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Procollagen-III peptide serum levels in Paget's disease of the bone.

A commercially available radioimmunoassay kit was used to determine aminoterminal procollagen-III peptide (pNcoll III) serum levels in patients with Paget's disease of the bone and control subjects. In patients with Paget's disease pNcoll III concentrations were significantly elevated. They decreased to varying degrees under chronic therapy with human and salmon calcitonin, disodium ethane 1-hydroxy 1,1-diphosphonate (EHDP), or a combination therapy of EHDP and human calcitonin. The results were compared with the effect on traditional biochemical markers of disease activity: serum alkaline phosphatase and urinary hydroxyproline excretion, both of which reacted more acutely to the various therapies than pNcoll III, although pretreatment correlations were close. The most probable source of pNcoll III is not the Pagetic bone per se, but the vascular, fibrous connective tissue replacing normal bone marrow.

Alkaline Phosphatase↗

Serum levels of carboxyterminal propeptide of type I procollagen in healthy children from 1st year of life to adulthood and in metabolic bone diseases.

Type I collagen is the major component of bone matrix; circulating carboxyterminal propeptide of type I procollagen (P-I-CP) levels reflect type I collagen synthesis in tissues and may be an useful index to investigate bone metabolism. We measured P-I-CP by a new radioimmunoassay in 300 healthy children and adolescents and in 40 healthy adults to provide reference data for P-I-CP values. In addition, 79 patients with diagnosed disorders of phospho-calcium metabolism (rickets, vitamin D deficient and vitamin D resistant, hyperparathyroidism, hypo- and pseudo-hypoparathyroidism, osteopenia) were evaluated. In the healthy subjects, serum P-I-CP values were higher in children than in adults; variations of P-I-CP levels were observed according to age and sexual maturation: higher values were found in the first years of life and during pubertal development; pubertal increase reflects the different timing of pubertal development in the two sexes. P-I-CP levels were increased in primary hyperparathyroidism and reduced in diseases related to impaired secretion or action of parathyroid hormone. Higher P-I-CP levels were found in vitamin D deficient and vitamin D resistant rickets. P-I-CP was reduced in anorexia nervosa and during chronic glucocorticoid treatment while it was increased in thyrotoxic osteoporosis. In idiopathic juvenile osteoporosis, P-I-CP values ranged from reduced to increased values. We conclude that P-I-CP may represent an additional biochemical marker of bone metabolism. Since age-related variations are present, reference data for the various ages are need for clinical application of this assay.

Adolescent↗

Interferon-alpha 2a increases serum concentration of hyaluronic acid and type III procollagen aminoterminal propeptide in patients with chronic hepatitis B virus infection.

Interferon-alpha (IFN-alpha) has become an important drug for the treatment of chronic viral liver diseases. However, the action of IFN-alpha remains unclear. We investigated whether human recombinant IFN-alpha modulates serum concentrations of hyaluronic acid (HA) and type III procollagen aminoterminal propeptide (P-III-NP) in 56 patients with chronic hepatitis-B under IFN-alpha therapy. IFN-alpha increased the HA serum level in 44 of 46 patients and, after cessation of treatment, HA serum levels returned to the pretherapy levels. The increase of HA serum level was higher in patients with active cirrhosis (aC) than in patients with chronic persistent hepatitis (CPH) and in patients with severe inflammation compared to those with moderate inflammation. Interestingly, HA serum concentration was unrelated to IFN dose and was of no predictive value for therapy response. In contrast, IFN-alpha increased significantly the P-III-NP serum level in patients with aC only. During follow-up, P-III-NP serum level decreased late in responders in parallel to the decrease of serum level of liver enzymes, in non-responders it was without significant change. The first dose of IFN induced a significant increase in HA serum level in each of 10 patients but in none of four healthy volunteers. In contrast, P-III-NP serum concentrations were not influenced by the first IFN-alpha dose. We conclude that: (1) immunstimulation with IFN-alpha induces a rapid increase of HA serum level in patients with chronic hepatitis B but not in normal persons; (2) IFN-alpha increases P-III-NP serum level only in patients with active liver cirrhosis; (3) measurement of HA and P-III-NP serum levels does not help predict response to IFN-alpha, and (4) HA serum level may be used as a compliance indicator.

Biopsy↗

Deletion of a Gly-Pro-Pro repeat in the pro alpha2(I) chain of procollagen I in a family with dominant osteogenesis imperfecta type IV.

We have investigated one member of a family with dominant osteogenesis imperfecta type IV through three generations. In protein-chemical studies of cultured fibroblasts derived from the proband, collagen I was overmodified, with normal processing of procollagen I, normal thermal stability, and a cyanogen bromide peptide map that suggested a C-terminal location of the structural abnormality in the collagen triple helix. Sequencing of the gene encoding the alpha2(I) chain of collagen I (COL1A2) indicated a nine base-pair deletion of nucleotides 3418-3426. When a polymerase chain reaction product containing the nucleotides in question was electrophoresed in a 12% polyacrylamide gel, two bands with a difference in size of nine base pairs could be shown. Sequencing of the molecular weight band confirmed the deletion of the nine base pairs involving codons 1003-1006 of COL1A2. The deletion introduced a SfiI restriction site that was used for confirmation of the deletion in genomic DNA from the proband. The deletion resulted in the removal of three amino acids (Gly-Pro-Pro), but this did not disrupt the Gly-X-Y sequence of the collagen triple helix, as is often the case in the more common glycine substitutions. We discuss the ways in which this deletion could result in osteogenesis imperfecta.

