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Biomedical subjects

F Linari

Publications and source records attributed to F Linari.

At least 37 records · Page 2Linked to original sources

Plasma and urine glycolate assays for differentiating the hyperoxaluria syndromes.

To differentiate hyperoxaluria syndromes we measured plasma and urine glycolate by a novel high performance liquid chromatographic procedure. Mean glycolate level was 7.9 +/- 2.4 mumol./l. in plasma and 422 +/- 137 mumol./24 hours in urine from 19 control subjects. Renal clearance was about 50% the glomerular filtration rate irrespective of the underlying disease. There was close correlation between glycolate and oxalate in plasma. Plasma glycolate was normal in all but 8 patients who had primary hyperoxaluria 1. Plasma assay detected the disease more efficiently than urine assay. Pyridoxine decreased oxalate biosynthesis in 2 of the 4 patients treated with it and glycolate assay confirmed this behavior. Glycolate excretion was significantly high in 3 of 8 patients of primary hyperoxaluria 1 patients. Idiopathic stone formers had mild increases in glycolate excretion but this was not related with oxalate excretion. Glycolate levels were normal in 5 patients with enteric hyperoxaluria. We conclude that glycolate assay is essential for identifying patients with primary hyperoxaluria 1 and may represent a valuable tool for differentiating hyperoxaluria.

Adolescent

Oxalate balance studies in patients on hemodialysis for type I primary hyperoxaluria.

Primary hyperoxaluria type I (PH1) always leads to end-stage renal failure (ESRF) due to deposition of calcium oxalate in the kidney. Regular dialysis therapy (RDT) can not overcome the excess production of oxalate, hence, systemic oxalate deposition occurs. The extent of tissue deposition and the rate at which oxalate accumulates influence the quality of life and survival of the patients. Therefore, an estimate of the oxalate balance needs to be made for patients on RDT. In this study, we suggest a simple model by which some of the main parameters of oxalate turnover can be assessed without using radioactive materials. Levels of oxalate, glycolate, and urea, and degrees of calcium oxalate saturation, were assessed on plasma ultrafiltrates from two patients with PH1, sampled before, at the end of a dialysis session, and over the entire interdialytic interval. In patients with PH1, oxalate increased linearly during the early phases and then the curve flattened at a concentration corresponding to approximately threefold saturation. The initial phase of the relationship was used to estimate generation rate of oxalate. The delayed phase was ascribed to the deposition of newly generated oxalate out of its miscible pool. Conversely, the relationship for glycolate and urea remained linear. This was also different from the values obtained in four patients with oxalosis-unrelated ESRF, whose oxalate levels increased linearly over the entire interdialytic interval. In the two patients with PH1, the overall oxalate generation was assessed at 4 to 7 mmol/d. The difference between generation and dialysis removal indicated that tissue deposition was greater than 50 mumol/kg body weight/d.(ABSTRACT TRUNCATED AT 250 WORDS)

Adult

Plasma profiles and dialysis kinetics of oxalate in patients receiving hemodialysis.

Regular dialysis treatment (RDT) does not obviate hyperoxalemia of chronic renal failure (CRF). However, there is emerging evidence suggesting that current dialysis prescription is not always associated to progressive oxalate accumulation. In view of the controversy still concerning this issue, we have investigated on plasma profiles and dialysis kinetics of oxalate in patients on RDT. Oxalate was determined by ion chromatography on serum ultrafiltrates and on the whole dialyzate in 23 stable patients on RDT for end-stage renal failure unrelated to primary hyperoxaluria. Nine patients were on traditional hemodialysis (HD) and 14 on soft hemodiafiltration (HDF). Dialysis prescription was set so as to obtain similar KT/V of urea. Mean dialyzer clearance of oxalate (KdOx) was calculated by standard procedures and was compared to urea (KdUrea) and creatinine (KdCr) clearances. Oxalate removal was measured on the whole spent dialyzate. Distribution volume of oxalate (VOx) was estimated by assuming a single-pool model and was used to estimate the oxalate appearance rate (OxAR). Plasma profiles showed that dialysis patients were virtually always hyperoxalemic. However, the threshold of supersaturation for calcium oxalate was exceeded in only 13 of 138 (9.4%) assayed ultrafiltrates, 13% on HD and 7.1% on HDF. Dialysis reduced plasma oxalate by more than 60%. There was a postdialysis oxalate rebound averaging 9.6% at 30 min from the end of dialysis. Plasma oxalate predialysis was independent of sex, age and time on dialysis. KdOx was mildly higher on HDF than on HD, and was lower than both KdUrea and KdCr, irrespective of the dialysis technique.(ABSTRACT TRUNCATED AT 250 WORDS)

