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Antigenic changes of the glomerular basement membrane after incorporation of hydroxyproline isomers.

We investigated antigenic changes of murine glomerular basement membrane (GBM) collagen type IV after oral feedings of 3-cis- or 4-cis-hydroxyproline. As shown by thin layer chromatography and gas chromatographic-mass spectrometric studies, 3-cis- and 4-cis-hydroxyproline were incorporated into the GBM collagen instead of the normal trans isomer and led to a considerable reduction of the collagen antibody reactivity on indirect immunofluorescence, passive hemagglutination and radioimmunoassay when compared to GBM collagen of control animals. We speculate that antigen modification due to the incorporation of stereoisomers could serve as a model for basement membrane immunopathology.

Animals↗

Renal tubular transport of proline, hydroxyproline, and glycine. 3. Genetic basis for more than one mode of transport in human kidney.

Impaired renal tubular transport of proline, hydroxyproline, and glycine was inherited as an autosomal recessive trait in two Ashkenazi-Jewish pedigrees and one French-Canadian family; the heterozygotes for the trait exhibited hyperglycinuria only. Intestinal transport of imino acids and glycine was not impaired in homozygotes. It is possible that more than one mutant allele may occur at a locus controlling tubular transport of the imino acids and glycine, since one subject with the imino-glycinuric phenotype had one parent who was not hyperglycinuric. More than 60% of the specific tubular transport function is still available in homozygotes for absorption of imino acids and glycine at endogenous substrate concentrations; however, this persistent transport is already saturated at these concentrations in contrast to the large capacity available in normal subjects. Furthermore, the glycine portion of this persistent transport is noninhibitable by imino acids in contrast to the normal situation. The imino acids can inhibit each other's uptake in mutant and normal phenotypes. Two modes of transport for the imino acids and glycine are proposed to explain these observations: (1) a common system with high capacity, and (2) two additional systems, each with low capacity (one-tenth or less of the common system). One of these systems is apparently shared by proline and hydroxyproline. The mutant allele(s) observed in this investigation occur at the locus for the common system.

Adolescent↗

Long-term effects of pulmonary damage in mice on lung weight, compliance, hydroxyproline content and formation of metastases.

The development of "metastatic" colonies of fibrosarcoma in the lungs of mice after IV injection of a single-cell suspension of tumour cells varied with time after irradiation or administration of bleomycin. Most notably, there was a late radiation-induced enhancement of colony-forming efficiency that coincided with an increase in lung weight and hydroxyproline content and a decrease in compliance. There was no radiation-induced enhancement of "spontaneous" metastases from a mammary carcinoma implanted in the thighs of the animal. The sequence of events after administration of bleomycin was similar to that after irradiation, but the duration of the treatment was different and the temporal relationship between enhanced colony-forming efficiency and changes in lung weight and hydroxyproline content was not as clear cut.

Animals↗

Automated analysis of total urinary hydroxyproline based on resin-catalysed hydrolysis.

An automated method for the analysis of total urinary hydroxyproline using strong cation resin tablets of Hypronosticon is described for use in a clinical laboratory and the results are compared with those obtained by other methods. Even though good recoveries are obtained using the technique described in the present work by adding the internal standards either before or after hydrolysis of urine, the present method gave consistently lower values of urinary hydroxyproline compared with a manual and an automated method.

Autoanalysis↗

Interrelationships between urinary sodium, calcium, hydroxyproline and serum PTH in healthy subjects.

The effect of sodium intake on the excretion of calcium, hydroxyproline (OHP) and serum parathyroid hormone (PTH) was examined. When seven healthy males increased their sodium intake by 200 mmol/day, excretion of calcium increased by 1.70 mmol/day and that of OHP by 27%. Serum PTH, however, did not increase significantly. In a cross-sectional study of 334 healthy female subjects there were strong positive correlations between fasting urinary sodium/creatinine (Na/Cr) and calcium/creatinine (Ca/Cr) ratios (r = 0.573) and between Na/Cr and urinary OHP/Cr ratios (r = 0.246). Serum PTH was negatively correlated with Na/Cr ratio (r = -0.161). We conclude that an increase in dietary sodium causes calciuria and an increase in hydroxyproline excretion. The results do not support the hypothesis that this is mediated by PTH.

