A novel quantification method for sulfur-containing biomarkers of formaldehyde and acetaldehyde exposure in human urine and plasma samples.

A novel method for the quantification of the sulfur-containing metabolites of formaldehyde (thiazolidine carboxylic acid (TCA) and thiazolidine carbonyl glycine (TCG)) and acetaldehyde (methyl thiazolidine carboxylic acid (MTCA) and methyl thiazolidine carbonyl glycine (MTCG)) was developed and validated for human urine and plasma samples. Targeting the sulfur-containing metabolites of formaldehyde and acetaldehyde in contrast to the commonly used biomarkers formate and acetate overcomes the high intra- and inter-individual variance. Due to their involvement in various endogenous processes, formate and acetate lack the required specificity for assessing the exposure to formaldehyde and acetaldehyde, respectively. Validation was successfully performed according to FDA's Guideline for Bioanalytical Method Validation (2018), showing excellent performance with regard to accuracy, precision, and limits of quantification (LLOQ). TCA, TCG, and MTCG proved to be stable under all investigated conditions, whereas MTCA showed a depletion after 21 months. The method was applied to a set of pilot samples derived from smokers who consumed unfiltered cigarettes spiked with 13C-labeled propylene glycol and 13C-labeled glycerol. These compounds were used as potential precursors for the formation of 13C-formaldehyde and 13C-acetaldehyde during combustion. Plasma concentrations were significantly lower as compared to urine, suggesting urine as suitable matrix for a biomonitoring. A smoking-related increase of unlabeled biomarker concentrations could not be shown due to the ubiquitous distribution in the environment. While the metabolites of 13C-acetaldehyde were not detected, the described method allowed for the quantification of 13C-formaldehyde uptake from cigarette smoking by targeting the biomarkers 13C-TCA and 13C-TCG in urine.Graphical abstract.

Urinary Excretion of Sulfur Metabolites and Risk of Cardiovascular Events and All-Cause Mortality in the General Population.

Aims: Thiosulfate and sulfate are metabolites of hydrogen sulfide (H2S), a gaseous signaling molecule with cardiovascular (CV) protective properties. Urinary thiosulfate excretion and sulfate excretion are associated with favorable disease outcome in high-risk patient groups. We investigated the relationship between urinary excretion of sulfur metabolites, and risk of CV events and all-cause mortality in the general population. Results: Subjects (n = 6839) of the Prevention of Renal and Vascular End-stage Disease (PREVEND) study were followed prospectively. At baseline, 24-h urinary excretion of thiosulfate and sulfate was determined. Median urinary thiosulfate and sulfate excretion values were 1.27 (interquartile range [IQR] 0.89-2.37) µmol/24 h and 15.7 (IQR 12.0-20.3) mmol/24 h, respectively. Neither thiosulfate nor sulfate excretion showed an independent association with risk of CV events. Sulfate, but not thiosulfate, was inversely associated with risk of all-cause mortality, independent of potential confounders (hazard ratio 0.73 [95% confidence interval 0.63-0.84], p < 0.001). This association appeared most pronounced for normolipidemic subjects (pinteraction = 0.019). Innovation: The strong association between sulfate excretion and mortality in the general population emphasizes the (patho)physiological importance of sulfate or its precursor H2S. Conclusion: We hypothesize that urinary sulfate excretion, which is inversely associated with all-cause mortality in the general population, holds clinical relevance as a beneficial modulator in health and disease. Antioxid. Redox Signal. 30, 1999-2010.

Novel sulphur-containing imatinib metabolites found by untargeted LC-HRMS analysis.

