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Circulating levels of 1,25 dihydroxyvitamin D, alkaline phosphatase, hydroxyproline, and osteocalcin associated with antler growth in white-tailed deer.

Alkaline phosphatase, hydroxyproline, osteocalcin, and 1,25(OH)2D were measured in biweekly serum samples obtained from 6 adult (greater than 4 years), 4 juvenile (1-4 years) and 4 fawn (less than 1 year) male white-tailed deer from Oct. 1983 to Oct. 1984. Antler length, from the pedicle to the tip, was measured at the time of serum sampling. Serum alkaline phosphatase activity and levels of hydroxyproline and osteocalcin were higher (P less than 0.05) in fawns compared with juveniles and adults reflecting increased bone metabolism in the younger deer. In adult deer serum alkaline phosphatase activity and hydroxyproline levels were elevated (P less than 0.05) during the period of antler growth, whereas serum osteocalcin and 1,25(OH)2D increased (P less than 0.05) during antler mineralization. Similar but less pronounced trends occurred in juvenile deer, possibly a reflection of skeletal growth in the younger animals. The data lend support for utilization of the deer antler cycle as a model for studies of bone disorders. Further work is needed to help clarify the role of hydroxyproline, osteocalcin, and 1,25(OH)2D in the antler cycle.

Age Factors↗

Relationships among intramuscular collagen, serum hydroxyproline and serum testosterone in growing rams and wethers.

Concentration and maturation of collagen and serum concentrations of hydroxyproline and testosterone were determined in growing rams and wethers to characterize developmental changes in collagen associated with a representative testicular steroid. Groups of eight rams and eight wethers were slaughtered at 12, 18, 24 and 30 wk of age. Concentrations of collagen in longissimus, supraspinatus and infraspinatus muscles and serum hydroxyproline were greater (P less than .05) in rams than in wethers at all ages. Collagen stability, as measured by collagen solubility and thermal shrinkage temperature, was greater (P less than .05) in wethers than in rams. Differences in collagen stability and serum hydroxyproline concentration indicated that collagen synthesis and turnover were more rapid in rams than in wethers. Serum hydroxyproline decreased (P less than .05) and collagen solubility decreased (P less than .05) with age, indicating that collagen turnover was occurring most rapidly in 12-wk-old lambs and that collagen maturation was predominant in 24- to 30-wk old lambs. Testosterone parameters measured in rams were unrelated within groups to collagen characteristics, possibly reflecting the high variability in testosterone secretion and the slow development of collagen. However, rams as young as 12 wk of age were under the influence of testosterone, and differences in collagen between rams and wethers were apparent at that time.

Animals↗

Urine hydroxyproline: a potential error in quantification and a proposed procedure for its measurement.

A modification of the colorimetric method of Bergman and Loxley for the measurement of urinary hydroxyproline using external aqueous hydroxyproline standards instead of individual internal standards is described. We show that this modification leads to an underestimation (average 32%) of hydroxyproline because suppression of colour development occurs in urine samples but not in aqueous standards. Use of an internal standard for each urine test corrects for this suppression. Recovery of hydroxyproline (250 mumol/L) added to 12 patient urine samples averaged 99% and the overall imprecision for the assay was then less than 5%. Modifications to the original hydrolysis and colour development procedures allow linearity to 1500 mumol/L. Details of our procedure are given.

Colorimetry↗

Increased content of neutrosoluble collagen and dialyzable hydroxyproline in panacinar emphysema and spontaneous pneumothorax.

Hydroxyproline transfer to an incubation medium was studied in ten emphysematous, nine relapsing pneumothorax, and eight control samples surgically removed from 2 females and 25 males, ranging in age from 17 to 69 years. Total hydroxyproline in the lung pieces, expressed as a fraction of DNA, was not modified. A large increase of dialyzable and nondialyzable hydroxyproline fraction was observed in the incubation medium of biopsies from emphysema dn pneumothorax groups. Although the pneumothorax group had average values between controls and emphysema, some pneumothorax patients exhibited very high content of soluble hydroxyproline, suggesting a common pattern between some relapsing pneumothorax and emphysema. These findings could not be due to extracellular degradation during the processing of the samples since the results were not affected by the presence of protease inhibitors. These observations suggest that individuals with emphysema and pneumothrax have abnormalities in lung collagen structure and/or biosynthesis resulting in increased degradation of this macromolecule.

