Biomarkers identified by urinary metabonomics for noninvasive diagnosis of nutritional rickets.

Nutritional rickets is a worldwide public health problem; however, the current diagnostic methods retain shortcomings for accurate diagnosis of nutritional rickets. To identify urinary biomarkers associated with nutritional rickets and establish a noninvasive diagnosis method, urinary metabonomics analysis by ultra-performance liquid chromatography/quadrupole time-of-flight tandem mass spectrometry and multivariate statistical analysis were employed to investigate the metabolic alterations associated with nutritional rickets in 200 children with or without nutritional rickets. The pathophysiological changes and pathogenesis of nutritional rickets were illustrated by the identified biomarkers. By urinary metabolic profiling, 31 biomarkers of nutritional rickets were identified and five candidate biomarkers for clinical diagnosis were screened and identified by quantitative analysis and receiver operating curve analysis. Urinary levels of five candidate biomarkers were measured using mass spectrometry or commercial kits. In the validation step, the combination of phosphate and sebacic acid was able to give a noninvasive and accurate diagnostic with high sensitivity (94.0%) and specificity (71.2%). Furthermore, on the basis of the pathway analysis of biomarkers, our urinary metabonomics analysis gives new insight into the pathogenesis and pathophysiology of nutritional rickets.

Dilated cardiomyopathy and adipic aciduria in nutritional rickets.

Hypocalcemia secondary to nutritional rickets is a rare cause of dilated cardiomyopathy. It is also not a recognized cause of dicarboxylic aciduria. We report the first case of adipic aciduria, presenting with dilated cardiomyopathy, secondary to hypocalcemia.

Aminoaciduria in calcium-deficiency rickets in northern Nigeria.

Generalized aminoaciduria is associated with vitamin D-deficiency rickets in humans, but there is little information regarding aminoaciduria in rickets caused by primary calcium deficiency. In contrast to rickets in other parts of the world, this bone disease in Nigeria is caused primarily by inadequate intake of dietary calcium. We conducted a clinical trial in Jos, Nigeria in 10 children with radiographically and biochemically proven rickets; an equal number of non-rachitic healthy children from the same area served as controls. Serum and 24 h urine samples were obtained at baseline and at 24 h, 1 week, 4 weeks, and 12 weeks after initiation of calcium supplementation (1000 mg/day) and were analysed for their content of amino acids. Serum calcium, phosphorus, intact parathyroid hormone (PTH), 25-hydroxyvitamin D, and 1,25-dihydroxyvitamin D were also measured at each time point. In the rachitic subjects urinary amino acid concentrations were elevated from 2- to 16-fold at baseline, while serum amino acid levels increased 1.5- to 3.8-fold compared to controls. After 12 weeks of calcium supplementation, serum and urine amino acids decreased. There was no correlation between the degree of aminoaciduria and serum PTH or 1,25-dihydroxyvitamin D concentrations. We conclude that the aminoaciduria in these rachitic children was related to their calcium status and not to their vitamin D or PTH status.

Calcium supplements increase the serum levels of crosslinked N-telopeptides of bone collagen and parathyroid hormone in rachitic Nigerian children.

OBJECTIVES: Biochemical markers of bone turnover were measured in the sera of 16 controls and 10 children with calcium-deficiency rickets, during a 12-week course of calcium supplementation (1 g CaCO3/d) that was effective in healing the bone lesions of the rachitic children. DESIGN AND METHODS: Serum levels of crosslinked N-telopeptides of bone collagen (NTx), parathyroid hormone (PTH), alkaline phosphatase (ALP), and urinary deoxypyridinoline (LP) were assayed at baseline and during the course of calcium therapy and compared with data of the 16 non-rachitic controls. RESULTS: Calcium therapy suppressed serum NTx and PTH levels in the rachitic children within 24 h; however, after the first week, PTH and NTx levels increased to the extent that at 12 weeks both were elevated when compared with controls or to baseline levels. Serum levels of NTx and PTH were correlated in the controls and experimental subjects (r = 0.63, p < 0.001). CONCLUSIONS: The rate of bone resorption, as estimated by serum NTx concentration, is increased during the healing of rachitic lesions.

Selective modulation by vitamin D of renal response to parathyroid hormone: a study in calcitriol-resistant rickets.