Untargeted metabolite profiling using high-resolution mass spectrometry coupled with liquid chromatography (LC-HRMS), followed by data analysis with the Compound Discoverer 2.0™ software, was used to study the metabolism of imatinib in humans with chronic myeloid leukemia. Plasma samples from control (drug-free) and patient (treated with imatinib) groups were analyzed in full-scan mode and the unknown ions occurring only in the patient group were then, as potential imatinib metabolites, subjected to multi-stage fragmentation in order to elucidate their structure. The application of an untargeted approach, as described in this study, enabled the detection of 24 novel structurally unexpected metabolites. Several sulphur-containing compounds, probably originating after the reaction of reactive intermediates of imatinib with endogenous glutathione, were found and annotated as cysteine and cystine adducts. In the proposed mechanism, the cysteine adducts were formed after the rearrangement of piperazine moiety to imidazoline. On the contrary, in vivo S-N exchange occurred in the case of the cystine adducts. In addition, N-O exchange was observed in the collision cell in the course of the fragmentation of the cystine adducts. The presence of sulphur in the cysteine and cystine conjugates was proved by means of ultra-high resolution measurements using Orbitrap Elite. The detection of metabolites derived from glutathione might improve knowledge about the disposition of imatinib towards bioactivation and help to improve understanding of the mechanism of its hepatotoxicity or nephrotoxicity in humans.

High sulfur content in corn dried distillers grains with solubles protects against oxidized lipids by increasing sulfur-containing antioxidants in nursery pigs.

Some sources of corn dried distillers grains with solubles (DDGS) contain relatively high amounts of oxidized lipids produced from PUFA peroxidation during the production process. These oxidized lipids may impair metabolic oxidation status of pigs. The objective of this study was to understand the effects of feeding corn-soybean meal diets (CON) or diets containing 30% highly oxidized DDGS with 1 of 3 levels of supplemental vitamin E (dl-α-tocopheryl acetate), none, the 1998 NRC level (11 IU/kg), and 10x the 1998 NRC level (110 IU/kg), on oxidative status of nursery pigs. The DDGS source used in this study contained the greatest thiobarbituric acid reactive substances (TBARS) value, peroxide value, and total S content (5.2 ng/mg oil, 84.1 mEq/kg oil, and 0.95%, respectively) relative to 30 other DDGS sources sampled (mean values = 1.8 ng/mg oil, 11.5 mEq/kg oil, and 0.50%, respectively). Barrows (n = 54) were housed in pens and fed the experimental diets for 8 wk after weaning and transferred to individual metabolism cages for collection of feces, urine, blood, and liver samples. Total S content was greater in DDGS diets than in CON (0.39 vs. 0.19%). Dietary inclusion of 30% DDGS improved apparent total tract digestibility of S (86.8 vs. 84.6%; P < 0.001) and S retained (2.94 vs. 2.07 g/d; P < 0.01) compared with CON. Although pigs were fed highly oxidized DDGS in this study, serum TBARS were similar between DDGS and CON treatments. There was an interaction between DDGS and dietary vitamin E level for serum concentrations of α-tocopherol. Serum α-tocopherol concentrations were greater (P < 0.001) in pigs fed DDGS diets than those fed CON when dl-α-tocopheryl acetate was not provided or provided at the NRC level but were similar when dl-α-tocopheryl acetate was supplemented at the 10x NRC level. Pigs fed DDGS diets had greater serum concentrations of S-containing AA, particularly Met (P < 0.001) and taurine (P = 0.002), compared with those fed CON. Liver glutathione concentration was greater in pigs fed DDGS diets than CON (56.3 vs. 41.8 nmol/g). Dietary inclusion of DDGS (P < 0.001) and vitamin E (P = 0.03) increased enzyme activity of glutathione peroxidase. The elevated concentrations of S-containing antioxidants (Met, taurine, and glutathione) in vivo may protect pigs against oxidative stress when feeding highly oxidized DDGS. Therefore, the increased S content in DDGS may be beneficial, and increasing concentrations of vitamin E in diets may not be necessary to protect pigs against metabolic oxidative stress when feeding high S and highly peroxidized DDGS.

Sulphur tracer experiments in laboratory animals using 34S-labelled yeast.