Adult

High-performance liquid chromatographic determination of plasma glycolic acid in healthy subjects and in cases of hyperoxaluria syndromes.

A liquid chromatographic procedure for the determination of glycolic acid in plasma is proposed. The system is based on pre-column derivatization of the alpha-keto acid by means of phenylhydrazine, coupled with the enzymatic oxidation of glycolate to glyoxylate. The phenylhydrazone formed is separated by reversed-phase liquid chromatography and detected by UV absorption. The measured within and between-batch CV imprecision was 2.6 and 11.3%, respectively, at 5.68 mumol/l glycolate concentration; the analytical recovery was 102.0 +/- 7.3% and the minimum detectable concentration of glycolate was 0.3 mumol/l. The reference interval for plasma glycolate was 4.51 to 12.20 mumol/l (n = 14). Results of determinations of plasma samples from uremic patients, patients with type I primary hyperoxaluria and patients with chronic renal failure secondary to systemic oxalosis are reported.

Chromatography, High Pressure Liquid

Glycolate determination detects type I primary hyperoxaluria in dialysis patients.

The detection of type I primary hyperoxaluria is based on the finding of exceedingly high oxalate excretion which is associated with increased glycolate excretion. The differential diagnosis of this disease may become a difficult task once end-stage renal disease (ESRD) and anuria have supervened. The various procedures thus far proposed to obviate this circumstance are complex, inaccurate or not reproducible. In this paper we propose the accurate liquid chromatographic determination of glycolate in blood and dialysate as a means to detect type I primary hyperoxaluria in patients on maintenance hemodialysis (RDT). The method is based on the enzymatic conversion of glycolate to glyoxylate coupled with alpha-keto acid derivatization with phenylhydrazine. The resulting phenylhydrazone is then resolved by high-performance liquid chromatograph (HPLC). With this method, plasma glycolate in 12 healthy controls was 7.8 +/- 1.7 mumol/liter, almost twentyfold less than previously reported. Five dialysis patients with high serum oxalate, of whom four with primary hyperoxaluria and one with Crohn's disease and presumed enteric oxalate hyperabsorption, were checked by this method and compared to nine patients with oxalosis-unrelated ESRD. The patients with hyperoxalemia were also evaluated for their response to pyridoxine therapy. The measurement of glycolate in blood drawn prior to and at the end of the dialysis session as well as in the dialysate soundly discriminated the patients with type I hyperoxaluria from all the other dialysis patients. Glycolate measurement was shown to be much more powerful than oxalate in that patients with oxalosis-induced ESRD exhibited an almost two hundred and fiftyfold increase compared to the oxalosis-unrelated patients.(ABSTRACT TRUNCATED AT 250 WORDS)

Adolescent

Plasma profiles and removal rates of inorganic sulphate, and their influence on serum ionized calcium, in patients on maintenance haemodialysis.

1. Regular dialysis treatment is reported to remove inorganic sulphate, but not to restore its level to normal. To evaluate the adequacy of modern dialysis techniques in maintaining the sulphate balance, intra- and interdialysis plasma profiles and removal rates of sulphate were studied in 20 stable patients on maintenance haemodialysis. The influence of sulphate levels on the distribution of calcium-complex species was also investigated. 2. Sulphate was determined by ion-exchange chromatography of both serum ultrafiltrates, taken at the start of, during, at the end of, and at 24 h and 48 h after a dialysis session, and whole diffusate collections. Dialyser clearances of sulphate were assessed by two independent procedures and compared with those of urea and creatinine, on two different methods of dialysis, i.e. traditional haemodialysis with Cuprophan hollow fibre filters, and haemodiafiltration with high-flux Polysulphone or polyacrylonitrile dialysers. Concentrations of the main serum ions were determined before and after dialysis and used to solve a multiple mass balance equation system by which concentrations of the calcium-complex species were calculated. 3. Before dialysis, sulphate levels were eight times those determined in 17 control subjects and remained higher than normal at the end of dialysis. These changes were independent of the dialysis procedure. There was a close correlation between serum levels of sulphate and creatinine. Dialyser clearances of sulphate were comparable with those of creatinine, but lower than those of urea. Clearances of all solutes were higher on haemodiafiltration.(ABSTRACT TRUNCATED AT 250 WORDS)

