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The hydroxyproline content of plasma of patients with impaired renal function.

The plasma concentration of hydroxyproline fractions were determined in 52 patients with diminished renal function. High levels were found in each case and a correlation was found between the serum creatinine concentration and the plasma concentrations of peptide-bound hydroxyproline. Bone biopsies was taken from 16 patients from renal insufficiency and the results of morphometric studies of the biopsies were correlated with the blood determinations. Free hydroxyproline levels correlated significantly both with bone formation and with bone resorption. However peptide-bound and protein-bound hydroxyproline levels did not correlate with bone histology. Hence the plasma concentration of free hydroxyproline in patients with renal insufficiency can be used as an index of bone collagen turn-over.

Alkaline Phosphatase↗

[The clinical value of the hydroxyproline excretion in 24 hour urine compared to other laboratory diagnostic parameters in ankylopoietic spondylarthritis].

Hydroxyproline provable in the urine of man is regarded as indicator of the collagen metabolism. In 40 patients with ankylopoietic spondylarthritis the excretion of hydroxyproline in the 24-hour-urine was determined with the help of Stegemann's method. With 27.5 mg for the total hydroxyproline and 2.6 mg for the free hydroxyproline in the group of patients statistically significantly higher values were established than in a comparative group of 52 persons (20.1 or 0.6 mg, respectively). In patients with higher activity of the process (anamnestically, clinically) the excretions were larger than in patients with lower activity. That in this reflects also the temporary factor is to be concluded from the higher excretion rate in patients with a shorter duration of the disease and a low rate with a longer course. In comparison to the insignificant diagnostic relevancy of other acute-phase-reactions (BSR, CRP, number of leukozytes, blood protein fractions) from this results the conclusion that the determination of hydroxyproline may be used for the diagnostics of activity and also for the early diagnostics. By the temporary and technical-material expenditure, however, its use is limited in the routine practice.

Adult↗

Effect of salt restriction on urine hydroxyproline excretion in postmenopausal women.

Fasting calcium and hydroxyproline excretion are related to fasting sodium excretion in postmenopausal women. We postulate that calcium excretion is sodium dependent and that hydroxyproline excretion is calcium dependent. Therefore, we sought to lower urinary hydroxyproline, which is a marker of bone resorption, by lowering urinary sodium. Fasting urine samples were obtained from 59 postmenopausal women before and after 2 to 7 days of dietary salt restriction. The urinary sodium-to-creatinine ratio fell from 16 to 7; calcium to creatinine, 0.30 to 0.26; and hydroxyproline to creatinine, 18.2 to 16.8. In the 28 subjects with starting sodium-to-creatinine ratios greater than 15, the hydroxyproline-to-creatinine ratio fell from 19.6 to 16.3. Salt restriction may be one way of reducing bone resorption in postmenopausal women, particularly in those whose sodium intake is high.

Aged↗

[Changes in the levels of total and protein-bound hydroxyproline in the serum and urine of patients with cancer of the larynx after radiotherapy].

The serum concentration of total and protein-bound hydroxyproline as well as urinary hydroxyproline excretion were measured in 52 laryngeal cancer patients before, during and after irradiation. The modified colorimetric method described by Prockop and Udenfriend was used. After irradiation the increased total hydroxyproline concentration and excretion were observed, whereas no changes in serum protein-bound hydroxyproline were stated. Any correlation between the results of therapy and serum/urinary hydroxyproline levels were observed.

Adult↗

[Hydroxyproline in breast cancer].

The content of hydroxyproline was determined in cancerous and noncancerous tissues obtained from 40 patients with primary breast cancer, and investigated from a clinicopathologic viewpoint. The level of hydroxyproline in cancerous tissue was higher than that in normal mammary gland, but was lower than that of the tissue of mastopathy. In relation to the histologic type, the level was the highest in scirrhous carcinoma, followed by papillotubular and solid tubular carcinoma. With regard to lymph node metastasis, the hydroxyproline level was significantly higher in positive than in negative metastatic cases. The corresponding level tended to be higher in patients at stages II-IV than in those at stage I. There was no obvious relationship to tumor size. However, a significant, negative correlation was found between the hydroxyproline level and patient age. Thus, the content of hydroxyproline was found to be closely related to age and clinicopathologic factors such as the histologic type and degree of lymph node metastasis.

Adenocarcinoma, Scirrhous↗

Bone isoenzyme of serum alkaline phosphatase and urinary hydroxyproline excretion in thyrotoxicosis.