We have evaluated the use of (34)S-labelled yeast to perform sulphur metabolic tracer experiments in laboratory animals. The proof of principle work included the selection of the culture conditions for the preparation of sulphur labelled yeast, the study of the suitability of this labelled yeast as sulphur source for tracer studies using in vitro gastrointestinal digestion and the administration of the (34)S-labelled yeast to laboratory animals to follow the fate and distribution of (34)S in the organism. For in vitro gastrointestinal digestion, the combination of sodium dodecyl sulphate-polyacrylamide gel electrophoresis and high-performance liquid chromatography and inductively coupled plasma mass spectrometry (HPLC-ICP-MS) showed that labelled methionine, cysteine and other low molecular weight sulphur-containing biomolecules were the major components in the digested extracts of the labelled yeast. Next, in vivo kinetic experiments were performed in healthy Wistar rats after the oral administration of (34)S-labelled yeast. The isotopic composition of total sulphur in tissues, urine and faeces was measured by double-focusing inductively coupled plasma mass spectrometry after microwave digestion. It was observed that measurable isotopic enrichments were detected in all samples. Finally, initial investigations on sulphur isotopic composition of serum and urine samples by HPLC-ICP-MS have been carried out. For serum samples, no conclusive data were obtained. Interestingly, chromatographic analysis of urine samples showed differential isotope enrichment for several sulphur-containing biomolecules.

Preservation of nutrients during long-term storage of source-separated yellowwater.

Source separation of human urine (yellowwater) enhances the sustainability of wastewater management and efficiency of nutrient recovery and recycling. Storage of source-separated yellowwater is recommended prior to agronomic reuse. At this point, it is of immense interest to determine the effect of storage time on quality of yellowwater. Therefore, this study focused on examining changes in some chemical properties of raw, undiluted, freshly collected, source-separated yellowwater stored for a period of 1 year under different temperature regimes: cold (4 °C), mild (10 °C) and warm (22 °C). Chemical parameters (biochemical oxygen demand (BOD(5)), N-tot, N-NO(2), N-NO(3), N-NH(4), P-tot, K, S, and pH), with the main focus on fertiliser nutrient compounds intended for agricultural utilisation, were tested. The outcomes revealed that both nitrification and denitrification processes took place in the stored yellowwater, and an increase in the pH level of up to pH greater than 9 was observed. The study found that the main macronutrients can be well preserved in yellowwater, as there were no substantial changes in the contents of these elements over a 1 year storage period at the three temperatures tested.

Profiling biological samples using ultra performance liquid chromatography-inductively coupled plasma-mass spectrometry (UPLC-ICP-MS) for the determination of phosphorus and sulfur-containing metabolites.

In this preliminary study UPLC-ICP-MS has been utilized to profile a range of different bio-fluids and tissue extracts for sulfur and phosphorus-containing metabolites. Particular attention has been given to the livers, plasma and urine from lean and obese Zucker rats, with a view to differentiating between them based solely on their respective sulfur or phosphorus profiles and/or their total sulfur and phosphorus content. In addition, bile and tumour extracts have been analysed to observe the nature of their profiles. To the best of our knowledge this is the first time ICP-MS has been used in a non-targeted metabonomic study. Results have shown lower limits of quantification for sulfur and phosphorus methods of 0.25 and 0.15 ng on column with CVs of 14.7% and 10.9% respectively. Total phosphorus analysis of the Zucker rat aqueous liver extracts, plasma and urine has shown the pattern of phosphorus concentrations to be statistically significantly different in the lean and obese Zucker rats. Chromatographic separation of the Zucker rat organic liver extracts and plasma allowed further differentiation between the lean and obese rats using their phosphorus profiles alone. In conclusion, this preliminary study has shown the potential of UPLC-ICP-MS to quantitatively discriminate between different species biofluids, fluids and tissues based solely on their phosphorus or sulfur concentrations and/or metabolomes.

The distribution of several elements in cat urine and the relation between the content of elements and urolithiasis.

