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Lever pressing for food reward and changes in dopamine turnover and uric acid in rat caudate and nucleus accumbens studied chronically by in vivo voltammetry.

The neurotransmitter dopamine (DA) is widely held to be involved in the maintenance of positively reinforced behaviour, on the basis of self-stimulation experiments and of the effects of drugs which alter DA transmission on self-stimulation, and on behaviours controlled by natural reinforcers. We have used the technique of linear sweep voltammetry at chronically implanted carbon paste electrodes to demonstrate that 30-min sessions of responding on a variable interval schedule for food pellet reward increases first uric acid and then homovanillic acid (HVA, the DA metabolite) in rat caudate. Ascorbic acid levels are reduced, but at later times. We have now established that very similar responses in magnitude and timing are seen in the nucleus accumbens. Furthermore, in individual animals, the response rates, the increases in dopamine turnover, and the changes in uric acid and ascorbic acid remain stable when studied periodically over a 3 month period. Our previous results are broadly consistent with those obtained by others using indirect single-point methods (ex vivo neurochemistry) or in vivo dialysis. However, our present extended observations made possible by the use of paste electrodes (rather than fibre electrodes or dialysis probes) open the possibilities of studying the evolution of neurochemical responses during learning, and the covariation of neurochemical and behavioural effects of drugs within individual animals.

Animals↗

Kinetics and fates of ammonia, urea, and uric acid during oocyte maturation and ontogeny of the Atlantic halibut (Hippoglossus hippoglossus L.).

Considering that amino acids constitute an important energy fuel during early life of the Atlantic halibut (Hippoglossus hippoglossus L.), it is of interest to understand how the nitrogenous end products are handled. In this study we focused on the kinetics and fates of ammonia, urea and uric acid. The results showed that ammonia (T(Amm): NH(3)+NH(4)(+)), and urea-N contents increased during final oocyte maturation. Urea-N excretion dominated the total nitrogenous end product formation in early embryos. Later, yolk T(Amm) levels increased in embryos and ammonia excretion was low. In the last part of the embryonic stage T(Amm) accumulation dominated, and was apparently due to yolk storage. Around hatching, the larval body tissues (larva with yolk-sac removed) accounted for 68% of whole animal urea-N accumulation, while T(Amm) levels increased predominately by yolk accumulation. Afterwards, ammonia excretion dominated and uric acid accumulation accounted for less than 1%. Urea, synthesised either through the ornithine-urea cycle, argininolysis or uricolysis, accounted for approximately 8% of total nitrogenous end product formation in yolk-sac larvae. The results suggested that a sequence occurred regarding which nitrogenous end products dominated and how they were handled. Urea excretion dominated in early embryos (<7 dPF), followed by yolk ammonia accumulation (7-12 dPF), and finally, ammonia excretion dominated in later embryonic and yolk-sac larval stages (>12 dPF).

Ammonia↗

[Effects of allopurinol on uric acid metabolism and lipid peroxidation in ischemic heart disease patients with stable angina].

The study was undertaken to examine 20 healthy individuals and 60 IHD patients with stable angina of various functional classes. All the examinees were males. By the random sampling method, all the patients were divided into 2 groups: (1) 30 were on erinit and (2) 30 on erinit and allopurinol in conventional therapeutical doses. The level of uric acid in the serum and daily urine, lipid peroxidation, the antioxidative system and central hemodynamics were examined in all the examinees. The examination was performed on admission and every 10-14 days of their hospital stay. A combined therapy with erinit and allopurinol led to a significant decrease in serum and daily urinary levels of uric acid and lipid peroxidation antioxidative system and an improvement of central hemodynamics. It is concluded that it is useful to include allopurinol into the combined therapy for IHD patients with stable angina.

Adult↗

A new microdissection technique for regional biochemical analysis of the rat spinal cord: serotonin, norepinephrine, dopamine and uric acid.