Adult↗

Urinary excretion of acid glycosaminoglycans and hydroxyproline in a patient with oculo-cerebro-renal syndrome.

The metabolism of ground substance in connective tissue of an 18-year-old boy with oculo-cerebro-renal syndrome was studied. He had characteristic clinical and laboratory findings described by Lowe et al. such as growth retardation, mental deficiency, glaucoma, cataracta, decreased muscle tone, metabolic acidosis, aminoaciduria and osteomalacia. The urinary excretion of acid glycosaminoglycans and of total hydroxyproline were 27 mg/day (as glucuronic acid) and 280 mg/day respectively on admission. Both values decreased to the upper limits of normal level transiently during treatment with alkali and vitamin D2. At that time, an improvement in bone abnormalities, a decrease of serum alkaline phosphatase, and an elevation of serum inorganic phosphate were observed. The therapy prevented him from progressive osteomalacia and cured him of it, but mucopolysacchariduria and hydroxyprolinuria did not disappear. Analytical electrophoresis on cellulose acetate sheets showed that urinary acid glycosaminoglycans were composed of undersulfated chondroitin 4-/6-sulfate and heparan sulfate with a ratio of 6:4, on admission. After oral administration of alkali, the excretion of heparan sulfate decreased and undersulfated chondroitin 4-/6-sulfate was determined as a main component of urinary acid glycosaminoglycans. The clinical and laboratory data in this case suggested that the increased excretion of acid glycosaminoglycans and total hydroxyproline was caused by abnormal metabolism in connective tissues, especially by the bone abnormalities, in this syndrome.

Adolescent↗

Effects of propranolol on changes in heart rate, heart weight, kidney weight, urinary hydroxyproline and weight gain induced by large doses of thyroxine in the rat.

The effects of the beta-adrenergic receptor blocking agent, DL-propranolol, and of the antithyroid drug, carbimazole, upon some manifestations of thyroxine (T4)-induced changes in peripheral metabolism were studied in rats. Propranolol lowered the heart rate, but did not alter the following changes induced by T4: increment in heart rate, increase in heart or kidney weight, increase in urinary hydroxyproline, decrease in body weight gain or increase in serum T4. Carbimazole administration lowered serum T4 and reduced weight gain, but had no effect upon heart rate or hydroxyproline excretion.

Animals↗

Effect of paraquat on plasma fibronectin, serum free hydroxyproline, serum ceruloplasmin and lung collagen content in monkeys.

The purpose of this study was to characterize paraquat toxicity in monkeys and to determine the feasibility of using the monkey as an animal model for the study of paraquat-induced pulmonary fibrosis in humans. Sixteen Japanese monkeys (Macaca fuscata), more than 3.5 years of age, with bodyweight ranging from 3.2 kg to 10.2 kg, were randomly divided into two groups. They were administered paraquat dichloride (PQ) by injection (s.c.) at a dose of 2.0 mg/kg bodyweight (12 monkeys) or s.c. saline, at a dose of 0.2 ml/kg bodyweight (control group, 4 monkeys), every two days for a period of four to five times. Eight monkeys (66.7%) from the PQ treatment groups expired due to subchronic PQ toxicity from days 11 to 35. Four monkeys (33.3%) from the PQ treatment group and all four monkeys from the control group survived the observation period of 66 days. On day 66, all of the surviving monkeys were sacrificed and examined for possible histopathological changes and the lung hydroxyproline content was determined. Our results indicate that the concentration of free hydroxyproline and plasma fibronectin did not vary significantly. The serum ceruloplasmin for the monkeys of the PQ treatment groups was significantly increased from day 14 to 21, compared to the control group. Also the total lung collagen of both the expired and surviving monkeys in the PQ treatment groups was elevated significantly, compared to the control group. The monkeys can provide extensive opportunities for research on the mechanism and the treatment of PQ poisoning in man.