The concentrations of elements in urine obtained from cats with urolithiasis were compared with those of healthy cats. The concentration of several elements, such as sodium (Na), phosphorus (P), sulfur (S), and potassium (K), in urine obtained from cats with urolithiasis was significantly higher than that of healthy cats. A significant correlation (p<0.01) was found between the concentration of magnesium (Mg) and that of other elements, such as P (r=0.8913), S (r=0.6817), and K (r=0.8391), in the urine obtained from healthy cats. A significant correlation (r=0.7422, p<0.05) was also obtained between the concentration of K and that of P in urine collected from cats with urolithiasis, but the slope of regression line was significantly different from that of the urine obtained from healthy cats. Other correlations observed in healthy cats were not obtained from cats with urolithiasis. However, a significant correlation between the concentration of magnesium (Mg) and that of calcium was obtained only from cats with urolithiasis. The results of the present study suggest that urinary concentrations of various elements in cats with urolithiasis are higher than those of healthy cats. Furthermore, the balance of elements in the urine of cats with urolithiasis was altered.

Studies of a urinary biomarker of dietary inorganic sulphur in subjects on diets containing 1-38 mmol sulphur/day and of the half-life of ingested 34SO4(2-).

OBJECTIVE: Sulphites are widely used food additives that may damage health, hence limits are set on their use. They are excreted in urine as sulphate, along with sulphate derived from sulphur amino acids. Dietary intakes of sulphites are hard to determine, so we have tested the utility of urinary nitrogen:sulphate ratio as a biomarker of inorganic sulphur (IS) intake. Additionally we determined the half-life of ingested (34)SO(4)(2-) from its urinary excretion. SUBJECTS: Twenty healthy adult subjects were recruited by poster advertisement, for a 24-h study where they ate specified foods, which were high in IS, in addition to their normal diet. The half-life of ingested (34)SO(4)(2-) was assessed in five healthy volunteers, given 5.9 mmols of Na(2)(34)SO(4) as a single dose and collecting all urine specimens for 72-96 h. Urine and duplicate diets from three previously conducted studies were analysed for nitrogen and sulphate content, thus expanding the range of IS intakes for evaluation. METHODS: Duplicate diets were analysed for IS content by ion exchange chromatography, while IS intake was predicted from urinary sulphate (g/day S)-(urinary nitrogen (g/day)/18.89). (32)S:(34)S ratios were determined by liquid chromatography mass spectrometry/mass spectrometry. RESULTS: The range of IS intake was 1.3-37.5 mmol S/day. Actual and predicted IS intakes were mmol/day+/-s.e. 9.2+/-0.65 and 7.0+/-0.45, respectively, and were correlated r=0.60 (n=108). The mean half-life of ingested (34)SO(4)(2-) was 8.2 h. CONCLUSIONS: From a 24-h urine collection, IS intake from the habitual diet can be determined for groups of individuals. To predict individual intakes of IS, which may include high sporadic amounts from beer and wine, at least 48 h of urine collection would be required.

Elemental sulfur identified in urine of cheetah, Acinonyx jubatus.

The urine of the cheetah, Acinonyx jubatus, is almost odorless, and probably for this reason, it has not attracted much attention from scientists. Using gas chromatography-mass spectrometry, we identified 27 and 37 constituents in the headspace vapor of the urine of male and female cheetah, respectively. These constituents, composed of hydrocarbons, short-chain ethers, aldehydes, saturated and unsaturated cyclic and acyclic ketones, 2-acetylfuran, dimethyl disulfide, dimethyl sulfone, phenol, myristic acid (tetradecanoic acid), urea, and elemental sulfur, are all present in the headspace vapor in very small quantities; dimethyl disulfide is present in such a low concentration that it cannot be detected by the human nose. This is only the second example of elemental sulfur being secreted or excreted by an animal. It is hypothesized that the conversion of sulfur-containing compounds in the cheetah's diet to elemental sulfur and to practically odorless dimethyl sulfone enables this carnivore to operate as if "invisible" to the olfactory world of its predators as well as its prey, which would increase its chances of survival.

Manipulating the dietary cation-anion difference via drenching to early-lactation dairy cows grazing pasture.