This report described a new microdissection procedure to evaluate the regional distribution of neuromediators in the rat spinal cord. Different segments are first divided into sagittal slices. From these, different grey and white matter regions can be microdissected. This permits selective biochemical measurements in different laminae, including the area around the central canal. White and grey matter can also be differentially analysed. Using HPLC with electrochemical detection, we report on the regional analysis of biogenic amines as well as uric acid. An increase in 5-hydroxytryptamine (5-HT) levels was observed from cervical to lumbar segments. 5-HT levels were highest in the motoneurone samples (lamina IX) of the lumbar cord. The next highest levels were found in lamina X, followed by the intermediate grey matter laminae and the dorsal horn. Small amounts of 5-HT were detected in the white matter. Interestingly, the ratio of 5-hydroxyindoleacetic acid (5-HIAA) to 5-HT was greatest in the white matter and least in the motoneurone sample. Norepinephrine (NE) levels were higher in the thoracic than in the cervical or the lumbar cord. The highest levels were found in the lamina X in thoracic segments. With this exception, no marked laminar difference in NE levels was observed. Dopamine (DA) levels were highest in the dorsal horn of the cervical and the thoracic cord, the next highest levels were found in the intermediate grey matter and lamina X in the same segments. The distribution of uric acid (UA) was comparable to that of NE: UA levels were highest in the thoracic cord, but no marked laminar difference was observed. On the other hand, UA levels in white matter generally exceeded those in the grey matter. These data indicate the value of a sagittal regional microdissection of the spinal cord. The ability to separately analyse different laminae of the cord (as well as differentiating grey and white matter) should prove useful in future studies of experimentally evoked changes in neurotransmitters within functionally distinct regions of the spinal cord.

Animals↗

Ultrafiltrable uric acid in serum of healthy humans and patients with renal calcium lithiasis.

Ultrafiltrable serum uric acid (u.a.) was determined by ultrafiltration under in vivo conditions in humans using a reliable technique described in detail. It could be demonstrated that u.a. binding to macromolecules occurs in healthy humans (controls) and in patients with renal calcium stones. The percentage of free u.a. in controls (n=60) averages 86.2 +/- 0.9 SEM. With increasing age, bound u.a. rises slightly. On the other hand, younger (less than 40 years) stone patients have significantly more bound u.a. than matched controls (80.7 +/- 1.0 SEM; p less than 0.001), whereas this is not found in elderly patients. The degree of binding is not related to concentration of plasma proteins but inversely related to free fatty acid concentration in healthy controls (r= -0.52; p less than 0.01). It is suggested that no augmentation of tubular u.a. filtered by the glomeruli could have occurred. The origin of fasting hyperuricosuria shown earlier to be a prominent feature of young renal calcium stone formers is yet unknown.

Adolescent↗

Evaluation of a quantitative solid phase reagent system for determination of blood analytes. Experiences with the analytes: LDH, bilirubin, BUN, glucose, and uric acid.

In the first part of the Seralyzer system evaluation the precision and accuracy was studied with a total of 1245 clinical specimens, various commercial control sera, and pooled human sera. The calculated overall precision (between-run, and within-run) and day-to-day precision (% CV) was found to be within 3.0 to 5.4 for glucose, 2.8 to 7.1 for BUN, 1.5 to 6.2 for uric acid, 5.3 to 7.5 for bilirubin, and 3.2 to 8.6 for LDH. The clinical values are in agreement with values from respective comparative methods, as indicated by the regression statistics. The analysis of Seralyzer accuracy data using quality control sera showed in some cases a between-method difference. Supporting studies simulating additional important clinical situations showed that the clinical values for BUN, glucose, and uric acid of approximately 200 specimens from the emergency ward correlated with the respective comparative method values. In this phase of the study we verified that the instrument calibration was stable for a 24-hour period and that there is no effect of module (test) change on precision of Seralyzer determinations. The intra- and inter-laboratory performance in general practitioners' laboratories could be demonstrated using quality control sera.

Bilirubin↗

Prooxidant and antioxidant properties of human serum ultrafiltrates toward LDL: important role of uric acid.

Oxidized LDL is present within atherosclerotic lesions, demonstrating a failure of antioxidant protection. A normal human serum ultrafiltrate of Mr below 500 was prepared as a model for the low Mr components of interstitial fluid, and its effects on LDL oxidation were investigated. The ultrafiltrate (0.3%, v/v) was a potent antioxidant for native LDL, but was a strong prooxidant for mildly oxidized LDL when copper, but not a water-soluble azo initiator, was used to oxidize LDL. Adding a lipid hydroperoxide to native LDL induced the antioxidant to prooxidant switch of the ultrafiltrate. Uric acid was identified, using uricase and add-back experiments, as both the major antioxidant and prooxidant within the ultrafiltrate for LDL. The ultrafiltrate or uric acid rapidly reduced Cu2+ to Cu+. The reduction of Cu2+ to Cu+ may help to explain both the antioxidant and prooxidant effects observed. The decreased concentration of Cu2+ would inhibit tocopherol-mediated peroxidation in native LDL, and the generation of Cu+ would promote the rapid breakdown of lipid hydroperoxides in mildly oxidized LDL into lipid radicals. The net effect of the low Mr serum components would therefore depend on the preexisting levels of lipid hydroperoxides in LDL. These findings may help to explain why LDL oxidation occurs in atherosclerotic lesions in the presence of compounds that are usually considered to be antioxidants.

