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PubMed1994

Chronic platelet-activating factor induces a decrease in pulmonary vascular compliance, hydroxyproline, and loss of vascular matrix.

We have previously demonstrated that chronic intravenous platelet-activating factor (PAF) induces morphologic remodeling of pulmonary arteries characterized by a decrease in internal and external elastic lamina circumference, pulmonary arterial contracture, and internal elastic lamina duplication. The mechanism of PAF-induced arterial contracture is unknown. In this study we determined whether PAF caused arterial contracture through cell loss by calculating the number of cell nuclei/total cross-sectional area of arteries. The nuclear ratio was increased in intra- and preacinar pulmonary arteries of PAF-treated rabbits. Hydroxyproline content of lungs stratified by anatomic region was significantly reduced in intra-acinar tissue of PAF-treated rabbits, indicating that PAF-induced vascular contracture was associated with loss of interstitial collagen. We next tested whether these morphologic alterations were associated with decreased pulmonary vascular compliance and increased resistance. Compliance and resistance were determined in isolated, perfused lungs from rabbits chronically treated with PAF. Compliance was calculated: (1) from the slope of the venous occlusion trace (CVO), (2) by increasing left atrial pressure (CLA), (3) by increasing flow (CHF), and (4) by the classic static technique (CAV) of adding volume (2 ml) to a passively drained lung. Vascular compliance was significantly reduced in PAF-treated lungs when measured by all four methods; however, pulmonary vascular resistance was unchanged. We conclude that structural changes that result from chronic intravenous PAF infusion affect the elastic modulus to a greater extent than factors that influence pulmonary vascular resistance.

Animals↗
PubMed1975

Isolation and characterization of two hydroxyproline-containing glycoproteins from normal animal lung lavage and lamellar bodies.

Two glycopeptides, present in particulate material obtained by pulmonary lavage from normal rabbits, were isolated and characterized. The same two glycopeptides were present in preparations of lamellar bodies from rabbit lung. The estimated molecular weights of the two glycopeptides by sodium dodecyl sulfate-acrylamide gel electrophoresis were found to be 62,000 and 36,000 and both were found to contain hydroxyproline and relatively high amount of glycine (11 and 15 per cent, respectively). Carbohydrate analysis of the two glycopeptides demonstrated the presence of glucosamine, sialic acid, mannose, Fucose, and galactose. Similar glycopeptides of the same molecular weights, and amino acid and carbohydrate compositions have been found in layage mateial isolated from lungs of patients with alveolar proteinosis. The data indicate that thses two collagen-like glycopeptides are major intra-alveolar proteins in many mammals, including humans.

Amino Acids↗
PubMed1985

Estimation of bone turnover evaluated by 47Ca-kinetics. Efficiency of serum bone gamma-carboxyglutamic acid-containing protein, serum alkaline phosphatase, and urinary hydroxyproline excretion.

Bone gamma-carboxyglutamic acid-containing (Gla) protein (BGP, osteocalcin) is a noncollagenous protein of bone present in plasma and removed by the kidney. Plasma BGP has been shown to be elevated in patients with certain bone diseases. The present study evaluates serum BGP (S-BGP), serum alkaline phosphatase (S-AP), and urinary hydroxyproline excretion (U-OHP) in diseases with differing bone turnover rates, and compares the accuracy of these measurements for estimating bone mineralization (m) and resorption (r) rates. S-BGP, S-AP, U-OHP, and creatinine clearance (Clcr) were measured in patients with primary hyperparathyroidism (n = 13), hyperthyroidism (n = 6), and hypothyroidism (n = 6). Bone mineralization and resorption rates were calculated from a 7-d combined calcium balance and 47Ca turnover study. A highly significant correlation (r = 0.69, P less than 0.001) was found between S-BGP and m. Multiple regression analysis disclosed a partial correlation between S-BGP and m when Clcr was taken into account (r = 0.82, P less than 0.001), and between S-BGP and Clcr when m was taken into account (r = -0.62, P less than 0.005). In accordance with this, a stronger correlation (r = 0.89, P less than 0.0001) was found between S-BGP X Clcr and m than between S-BGP and m. A less significant correlation was found between S-AP and m (r = 0.45, P less than 0.05). Furthermore, U-OHP showed a highly significant positive correlation to r (r = 0.78, P less than 0.001). Thus, in the studied disorders of calcium metabolism, individual serum levels of BGP depend on both mineralization rate and renal function. Serum levels of BGP corrected for alterations in renal function are superior to uncorrected S-BGP and to S-AP levels in the estimation of bone mineralization rates.