Studies were performed in 28 patients with proven active untreated thyrotoxicosis and 20 healthy age and sex matched adult subjects. In all subjects serum levels of thyroxine, calcium, phosphate and total activity of serum alkaline phosphatase and its bone isoenzyme were examined. Total urinary hydroxyproline excretion was measured after hydroxyproline free diet. Significant differences were found in the mean serum alkaline phosphatase and its bone isoenzyme levels and in the excretion of urinary hydroxyproline between a group of patients with thyrotoxicosis and a control group. In this study increased bone isoenzyme of alkaline phosphatase activity above the normal range was found in 24 patients with thyrotoxicosis and increased the excretion of urinary hydroxyproline in all patients with thyrotoxicosis. The relationship between the increased bone alkaline phosphatase and urinary hydroxyproline values indicated and increased bone turnover in the early period of disease.

Adult↗

Liquid chromatography of free hydroxyproline in serum.

We report a "high-performance" liquid-chromatographic method for measuring free hydroxyproline in serum. After separating the free hydroxyproline from the protein- and peptide-bound forms by centrifugal ultrafiltration, we derivatize the free form with 4-dimethylaminoazobenzene-4'-sulfonyl ("dabsyl") chloride and separate and quantify the derivative on a C18 chromatographic column. The mobile phase is sodium acetate buffer and acetonitrile, in a multilinear solvent gradient. The derivative is detected at 436 nm. Mean analytical recovery was 73%. The assay curve is linear from 14 to 213 pmol of hydroxyproline per 5-microL injection. The within- and interassay CVs were 8% at 10.7 mumol/L hydroxyproline and 10% at 13.5 mumol/L hydroxyproline, respectively.

Adult↗

Epidemiological study on the hydroxyproline: creatinine ratio as an index of growth rate.

A number of studies has suggested that urinary hydroxyproline can be used as an index of collagen metabolism and urinary hydroxyproline excretion is closely related to growth hormone. This report is a cohort study to demonstrate the relationship between the urinary hydroxyproline: creatinine ratio and growth rate for four years. The pattern of increased urinary hydroxyproline: creatinine ratio in adolescence appeared to be closely ralated to the general pattern of growth rate measured in school children. In has been shown that the urinary hydroxyproline: creatinine ratio is the most suitable index of the growth rate.

Body Height↗

Relevance of urinary excretion of alcian blue-glycosaminoglycans complexes and hydroxyproline to disease activity in rheumatoid arthritis.

Urinary excretion of glycosaminoglycans (GAG) and hydroxyproline was studied in 42 adult patients with rheumatoid arthritis (RA). A significant increase of GAG and hydroxyproline was found in RA patients compared to normal controls. RA patients fulfilling the criteria of disease activity were found to have urinary excretion of GAG and hydroxyproline higher than of RA patients not fulfilling these criteria. No significant correlation however was observed between urinary GAG and hydroxyproline. GAG and hydroxyproline excretion might be the expression of cartilage destruction and bone involvement and both may be related to the activity of the rheumatoid process.

Adult↗

Hydroxyproline-rich glycoprotein in the seed coat of field bean.

1. The seed coats of field bean (Dolichos lablab var. lignosus) were found to contain bound hydroxyproline (0.26 g per 100 g dry weight) extractable in 30% with 5% trichloroacetic acid or in 90% with 10% potassium hydroxide. 2. The alkaline extract was fractionated by DEAE-cellulose chromatography into several hydroxyproline-containing fractions, composed of protein and carbohydrate in varying proportions. From the major fraction, a hydroxyproline-containing glycoprotein fraction was isolated by Sephadex G-200 chromatography. It gave a single band on agar-gel electrophoresis, and contained: protein (23.1%), arabinose (11.2%), galactose (13.4%), glucose (17.6%), mannose (22.6%) and uronic acids (11.3%). The content of hydroxyproline in the protein moiety was about 9%. Hydrolysis of the glycoprotein fraction with barium hydroxide yielded three components containing hydroxyproline and arabinose at the ratios of 1:2, 1:3 and 1:4.

Chromatography, DEAE-Cellulose↗

Estimation of central activity of hydroxyproline--the amino acid characteristic for collagen degradation products.