Diets offered to grazing dairy cows can vary considerably in their dietary cation-anion difference (DCAD) and are often well in excess of what has been considered optimal. The effects of a range of DCAD on the health and production of pasture-based dairy cows in early lactation was examined in a randomized block design. Four groups of 8 cows were offered a generous allowance of pasture (45 +/- 6 kg/d of dry matter (DM) per cow) for 35 d and achieved mean pasture intakes of approximately 17 kg/d of DM per cow. Cows were drenched twice daily with varying combinations of mineral compounds to alter the DCAD. Dietary cation-anion difference ranged from +23 to +88 mEq/100 g of DM. A linear increase in blood pH and HCO(3)(-) concentration and blood base excess, and a curvilinear increase in the pH of urine with increasing DCAD indicated a nonrespiratory effect of DCAD on metabolic acid-base balance. Plasma concentrations of Mg, K, and Cl declined as DCAD increased, whereas Na concentration increased. Urinary excretion of Ca decreased linearly as DCAD increased, although the data suggest that the decline may be curvilinear. These results in conjunction with the increased concentrations of ionized Ca suggest that intestinal absorption of Ca or bone resorption, or both, increased as DCAD declined. Dry matter intake, as measured using indigestible markers, was not significantly affected by DCAD. However, the linear increase in the yield of linolenic acid, vaccenic acid, and cis-9, trans-11 conjugated linoleic acid in milk, as DCAD increased is consistent with a positive effect of DCAD on DM intake. Increasing DCAD did not significantly affect milk yield or milk protein, but the concentration and yield of milk fat linearly increased with increasing DCAD. The increased milk fat yield was predominantly a result of increased de novo synthesis in the mammary epithelial cells, although an increase in the yield of preformed fatty acids also occurred. Milk production results suggest that DCAD for optimal production on pasture diets may be higher than the +20 mEq/100 g of DM previously identified for total mixed rations.

Urinary sulfur excretion and the nitrogen/sulfur balance ratio reveal nonprotein sulfur amino acid retention in piglets.

We evaluated the use of urinary sulfur (S) excretion as a measure of sulfur amino acid (SAA) catabolism and the nitrogen/sulfur (N/S) molar balance ratio as an indicator of nonprotein SAA storage in growing piglets. After confirming that an intravenous dose of sulfate is fully recovered in urinary sulfate, we measured urinary S recovery after an intravenous dose of methionine in 6 piglets fed an adequate protein (AP) diet and 6 piglets fed a low protein (LP) diet with normal energy provision. As measured over 48 h, recoveries of the methionine load as urinary total S was 106% in the AP group but only 69% in the LP group (P < 0.05). On the baseline diets the N/S balance ratio in the AP group was 36, whereas that in the LP group was 30 (P < 0.05); immediately after the methionine load, this ratio remained constant in the AP group but decreased further, to 26 (P < 0.05) in the LP group. These results indicate that protein-deficient piglets accumulate relatively more S than N from their diet, and under these conditions a significant portion of the S derived from a methionine load is retained in nonprotein compounds. Urinary S excretion, a simple nontracer measurement, can provide an accurate measure of SAA catabolism, and the N/S balance ratio is a potentially useful indicator of changes in nonprotein SAA stores of growing piglets.

Directly coupled liquid chromatography with inductively coupled plasma mass spectrometry and orthogonal acceleration time-of-flight mass spectrometry for the identification of drug metabolites in urine: application to diclofenac using chlorine and sulfur detection.

We report the application of high-performance liquid chromatography (HPLC) linked to inductively coupled plasma mass spectrometry (ICPMS) and orthogonal acceleration time-of-flight mass spectrometry (oa-TOFMS) for the identification of phase I and II urinary metabolites of diclofenac. The metabolites were separated by reversed-phase HPLC monitored with a UV diode array detector (UV-DAD) after which 90% of the eluent was directed to an ICPMS source, with the remainder going to an oa-TOF mass spectrometer. Compounds containing (35)Cl, (37)Cl and (32)S were detected specifically using ICPMS and identified by oa-TOFMS. The metabolites detected and identified in this way included glucuronic acid and sulfate conjugates, mono- and dihydroxylated and free diclofenac. In addition a previously unreported in vivo metabolite, an N-acetylcysteinyl conjugate of diclofenac, was also characterised. This is the first application of the combination of HPLC/UV-DAD/ICPMS/oa-TOFMS for the investigation of the metabolic fate of chlorinated xenobiotics by direct biofluid analysis.