Adult↗
PubMedDate not supplied

Effect of serum zinc level on amount of collagen-hydroxyproline in the healing gut during total parenteral nutrition: an experimental study.

Changes of serum zinc level in dogs receiving either a zinc-free solution or a zinc-supplement solution during total parenteral nutrition (TPN) and the influence of variety of serum-zinc level on serum albumin and collagen-hydroxyproline (C-Hyp) in the incised intestine were investigated. The serum zinc level of zinc-free dogs that was 126.0 +/- 34.3 micrograms per deciliter at the initiation of TPN decreased significantly to 80.0 +/- 15.0 micrograms per deciliter during the one week of TPN, whereas that of zinc-supplement dogs did not decrease during the same period of TPN. Lowering of the serum albumin value was more evident in zinc-free dogs than in zinc-supplement dogs. The amount of C-Hyp in the incised area of the intestine of zinc-free dogs was 2.63 +/- 0.66 micrograms per milligram and that of zinc-supplement dogs was 4.26 +/- 1.04 micrograms per milligram and this difference was significant. It was found that serum zinc level during TPN sharply reflects a supplement of zinc, and C-Hyp in the incised gut and serum albumin of zinc-free dogs are lower than those of zinc-supplement dogs.

Animals↗
PubMed1984

Urinary hydroxyproline in multiple myeloma: correlation with clinical stages and bone disease.

Twenty-four-hour urinary hydroxyproline excretion (HOP) (normal values: 6-22 mg/day/m2) was measured by the Hypronosticon test in 50 untreated patients with plasma cell myeloma. At diagnosis, HOP was elevated in 36 of 50 patients (72%) with a mean value of 35.9 mg/day/m2. Extent of bone lesions and clinical stage were accurately assessed in all patients. Higher HOP values were found in patients with a higher degree of bone lesions (multiple lytic areas and/or destruction of skeletal segments). According to clinical stages, HOP was very elevated only in stage III (mean value: 43.7); in stages I and II the mean value (25.2) was just above the normal range. Our data indicate that HOP in multiple myeloma at diagnosis is closely related to the extension of skeletal lesions and that during the clinical course it may be useful in the follow-up of bone disease.

Adult↗
PubMedDate not supplied

Comparison between CEA, TPA, CA 15/3 and hydroxyproline, alkaline phosphatase, whole body retention of 99mTc MDP in the follow-up of bone metastases in breast cancer.

The development of bone metastases in cancer can be monitored easily using three markers: 24 h urinary hydroxyproline excretion (HOP) (an index of osteoclastic activity), serum alkaline phosphatase (Alk.Ph.) (an index of osteoblastic activity) and 24 h whole body retention of 99mTc-methylene diphosphonate (WBR%) (an index of bone turnover). To evaluate the effectiveness of this group of bone tumor markers in breast cancer we compared it with the following group of three markers which are commonly used in the monitoring of breast cancer and in the follow-up of advanced disease with or without bone metastases: carcinoembryonic antigen (CEA), tissue polypeptide antigen (TPA) and breast carcinoma antigen (CA 15/3). In 48 patients with bone metastases CEA, TPA and CA 15/3 were shown to be sensitive (79%, 85%, 90% respectively), while HOP, Alk.Ph. and WBR%, which are commonly accepted as reliable markers of bone activity, showed a lower sensitivity (67%, 46%, 75% respectively). These results may be explained by the lack of osteoclastic or osteoblastic (or both) activity at the time of diagnosis. This explanation is supported by the fact that the bone markers HOP, Alk.Ph. and WBR% were found to be more sensitive than the others in the subsequent follow-up study. We conclude that in our study, CEA, TPA and CA 15/3 are at first more sensitive than Alk.Ph., HOP and WBR% but during the follow-up Alk.Ph., HOP and WBR% are possibly both more specific and more sensitive.

Adult↗
PubMed1996

Determination of total hydroxyproline and proline in human serum and urine by HPLC with fluorescence detection.