Adolescent↗

Nerve regeneration and pharmacological suppression of the scar reaction at the suture site. An experimental study on the effect of estrogen-progesterone, methylprednisolone-acetate and cis-hydroxyproline in rat sciatic nerve.

Pharmacological suppression of the scar reaction was tested as a possible method for improving axonal regeneration after transection and suture of the sciatic nerve of rats. The animals in the experimental groups were systemically treated post-operatively with estrogen-progesterone, methylprednisolone-acetate (Depomedrone) or cis-hydroxyproline. Only in the cis-hydroxyproline treated animals was there a tendency towards a decrease in fibroblastic activity at the suture site after 4 weeks. However, there was no difference in the growth of axons to the distal nerve segment after pharmacological treatment as compared to non-treated animals, when tested 12 weeks post-operatively. The motor nerve conduction velocity was however significantly greater in both the cis-hydroxyproline and the methylprednisolone-acetate groups as compared with the estrogen-progesterone and the control groups. In conclusion, cis-hydroxyproline and also methylprednisolone-acetate may improve the function of regenerating peripheral nerve, while estrogen-progesterone does not seem to have this effect.

Animals↗

Hydroxyproline levels and collagenolytic activity in synovial fluids of patients with rheumatic diseases.

Free, total, and peptide hydroxyproline levels were determined in synovial fluid obtained from the knee joints of 60 patients with theumatoid arthritis (RA), and 26 patients with degenerative joint disease. In addition, in 160 synovial fluid samples obtained from 121 patients including 50 with degenerative joint disease, 60 with RA, 3 with Reiter's syndrome, 3 with hydarthrosis intermittens and 5 with ankylosing spondylitis, the collagenolytic activity was determined. The mean values of free and peptide hydroxyproline in the inflammatory and degenerative fluids were the same, but slight differences were found in the mean values of total hydroxyproline. No effect on the level of free and bound hydroxyproline was observed after treatment with intra-articular hydrocortisone and gold salts. The collagenolytic activity of synovial fluid was registered in 38% of cases of RA and in some cases of Reiter's syndrome and hydrarthrosis intermittens, but it was not found in 50 cases of degenerative joint disease or in cases of ankylosing spondylitis. During a longer observation of patients with inflammatory forms of RA a variability in the collagenolytic activity was observed in repeated examinations of the fluid obtained from the same patient; this activity appeared and disappeared. The incidence of collagenolytic activity and its values were higher in patients with active rheumatoid process and this activity was present more frequently in patients with a short history of the disease (up to 3 years). The collagenolytic activity of rheumatoid fluids was, to a high degree, inhibited by normal human serum. The problem of presence or lack of collagenolytic activity in rheumatoid fluids is discussed.

Adult↗

Synovial fluid and serum hydroxyproline fractions in rheumatoid arthritis.

In rheumatoid arthritis (RA) the synovial fluid lymphocyte count closely parallels the synovial fluid dialysable hydroxyproline, a marker of collagen resorption. This observation stresses the primordial role of lymphocytes in RA joint injury. The actual and eventually potential destruction of any single joint, as expressed by synovial fluid dialysable hydroxyproline levels, seems to reflect the general picture of disease activity as evaluated by the number of active joints (joint count), ESR and rheumatoid factor titres (latex fixation). Although urinary hydroxyproline levels are generally within the normal range in RA, they can be used as index of articular tissue destruction and as a parameter of overall disease activity when groups of patients are studied longitudinally in detail. The present study sheds no further light on the significance of synovial fluid and blood non-dialysable hydroxyproline levels.

Arthritis, Rheumatoid↗

Urinary hydroxyproline excretion in male and female chickens.

Hydroxyproline was measured in 24-hr urine samples at 60 and 120 days of age in male and female broiler chickens that were prepared by the "anus praeternaturalis" technique. Twenty-four-hour urine output did not differ between females and males (32.82 +/- 1.02 ml and 33.98 +/- .54 ml, average +/- SD, respectively), whereas 24-hr urinary hydroxyproline excretion was greater in females than in males after 60 days of age. Urinary hydroxyproline excretion tended to decrease with age in males and increase with age in females. Because most of the hydroxyproline is probably derived from bone collagen, the results suggest that there are differences between mineral exchange processes in female and male chickens.