Massive loss of sulfur in HIV infection.

Skeletal muscle tissue from SIV-infected macaques was previously found to contain abnormally high sulfate and low glutathione levels indicative of an excessive cysteine catabolism. We now confirm the peripheral tissue as a site of massive cysteine catabolism in HIV infection and have determined the urinary loss of sulfur per time unit. The comparison of the sulfate concentrations of the arterial and venous blood from the lower extremities of 16 symptomatic HIV+ patients and 18 HIV- control subjects (study 1) revealed (1) that the peripheral tissue of HIV+ patients with or without highly active antiretroviral therapy (HAART) releases large amounts of sulfate and (2) that plasma sulfate, thioredoxin, and interleukin-6 levels are elevated in these patients. A complementary investigation of 64 asymptomatic HIV+ patients and 65 HIV- subjects (study 2) revealed increased plasma sulfate levels in the asymptomatic patients. The analysis of the daily urinary excretion of sulfate and urea of another group of 19 HIV+ patients and 22 healthy HIV- subjects (study 3) confirmed (1) that HIV+ patients experience a massive loss of sulfur and (2) that this loss is not ameliorated by HAART. The sulfur loss of asymptomatic patients was equivalent to a mean loss of about 10 g of cysteine per day. If extrapolated, this would correspond to an alarming negative balance of approximately 2 kg of cysteine per year under the assumption that the normal sulfate excretion equivalent to approximately 3 g of cysteine per day is balanced by a standard Western diet. The abnormally high sulfate/urea ratio suggests that this process drains largely the glutathione pool.

Some sulfur-containing metabolites of tri-n-butyltin chloride in male rats.

In an attempt to elucidate metabolic destination of TBTO, sulfur-containing metabolites were investigated in the urine. Tri-n-butyltin chloride (TBTC), tri-n-butyltin oxide (TBTO), and their in vitro metabolites in rat liver microsomal enzyme systems, di-n-butyl(3-hydroxybutyl)tin chloride (T3OH), di-n-butyl(3-oxobutyl)tin chloride (T3CO), dibutyltin dichloride (DBTC), and monobutyltin trichloride (MBTC), were intraperitoneally administered to rats. In particular, administration of T3OH and T3CO gave higher amounts of mercapturic acid derivatives, such as N-acetyl-S-(3-oxobutyl)-L-cysteine (3CO-MA) and N-acetyl-S-(3-hydroxybutyl)-L-cysteine (3OH-MA), than TBTC or TBTO. On the other hand, DBTC and MBTC did not yield measurable amounts of 3CO-MA and/or 3OH-MA. The appearance of organotin metabolites in urine indicates that T3OH, T3CO, and hypothesized secondary metabolites, such as n-butyl(3-hydroxybutyl)(3-oxobutyl)tin chloride, n-butyl(3-hydroxybutyl)(4-hydroxybutyl)tin chloride, etc., are subject to the action of glutathione S-transferase to give mercapturic acid derivatives. These sulfur-containing metabolites (3CO-MA and 3OH-MA) were also found in control rat urine.

The effect of dietary sulfur-containing amino acids on calcium excretion.

The relationships between dietary protein and sulfur amino acid (methionine and cystine or taurine) intakes and urinary calcium excretion were examined both in animals and in young men. Thirty-two adult Wistar rats were divided into 4 groups, i.e., basal diet (group I), supplemented with albumin (II), methionine and cystine (III), or taurine (IV). During the 5-week feeding period, food consumption was recorded and 48 h urine samples were collected 4 times for each rat. Urinary calcium, creatinine and sulfate were measured. The results showed that the calcium and sulfate excretion in rats in group II and III were significantly higher than rats in the basal diet group. In contrast, supplementing a basal diet with taurine did not increase sulfate excretion and failed to induce hypercalciuria. The same result was also observed in the study carried out in Chinese young men. An increase in protein intake from 67 g to 107 g caused an increase in urinary calcium and sulfate. Supplementation with methionine and cystine in an amount to simulate those in the high protein diet had a similar effect. Adding taurine to the diet had no effect on urinary calcium and sulfate excretion. About 60 percent of the supplemented taurine in the diet was detected in the urine.