Calcitriol-resistant rickets (CRR) is an autosomal recessive disease caused by mutated nonfunctioning vitamin D receptors. Because of their lack of biological activity of vitamin D, CRR patients were studied to investigate whether vitamin D modulates the effects of PTH on renal tubules. Five patients with CRR and three controls were studied. After normalization of serum calcium, phosphorus, and PTH levels by oral and i.v. administration of calcium, exogenous PTH-(1-34) was infused, and timed fractions of urine were collected for measurements of cAMP, sodium, potassium, phosphorus, calcium, and bicarbonate. Serum 1,25-dihydroxyvitamin D3 [1,25-(OH)2D3] was measured before and after PTH-(1-34) infusion. Urinary cAMP and fractional excretion of potassium, phosphorus, and bicarbonate were similar in CRR patients and controls, as was the rise in the serum 1,25-(OH)2D3 concentration after PTH-(1-34) infusion. However, urinary excretion of calcium and sodium decreased after PTH-(1-34) infusion in controls, but not in CRR patients. These results suggest a selective modulation by vitamin D of the renal response to PTH; 1,25-(OH)2D3 facilitates PTH-induced calcium and sodium reabsorption, but does not influence PTH-induced cAMP excretion; phosphorus, potassium, and bicarbonate tubular transport, or 1 alpha-hydroxylation of 25-hydroxyvitamin D3.

Vitamin D dependent rickets type II and normal vitamin D receptor cDNA sequence. A cluster in a rural area of Cauca, Colombia, with more than 200 affected children.

Vitamin D dependent rickets type II is an autosomal recessive disease caused by the vitamin D defective receptor. More than 200 patients with different types of lower limb deformities were detected in a rural area of the Cauca department in the southwest part of Colombia. Patients were well nourished and in good physical condition in spite of their deformities. None of them presented alopecia, myopathy, seizures or aminoaciduria. Serum analysis showed significantly lower serum calcium as compared to normal relatives, though in the normal low range, normal phosphorus, high alkaline phosphatase, normal 25-hydroxyvitamin D3 and high 1,25-dihydroxyvitamin D3, indicating target organ resistance. The cDNA analysis showed normal nucleotide sequence. We suggest that our patients represent a distinct form of receptor-positive resistance to vitamin D. This report is the first extensive study on this class of patients.

Evidence suggesting hyperoxaluria as a cause of nephrocalcinosis in phosphate-treated hypophosphataemic rickets.

Urinary excretion of oxalate and phosphate was measured in twelve vitamin-D-treated, phosphate-supplemented patients with X-linked hypophosphataemia (XLH; four children, eight adolescents and adults) to investigate possible causative factors of nephrocalcinosis other than calcium. Oxalate excretion correlated highly with urinary phosphate excretion and with intake of phosphate supplements corrected for body surface area. Young children received the highest relative doses of phosphate (range 2.27-10.8 g/1.73 m2 daily) and their urinary oxalate excretion was very high (0.94-3.38 mmol/1.73 m2 daily). The urinary oxalate excretion of untreated adults with XLH was within normal limits. Six patients had evidence of nephrocalcinosis on ultrasound. The high urinary oxalate excretion in phosphate-supplemented XLH may be seen as a special type of enteric hyperoxaluria, in which the conditions of calcium-oxalate crystal precipitation could be reached even at normal levels of urinary calcium excretion. Urinary excretion of both calcium and oxalate should therefore be monitored during treatment in young XLH patients.

Urinary hydroxyproline in infants with and without fractures/rickets.

Molar ratios of peptide-bound and free hydroxyproline:creatinine (OHPr:Cr) in urine were measured at 3, 6, 9, and 12 months of age in two groups of very-low-birthweight (VLBW less than or equal to 1500 g) infants. Group A (15 infants) had radiographically confirmed fractures and (or) rickets (F/R); Group B (17 infants) did not. The urinary peptide-bound OHPr:Cr ratio varied widely within groups and was greatest at three months in both groups: A = 0.81 +/- 0.45 and B = 0.55 +/- 0.32 (mean +/- SD). The ratio decreased with increasing postnatal age for each group but was not statistically different between groups throughout the study. The urinary free OHPr:Cr ratio also was greatest at age three months (A = 0.32 +/- 0.15 and B = 0.53 +/- 0.46), rapidly decreasing afterwards, and was not statistically different between groups throughout the study. We conclude that, in VLBW infants, bone turnover as indicated by the urinary peptide-bound OHPr:Cr ratio is highest during early infancy; however, the wide range of values for this ratio suggests that its use alone is not sufficient for detection of F/R in VLBW infants. The rapid decrease in free OHPr:Cr ratio is presumably related to the maturation of renal tubular function.

