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Intra-individual variation in fasting urinary calcium- and hydroxyproline-creatinine ratios measured in metabolic bone clinic patients as both outpatients and inpatients.

The total intra-individual variation of fasting urinary calcium- and hydroxyproline-creatinine ratios were determined both as outpatients and as inpatients in 46 patients with metabolic bone disorders, including 20 women with osteoporosis. The intra-individual variation of both measurements was reduced considerably by collecting samples in the ward rather than in the outpatient setting, although this was not the case for a sub-group of osteoporotic women. The large intra-individual variation observed for calcium and hydroxyproline (30-40% in osteoporotic women) can give rise to misclassification in screening for fast bone losers. The very large critical difference required (109% and 94%, respectively, for calcium and hydroxyproline) for two results to reflect a significant change, is a major limitation in the use of these analytes for monitoring bone resorption in menopausal and osteoporotic women.

Aged↗

Serum total hydroxyproline assay: effects of age, sex and Paget's bone disease.

An assay was developed for total hydroxyproline (protein, peptide and free) in 12 h fasted human serum. Values obtained from 80 adults judged normal by laboratory and physical examinations showed age and sex effects; values increased with age and were higher for men. Assay of serum samples from 22 patients with Paget's bone disease revealed values significantly above normal taking age and sex into consideration (p = 0.001). Serum alkaline phosphatase and serum hydroxyproline values for these patients were correlated (r = 0.91). Serum total hydroxyproline may be a useful assay for evaluating metabolic bone diseases but age and sex must be taken into consideration in the interpretation.

Aged↗

Urinary hydroxyproline excretion as a marker of bone metastasis in prostatic cancer.

In 25 patients with prostatic cancer and 26 patients with benign prostatic hypertrophy, urinary hydroxyproline excretion as estimated by the method described by Cleary and Saunders. The patients were kept on a diet free of gelatin. Urinary creatinine was measured in all patients and serum acid phosphatase, prostatic acid phosphatase, alkaline phosphatase and calcium were measured in patients with prostatic cancer. Urinary hydroxyproline excretion and the hydroxyproline/creatinine ratio were both elevated in patients who had prostate cancer with bone metastasis when compared to values in patients who had benign prostatic hypertrophy or prostatic cancer without bone metastasis. These two were more sensitive indicators of bone metastasis than serum acid phosphatase, prostatic acid phosphatase, alkaline phosphatase and calcium. These results suggest that urinary hydroxyproline is a valuable index of bone metastasis in prostatic cancer.

Bone Neoplasms↗

Age-related changes in glomerular volume and hydroxyproline content in rat and human.

Total 4-hydroxyproline content and volume were measured in the same sample of microdissected glomeruli obtained fro rat and human outer or inner cortex. Glomerular volume was determined by computer-assisted image analysis, and 4-hydroxyproline was measured by a highly sensitive gas-liquid chromatographic method. Results were expressed as weight of basement membrane material by comparison with the amount of 4-hydroxyproline in purified basement membrane/mesangial matrix preparations. Microanalyses were possible in samples containing as few as eight human glomeruli. Rat glomerular size increased sevenfold between 5 wk and 2 yr of age, with volume being consistently 36 to 45% greater in inner than in outer cortex glomeruli. Basement membrane material content per glomerulus markedly increased with age (12-fold); however, when expressed per unit volume, this change was greatly reduced (2-fold). Expressed per volume, inner and outer cortex glomerular content of basement membrane material was always similar, regardless of age. Therefore, a greater glomerular size, in itself, does not accelerate the rate of basement membrane material deposition. Glomerular size distributions (measured by skewness and kurtosis) did not change, indicating that, although glomerular volume increases with age, aging does not appear to cause the emergence of distinct glomerular populations within an age group. Basement membrane material accumulation is probably a generalized change. Human glomeruli increased sevenfold in size from infancy to adulthood and then declined during senescence. Contrary to that in the rat, glomerular basement membrane material content appeared to closely follow size changes, thus, varying little from infancy to senescence if expressed per unit of glomerular volume.

Adolescent↗

Urinary concentration of a specific peptide of type I collagen of bone (CrossLaps): correlation to hydroxyproline.

Urinary bone resorption markers, CrossLaps and hydroxyproline are compared in a non-selected group of 93 women. The correlation between CrossLaps and hydroxyproline is satisfactory. The r value is 0.79. Furthermore, it is investigated whether CrossLaps can substitute for hydroxyproline in the estimation of bone loss, using a model based on the combination of several biochemical markers. The results indicate that the two systems reflect related or parallel events, and show that CrossLaps is suitable for use in a normal clinical chemistry laboratory.