Age Factors↗

[A study on markers of bone metabolism in patients with non hypercalciuric calcium stone--measurement of gamma-carboxyglutamic acid and hydroxyproline].

Thirty-one patients (26 male and 5 female patients) of non-hypercalciuric Ca stone formers were studied concerning their bone metabolism. We measured plasma free gamma-carboxyglutamic acid, plasma hydroxyproline, urinary free gamma-carboxyglutamic acid and urinary hydroxyproline as markers of bone metabolism. Plasma free gamma-carboxyglutamic acid level showed significantly higher in stone formers than normal controls (1.01 +/- 0.37 nmol/ml vs 0.39 +/- 0.13 nmol/ml). But no significant difference were observed in plasma hydroxyproline level (1.99 +/- 0.73 micrograms/ml vs 2.27 +/- 0.92 micrograms/ml), urinary free gamma-carboxyglutamic acid (52.0 +/- 17.6 nmol/mg.Cr vs 45.7 +/- 9.8 nmol/mg.Cr), and urinary hydroxyproline (3.87 +/- 2.02 micrograms/mg.Cr vs 3.33 +/- 1.63 micrograms/mg.Cr). In conclusion, these results suggest that abnormality of bone metabolism may be not a primary event in causing stone formation in these patients.

1-Carboxyglutamic Acid↗

Hydroxyproline index: an indicator of nutritional status. Study of Mississippi children.

The hydroxyproline index was used to evaluate nutritional status of four- to five year-old black children in two Mississippi counties. Positive partial regression coefficinets were highly significant for weight in both counties and significant for height in one. These data seem to confirm results of other workers that changes in the index may reflect change in body weight rather than in hydroxyproline; creatinine ratio. Eighty per cent of the children had high serum gamma globulin levels suggesting infection, which has been shown to cause a high degree of variability in hydroxyproline excretion. Results indicate that hydroxyproline index may not be a valid indicator of nutritional status for this popualtion group.

Anthropometry↗

Seprafilm reduces adhesions to polypropylene mesh and increases peritoneal hydroxyproline.

Polypropylene mesh is an effective and widely used material in repairing abdominal wall defects, but it causes dense adhesions when in contact with abdominal viscera directly. As a consequence of this process intestinal obstruction and enterocutaneous fistula may develop. The purpose of the present study was to determine whether Seprafilm, a bioresorbable translucent membrane, reduces abdominal visceral adhesions to polypropylene mesh and whether Seprafilm has any effects on peritoneal tissue hydroxyproline levels. Twenty-six adult Wistar rats were used. A full-thickness abdominal wall defect was created and cecal abrasions were created to induce adhesion formation in each animal. All of the abdominal defects were repaired with polypropylene mesh. In addition a Seprafilm membrane was laid over the abdominal viscera in the Seprafilm group (n = 13). The abdominal cavity was evaluated for adhesion formation, and peritoneal biopsies were taken for the measurement of tissue hydroxyproline levels at the 14th day. The use of Seprafilm resulted in significant reduction in the adhesion formation (P = 0.002) and a significant increase in peritoneal hydroxyproline level (P < 0.0001). These findings demonstrate that the increase of peritoneal hydroxyproline levels caused by Seprafilm might play a role on the antiadhesive effects of Seprafilm.

Abdominal Muscles↗

Urine products of bone breakdown as markers of bone resorption and clinical usefulness of urinary hydroxyproline: an overview.

PURPOSE: The purpose of this study is to review the urine products of bone breakdown as markers of bone resorption and usefulness of urinary hydroxyproline. DATA: Related researches published in 1985 - 2000 were systematically reviewed. RESULTS: Bone markers could be used for early diagnosis of bone metabolic diseases. Biochemical markers of bone resorption that reflect osteoclast activity and/or collagen degradation provide a new and potentially important clinical tool for the assessment and monitoring of bone metabolism. Assessment of bone resorption can be achieved with measurement of urinary hydroxylysine glycosides, urinary excretion of the collagen pyridinium cross-links, urinary excretion of type I collagen telopeptide breakdown products (cross-linked telopeptides) and urinary hydroxyproline. CONCLUSION: Urinary hydroxyproline has been in use as a marker of bone resorption, but it lacks sensitivity and specificity. It is a modified amino acid that is a metabolic product of collagen breakdown. Hydroxyproline may be released either free or with fragments of the collagen molecule attached during bone resorption, and it is also liberated by the breakdown of complement and nonskeletal collagen.