A method to measure total hydroxyproline (Hyp) and proline (Pro) in human serum and urine by HPLC was developed. Hyp and Pro in acid hydrolysates of serum and urine were derivatized with 4-(5,6-dimethoxy-2-phthalimidinyl)phenysulfonyl chloride after treatment with o-phthaladehyde and cleanup on Bond Elut C18 column. The derivatives of imino acids were separated on a reversed phase column by gradient elution with acetonitrile and phosphate buffer (1 mmol/l, pH 7) and detected by fluorescence measurement at 315 nm (excitation) and 385 nm (emission). Detection limits for both Hyp and Pro were 10 fmol per injection. The within-day and day-to-day relative standard deviations for Hyp and Pro in serum and urine were less than 3.19%. The recoveries of Hyp and Pro added to serum and urine were about 100%. The present method was applied to determine total Hyp and Pro in serum and urine from normal subjects and patients with chronic renal failure.

Adult↗
PubMed2000

Free 4-hydroxyproline content in serum of bedridden aged people is elevated due to fracture.

Hydroxyproline (Hyp) content in biological fluids is used as a parameter of collagen catabolism, especially bone resorption. In this paper, we examined the relationship between aging and serum free 4-Hyp and proline (Pro) content using a newly introduced analytical technique which is chemiluminescence determination with electrogenerated tris (2,2'-bipyridine) ruthenium(III). Serum was collected from 50 bedridden aged people and 131 normal subjects living in Yamato-son, Amamioshima, Kagoshima. In the free 4-Hyp content of normal serum, the change in females is more gentle than that males with aging. The free 4-Hyp content of bedridden aged serum is significantly (p<0.05) elevated compared to normal aged serum. Bedridden aged females and males also show levels significantly (p<0.05) elevated in comparison to normal. These changes are due to bone resorption based on bedridden patients' atrophy. In bedridden aged people, free 4-Hyp of those with a fracture was significantly (p<0.05) elevated than in people without a fracture. No relationship between low bone density and high free 4-Hyp in serum was observed. It seems that bone resorption or bedridden atrophy may be enhanced in bedridden aged people by a fracture. In free Pro content in serum, no significant change was observed. These results show that the free 4-Hyp content in serum is useful in the clinical or biological inspection of bone resorption, especially in aged people.

Adult↗
PubMed1986

Drug metabolizing activity in rats with chronic liver injury induced by carbon tetrachloride: relationship with the content of hydroxyproline in the liver.

The drug metabolizing activity of rat liver during long-term administration of carbon tetrachloride (CCl4), and its relationship with the content of hydroxyproline (Hyp) in the liver were examined. The contents of cytochrome P-450 (P-450) and cytochrome b5 (b5) and the metabolization of aniline, aminopyrine, 7-ethoxycoumarin (7-EC) and benzo(a)pyrene (B(a)P) in the microsomal fraction were examined five days after the final administration of CCl4. The contents of P-450 and b5 and the activity to metabolize the four substrates were gradually reduced as the Hyp content in the liver increased. However, aminopyrine N-demethylation and B(a)P hydroxylation, particularly the latter, was more reduced than aniline hydroxylation and 7-EC O-deethylation in the early stage of hepatic fibrosis. Such differences may be due mainly to the different P-450 subtypes affected by CCl4.

Animals↗
PubMed1988

Biological variations in serum total hydroxyproline concentration in the beagle dog.

A newly developed assay for hydroxyproline (Hyp) in physiological samples was used for determining the biological variations in serum total Hyp in the beagle dog. The results showed that dietary Hyp restriction in the beagle dog results in a significant decrease in serum total Hyp over the first 24 h and that there are no obvious circadian variations in serum total Hyp concentrations in beagle dogs on dietary Hyp restriction.

Analysis of Variance↗
PubMed1994

[Effects of aging on urinary secretion of 3-hydroxyproline and its importance for cancer screening in the elderly with ROC analysis].

OBJECTIVE: to observe the influence of aging on urinary secretion of 3-hydroxyproline (3-Hyp) in normal subjects and in cancer patients and study the effects of aging on the accuracy of urinary 3-Hyp as a general-purpose cancer screening test. DESIGN: We reanalyzed, from the aspect of gerontology, the amounts of 3-Hyp in the urine samples of the 211 healthy persons and 94 patients with cancer reported earlier. The two groups were divided in 3 age subgroups: adult, 30 to 44 years, middle-aged, 45 to 64 years, and aged, 65 years and older. The cancer stage was separated into 3 categories: stage 1, the early stage without any invasion, stage 2, locally invasive with no distant metastasis, and stage 3, advanced cancer. The correlation of age and urinary 3-Hyp was analyzed in normal subjects and cancer patients. Sensitivity and specificity were calculated for different cutoff points in the diverse groups to construct the receiver operating characteristic (ROC) curves. MAIN RESULTS: We observed that aging decreases the urinary secretion of 3-Hyp in normal subjects, with a correlation coefficient of -0.250 (p < 0.001). The mean levels of urinary 3-Hyp in patients with cancer were significantly higher than in the normal subjects, p = 0.005. The correlation coefficient of urinary 3-Hyp and age was not significant (0.100) in cancer patients. The cutoff points of 0.800 mg/g creatinine of urinary 3-Hyp for persons under 65 years of age and 0.600 mg/g creatinine for persons 65 years or older were the best thresholds for cancer screening. In the ROC analyses, we observed that 3-Hyp has higher accuracy for cancer screening in the aged group for all stages together and in the aged and middle-aged for early stages of cancer. CONCLUSIONS: The urinary 3-Hyp test was more effective for cancer screening in old persons (65 years or older), where cancer is more frequent and more difficult to discover. The authors stress the necessity and importance of verifying the effectiveness of urinary 3-Hyp for general cancer screening in a larger population and in a community set.

