desmosine and deoxypyridinoline

Chronic graft-versus-host disease (cGVHD) is a common and potentially lethal complication of allogeneic hematopoietic stem cell transplantation (HSCT). cGVHD as well as the transplant procedure itself (chemotherapy with or without radiotherapy) can lead to the degradation of connective tissue components such as elastin and collagen. The catabolism of these structural proteins releases desmosine (DES), lysylpyridinoline (LP), hydroxylysylpyridonoline (HP), and related pyridinium-based cross-linkers analogues that could represent potential biomarkers for cGVHD. This study reports the development of a sensitive liquid chromatography/tandem mass spectrometry method for the simultaneous analysis of N-propyl derivatives of DES, HP, and LP. The concentrations of free and total forms of urinary DES, HP, and LP were determined using synthetic deuterated internal standards. This method enabled accurate quantitation of these pyridinium-based cross-linkers from as little as 100 microL of urine with detection limits of 0.03-0.10 ng/mL. These compounds were analyzed in urine samples from three groups of patients: (1) Healthy volunteers, (2) Autologous HSCT recipients (who cannot develop cGVHD), and (3) Allogeneic HSCT recipients at onset of cGHVD. These analyses revealed that the urinary concentrations of DES, HP, and LP in the autologous recipients were greater or equal to the cGVHD group although both groups showed marked increase in the levels of these compounds compared to healthy individuals. These results suggest that the chemotherapy treatment has significant effects on the turnover of elastin and collagen, and that these biomarkers could be effective during prospective analyses to determine the onset of cGVHD.

Topics: Adult; Aged; Amino Acids; Biomarkers; Chromatography, Liquid; Desmosine; Female; Graft vs Host Disease; Hematopoietic Stem Cell Transplantation; Humans; Male; Middle Aged; Tandem Mass Spectrometry

Urinary levels of collagen- and elastin-crosslink amino acids have been used as biologic markers for degradation of collagen and elastin in the body. Circadian variation of collagen-crosslink amino acids is well known. The current study was undertaken to determine whether there is also circadian variation in excretion of elastin-crosslink amino acids. We used an isotope dilution-HPLC assay to measure the elastin-crosslink amino acids, desmosine (DES) and isodesmosine (IDES), and the collagen-crosslink amino acids, hydroxylysyl pyridinoline (HP) and lysyl pyridinoline (LP), in urine. Sixteen apparently healthy subjects collected urine from 5:00 to 7:00 AM, and from 5:00 to 7:00 PM. Mean urinary excretion of DES and IDES in women was 56% and 41% higher (P < 0.001), respectively, in AM versus PM specimens when normalized by the creatinine content of the urine specimen. For men, the corresponding values were 11% and 13% higher (not statistically significant). Mean urinary excretion of HP and LP in women was 61% and 71% higher (P < 0.001), respectively, in AM versus PM specimens. For men, the corresponding values were 11% and 19% higher (not statistically significant). Differences were not found in the AM versus PM rates of excretion of creatinine in men or women. These findings demonstrate the occurrence of circadian variation in HP, LP, DES and IDES in women but not in men. We conclude that the time of collection of urine specimens, especially from women, must be taken into consideration in using the urinary levels of these crosslink amino acids as biologic markers for collagen or elastin degradation.

Topics: Adult; Amino Acids; Biomarkers; Circadian Rhythm; Collagen; Cross-Linking Reagents; Desmosine; Elastin; Female; Humans; Isodesmosine; Male; Middle Aged; Reference Values; Sex Characteristics