Biomarkers↗

Total urinary hydroxyproline determined with rapid and simple high-performance liquid chromatography.

A precolumn derivatization method was optimized for rapid and specific analysis of total urinary hydroxyproline by HPLC. After an overnight hydrolysis, urine samples dried and reconstituted with the internal standard cysteic acid (in sodium hydrogen carbonate, pH 9.3) were derivatized with N,N-diethyl-2,4-dinitro-5-fluoroaniline (FDNDEA) at 100 degrees C for 20 min. The DNDEA-hydroxyproline adduct was separated on an Ultrasphere ODS column with a mobile phase of acetate buffer (containing triethylamine, 6 mL/L, pH 4.3) and acetonitrile (80/20, by vol), and was detected at 360 nm. A single run took 18 min with a hydroxyproline retention time of 7.3 min. The assay showed a linear response to hydroxyproline concentrations from 5 to 100 mg/L with a detection limit of 0.8 ng injected, corresponding to 2 mg/L in urine. Mean (SD) analytical recovery was 94.2 (13)% and 104 (9)% at 10 and 50 mg/L, respectively. Within-run and between-run CVs (n = 10) were 3.74% and 4.33%, respectively, for 25 mg/L. Results for samples (n = 50) analyzed by HPLC (y) vs ion-exchange chromatography with postcolumn ninhydrin reaction (x) correlated well: y = 0.98x + 1.02 (r = 0.985, Sxy = 3.13). In another comparison, involving 173 samples, a colorimetric procedure (Hypronosticon, x) gave slightly higher values than the HPLC method (y): y = 0.83x + 2.21 (r = 0.937, Sxy = 4.6).

Aniline Compounds↗

[The effect of STAT1 antisense oligonucleotides on lung fibroblast proliferation and hydroxyproline secretion].

OBJECTIVE: To investigate the effect of signal transducer and activator of transcription 1 (STAT1) antisense oligonucleotides on lung fibroblast proliferation and hydroxyproline secretion. METHODS: Ten adult female Wistar rats were randomly divided into two groups: one group was intratracheally instilled with bleomycin (BLM), while another group with 0.9% NaCl solution (NS). After 7 days, the rats were killed by right ventricle of heart exsanguinations under ketamine anaesthesia, and bronchoalveolar lavage (BAL) was performed to obtain alveolar macrophage (AM). AMs from the BLM group were divided into four groups, treated with STAT1 antisense oligonucleotides, STAT1 sense oligonucleotides, dexamethasone and medium alone (control), respectively. AMs and media were collected after culture for 36 h. The mRNA and protein expression of STAT1 and ICAM-1 in AMs were detected by RT-PCR and Cell-ELISA, respectively. The conditioned media were co-cultured with lung fibroblasts for 30 h, and then the cell proliferation and the concentration of hydroxyproline were examined. RESULTS: (1) The STAT1 mRNA expression by AMs in the STAT1 antisense oligonucleotides group (31.8 +/- 3.5) was lower than those of AMs in the STAT1 sense oligonucleotides group (64.2 +/- 4.3), the dexamethasone group (44.1 +/- 4.6) and the control group (65.5 +/- 4.6) (P < 0.05). Moreover, the STAT1 mRNA expression by AMs in the dexamethasone group was also lower than those of AMs in the STAT1 sense oligonucleotides group and the control group (P < 0.05), but the STAT1 mRNA expression by AMs in the STAT1 sense oligonucleotides group was not different from that of the control group (P > 0.05). The STAT1 mRNA expression by AMs in the NS group (14.9 +/- 3.1) was lower than those of AMs in the STAT1 antisense oligonucleotides group, the STAT1 sense oligonucleotides group, the dexamethasone group and the control group (P < 0.05). (2) The mRNA expression of ICAM-1 showed similar changes to the STAT1 mRNA expression by AMs. (3) The STAT1 protein expression by AMs in the STAT1 antisense oligonucleotides group (4.4 +/- 0.6) or in the NS group (3.7 +/- 0.4) was lower than those of AMs in the STAT1 sense oligonucleotides group (7.7 +/- 0.7), the dexamethasone group (5.9 +/- 0.4) and the control group (7.6 +/- 0.6) (P < 0.05); and the STAT1 protein expression by AMs in the dexamethasone group was also lower than those of AMs in the STAT1 sense oligonucleotides group and the control group (P < 0.05), but the STAT1 protein expression by AMs in the STAT1 sense oligonucleotides group was not different from that of the control group (P > 0.05). (4) The changes of ICAM-1 protein expression, lung fibroblast proliferation and hydroxyproline concentration were consistent with the changes of STAT1 protein expression by AMs. CONCLUSIONS: STAT1 antisense oligonucleotides could inhibit the mRNA and the protein expression of STAT1 and ICAM-1 in AMs. STAT1 antisense oligonucleotides also inhibited lung fibroblast proliferation and hydroxyproline secretion.