Adult↗
PubMed1983

Suppressive effect of elcatonin, an eel calcitonin analogue, on excessive urinary hydroxyproline excretion in polyostotic fibrous dysplasia (McCune-Albright's syndrome).

A 12-year-old Japanese girl with polyostotic fibrous dysplasia and endocrine concomitants, was treated with elcatonin, a synthetic eel calcitonin analogue, 10 MRC unit/twice a week given by intramuscular injection. Significant decreases in 24 hr urinary content of hydroxyproline and other amino acids from bone collagen were observed during the course of treatment over 5 months. This biochemical result suggests that the synthetic eel calcitonin analogue exhibits the therapeutic effect in patients with polyostotic fibrous dysplasia by inhibiting bone resorption.

Alkaline Phosphatase↗
PubMed2005

Triple helical structure and stabilization of collagen-like molecules with 4(R)-hydroxyproline in the Xaa position.

In this study, we examine the relationships between the structure and stability of five related collagen-like molecules that have hydroxyproline residues occupying positions not observed in vertebrate collagen. Two of the molecules contain valine or threonine and form stable triple helices in water. Three of the molecules contain allo-threonine (an enantiomer of threonine), serine, or alanine, and are not stable. Using molecular dynamics simulation methods, we examine possible explanations for the stability difference, including considering the possibility that differences in solvent shielding of the essential interchain hydrogen bonds may result in differences in stability. By comparing the structures of threonine- and allo-threonine-containing molecules in six polar and nonpolar solvation conditions, we find that solvent shielding is not an adequate explanation for the stability difference. A closer examination of the peptides shows that the structures of the unstable molecules are looser, having weaker intermolecular hydrogen bonds. The weakened hydrogen bonds result from extended Yaa residue Psi-angles that prevent optimal geometry. The Phi-Psi-maps of the relevant residues suggest that each residue's most favorable Psi-angle determines the corresponding collagen-like molecule's stability. Additionally, we propose that these molecules illustrate a more general feature of triple-helical structures: interchain hydrogen bonds are always longer and weaker than ideal, so they are sensitive to relatively small changes in molecular structure. This sensitivity to small changes may explain why large stability differences often result from seemingly small changes in residue sequence.

Collagen↗
PubMed1976

The degree of bone mineralization in hyperthyroidism estimated from the phosphorus to hydroxyproline ratio in bone.

The proportion between mineral and collagen in bone tissue, "the degree of mineralization", can be evaluated from the phosphorus to hydroxyproline ratio (P/Hypro ratio), determined in bone biopsies. The ratio was measured in 27 unselected hyperthyroid patients. No significant correlation was found between the P/Hypro ratio and the age of the patient, the duration of the disease or the value of serum thyroxine. The mean value of the P/Hypro ratio for the group studied was not significantly different from the normal mean. The results of the study indicate a normal degree of mineralization in the bone of patients with hyperthyroidism.

Adult↗
PubMed1979

Urinary excretion of calcium, hydroxyproline and 3',5'-cyclic adenosine monophosphate in primary hyperparathyroidism.