Animals↗

Fluctuation of the mucosal hydroxyproline content in compound 48/80-induced gastric lesions in rats.

Extensive gastric mucosal lesions were observed after i.p. administration of compound 48/80 (0.75 mg/kg) for 3 days to rats. The mucosal hydroxyproline content was significantly increased at 4 h after the last injection of compound 48/80 and subsequently decreased as the lesions were healed. Treatment with cimetidine (50 mg/kg, i.p.) did not prevent compound 48/80-induced gastric lesions. The increase in the mucosal hydroxyproline content was significantly reduced by treatment with cimetidine. This fact suggested that cimetidine might have an inhibitory effect on the regeneration of connective tissue.

Animals↗

The aortic content of glycosaminoglycans, hydroxyproline and calcium in experimental uraemia with special reference to parathyroidectomy and vitamin-D treatment.

The contents of the various glycosaminoglycan (GAG) fractions, hydroxyproline and calcium in the thoracic aorta from the rat were studied. The effect on these contents of induced uraemia (3/4 kidney resection) combined with parathyroidectomy and/or 1-alpha-hydroxycholecalciferol (1-alpha-OH-D3) treatment was evaluated. Confirming earlier results increased levels of GAGs (predominately of chondroitin and dermatan sulphate) were found in the aortae of uraemic rats. The present study showed that parathyroidectomy or treatment with 1-alpha-OH-D3 did not significantly further alter the aortic content of GAGs in uraemic rats. Arteriosclerotic changes (calcifying medial necrosis) were only found in non-parathyroidectomized uraemic rats given 1-alpha-OH-D3. The aorta from these rats had a lower total content of GAGs, mainly due to a decrease in hyaluronic acid, than that from uraemic rats without calcifications. The aortic content of calcium was lowest in uraemic parathyroidectomized rats. Treatment with 1-alpha-OH-D3 increased this content, which was augmented 100-200-fold when calcifications were present. No correlation between the aortic contents of calcium and GAGs was found. There was a statistically significant correlation between the aortic content of calcium and the serum calcium X phosphate product. A significantly lower content of hydroxyproline was found in uraemic non-parathyroidectomized rats given 1-alpha-OH-D3, especially in those with arteriosclerosis.(ABSTRACT TRUNCATED AT 250 WORDS)

Animals↗

Hydroxyproline turnover in dermal collagen grafts in reconstituted skin wounds in the rat.

Using donor skin labelled with tritiated hydroxyproline and proline, a comparative study has been made of hydroxyproline (and by extrapolation collagen) turnover in skin, dermal collagen and glutaraldehyde crosslinked dermal collagen grafts in unlabelled isogenic recipient rats. In contrast with non-crosslinked collagen and skin grafts, collagen grafts stabilized by crosslinking with 0.01% glutaraldehyde underwent little loss of original graft collagen and maintained their original collagen mass over a 22 week period. As the skin grafts acquired a similar amount of new collagen to that lost, they also maintained their original mass. However, the non-crosslinked collagen grafts showed only a small gain of new collagen and lost 57% of their original mass.

Animals↗

Observation on the use of 1 alpha-hydroxyvitamin D3 in the prevention of bovine parturient paresis: the effect of a single injection on plasma 1 alpha-hydroxyvitamin D3, 1,25-dihydroxyvitamin D3, calcium, and hydroxyproline.