Animals↗

[Excretion of hydroxyproline in the urine of workers engaged in the production of iron-manganese alloys].

Statistically significant increase of the excretion of hydroxyproline with urine has been found to occur in workers employed in iron-manganese alloy manufacture, compared with the excretion of hydroxyproline in urine of the control group. Besides, the workers employed for 10 years and longer were characterized by statistically significantly increased excretion of hydroxyproline with urine as compared to the workers employed for shorter periods. No significant differences in excretion of hydroxyproline with urine, however, were detected among workers with chronic bronchitis and among those with smoking factor less than or equal to 10 and smoking factor greater than 10. (Smoking factor = number of packages of cigarettes smoked per day times years of smoking).

Adult↗

[Changes of the urinary excretions of hydroxyproline and fibronectin fragment in acute pancreatitis].

Urinary excretions of hydroxyproline and fibronectin fragment (FN fragment) were serially investigated in the patients with acute pancreatitis or acute exacerbation of chronic pancreatitis. While urinary excretion of FN fragment showed the maximal level on the first day of admission, high levels of urinary hydroxyproline were observed on the second to fifth day. As to the changes in the individuals, peak level of urinary FN fragment always preceded that of hydroxyproline. And it was assumed that the elevation of FN fragment excretion on the early phase of pancreatitis reflected tissue damages of pancreas itself and complicated organs, and following elevation of hydroxyproline showed enhanced collagen metabolism induced by acute inflammation and tissue damage. According to the severity of pancreatitis, urinary excretion of FN fragment on the first day increased, and it was therefore suggested that urinary FN fragment would be one of the parameters for the assessment of the severity of acute pancreatitis.

Acute Disease↗

Separation of hydroxyproline-containing protein from C1q (a subcomponent of complement) in serum.

Precipitation of the euglobulin fraction from serum separates C1q from another hydroxyproline-containing protein(s), which remains in the serum-minus-euglobulin fraction. The presence of C1q in the euglobulin fraction has been verified by radial immunodiffusion assay, by sensitivity to collagenase, by hydroxyproline content, and by electrofocusing. The presence of another hydroxyproline-containing protein(s) in the serum-minus-euglobulin fraction has been verified by the same criteria. The isoelectric pH range of C1q is 6.1-7.0; the isoelectric pH range of the other hydroxyproline-containing protein(s) is 4.2-5.5.

Blood Protein Electrophoresis↗

[Use of urinary hydroxyproline excretion as a marker of bone metastases in prostatic cancer (1)].

Twenty-four hour urinary hydroxyproline and urinary hydroxyproline creatinine ratio was measured without prior dietary restriction in 24 patients with prostatic cancer and 16 patients with benign prostatic hypertrophy. Both were elevated in patients with prostatic cancer with active bone metastases compared to the values of prostatic cancer without bone metastasis and benign prostatic hypertrophy. In these cases, the values of urinary hydroxyproline creatinine ratio were more reliable. The results show that urinary hydroxyproline creatinine ratio is a very sensitive indicator of active bone metastases of prostatic cancer without dietary restriction.

Bone Neoplasms↗