Adult↗

Dose-dependent effects in the subacute response of the rat lung to quartz. II. Protease activities and levels of soluble hydroxyproline in lung lavage.

The response of the rat lung to a range of doses of quartz at 50 and 100 days after its administration by intratracheal instillation has been assessed by bronchopulmonary lavage. The effects on the levels of protein and hydroxyproline and on the PZ peptidase and collagenolytic activities of lavage fluid supernatants are described and an assessment of the hydroxyproline content of recovered cells is made. In addition, assays for PZ peptidase and collagenase activity were carried out on polymorph enriched fractions and on samples obtained from short-term culture of recovered macrophages. Both protein and hydroxyproline measurements did show dependence on the amount of quartz instilled and the time after its administration. Measurements of PZ peptidase and collagenase activities suggest that the polymorphonuclear leukocytes, not the macrophages, are the major source of these degradative enzymes. The relevance of these findings with regard to the importance of the PMN in quartz-induced fibrosis, is discussed.

Animals↗

Hydroxyproline-containing structural glycopeptide fractions from subacute inflammation connective tissue.

Hydroxyproline-containing structural glycopeptide fractions were isolated from collagenase-digested neutral salt-insoluble collagen of five-day sponge-implant connective tissue of the rat. The glycopeptide fractions characterized migrate as a single, strongly anionic band on disc gel electrophoresis at pH 9.5, are eluted on gel filtration as a small molecular weight peak, approximately 2000, and are resolved into thirteen glycopeptide fractions by DEAE-cellulose chromatography. Amino acid analyses of some of these fractions indicate a similarity in composition, the principal ones being aspartic and glutamic acids, serine, glycine, alanine, valine, proline and hydroxyproline. Three neutral carbohydrates, glucose, mannose and xylose, in different relative proportions and hexosamine are also present in the fractions. Amino-terminal amino acid determinations indicate a microheterogeneity of the glycopeptides. The electrophoretic behaviour and non-diffusibility of the small molecular weight glycopeptides suggest an intimate association between acidic hydroxyproline-containing peptides and carbohydrate components of developing connective tissue.

Amino Acids↗

Changes in calcium and hydroxyproline content of cortical bone after compression and neutral plate fixation.

Calcium and hydroxyproline content of intact rabbit tibio-fibular bone was assay chemically 1 day to 36 weeks after compression and neutral plate fixation with a 4-hole stainless steel ASIF/DCP plate. During the first three postoperative weeks calcium content of the cortical bone decreased by an almost significant amount (P less than 0.05); thereafter, calcium values remained subnormal but did not differ significantly from initial postoperative values. Hydroxyproline content increased steadily throughout the experiment (P less than 0.001). The changes in chemical composition of the bone were of the same magnitude after compression and neutral plate fixation. The results suggest that the cancellous transformation of cortical bone, known to take place under rigid plates, reflects an active remodelling process of the bone in which the calcium and hydroxyproline content changes only slightly.

Animals↗

Serum somatomedin A and non-dialyzable urinary hydroxyproline in girls with idiopathic scoliosis.

Serum somatomedin A was determined by radioreceptor assay in 31 girls with idiopathic scoliosis and 30 controls, with ages ranging from 9.7-16.2 years. In the material as a whole no significant difference could be found between scoliotics and controls. Similarly no differences were found in the girls under 13 years of age. However, in girls older than 13 years the controls had significantly higher somatomedin A levels than the scoliotics. In the second part of the study 26 girls with idiopathic scoliosis and 22 controls were examined with regard to the urinary excretion of non-dialyzable hydroxyproline,. which has been suggested to be correlated with collagen synthesis. The percentage non-dialyzable fraction of the total hydroxyproline was found to be significantly higher in the control girls. The mean age was 12.8 years for both groups. The results suggest a higher growth rate in the controls than in the scoliotics for girls over 12 years. There is, however, no definite evidence as to the function of the somatomedins or of the relationship between non-dialyzable hydroxyproline and growth.

Adolescent↗

Changes in the fatty acid composition and hydroxyproline content in rat lung in relation to collagen synthesis after paraquat administration.