Adult

Serum calcium oxalate saturation in patients on maintenance haemodialysis for primary hyperoxaluria or oxalosis-unrelated renal diseases.

1. The serum oxalate concentration rises in chronic renal failure and it is only partially eliminated by regular dialysis treatment. However, the recent literature is not conclusive on whether progressive oxalate retention and secondary oxalosis should be expected in patients on regular dialysis treatment. 2. To further investigate this, we have estimated the state of saturation with respect to calcium oxalate monohydrate in plasma ultrafiltrates from 28 patients on maintenance haemodialysis and eight healthy control subjects, matched for sex and age. Five patients had type I primary hyperoxaluria and histologically proven oxalosis, whereas 23 had oxalosis-unrelated renal diseases. Dialysis efficiency was quantified as the KdTd/V of urea. Samples were obtained from each patient before, immediately after and 48 h after a dialysis session. Fasting samples were obtained from the control subjects. Oxalate was determined in both plasma ultrafiltrates and the whole dialysate by ion-exchange chromatography, after a non-delayed and [14C]oxalate-recovery-controlled procedure. The state of saturation with calcium oxalate monohydrate was estimated by means of a computer system which solved the interactions among 45 complex species. 3. The fasting plasma oxalate concentration (means +/- SD) in ultrafiltrates from healthy subjects was 3.8 +/- 1.5 (range 1.4-5.8) mumol/l, and the state of saturation with calcium oxalate monohydrate was 0.096 +/- 0.04.(ABSTRACT TRUNCATED AT 250 WORDS)

Adolescent

Renal handling of citrate in chronic renal insufficiency.

Citrate is a relevant component of the inhibitory potential of the urine environment. Its excretion and renal handling have been widely studied in subjects with normal renal function, but little is known about changes induced by chronic renal insufficiency. We have investigated renal handling of citrate in 50 patients with different degrees of renal insufficiency as compared to 30 healthy subjects with normal renal function. Among patients 34 were defined as having mild renal insufficiency based on a GFR of 80 through 40 ml/min/1.73 m2 BSA, while 16 had moderate-to-severe renal failure, defined by a GFR ranging from 40 to 20 ml/min/1.73 m2 BSA. Serum citrate increased in mild renal insufficiency, while it tended to be restored to normal values at more advanced renal failure. There was a stepwise decrease in the filtered load of citrate as GFR decreased, while its renal clearance was significantly reduced only at higher degrees of renal failure. This behavior was due to an increase in the fractional excretion of citrate which was inversely related to the decrease in GFR (p = 0.015). These data suggest that serum citrate levels and excretion are governed by renal mechanisms at mild degrees of renal insufficiency; in these conditions citrate is shown to behave conformly to other poorly reabsorbable anions such as sulfate. At more advanced renal failure the ensuing metabolic acidosis plays a crucial role as a regulatory factor of both serum concentration and renal handling of citrate, by increasing cellular uptake and metabolism as well as tubular reabsorption of this anion.(ABSTRACT TRUNCATED AT 250 WORDS)

Acid-Base Imbalance

Ion-chromatographic determination of L-tartrate in urine samples.

