the kidney. The regime consists of allopurinol 100 mg and sodium bicarbonate 2 to 4 grams given orally three times a day. Treatment period lasted up to six months, with dissolution of stones.

Adult↗
PubMed1985

Plasma uric acid: mass spectrometric method for control and comparison of intra- and interlaboratory studies.

We propose a preliminary study for the utilization of isotope dilution and mass fragmentography for the measurement of plasma uric acid. Using stable isotopes it is possible to measure this compound in comparison with the molecule labelled with three 15N atoms (1,3,9). We achieved the determination at the molecular peak. A linear calibration curve is obtained from standard solutions between 0 and 600 mumol/l. The method has good precision and the results obtained with different quality control materials demonstrate the need for better accuracy. This method could, after more detailed studies, be a good candidate for a definitive method.

Gas Chromatography-Mass Spectrometry↗
PubMed1980

Ticrynafen in anephric patients: effects on uric acid and pharmacokinetics.

Ticrynafen was given to 6 anephric patients undergoing maintenance hemodialysis. Ticrynafen was given daily for the 3 days between hemodialyses. Ticrynafen had no effect on the interdialysis rise in serum uric acid levels. Ticrynafen did not accumulate in serum, but levels of metabolites continued to rise over the 3 days. Hemodialysis (5 hr) reduced levels of ticrynafen by 38% but had less effect on metabolite levels. There was no effect on serum cholesterol or triglycerides.

Adult↗
PubMed1979

Oxygen binding of fetal and adult bovine hemoglobin in the presence of organic phosphates and uric acid riboside.

The P50 values of "stripped" fetal and adult bovine hemoglobin were 18.4 and 28.9 respectively. Neither the oxygen-hemoglobin dissociation curve nor the Hill coefficient, n, of fetal or adult bovine hemoglobin was affected by uric acid riboside (UAR), 2,3-diphosphoglyceric acid (2,3-DPG), adenosine triphosphate (ATP), or inositol pentaphosphate (IPP). Combinations of UAR and ATP with adult bovine hemoglobin or 2,3-DPG and ATP with fetal hemoglobin also had no effect. It was concluded that neither adult nor fetal bovine red cells contained an identifiable compound which affects the binding of oxygen to hemoglobin.

Adenosine Triphosphate↗
PubMed1992

Possible mechanism of inhibition of nitrite-induced oxidation of oxyhemoglobin by ergothioneine and uric acid.

The time course of oxyhemoglobin oxidation by nitrite consisted of a kinetic lag followed by a transition phase which progressed into a rapid autocatalytic phase. The imidazolthione and imidazolone derivatives, ergothioneine and uric acid, respectively, caused an increase in the duration of the lag phase in a concentration-dependent manner, without affecting the onset and rate of the autocatalytic phase. Neither compound reacted with H2O2 or nitrite, oxidizing species required in the initiation steps of oxyhemoglobin oxidation. On the other hand, both compounds reduced effectively and at comparable rates the high oxidation state of hemoglobin, i.e., ferrylhemoglobin, which is an intermediate species occurring in the autocatalytic phase. In addition, the rate of ergothioneine oxidation, upon its reaction with ferrylmyoglobin, was accelerated by nitrite, thus suggesting a reaction between the thione and nitrogen dioxide. Nitrogen oxide and ferrylhemoglobin are key species in the free radical chain propagation leading to oxyhemoglobin oxidation by nitrite. These data support the view that ergothioneine and urate delay oxyhemoglobin oxidation by nitrite upon the temporary removal of the propagating species, i.e., nitrogen dioxide and, secondarily, ferrylhemoglobin, and within a mechanism encompassing alterations of the nitrite in equilibrium with nitrogen dioxide and ferrylhemoglobin in equilibrium with methemoglobin redox transitions.

Adult↗
PubMed1980

[Uric acid and oxalate lithiasis. Physico-chemical and crystallo-chemical explanation of the relationship].

It is tried to explain physico-chemically or crystallo-chemically the so-called "salting out effect", i.e. the influence of the Ca-oxalate precipitation by uric often discussed in urolith research which, indeed, is regarded as real by several researchers and by other investigators refused with emphasis, in order to understand better the connections between hyperuricosuria and oxalate lithiasis which obtrude themselves from the observation material. However, this phenomenon of the facilitated oxalate precipitation at increased uric acid level in the urine further needs directed research.

Calcium Oxalate↗