Urinary excretion of calcium (Ca), hydroxyproline (Hyp) and 3',5'-cyclic adenosine monophosphate (cAMP) was measured during fasting, and in the afternoon, over a 3 day period. Twelve hyperparathyroid patients, of whom 6 were re-studied after successful parathyroid surgery, and 10 control subjects participated, and were maintained on a collagen free diet for the duration of the study. Expressed as creatinine ratio values, Hyp was significantly higher in the morning than during the afternoon, whereas the Ca excretion pattern showed low morning and high afternoon values for all groups. cAMP excretion did not change during the two sampling periods. Large day to day variations for each parameter were observed in the individual patient. The value of cAMP measurements in the diagnosis of primary hyperparathyroidism was confirmed. The results may imply that a diurnal variation in Hyp excretion exists in primary hyperparathyroidism and that food intake produces a suppression of Hyp excretion, possibly secondary to suppression of parathyroid function or, in our view, to increased calcitonin excretion.

Adult↗
PubMed1986

[Behavior of urinary hydroxyproline and the effect of cigarette smoking in silicosis].

This study was conducted through regular pneumonoconiosis examination according to the law on 1,096 employees of medium and small-sized ceramic enterprises in Tokai district in 1981-82. Interview examination with BMRC questionnaire, X-ray examination and measurement of urinary hydroxyproline to creatinine ratio (HOP ratio) were carried out in order to elucidate the relationship between silicosis and urinary HOP ratio and to demonstrate the effect of smoking on pneumofibrosis. Grade of silicosis was classified into five types (0 to 4) based on the Japanese Classification of Radiographs of Pneumoconioses. In evaluating the behavior of urinary HOP ratio, when smoking factor is added in the early grade of pneumofibrosis (type 1 and type 2), collagen decomposition rate is rapidly repressed and fibroplastic conditions develop to the final grade as type 3 and 4, although smoking itself does not seem to induce pneumofibrosis. To exclude the effects of smoking, nonsmoking group was used for measurement of HOP ratio by grade. The HOP ratio in type 0 was lowest and HOP ratio increased in the order of type 1 and type 2. The turning point was found in type 2 and their HOP ratio decreased one after another. The turning point shifted from type 2 to type 3 in the case of non-smokers without any index symptoms by BMRC questionnaire and also shifted to type 1, in the case of non-smokers with them. Shifting of turning point suggests that index symptoms also promote fibroplastic activities.

Adult↗
PubMed1993

Hydroxyproline and total protein levels in gingiva and gingival crevicular fluid in patients with juvenile, rapidly progressive, and adult periodontitis.

The aim of this study was to compare hydroxyproline (Hyp) and total protein levels both in the gingiva and gingival crevicular fluid (GCF) from juvenile (JP), rapidly progressive (RPP), and adult periodontitis (AP) patients and periodontally healthy controls (C). A total of 60 individuals, 15 from each group, were studied. GCF was obtained before gingival sampling. Clinical measurements were recorded. The gingival samples were harvested by full thickness flap operation from patients and immediately after the extraction of the teeth for orthodontic reasons from controls. The samples were analyzed biochemically. GCF Hyp levels were significantly higher in the AP group than those of the RPP and C groups. The strong positive correlations between gingival and GCF Hyp levels were determined in the disease groups. Total protein levels in both the gingiva and GCF were significantly higher in disease groups. However, the differences between the disease groups were not statistically significant. Correlations between the clinical parameters and Hyp levels in gingiva and GCF were determined. The findings indicate the Hyp and total protein levels both in the gingiva and GCF appeared to be increased in the disease groups. These findings suggest that both the synthesis and degradation of collagen and total protein are elevated in periodontal disease. However, our findings do not support the concept that measurement of either Hyp or total protein levels in gingiva or GCF is a suitable or reliable criterion in determination of disease activity.

Adolescent↗
PubMed1993

[Relationship between aging and hydroxyproline content of serum in human being].

The authors have studied the effects of physical exercise on extracellular substances of several tissues in animals. Hydroxyproline (Hyp) exist in the form of peptide and the free state in blood. It is generally thought that Hyp in blood derived from the decomposition of collagen in tissue. To clarify the influence on collagen metabolism due to aging and exercise training, we attempted to see the changes of Hyp content of serum before and after exercise in human being. Subjects were 51 adult aged 20 to 74 years, and 10 nonathletes and 51 athletes, aged 18 to 22 years. The Hyp content in serum was measured by woessner's method. The Hyp content of serum decreased with age. This fact suggests that the formation of intramolecular and intermolecular cross-links in collagen of tissue is accelerated by aging. And there is a possibility that the proportion of insoluble collagen increased in tissue of aged human being. Before the exercise, Hyp content of serum in athlete was lower than that in nonathletes. Hyp content of serum increased remarkably after the exercise in athlete. These results suggest that the collagen metabolism may be accelerated by continuous exercise training.

Adolescent↗