Prenatal deficiency of phosphate, phosphate supplementation, and rickets in very-low-birthweight infants.

A preliminary study showed that placental histology was abnormal for babies who subsequently had rickets. The findings--low plasma phosphate concentration; maximum percentage tubular reabsorption of phosphate; and high urinary calcium loss--are those of renal conservation of phosphate in the presence of phosphate deficiency. In a controlled trial no baby receiving phosphate supplements (50 mg per day) had radiological evidence of rickets whereas bone changes were apparent in 42% of the control group. Prenatal deficiency of phosphate, due to placental insufficiency, can be corrected by phosphate supplementation thereby preventing rickets of prematurity.

[Serum concentrations of 25 hydroxyvitamin D in hospitalized infants. Relation to calciuria]. / Taux sériques de 25 hydroxyvitamine D chez les nourrissons hospitalisés. Rapport avec la calciurie.

In attempt to evaluate the vitamin D status of the infants of our area under the mode of prophylaxis of carential rickets actually used in France, serum 25 hydroxyvitamin D (25 OHD) levels were measured in 65 infants (age 3 - 32 months) during their hospitalisation for acute illness. Most infants were receiving vitamin D either in daily doses (1,200 - 1,600 u) or in unique loading doses (200,000 - 600,000 u every 4 - 6 months). With this prophylaxis serum concentrations of 25 OHD were elevated, i.e. above 75 nmol/l, in more than 50% of the infants, reaching 474 nmol/l in one case. Calciuria estimated by the calcium/creatinine urinary ratio tended to increase in parallel with the serum 25 OHD level. From these data it is concluded that the actual prophylaxis of carential rickets in France frequently uses excessive doses of vitamin D and that new rules have to be established.

Phosphaturic mesenchymal tumors. A polymorphous group causing osteomalacia or rickets.

Reported are the pathologic features of 17 mesenchymal tumors documented as causing osteomalacia or rickets. Although these tumors were histologically polymorphous, they were classifiable into four morphological groups. In the first group there were ten unique tumors showing mixed connective tissue features and containing variably prominent vascular and/or osteoclast-like giant-cell components. Tumors of this group also displayed focal microcystic changes, osseous metaplasia, and/or poorly developed cartilaginous areas. The cartilaginous areas sometimes showed considerable dystrophic calcification. With one exception, all tumors of this group occurred in soft tissue and demonstrated benign clinical behavior. The single malignant tumor originated in bone, recurred locally, and metastasized to lung. The tumors comprising the remaining three groups (six tumors) occurred in bone, demonstrated benign clinical behavior, and were grouped according to their close resemblance to tumors known to occur in bone, that is osteoblastoma-like (four tumors), nonossifying fibroma-like (two tumors), and ossifying fibroma-like (one tumor).

[Hydroxyproline in morning urine. A reliable parameter for bone turnover in childhood]. / Die Hydroxyprolin-Ausscheidung im Morgen-Urin. Ein geeigneter Parameter des Knochen-Umsatzes im Kindesalter.

OH-P/Cr was measured in morning fasting urine specimens of 300 healthy subjects and of children with disorders of calcium metabolism receiving no diet. In healthy children the values were sex and age dependent reflecting the different height velocities. The OH-P excretion was not different in schoolchildren receiving a OH-P-poor diet for at least two days in comparison to subjects of the same age group with unrestricted diet. OH-P/Cr correlated well with serum alkaline phosphatase (AP) activity and decreased rapidly after a calcium load. OH-P/Cr and AP were elevated in patients with increased bone turnover (hyperparathyroidism and hypophosphatemic rickets). In contrast, the OH-P excretion was normal in children with permanent or transient isolated hyperphosphatasemia. In children with vitamin D deficiency rickets there was a further increase of OH-P/Cr in response to vitamin therapy, while the AP activity, which reflects osteoblastic activity, tended to fall. This indicates that the observed increment of the OH-P excretion in these children is due to a temporary resorption of osteoid caused by the increasing levels of vitamin D metabolites. It is concluded that the measurement of OH-P/Cr provides a useful tool of bone turnover in children, in that it makes complete 24 h-urine collections and a OH-P free diet unnecessary. In combination with other indices of calcium metabolism the determination of the OH-P ratio is considered to be a valuable measure for the diagnosis and follow-up of bone disorders.(ABSTRACT TRUNCATED AT 250 WORDS)