We propose using ion chromatography to determine tartrate concentration in urine. A 100-microL sample of urine is diluted and injected into the chromatograph. Tartrate is eluted within 11.5 min as a distinct and well-resolved peak. The sensitivity of the standard procedure (signal-to-noise ratio, 3/1) is 30 mumol/L. The intra-run and interrun coefficients of variation are 2.5% and 4.1%, respectively. Mean analytical recovery of known amounts of added tartrate ranges between 94.2% and 104.0%. We investigated the specificity of the procedure by analyzing urine containing added dicarboxylic acids structurally related to tartrate. The reliability of the procedure makes it suitable for investigating tartrate metabolism, e.g., the potential role of tartrate as an inhibitor of crystallization in calcium nephrolithiasis.

Adult

Ion chromatographic determination of plasma oxalate in healthy subjects, in patients with chronic renal failure and in cases of hyperoxaluric syndromes.

An ion chromatographic procedure for the determination of plasma oxalate is proposed, in which the ultrafiltered sample is injected into an ion-chromatographic system. Sample processing appears effective in avoiding spontaneous oxalogenesis. Sensitivity (down to 1.0 mumol/l) allows determinations in normal and pathological samples; recoveries from plasma ultrafiltration are 94.6 +/- 11.7%. Protein binding was investigated and precautions to improve recoveries from plasma ultrafiltration are proposed. The technique is simple to perform from healthy controls averaged 6.75 +/- 2.62 mumols/l (mean +/- S.D. n = 18); samples from patients with primary hyperoxaluria and chronic renal failure undergoing regular dialysis were also analysed and some of the data obtained are reported and discussed.

Adult

Prevalence of chronic renal insufficiency in the course of idiopathic recurrent calcium stone disease: risk factors and patterns of progression.

The occurrence of chronic renal insufficiency was investigated in 171 patients with severe idiopathic calcium stone disease. Ninety healthy subjects matched for age and sex were used as controls. The patients were thereafter subclassified into two subgroups, assuming a GFR of 80 ml/min/1.73 m2 body surface area as a cut-off value: the normal GFR, 141 patients, and the impaired GFR, 30 patients. The normal GFR group included more males and the patients were younger both at onset and at presentation. In the impaired GFR group the disease lasted longer, but the overall stones and the stone recurrence rate were as high as those of the normal GFR patients. The single stone episodes were more severe in the former group as suggested by the occurrence of more surgery and complications. The GFR level was in part predicted by the age of patients; however, stone disease was shown to induce a clear-cut influence in accelerating the natural worsening of GFR with age. The onset of renal insufficiency causes multiple changes in renal pathophysiology, which result in a sharp decrease in the urine saturation with respect to calcium salts. These changes account for the decrease in the stone recurrence rate in the impaired GFR group. Thus, unless factors independent of or complicating the calcium stone disease supervene, the renal insufficiency of treated patients remains mild and relently progressive.

Adult

Preventing ascorbate interference in ion-chromatographic determinations of urinary oxalate: four methods compared.

In an attempt to decrease ascorbate interference on the ion-chromatographic determination of urinary oxalate, we compared the effectiveness of four different methods for ascorbate elimination by analyzing a representative urine pool supplemented with successive ascorbate additions. Two of the methods--treatment with ferric ions or boric acid--have been described elsewhere; treatments with nitrites or ascorbate oxidase (EC 1.10.3.3) are investigated here as possible alternatives. Consideration of the main features, advantages, and drawbacks of the four procedures leads us to conclude that boric acid dilution is a good routine method and that pre-incubation with ascorbate oxidase reliably prevents ascorbate interference in assays of urinary oxalate.

Adult

Derivatization and high-performance liquid chromatographic determination of urinary glycolic acid.

A high-performance liquid chromatographic method for the determination of urinary glycolic acid is proposed, based on pre-column derivatization with phenylhydrazine coupled with the enzymatic oxidation of glycolate to glyoxylate. The phenylhydrazone formed is separated by liquid chromatography and detected at 324 nm. The minimum detectable concentration of glycolate was 10.0 mumol/l. The recovery of glycolate added to urine averaged 96.1%. The day-to-day coefficients of variation calculated by analysis of two urine samples with normal and high glycolate contents were 4.6 and 7.5%, respectively. Results of analyses of urine samples from healthy persons, idiopathic calcium stone formers and Type I primary hyperoxaluria patients are reported.

Chromatography, High Pressure Liquid