Keloid is a tissue with an excessive accumulation of collagen. In this study, we have partially characterized post-translational modifications of type I collagen in human keloid in order to pursue their potential involvement in this pathology. The levels of lysyl hydroxylation of the helical portions of alpha 1 and alpha 2 chains of type I collagen in keloid were significantly higher than those of normal, while the levels of prolyl hydroxylation were identical between these two groups. The contents of the major reducible cross-links in dermal collagen, dehydro-hydroxylysinonorleucine and dehydro-histidinohydroxymero-desmosine, were both significantly higher in keloids (up to sixfold) than those of normal. In addition, significant amounts of hydroxylysine-aldehyde derived cross-links that are characteristic of skeletal tissue collagens, dehydro-dihydroxylysinonorleucine (about 0.3 mole/mole of collagen) and pyridinoline (about 0.1 mole/mole of collagen), were found in keloids. These results indicate that keloid-forming cells are phenotypically different from those in normal dermis and that the collagen produced is highly cross-linked. The increased cross-linking provides the fibrils with more stability that may result in an accumulation of collagen.

Topics: Adult; Amino Acids; Collagen; Cross-Linking Reagents; Desmosine; Dipeptides; Histidine; Humans; Hydroxylation; Hydroxylysine; Hydroxyproline; Keloid; Middle Aged; Protein Processing, Post-Translational; Protein Structure, Secondary; Skin

Elastin degradation has been reported to be increased in patients with cystic fibrosis (CF). In order to further explore evidence for elastin degradation in a group of 18 patients with CF with a wide range of disease severity, we used an isotope dilution method to measure urinary desmosine (DES) and isodesmosine (IDES), amino acids derived exclusively from cross-linked elastin, and hydroxylysylpyridinoline (HP) and lysylpyridinoline (LP), amino acids derived exclusively from cross-linked collagen. Urinary DES and IDES (mean +/- SD) were 23.9 +/- 30.7 and 18.5 +/- 22.4 micrograms/g creatinine, respectively, in the patients with CF versus 7.5 +/- 1.7 and 6.8 +/- 1.4 micrograms/g creatinine, respectively, in 10 healthy control subjects (p < 0.001); only two patients with CF had DES values within the control range. The values of urinary HP and LP in the CF group were 54.9 +/- 39.1 and 12.3 +/- 8.6 nmol/mmol creatinine, respectively, versus 24.5 +/- 5.8 and 5.1 +/- 2.7 nmol/mmol creatinine, respectively, in the controls (p < 0.005). Both HP and LP were highly correlated (r = 0.71, p < 0.0001). Patients with CF had active pulmonary inflammation; neutrophils were abundant in the bronchoalveolar lavage fluid of the CF group and correlated with elastase activity measured with methoxysuccinyl Ala-Ala-Pro-Val paranitroanilide (r = 0.61, p < 0.05). Airway neutrophils had decreased expression of the complement receptor CR1 (CR1/CR3 of 0.17 +/- 0.15 versus 1.0 for blood neutrophils), a change known to be caused by uninhibited neutrophil elastase. We conclude that lung elastin is the most likely source of the increased DES and IDES in CF.

Topics: Adult; Amino Acids; Bronchoalveolar Lavage Fluid; Case-Control Studies; Collagen; Cystic Fibrosis; Desmosine; Elastin; Female; Humans; Isodesmosine; Leukocyte Elastase; Male; Neutrophils; Pancreatic Elastase; Receptors, Complement

One of the most rapid changes in collagen and elastin content of a tissue occurs in the uterus following postpartum involution. We measured the urinary excretion of specific amino acid markers for mature elastin (desmosine [DES] and isodesmosine [IDES]) and fibrillar collagen (hydroxylysyl pyridinoline [HP] and lysyl pyridinoline [LP]) before and after parturition in three gravid subjects. For that purpose, we used an isotope dilution method coupled with gel filtration and HPLC. The highest DES values were found 2-5 weeks postpartum and were 18-45 micrograms/g creatinine or two to six times those found for healthy neversmoking nongravid females (7.7 +/- 0.3 micrograms/g creatinine, mean +/- SE). The highest levels of urinary HP for each subject were found 2-3 weeks after parturition and were 115-607 nmol/mmol creatinine or 4-21 times those found for healthy neversmoking nongravid females (28.1 +/- 1.3 nmol/mmol creatinine). For the gravid subjects as a group and also for each subject, the mean values for urinary DES, IDES, HP, and HP/LP during the first 6 weeks postpartum were significantly greater than the mean baseline values beginning 27 weeks postpartum. For the gravid subjects as a group, the mean value for urinary HP/LP during the first 6 weeks postpartum was significantly greater than the value during the 20 weeks preceding parturition. This suggested that the tissue(s) of origin of the excess HP, during the 6 weeks following parturition, was not bony and was consistent with a uterine origin.