Israeli Friesian cows in the third or later lactation, were injected intramuscularly with 1 alpha-hydroxyvitamin D3 in propylene glycol. Plasma 1 alpha-hydroxyvitamin D3 peaked 24 to 48 h after 700 micrograms of the drug was injected and decreased to the initial concentration 4 d later. The disappearance rate of 1 alpha-hydroxyvitamin D3 from the blood was .330 d-1 (biological half-life of 2.1 d). Plasma 1,25-dihydroxyvitamin D3 peaked 24 to 48 h after the injection and approached the initial concentration 5 to 7 d later. Plasma calcium increased within 24 h, reached a plateau at 4 to 6 d, and then decreased. Administration of 700 micrograms 1 alpha-hydroxyvitamin D3 resulted in higher plasma 1,25-dihydroxyvitamin D3 and calcium concentrations than when 350 micrograms of the drug was injected. 1 alpha-Hydroxyvitamin D3 did not affect plasma hydroxyproline. The injection of 700 micrograms 1 alpha-hydroxyvitamin D3 52 to 36 h before parturition resulted in a greater increase in plasma 1,25-dihydroxyvitamin D3 than that obtained with 350 micrograms. In both groups 1,25-dihydroxyvitamin D3 peaked 24 h after injection, as opposed to the controls, which peaked 72 h postpartum. At parturition, the 700-micrograms dose resulted in a slight hypercalcemia; the 350-micrograms dose moderated the hypocalcemia prevalent during this period, whereas in controls hypocalcemia was more pronounced. In cows injected with 700 micrograms, hypercalcemia was observed until the 15th d postpartum. Plasma hydroxyproline was elevated in all cows 48 to 72 h after parturition, but the increase was smaller in animals injected with 1 alpha-hydroxyvitamin D3.(ABSTRACT TRUNCATED AT 250 WORDS)

Animals↗

The effects of manganese, biotin, and choline on hexosamine and hydroxyproline content as related to leg weakness.

Three experiments were conducted with day-old broiler chicks to determine the effects of dietary choline, biotin, and manganese on cartilage hexosamine and hydroxyproline content. The incidence of leg weakness was 75% or higher on all basal diets. The levels of supplementation found to minimize leg weakness were 1540 mg choline chloride/kg diet, .2 mg biotin/kg diet, and 100 mg manganese/kg diet. The levels required to maintain normal epiphyseal cartilage hexosamine content were 385 mg of choline chloride, .05 mg of biotin, and 25 mg of manganese/kg of diet. Substantially higher levels of nutrients were needed to reduce the incidence of leg weakness. Only with manganese supplementation was the incidence of leg weakness significantly reduced at the level of supplementation required for normal hexosamine content. Hydroxyproline content was not affected by varying the levels of supplemental choline, biotin, or manganese.

Animals↗

Investigation into the distribution of total, free, peptide-bound, protein-bound, soluble- and insoluble-collagen hydroxyproline in various bovine tissues.

Collagen is a family of proteins which consists of several genetically distinct molecular species and is intimately involved in tissue organization, function, differentiation and development. The purpose of this study was to investigate the concentration of different hydroxyproline (Hyp) fractions viz., total, free, peptide-bound, protein-bound, soluble- and insoluble-collagen hydroxyproline (Hyp) in various bovine tissues. Results showed that liver had the highest concentration of free Hyp followed by kidney, brain, spleen, lungs, muscle and heart. Liver also had the highest concentration of peptide-bound collagen Hyp followed by kidney, heart, spleen, lungs, brain and muscle. The concentration of protein-bound collagen Hyp was highest in the liver, followed by kidney, spleen, lungs, muscle, brain and heart. Total Hyp was highest in the liver, followed by kidney, spleen, brain, heart, muscle and lungs. Liver also had significantly high concentration of collagen as compared to other tissues examined (P<0.001). Spleen had the significantly higher concentration of soluble-collagen Hyp when compared to other tissues (P<0.001). This was followed by heart, muscle, lungs, brain, kidney and liver. Heart had the highest concentration of insoluble-collagen Hyp followed by lungs, kidney, liver, muscle, spleen and brain. The variation among the insoluble-collagen Hyp concentration of heart and muscle, spleen and brain was significant (P<0.001). We speculate that these differences could be due to the variation in turn over of rate of collagen metabolism in this species.

