Use of a standard urine assay for measuring the phosphate content of beverages.

Hyperphosphatemia is strongly associated with cardiovascular morbidity and mortality in patients with chronic kidney disease. Phosphate in beverages is readily absorbed and could have a significant impact on serum phosphate levels. Patients are routinely warned about the phosphoric acid in colas, but information on the phosphate content of other beverages is difficult to find. We have shown that the phosphomolybdate method, which is used in the vast majority of hospital laboratories for measuring phosphate in urine, can give an accurate measurement of the phosphate content of beer, cider, wine, and soft drinks. No change to the standard assay protocol is required. There was considerable variation between different types of wine and beer, probably due to the methods of production. The information the assay provides could enable staff providing dietary advice to compare locally available beverages and help patients to avoid or limit their intake of those with high phosphate content.

Titratable acidity: a Pitts concept revisited.

Titratable Acidity (TA) in urine can be measured directly or calculated from actual and reference pH, by using the pKa2 6,8 for phosphate. In urine, H2PO4(-) represents the excretion of filtered H2PO4(-), filtrated HPO4(2-) being completely reabsorbed by the proximal tubule (the Van Slyke approach). Since excretion of H2PO4(-) frequently exceeds its glomerular filtration, this approach is considered inadequate by Pitts. He claimed that it is the tubular H(+) secretion which converts filtered HPO4(2-) to H2PO4(-), thereafter excreted in urine. This is only true under conditions of inorganic acid or neutral phosphate loading, when the maximum tubular phosphate reabsorption (TmPi) is overcharged. In controls, H2PO4(-) excretion is lower than its glomerular filtration, provided that acid-base status is normal and tubular phosphate reabsorption is below the TmPi. The TmPi is lower than its glomerular filtration, provided that acid-base status is normal and tubular phosphate reabsorption is below the TmPi. When the TmPi is exceeded, a portion of HPO4(2-) escapes proximal reabsorption, reaching the distal tubule where its absorption is precluded, while tubular H(+) secretion converts HPO4(2-) to H2PO4(-). In man and dog, the attainment of TmPi is evidenced by a FE% of 20%, and only beyond this limit H2PO4(-) excretion exceeds glomerular filtration. When FE% is lower than 20%, H2PO4(-) filtration exceeds excretion, HPO4(2-) being completely reabsorbed at the proximal tubule by NaPi-2a and 2c cotransporters. While Van Slyke's approach is always valid, Pitts' approach is only valid under loading conditions, when the two processes of H2PO4(-) excretion overlap each other. NH (+4) increases inversely to TA excretion in conditions of acidosis and tP restriction, but is independent of TA in Pi-replete dogs, independently of acidosis.

Evaluation of sodium bisulphate and phosphoric acid as urine acidifiers for cats.

Eighteen cats were used to compare the urine acidifying properties of sodium bisulphate to phosphoric acid. Acidifying agents were added at one of three concentrations (0.4, 0.6, or 0.8%, as-is basis). Cats were offered a commercial diet to determine basal urinary pH, and then again for a 1 week period between blocks 1 and 2. Cats were acclimated to the diets for 6 days, and urine samples were collected on day 7 at 0, 4, and 8 h postfeeding to obtain pre- and postprandial urinary pH. Intakes of diets containing sodium bisulphate tended (P < 0.07) to be lower than intakes of diets containing phosphoric acid. Cats consuming the 0.8% phosphoric acid diet had higher (P < 0.05) food intakes than cats consuming either the 0.4 or 0.6% phosphoric acid-containing diets. There was significant (P = 0.01) linear and quadratic response for food intake in cats consuming the sodium bisulphate-containing diet. Cats consuming the 0.4 and 0.8% phosphoric acid-containing diets tended (P = 0.07) to have higher water intakes than cats consuming the 0.6% phosphoric acid-containing diet. There were no differences (P > 0.05) in urine pH and specific gravity between cats fed the different acidifier types. Cats consuming the 0.6% phosphoric acid-containing diet tended (P = 0.07) to have a higher urine pH 8 h post-feeding than cats consuming the 0.4 and 0.8% phosphoric acid-containing diets. Urine pH was highest at 4 h post-feeding except for cats fed the 0.4% sodium bisulphate- and the 0.6% phosphoric acid-containing diets. No differences (P > 0.05) between acidifiers were found in faecal score or in faecal dry matter and organic matter concentrations. A quadratic response was detected in faecal score for cats consuming the phosphoric acid-containing diets. Cats consuming the 0.6% phosphoric acid diet tended (P = 0.06) to have a lower faecal score than cats consuming the 0.4 and 0.8% phosphoric acid diets. For faecal dry matter, a linear trend was detected in cats consuming the sodium bisulphate (P = 0.08) and phosphoric acid-containing (P = 0.04) diets. Sodium bisulphate and phosphoric acid generally behaved in a similar fashion when incorporated in dry cat diets.

Indices of intact serum parathyroid hormone and renal excretion of calcium, phosphate, and magnesium.

Up to date reference ranges were established for fasting renal excretion of calcium, phosphorus, and magnesium on 101 healthy children aged 2-15 years. A normal range for intact parathyroid hormone was also measured. The indices of calcium and magnesium excretion showed no correlation with age or sex so that a common range for all children could be established. The 97th centile values for urinary calcium:creatinine and magnesium:creatinine ratios were 0.69 mmol:mmol and 1.05 mmol:mmol respectively. The calculated tubular maximum for phosphate/litre of glomerular filtrate (TmPO4/GFR) showed no correlation with age with a geometric mean value of 1.67 mmol/l. The normal range for intact serum parathyroid hormone for the age group was 11-35 ng/l, which is lower than the adult normal range using the same assay. There was an inverse correlation between TmPO4/GFR and intact parathyroid hormone in this group of normal children.