OBJECTIVES: Effect of paraquat on the fatty acid composition (weight percentage) of rat lung was studied with particular reference to the change of hydroxyproline content in the course of paraquat-induced dysfunction and subsequent repair. METHODS: Eight-week-old male Wistar rats were administered paraquat at 20 mg/kg body weight subcutaneously, and the wet weight, hydroxyproline content and fatty acid composition of lungs of each group rats were analyzed at 2, 7, 14 or 28 days after treatment, respectively. RESULTS: The percentage of palmitic acid (C16:0), arachidonic acid (C20:4) and docosahexaenoic acid (C22:6) significantly increased, and the percentage of oleic acid (C18:1) and the ratio of monounsaturated fatty acids/saturated fatty acids (M/S) significantly decreased comparing to control on day 28 after paraquat administration. The time-course of each fatty acid was observed for 28 days after paraquat administration. M/S ratio decreased after paraquat administration up to the 28th day, but the polyunsaturated fatty acids/saturated fatty acids (P/S) ratio decreased during the first 7 days, followed by a increase, and then reached higher level than the 0 day control at the 28th day. Hydroxyproline also increased between the 14th and the 28th days. Eicosapentaenoic acid (C20:5) had once increased during the first 2 days and decreased gradually, while C20:4 maintained high level in this period. C22:6 increased after paraquat administration and maintained high level up to the 28th day. This result indicated that desaturation and elongation in n-3 series fatty acids were accelerated after paraquat treatment, and consequently C20:5 was rapidly converted into C22:6 and decreased. CONCLUSIONS: Paraquat might cause elevation of unsaturated fatty acids, espe- cially C20:4 but not C20:5 by the stimulation of the fatty acid desaturase system, and could consequently stimulate local collagen synthesis by C20:4 metabolites in the healing stage.

Animals↗

[Effects of supplement Zn on levels of Zn in serum, growth hormone and hydroxyproline].

OBJECTIVE: In order to research the effects of supplement Zn on wound healing, the changes of Zn in serum and skin, growth hormone and hydroxyproline of skin were observed in burn rats. METHOD: 80 SD rats were randomly divided into four groups: Control group of 8 rats, N group (diet containing Zn 40 micrograms/g), W group (application of Zn on burn skin), H group (diet containing Zn 80 micrograms/g). 8 rats of each group were killed on 1, 3, 7 days after burn. RESULTS: The levels of serum Zn declined on 1 day postburn in N and W groups, whereas the content of serum Zn in H group elevated and were 2.4, 2.0 times as much as that of N and W groups respectively (P < 0.01). Zn in burn skin increased significantly on 3, 7 days in W group and was 1.3, 1.5 times as much as that of N group. Serum growth hormone increased in all groups postburn. The highest level appeared in H group. Hydroxyproline in burn skin decreased in all groups, but it began to increase on 7 day postburn in W group. CONCLUSION: Supplementation of Zn by diet rapidly raises the levels of Zn in serum, burn skin and growth hormone. It is beneficial to apply Zn to the wound to improve wound healing. Hydroxyproline increased significantly with application of Zn to burn wound. It is better to supply Zn by diet and topical application simultaneously.

Animals↗

A chemical evaluation of collagen and hydroxyproline after treatment with GK-101 (N-Chloroglycine).

The reactions of GK-101 (N-chloroglycine) with collagen and hydroxyproline, were studied with regard to caries removal. After reaction with N-chloroglycine, collagen was analyzed for bound available chlorine by iodiometric titration, and hydroxyproline conversion to pyrrole-2-carboxylic acid was measured by they production of a red chromogen after PAB treatment. The results indicated that collagen was chlorinated by N-chloroglycine and hydroxyproline was converted to pyrrole-2-carboxylic acid after N-chloroglycine reaction. These findings tend to indicate two possible pathways of N-chloroglycine protein interaction.

Carboxylic Acids↗

Serum free proline and free hydroxyproline in patients with chronic liver disease.

Serum free proline and free hydroxyproline were determined in 71 patients with liver disease and in 62 control subjects. The group with liver disease included 60 patients with liver cirrhosis and 11 with chronic active liver disease. Forty-five of the cirrhotic patients were alcoholics, 9 of which were studied during an episode of alcoholic hepatitis. The control group consisted of 24 healthy volunteers, 19 primary malnourished patients, and 19 severely ill patients without liver disease. The values obtained in normal subjects were quite constant; no differences related to sex or age were detected. In patients with malnutrition, and especially in severely ill patients, the proline values were always below the normal limit. Patients with nonalcoholic cirrhosis or chronic active liver disease had serum proline and hydroxyproline values similar to those of normal subjects. However, the patients with alcoholic liver cirrhosis had proline and hydroxyproline values significantly higher than the normal group. Furthermore, in patients with alcoholic hepatitis the serum free proline values were significantly higher than in the other groups. The results suggest that alcohol might have a direct effect on proline metabolism or facilitate its release from the liver cell.

Adolescent↗

Excretion of urinary hydroxyproline in correlation with severity of induced osteomyelitis in rabbits.