The central activity of hydroxyproline - the amino acid characteristic for collagen degradation products was studied. The levels of urinary and serum hydroxyproline are elevated in many disturbances, among them is a mental deficiency connected with hydroxyprolinemia. Previous studies showed that the mixture of collagen degradation products is biologically active and exerts and effect on central nervous system. In our study hydroxyproline significantly decreased the psychomotor activity of rats in Lat's test, enhanced amphetamine stereotypy in lower doses, and enhanced catalepsy in one of used doses. It prolonged the time of thiopental sleeping. The pentylenetetrazole seizures latency was not significantly prolonged and duration of seizures was shorter after injection (icv) of hydroxyproline. Our investigations show, that hydroxyproline is the active amino acid in the mixture of collagen degradation products and may be responsible for central activity of this mixture.

Animals↗

[Hydroxyproline in the diagnosis of bone metastases in breast cancer].

15 patients with breast cancer and proven bone metastases have been studied. Free serum and total urinary hydroxyproline were measured and the ratio (hydroxyproline/creatinine) - 100 in a morning urine specimen was calculated. The preliminary results show elevated free serum hydroxyproline in 8 of 15 patients; total urinary hydroxyproline in a 24-h collection and the ratio hydroxyproline/creatine in a morning urine specimen were elevated in all cases. These results agree with those of the literature. Further studies of free hydroxyproline and of the ratio hydroxyproline/ creatinine in a morning urine specimen in similar patients with provide more information on the possibility of using these measurements, alone or in combination, for the early detection or follow-up under therapy of bone metastases.

Adult↗

Effect of antiphlogistic steroids on urinary hydroxyproline excretion in systemic lupus erythematosus.

Total hydroxyproline excretion in 24-hour urine was determined in 20 cases of systemic lupus erythematosus (SLE) before and during treatment. Additionally the excretion of hydroxyproline was determined in 14 patients receiving steroids for various other diseases (endocarditis, thrombocytopenia, acute leukemia, apofocal polyarthritis, ischialgia, spastic bronchitis, pleuritis and Dressler's post-infarction syndrome). The normal hydroxyproline value was established in 40 healthy persons. The normalization of pathologically increased hydroxyproline excretion in patients with SLE during treatment with prednisone in doses from 1.0 to 1.35 mg/kg/daily was associated with remission of clinical symptoms. Large doses of prednisone in pathological processes not primarily affecting connective tissue increased the 24-hour excretion of hydroxyproline and small doses of prednisone had no effect on hydroxyprolinuria.

Adult↗

Hydroxyproline Formation and Its Relation to Auxin-induced Cell Elongation in the Avena Coleoptile.

A study has been made of the effects on hydroxyproline formation of 4 factors that influence the rate of cell elongation in the Avena coleoptile; auxin, sugars, an external osmoticum, and actinomycin D. Hydroxyproline formation is increased by a combination of auxin and sucrose, but is affected to a much lesser extent by either factor alone. Its formation is inhibited by an external osmoticum but is scarcely affected by actinomycin D. The lack of correlation between the amount of hydroxyproline synthesis and the growth rate suggests that hydroxyproline formation is not involved in the actual process of wall loosening. It is suggested, instead, that if the wall is to retain its capacity for rapid extension, those hemicelluloses which are incorporated into it by intussusception rather than by apposition must be attached to a hydroxyproline-protein.

Journal Article↗

Two different families of hydroxyproline-rich glycoproteins in melon callus: biochemical and immunochemical studies.