Identification of new metabolites of phosphoramide and nor-nitrogen mustards and cyclophosphamide in rat urine using ion cluster techniques.

The metabolism of nor-nitrogen (NNM) and phosphoramide mustards (PM) and cyclophosphamide (CP) was investigated in the Sprague-Dawley rat using chemical ionization mass spectrometry and ion cluster techniques. Following administrations of a 1:1 mixture of the non-labeled and the corresponding side-chain deuterium-labeled compounds to separate rats, the urinary extracts were screened for cluster ions which were characteristic of the administered compounds and their derived metabolites, and on this basis, tentative identifications of known and new metabolites were accomplished. Combining derivatization, deuterium labeling on strategic locations, gas chromatography/mass spectrometry, and chemical synthesis in some cases, 3-(2-chloroethyl)-1,3-oxazolidin-2-one was identified as a major metabolite for NNM, PM and CP and 3-(2-chloroethyl)-4-hydroxy-1,3-oxazolidin-2-one was identified as a major metabolite for NNM and PM, but a minor metabolite for CP. A new dechlorinated metabolite for CP, 3-(2-hydroxyethyl)-1,3-oxazolidin-2-one, was also identified.

[Standard procedure and the diagnostic criteria for the Ellsworth-Howard test using human PTH-(1-34)].

The present study was undertaken to establish a standard method to perform the Ellsworth-Howard test using human PTH-1-34). For this purpose we made a survey of literature concerning the Ellsworth-Howard test and then examined the data of the Ellsworth-Howard tests performed on 178 hypoparathyroid patients using human PTH-(1-34). The main items of investigation were: (i) to determine the appropriate dose of PTH for administration to adults and children; and (ii) to define the criteria of practical usefulness for the differential diagnosis of the types of hypoparathyroidism. From the analysis of the data, the following findings and conclusions were obtained. The dose of human PTH-(1-34) appropriate for diagnostic use is 100 U per person for adults and 100 U per body surface area of one square meter (100 U/m2) for children. The criteria of positive response in the Ellsworth-Howard test are defined as follows. a) phosphaturic response: (U4 + U5) - (U2 + U3) = more than 35 mg/2 h b) cyclic AMP response: U4 - U3 = more than 1 mumol/h, and U4/U3 = more than 10 times. In the above formula, U2 - U5 represent the urine samples collected hourly in order. PTH is injected at the time between U3 and U4. For the application of the criteria in children, one should use the values corrected for body surface area of one square meter. It is necessary to confirm the following conditions before the application of the criteria: the presence of hypocalcemia and hyperphosphatemia; the lack of phosphate deficiency (basal urinary phosphate excretion more than 10 mg/2 h); the accuracy of timed urine collections (ratio of creatinine excretion during 2 hours before PTH to that after PTH administration in the range from 0.8 to 1.2); and the absence of marked diurnal variation in phosphate excretion (difference in phosphate excretion between the two basal hourly urine less than 17.5 mg/h). To ensure the above conditions, medications such as phosphate-binding antacids should be withheld for at least 1 week before the test, and the test should be performed according to the standard procedure described in this paper. The diagnosis of pseudohypoparathyroidism Type II should be done cautiously. It is necessary to take account of the high basal urinary cyclic AMP excretion and the elevated serum PTH level along with the results of the Ellsworth-Howard test (positive cyclic AMP response and negative phosphaturic response) for a definite diagnosis of this entity.

Approach to acid-base problems in the critically ill and injured.

The use of the Henderson-Hasselbalch equation and the relationships between bicarbonate levels and the pCO2 or carbonic acid concentration in evaluating acid-base abnormalities are explained. The etiology, pathophysiology, diagnosis and treatment of respiratory alkalosis and acidosis and metabolic alkalosis and acidosis are discussed. The results of laboratory tests should be examined in relation to the patient's condition and consistency with other laboratory tests. Therapy is directed at correcting the underlying problems and, secondarily, at correcting the numbers. Patients respond primarily to rate of change and not absolute numbers. Therefore, problems should be corrected at approximately the rate they develop. Treatment should be guided by continued patient observation and serial laboratory studies.

Use of high-resolution open tubular glass capillary columns to separate acidic metabolites in urine.

We used high-resolution glass capillary columns to study the trimethylsilyl derivatives of some acidic metabolites found in pooled urine specimens form control and postpartum subjects. About 30 compounds were identified by gas chromatography-mass spectrometry-computer techniques. In general, open tubular capillary columns effect better resolution of trimethylsilyl derivatives of organic acids than do conventional packed columns. GESE-30 proved to be a good general-purpose stationary phase, whereas OV-17 offered certain advantages in resolving aromatic acid components.

Calcification of collagen by urine in vitro: dependence on the degree of saturation of urine with respect to brushite.

A state of supersaturation of urine with respect to brushite is considered to be important in the formation of renal stones composed of calcium phosphate. 56 supersaturated urine specimens and 44 undersaturated specimens were incubated with collagen (Sigma collagen). Most of the supersaturated specimens calcified the collagen, whereas none of the undersaturated ones did so. Among samples which calcified the collagen, whereas none of the undersaturated ones did so. Among samples which calcified the collagen, the activity product of Ca(++) and HPO(4) (=) after incubation with collagen was essentially the same as that after incubation of the same specimen with brushite; it usually differed from that obtained after incubation with octacalcium phosphate or hydroxyapatite. The molar calcium-to-phosphorus ratio of the solid phase in collagen was approximately 1. These results suggested that the solid phase formed in collagen is brushite. This conclusion was confirmed by the direct identification of brushite in collagen by X-ray diffraction.