Osteomyelitis was induced artificially by injecting Staphylococcus aureus culture in the tibiae of young rabbits. Weekly estimations of hydroxyproline in urine were done for six weeks. It was found that the osteomyelitic rabbits excreted more hydroxyproline (about 96%) two weeks after the infection in comparison to the control animals and it continued to be very high (about 138%) six weeks after the infection. The results indicate that urinary hydroxyproline reflects degradation of collagen fibers in the bone, and may be an indicator of the severity of the disease.

Animals↗

[Effect of secondary hyperparathyroidism upon serum hydroxyproline in patients with renal failure].

Diagnosis of secondary hyperparathyroidism in 12 patients treated with repeated hemodialysis was established on the basis of clinical symptoms and biochemical studies. Following subtotal parathyroidectomy the resected parathyroid specimens were examined histologically. The diagnostic value of total and free hydroxyproline in the serum in secondary hyperparathyroidism was found to be similar to the diagnostic value of the parathyroid hormone. The correlation with histological diagnosis was from 93% to 100%. After parathyroidectomy a significant decrease in the concentrations of total and free hydroxyproline in the serum was noted being the quickest within the first six months following the resection. The decrease in the concentrations of total and free hydroxyproline in the serum in patients with secondary hyperparathyroidism are valuable diagnostic markers of bone collagen metabolism and bone repair following parathyroidectomy.

Adult↗

Effects of three different calcium preparations on urinary calcium and hydroxyproline excretion in postmenopausal osteoporotic women.

Some authors have claimed that 'solubilized' calcium preparations are better absorbed than calcium carbonate, while others have reported that all forms are equally well absorbed. We measured radiocalcium absorption in 35 postmenopausal osteoporotic women and then gave them on three successive evenings, in random order, three different proprietary calcium preparations (Sandocal containing 1 g of effervescent calcium, Calsup containing 1 g of calcium as the carbonate, and Caltrate containing 1.2 g of calcium as the carbonate). The daily urinary calcium excretion rose significantly and similarly on all three supplements and was greater in the high calcium absorbers than the low calcium absorbers. The fasting urinary hydroxyproline excretion was significantly decreased the morning after administration of each preparation, and one-way analysis of variance showed no significant difference between the days of administration or the type of supplement. The decrease was greater for high absorbers than for low absorbers on all three supplements but the differences did not reach statistical significance. By 36 hours after the last calcium supplement the urinary hydroxyproline had returned to baseline. The response of hydroxyproline excretion (and by implication bone resorption) appears to be rapid in onset and short lived. Strict compliance is therefore important in patients on calcium therapy.

Absorption↗

The hydroxyproline content in the lung tissue of young pigs after fractionated irradiation with photons or neutrons.

The radiation response of normal lung tissue was studied in 99 young pigs. Five fractions of Co-60-photons and of neutrons with an average energy of 6.2 MeV were applied during overall treatment times of five and 35 days respectively. The irradiation field comprised the whole of the right lung. At eight to ten weeks and at eight to 13 months animals were sacrificed and the removed lungs were investigated histopathologically and biochemically. Radiation induced fibrosis was evaluated by determining the ratio of hydroxyproline concentration of irradiated and non-irradiated lung in the same animal. Above a threshold there was a significant increase of the hydroxyproline ratio with increasing doses. Histopathological findings and hydroxyproline ratio were in close agreement. RBE values between 3.8 and 4.5 were calculated in accordance with the results using the other criteria of lung damage in our studies on pigs.

Animals↗

Phosphorylation of hydroxyproline in a synthetic peptide catalyzed by cyclic AMP-dependent protein kinase.

The cyclic AMP-dependent protein kinase catalyzes the phosphorylation of hydroxyproline present in the heptapeptide, Leu-Arg-Arg-Ala-Hyp-Leu-Gly. The Km value for the reaction with this substrate was high (approximately 18 mM) compared to the Km values reported for the analogous threonine and serine-containing peptides, which were 0.59 mM and 0.016 mM, respectively (Kemp, B.E., Graves, D.J., Benjamini, E., and Krebs, E.G. (1977) J. Biol. Chem. 252, 4888-4894). The Vmax value with the hydroxyproline-containing peptide was 1 mumol . min-1 mg-1 in contrast to Vmax values of 6 mumol . min-1 mg-1 and 20 mumol . min-1 mg-1 for the threonine- and serine-containing peptides, respectively. Phosphate esterified to hydroxyproline present in the peptide was relatively stable in hot alkali, only 10% being released as Pi within 30 min in 0.1 N NaOH at 100 degrees C, whereas all of the phosphate was released from the phosphoserine peptide analogue under these conditions. Phosphohydroxyproline in the peptide was also more stable to acid (5.7 N HCl, 110 degrees C) than phosphoserine, the time for 50% release as Pi being 15 h in contrast to 6 h for the latter.

Animals↗