Amidines↗

[Role of uric acid in hypertension and in the progression of chronic renal disease].

In humans, uric acid (UA) is the main urinary metabolite of purines, whereas in other species (avians) it is the chief nitrogen compound excreted in the urine. The absence of uricase, due to nonsense or splice mutations occurring during the evolution of primates, results in blood urate levels much higher in humans than in other mammalians. This could have favorable implications, including protection from oxidative damage, as well as possibly allowing better blood pressure (BP) control in settings of low dietary Na+ intake. UA is a stimulus of the renin-angiotensin system (RAS). Moreover, blood UA levels are a major marker of proximal tubular function and overall volume status. On the other hand, hyperuricemia is a co-factor in Na+ -sensitive arterial hypertension, a marker and perhaps itself responsible for microvascular and systemic damage through RAS stimulation, inhibitory effects on endothelial cells, and the proliferation of perivascular smooth muscle cells. Recent studies in rats rendered hyperuricemic through administration of oxonic acid demonstrate the induction of hypertension and severe microvascular damage when associated with subtotal nephrectomy or chronic cyclosporine treatment. The opportunity to pharmacologically manipulate blood UA levels through inhibitors of synthesis (allopurinol), uricosuric agents (benziodarone), or recombinant urate oxidase (rasburicase) provides a relevant therapeutic potential in UA metabolism disorders, tumor lysis syndrome, and possibly in essential hypertension, as well as chronic progressive renal disease.

Animals↗

Pharmacologic evaluation of the renal handling of uric acid and oxypurines.

The effect of drugs that alter the renal tubular transport of urate in the renal excretion of the oxypurines, hypoxanthine (Hx) and xanthine (X), was used to analyze the tubular mechanisms involved in oxypurine excretion in normal subjects and in patients with idiopathic renal hypouricemia. In healthy subjects, administration of the drugs, benzbromarone (Bb) and probenecid (Pb), brought a marked uricosuric effect, but did not alter oxypurine excretion. One of 2 hypouricemic patients exhibited no uricosuric effects with Bb and the other showed a slight uricosuric effect with the drug. Bb administration, however, produced no increase in oxypurine excretion in either patient. In healthy subjects, administration of pyrazinamide (PZA) suppressed the excretion of urate and oxypurines to varying degrees: relative to the baseline values, the fraction excretion of urate was reduced by 95% or more, that of Hx by about 24%, and that of X by about 64%. In patients with hypouricemia, the effects of PZA on urate and oxypurine excretion were impaired. According to these findings, we can speculate that the oxypurines have the same renal secretory mechanism as uric acid. However, the reabsorptive mechanisms of oxypurines at the postsecretory site are likely to differ from that of uric acid.

Adult↗

Chemiluminescence biosensor chip based on a microreactor using carrier air flow for determination of uric acid in human serum.

A chemiluminescence biosensor on a chip coupled to a microfluidic system and a microreactor is described in this paper. The chemiluminescence biosensor measured 25 x 75 x 6.5 mm in dimension, and was readily produced in an analytical laboratory. The sol-gel method is introduced to co-immobilize horseradish peroxidase (HRP) and luminol in the microreactor, and to immobilize uricase in the enzymatic reactor. The main characteristic of the biosensor was to introduce air as the carrier flow instead of the more common solution carrier for the first time. The uric acid was determined by a chemiluminescent (CL) reaction between the hydrogen peroxide produced from the enzymatic reactor and luminol under the catalysis of HRP in the microreactor. The linear range of the uric acid concentration was 1 to 100 mg L(-1) and the detection limit was 0.1 mg L(-1) (3sigma).

Biosensing Techniques↗

[Outpatient care in uric acid disorders].

In 35 patients with uricopathy (arthritis urica, essential hyperuricaemia) the serum uric acid values, consumption of alcohol, overweight, hypertension and liver enzyma under dietary and medicamentous therapy (allopurinol) were investigated. Here during an observation lasting 12--48 months allopurinol proved as a well tolerable uricostatic drug which was sufficiently effective also with a considerable exogenic purine application. We only insufficiently succeeded in influencing overweight and alcohol consumption as well as hypertension by rheumatic dispensary care and family doctor.