Two different families of hydroxyproline-rich glycoproteins, HRGP(1) and HRGP(2), have been isolated from melon callus and separated by ion exchange chromatography on CM-sepharose. HRGP(1) corresponds to an arabinogalactan protein. The sugar portion of HRGP(1) accounts for 94% of the molecule and contains galactose (66%) and arabinose (34%); these residues are present as polysaccharide side chains attached to hydroxyproline. Hydroxyproline is the main amino acid residue (46%) of the protein moiety. The arabinogalactan protein nature of HRGP(1) has been checked by its ability to positively react with the beta-glucosyl Yariv antigen; the (3)H-labeled deglycosylated HRGP(1) also called HRP(1) migrates upon electrophoresis as a single band of molecular weight 76,000. HRGP(2) was fractionated by affinity chromatography on heparin-Ultrogel into three different glycoproteins, HRGP(2a,2b) and (2c). Two of these glycoproteins behave as polycations (HRGP(2b) and (2c)) and are chemically distinct from HRGP(2a). HRGP(2b) is the most abundant component and contains 41% protein and 50% sugar. Hydroxyproline, lysine, tyrosine, and arabinose are the most prominent residues of their respective moiety. The glycosylation pattern of hydroxyproline indicates that HRGP(2b) is related to and possibly a precursor of the wall HRGP; as in melon cell wall HRGP, Hyp-Ara(3) predominates, and small amounts of a putative Hyp-Ara(5) a hitherto unreported hyp-arabinoside, are recorded. The molecular weight of HRP(2b), the protein portion of HRGP(2b) is 55,000 +/- 5,000, as estimated after deglycosylation of the molecule with trifluoromethane sulfonic acid. Antibodies have been raised against HRGP(2b) and HRP(2b). Immunodiffusion shows that each antigen (HRGP(2b) or HRP(2b)) reacts with its own IgG, and cross-reacts with the heterologous IgG, thereby indicating the presence of common (unglycosylated) and specific (glycosylated and deglycosylated) epitopes. The arabinogalactan protein HRGP(1) is not recognized by either antibody and HRGP(2b) does not react with the Yariv antigen. Immunoprecipitation of (3)H-labeled HRP(1) and HRP(2b) in the presence of goat antirabbit IgG, followed by gel electrophoresis, allows to recover HRP(2b) only. Again, HRP(2b) is immunoprecipitated by the two antisera.

Journal Article↗

A developmentally regulated hydroxyproline-rich glycoprotein in maize pericarp cell walls.

We have studied the accumulation of peptidyl hydroxyproline in the pericarp of developing maize (Zea mays L., Golden cross Bantam sweet corn) kernels. Although this hydroxyproline accumulates throughout development, it is most soluble and its content per milligram dry weight greatest at midmaturation stages of development. Salt-soluble proteins containing this hydroxyproline from isolated cell walls of developing kernels were fractionated on a CsCl density gradient and on a Chromatofocusing column, resulting in the purification of an hydroxyproline-rich glycoprotein, PC-1. PC-1 is a basic protein of approximately 65 to 70 kilodaltons in molecular weight with an isoelectric point of at least 10.2 and a density of 1.38 to 1.39 in CsCl. Amino acid composition data indicate that it is rich in hydroxyproline, threonine, proline, lysine, and glycine. Its relation to dicot extensin is discussed.

Journal Article↗

Hydroxyproline peptides and bone mass in postmenopausal and osteoporotic women.

Total and nondialyzable hydroxyproline excretion was measured in 59 postmenopausal women and 68 women with spinal osteoporosis. Hydroxyproline excretion was similar in both groups of women and the hypothesis that hydroxyproline excretion is normally distributed could not be rejected for either group. No relationship was found between hydroxyproline excretion (total and percentage of nondialyzable) and body weight, height, body surface area, or total body calcium or bone mineral content of the radius, or these latter values normalized for age, sex, and body size. There was no difference in hydroxyproline excretion in osteoporotic women who took supplemental calcium as compared to those that did not. These data fail to provide any evidence that bone turnover in osteoporotic women differs from that in younger postmenopausal women, or that osteoporosis arises from a subpopulation of women with rapid bone loss.

Body Height↗

A study of adriamycin-reduced wound breaking strenght in rats. An evaluation by light and electron microscopy, induction of collagen maturation, and hydroxyproline content.

The cause of the reduced wound breaking strength (WBS) in adriamycin-treated animals is unknown. Differential staining of histologic sections, collagen fiber diameter measurements by electron microscopy (EM), induction of maturation, and hydroxyproline content analysis were used to examine this defect. Differential staining and formaldehyde induction of maturation revealed no differences between mature and immature collagen content in scars of adriamycin-treated and control animals. Mean collagen fiber diameters in control rats were 76 +/- 27 nm and, in adriamycin treated rats, 62 +/- 10 nm (mean +/- SD) at 21 days. There was no difference in total hydroxyproline content between ADR and C animals. However, significant differences existed between the specific activity of new or 3H-hydroxyproline (scar collagen) at 14 days (controls, 372 +/- 9 cpm; adriamycin-treated, 129 +/- 5 cpm) and at 21 days (controls, 365 +/- 11 cpm; adriamycin treated, 172 +/- 14 cpm; mean +/- SEM; P less than 0.001). It appears that the defect contributing to reduced WBS in adriamycin treated animals is not due to a collagen maturation defect but rather to a reduction in scar collagen accumulation as measured by new hydroxyproline content and reduced fiber diameter as determined by EM measurements.

Animals↗