Genetic and environmental effects on urinary kallikrein, catecholamines, sodium, potassium, urea nitrogen and inorganic sulfate sulfur levels in school-age twins.

A genetic analysis of twins at school was undertaken using as variables urinary concentrations of kallikrein, catecholamines, sodium and potassium which have been demonstrated to be associated with blood pressure levels. In addition to these variables, urinary concentrations of urea nitrogen and inorganic sulfate sulfur which are indices of protein intake were investigated. 35 pairs of monozygotic twins and 19 pairs of dizygotic twins aged from 6 to 14 years were examined. Variance and correlation tests for genetic analysis indicated that in school children, hereditary factors play a role in the control of urinary potassium, sodium and kallikrein excretion. However, with regard to the urinary excretion of catecholamines, urea nitrogen and inorganic sulfate sulfur, hereditary control is not so apparent.

Re-evaluation of the "week-end effect" data: possible role of urinary copper and phosphorus in the pathogenesis of renal calculi.

Early morning urinary concentrations of 10 elements which had demonstrated a "week-end effect" in a previous study, were subjected to a normalization procedure thereby allowing a re-assessment of their potential role in urolithiasis. After transformation of each concentration to a weighted proportion of the total concentration on each day, only Cu and P values were significantly different for kidney stone formers and healthy controls on all three days indicating that these elements may play a role in the pathogenesis of renal calculi. The results obtained in this study demonstrate that a more meaningful picture of the possible differences in the urinary concentrations of stone formers and normal controls might emerge if "proportional" rather than "raw" concentrations are compared.

Relative rates of appearance of nitrogen and sulphur: implications for postprandial synthesis of proteins.

The purpose of this study was to gain insights on the temporal fate of proteins based on the rate of appearance of waste products of nitrogen (urea) and sulphur (sulphate) metabolism. Urine was collected every 2 h from 25 normal subjects to measure the rates of excretion of urea, creatinine, and sulphate throughout the 24-h cycle. Samples of blood and urine were also obtained over a 4-h period from 10 subjects who consumed a mixed meal containing 0.4 g protein/kg body weight to obtain information on the relative rates of degradation of amino acids with and without sulphur in a noninvasive fashion. The daily excretion (mean +/- SEM) of urea, creatinine, and sulphate was 396 +/- 28, 14 +/- 0.4, and 15 +/- 0.6 mmol, respectively; the molar sulphate/nitrogen (S/N) ratio was 2.0 +/- 0.1%. There were relatively minor (< 20%) excursions in the rate of excretion of urea and creatinine in any 2-h period as compared with the corresponding 24-h rate; the concentrations of urea and creatinine in plasma also varied < 20% throughout the day. Only 23% of the nitrogen in protein in the standard meal appeared as urea in the 210 min after this meal was consumed. The small changes in the rate of appearance of urea and creatinine imply that the oxidation of amino acids was spread out over the day.(ABSTRACT TRUNCATED AT 250 WORDS)

Effect of dietary salt (NaCl) supplementation on urinary profile of guinea pigs (Cavia porcellus).

Sodium chloride supplementation (120 mg/kg of body weight/day) for 12 days increased the urinary excretion of calcium from 91.6 +/- 9.0 to 159.4 +/- 16.0 mumol/day and of sulphate from 266.8 +/- 24.5 to 1176.9 +/- 87.2 mumol/day in guinea pigs. The stone risk due to increased urinary calcium excretion could possibly be counterbalanced by increasing urinary sulphate excretion. High salt intake, thus, could not increase the risk of stone formation.