Adult↗

Serum uric acid concentrations in a Xhosa community in the Transkei of Southern Africa.

Serum uric acid (SUA) concentrations were measured in specimens obtained from 479 respondents aged 15 and over during an epidemiological survey of bone and joint conditions in a tribal Xhosa community living in the Transkei, Southern Africa. Population SUA levels in the Xhosa were significantly lower than those previously observed in South Africa Negro groups in rural and urban environments. This finding agrees with the observations that population SUA concentrations tend to rise with increasing complexity of life style.

Adolescent↗

Recapture of [S]-allantoin, the product of the two-step degradation of uric acid, by urate oxidase.

Urate oxidase from Aspergillus flavus catalyzes the degradation of uric acid to [S]-allantoin through 5-hydroxyisourate as a metastable intermediate. The second degradation step is thought either catalyzed by another specific enzyme, or spontaneous. The structure of the enzyme was known at high resolution by X-ray diffraction of I222 crystals complexed with a purine-type inhibitor (8-azaxanthin). Analyzing the X-ray structure of urate oxidase treated with an excess of urate, the natural substrate, shows unexpectedly that the active site recaptures [S]-allantoin from the racemic end product of a second degradation step.

Allantoin↗

Effect of nifedipine on tubular handling of uric acid in transplanted kidney on cyclosporine A treatment.

Hyperuricemia resulting from tubular urate transport defects is a well-known nephrotoxic effect of cyclosporine A (CyA) on renal blood perfusion in organ recipients. The aim of this study was to define the mechanism of the tubular urate transport defect in hyperuricemic renal graft recipients on CyA and to evaluate the effect of nifedipine retard administration on this tubular dysfunction. Tubular uric acid transport was evaluated by the probenecid test in 17 hyperuricemic (group 1) and 6 normouricemic (group 2) renal graft recipients treated with CyA. Maximal urate excretion after probenecid administration was 25.5 +/- 4.6 versus 42.5 +/- 7.7% (p < 0.001) and postsecretory urate reabsorption was 79.2 +/- 8.3 versus 78.5 +/- 9.7% (NS), respectively. The effects of nifedipine retard on renal urate transport were evaluated in 6 hyperuricemic patients. Seven days of nifedipine therapy did not significantly decrease mean serum uric acid levels (7.7 +/- 1.6 to 7.1 +/- 1.1 mg/dl) nor increase urate clearance (3.8 +/- 1.3 to 4.7 +/- 1.6 ml/min/1.73 m2). The uricosuric effect of probenecid was manifested by an increase in tubular urate transport from 25.5 +/- 4.6 to 37.3 +/- 7.2%, p < 0.01, paralleled by an increase in postsecretory urate reabsorption from 20.5 +/- 3.7 to 31.4 +/- 5.7% (p < 0.003). Postsecretory reabsorption expressed as a percentage of secreted urate in both evaluations did not differ significantly (80.3 +/- 6.2 vs. 84.4 +/- 3.1%).(ABSTRACT TRUNCATED AT 250 WORDS)

Adolescent↗

Longitudinal study of urinary excretion of phosphate, calcium, and uric acid in mutant NHERF-1 null mice.

NHERF-1 binds numerous renal protein targets, including the proximal tubule transporters Na(+)/H(+) exchanger 3 (NHE3) and Na(+)-phosphate cotransporter 2a (Npt2a). Young NHERF-1(-/-) male mice display defective targeting of Npt2a to apical membranes in the renal proximal tubule and manifest hypophosphatemia and increased urinary excretion of phosphate. The present studies describe the changes in the urinary excretion of phosphate, calcium, uric acid, and sodium in male and female wild-type and NHERF-1 null mice over a time period from 12 to 54 wk of age. Young male and female NHERF-1(-/-) mice demonstrated increased urinary excretion of phosphate and urine phosphate/creatinine ratios. There was an age-related decline in the phosphate/creatinine ratio in mutant mice such that there were no differences between wild-type and NHERF-1(-/-) by 24 to 30 wk of age despite the continued presence of hypophosphatemia. Male and female NHERF-1 null mice also demonstrate increased urine calcium/creatinine and uric acid/creatinine ratios compared with wild-type controls. These studies indicate defects in the renal tubule transport of phosphate, calcium, and uric acid in NHERF-1(-/-) male and female mice that could account for the increased deposition of calcium in the papilla of null mice.

Age Factors↗