Animals↗

Response of urinary hydroxyproline to dietary protein and fasting in white-tailed deer.

The effects of dietary protein, fasting, and refeeding on urinary hydroxyproline of nine captive female white-tailed deer (Odocoileus virginianus) were examined from 23 February to 3 May 1984 in northern Minnesota. In the fasted group, mean hydroxyproline:creatinine (OHP:C) was greater (P less than 0.05) at week 4 compared to baseline at week 0. Between fasted deer and deer fed high protein-high energy (HPHE) and low protein-high energy (LPHE) diets, no difference in OHP:C ratios was detected during the initial 4 wk of the study. Urinary OHP:C ratios were significantly (P less than 0.05) greater in the fasted group during refeeding, concomitant with greater feed consumption and weight gain. There was also a significant (P less than 0.02) time effect in the fasted-refed group; OHP:C ratios increased during these two phases of the study. There was no difference between the HPHE and LPHE fed deer in renal OHP excretion. However, mean OHP:C ratios in yearlings (16.8 +/- 2.2) were greater (P less than 0.001) than in the adults (7.5 +/- 1.2) of those groups, indicating a higher collagen turnover rate. Urinary OHP:C shows potential as an indicator of growth and starvation, and the data presented may serve as reference values.

Animal Feed↗

Tissue engineering of a bioprosthetic heart valve: stimulation of extracellular matrix assessed by hydroxyproline assay.

Creation of an autologous heart valve by tissue engineering offers a promising approach to cardiac surgery. Although we have demonstrated successful formation of native valve analogous tissue in vitro, hemodynamic competence remains a serious problem. The aim of this study was to optimize in vitro formation of collagen as a precondition for mechanical stability of new tissue. Human myofibroblasts were seeded on square sheets of biodegradable scaffolds (control). To stimulate collagen production, one series was cultured with L-ascorbic acid 2-phosphate. In a second series, the seeded scaffolds were subjected to tension by mounting them on a frame. After 4 weeks of culture time, the collagen content of the different series was assessed by hydroxyproline assay. Light and scanning electron microscopy were performed. Hydroxyproline content of the framed scaffolds was 10 times higher than that of the control group (p < 0.05) and 6 times higher than in the unframed scaffolds grown with ascorbic acid (p < 0.05), respectively. Scanning electron microscopy proved extensive formation of solid tissue in the framed samples. These results demonstrate that supplementation of myofibroblast cultures with ascorbic acid, especially if grown on strained scaffolds, significantly increases collagen content, which is crucial for mechanical stability. This concept is a further step toward the creation of a hemodynamically competent autologous heart valve.

Ascorbic Acid↗

The degree of mineralization in normal bone tissue estimated as the phosphorus to hydroxyproline ratio.

The degree of mineralization can be evaluated from the phosphorus to hydroxyproline ratio in bone biopsies. A small but significant rise in the ratio with increasing age was observed in 137 persons without bone diseases, aged from 20 to 89 years. No significant difference between men and women was found with regard to the phosphorus to hydroxyproline ratio. The study indicates that with increasing age, in addition to senile osteopenia, there is also a change in the form of an increase in the degree of mineralization.

Adult↗

Effect of alcohol on total urinary hydroxyproline excretion.

Urinary hydroxyproline excretion was measured in 33 patients regularly drinking over 80 gm. alcohol daily. Those with changes of cirrhosis on liver biopsy excreted significantly more hydroxyproline than those with normal appearances, fatty change or hepatofibrosis. In 16 patients, the excretion was measured both when they were drinking and when abstaining from alcohol, in the form of several beverages. The rate of excretion was significantly higher after alcohol has been withdrawn. This result raises the possibility that alcoholic beverages may inhibit collagen degradation. If substantiated, it further suggests that the effect of alcohol on collagen metabolism may be of importance in the pathogenesis of alcoholic cirrhosis.

Alcoholism↗