Topics: Amino Acids; Collagen; Creatinine; Desmosine; Elastin; Female; Humans; Kinetics; Postpartum Period; Pregnancy; Reference Values; Smoking; Uterus

It has been hypothesized that emphysema results from damage to the elastic fiber network of the lungs as a result of elastase-antielastase imbalance. We used a new assay for urinary desmosine (DES) and isodesmosine (IDES), specific markers for the degradation of mature crosslinked elastin, and hydroxylysylpyridinoline (HP) and lysylpyridinoline (LP), specific markers for the degradation of mature crosslinked collagen, in order to examine elastin and collagen degradation in relation to current cigarette smoking and the presence of chronic obstructive pulmonary disease (COPD). The study sample consisted of 22 never-smokers (NSM group), 13 current smokers without airflow obstruction (SM group), and 21 patients with COPD (COPD group), including both current and former smokers. The relation between the creatinine-height index and FEV1 was used to correct for possible loss of muscle mass and decreased excretion of creatinine in the COPD group. Mean urinary excretion of elastin-derived crosslinks in the COPD group (DES, 11.8 +/- 5.1 [mean +/- SD]; IDES, 11.3 +/- 5.0 micrograms/g creatinine) and in the SM group (DES, 11.0 +/- 4.2; IDES, 10.2 +/- 2.5 micrograms/g creatinine) was significantly higher than in the NSM group (DES, 7.5 +/- 1.4; IDES, 6.9 +/- 1.3 micrograms/g creatinine). In multivariate analysis, current smoking and the presence of COPD were significantly and independently associated with higher urinary excretion of elastin degradation products, and there was no significant interaction between current smoking and the presence of COPD.(ABSTRACT TRUNCATED AT 250 WORDS)

Topics: Adult; Amino Acids; Biomarkers; Collagen; Desmosine; Elastin; Female; Humans; Isodesmosine; Lung Diseases, Obstructive; Male; Middle Aged; Prospective Studies; Smoking

To measure the urinary excretion of specific cross-link amino acid markers for mature elastin (desmosine [DES] and isodesmosine [IDES]) and fibrillar collagen (hydroxylysylpyridinoline [HP] and lysylpyridinoline [LP]) in systemic sclerosis (SSc) patients and healthy controls.. Urine specimens from 20 patients with SSc and 22 controls were assessed for DES, IDES, HP, and LP using high performance liquid chromatography and ultraviolet absorption spectroscopy, in combination with an isotope dilution technique in which the urine specimen was spiked with isotopically labeled cross-link amino acids.. Mean +/- SD levels of urinary DES and IDES were elevated in SSc patients by 2-3-fold, and urinary HP and LP by 3-4-fold, compared with controls (DES 21.0 +/- 9.4 versus 7.5 +/- 1.4 micrograms/gm creatinine; HP 109.0 +/- 72.9 versus 24.9 +/- 5.7 nmoles/mmole creatinine). Nineteen of the 20 SSc patients had urinary DES and HP values that were > 3 SD above the control mean. A significant elevation in the HP:LP ratio in SSc patients as compared with controls (mean +/- SD 6.9 +/- 1.5 versus 5.5 +/- 1.3) indicated a soft tissue origin for much of the increased HP.. Patients with SSc have higher levels of urinary cross-link amino acids specific for the degradation of mature collagen and elastin. These markers distinguish most SSc patients from healthy controls.

Topics: Adult; Aged; Amino Acids; Collagen; Desmosine; Elastin; Female; Humans; Isodesmosine; Male; Middle Aged; Reference Values; Scleroderma, Systemic