Adult↗

Serum N terminal procollagen III fragment: a predictive marker of joint destruction in rheumatoid arthritis?

The level of the N terminal fragment of procollagen III (P3NP) levels of 100 consecutive patients affected by rheumatoid arthritis (RA) were evaluated in comparison with disease activity markers (erythrocytes sedimentation rate, C reactive protein), immunological status (rheumatoid factors, immune complexes) and joint destruction (assessed according to the Steinbrocker index). P3NP levels showed no significant relationship either to disease activity or to immunological status; however, a strong association was fond between X-ray grade and P3NP values. A two-year retrospective study of 32 patients belonging to the original 100 patient population allowed us to assign a predictive value for joint destruction to the P3NP level in the early stage of the disease.

Adult↗

Serum aminoterminal type III procollagen peptide in inflammatory and degenerative rheumatic disorders.

Measurement of the aminoterminal type III procollagen peptide in serum has been suggested as a marker of the biosynthesis of collagen type III, a major connective tissue component in repair processes. In the present study the propeptide level correlated with the inflammatory synovial mass in rheumatoid arthritis and osteoarthritis. This implies that the propeptide level reflects the collagen type III synthesis occurring in the synovial repair processes, whether they were caused by inflammatory or degenerative rheumatic disorders. Physical activity did not enhance the transition of the propeptide from the synovial fluid or the inflamed synovial membrane to the blood. Normal serum propeptide values were observed in most patients with ankylosing spondylitis and degenerative diseases of the spine. This may reflect the lower amount of inflammatory tissue in these diseases and hence the sensitivity of the assays.

Adult↗

Clinical studies of type-I procollagen carboxyterminal peptide in serum of patients with gastric cancer: comparison with CEA and CA19-9.

The serum level of a newly developed monoclonal antibody against type-I procollagen carboxyterminal peptide (P-1-P) was determined in patients with gastric cancer. The location of P-1-P in gastric cancer tissue was also investigated. We found that: (1) The serum P-1-P level and the positivity rate in patients with gastric cancer were similar to those in patients with other malignant or benign GI diseases and healthy individuals. (2) In patients with gastric cancer, the P-1-P positivity rate was significantly lower than that of CEA or CA19-9. (3) In patients with gastric cancer, the P-1-P positivity rate increased as the disease stage advanced. (4) Among patients with gastric cancer, the P-1-P positivity rate was significantly higher in those with scirrhous type than in those with medullary or intermediate type. (5) P-1-P was detected in the cytoplasm of cancer cells. P-1-P staining was stronger in scirrhous type and histologically undifferentiated gastric cancer. These results show that P-1-P can serve as a good marker for scirrhous type gastric cancer. The production of collagen by cancer cells themselves seems to be involved in collagen production in scirrhous type gastric cancer.

Adenocarcinoma, Scirrhous↗

Serum type IV collagen in various liver diseases in comparison with serum 7S collagen, laminin, and type III procollagen peptide.

The clinical significance of the immunoreactive triple helical domain of type IV collagen in serum was evaluated in 73 healthy controls and 161 patients with various biopsy-proven liver diseases. Although serum levels of type III procollagen peptide were increased in all liver diseases, those of type IV collagen, 7S collagen, and laminin were principally increased in chronic liver diseases associated with hepatic fibrogenesis/fibrosis. In both non-alcoholic and alcoholic liver diseases, 7S collagen was increased in serum, while type IV collagen and laminin in serum were particularly increased in alcoholic liver diseases and in hepatocellular carcinoma, in which latter the sensitivity was greater for type IV collagen than for laminin. Gel filtration analysis in Sephacryl S-400 revealed type IV collagen in serum to be a single molecular form with a molecular weight that correspond to type IV collagen, whereas 7S collagen was recognized as several heterogeneous macromolecules. These findings indicate that serum type IV collagen is derived from the type IV protocollagen pool, and is a sensitive marker for the fibrogenetic process in hepatic basement membranes.

Adult↗

Serum level of carboxyterminal propeptide of type I procollagen in haemodialysis patients.

Serum concentrations of carboxyterminal propeptide of type I procollagen (PICP) and carboxyterminal parathyroid hormone (C-PTH) were measured in 33 haemodialysis patients. Serum levels of PICP and C-PTH in the haemodialysis group were significantly higher than those in the controls. Although there was no correlation between serum C-PTH and creatinine, a positive correlation was found between PICP and creatinine. This indicates that the measurement of PICP in serum is useful for monitoring metabolic bone diseases in haemodialysis patients.

Adult↗

Post-traumatic dynamic change of carboxyterminal propeptide of type I procollagen, alkaline phosphatase and its isoenzymes as predictors for enhanced osteogenesis in patients with severe head injury.

Patients suffering from severe head injury and fractures of long bones or large joints often show enhanced osteogenesis, with hypertrophic callus formation and/or heterotopic ossifications. The advantage of this phenomenon is early consolidation of the fractures. An extreme disadvantage is extensive periarticular calcification, resulting in complete ankylosis of the affected joint. In spite of numerous efforts aimed at clarifying the way in which severe head injury can influence osteogenesis at a distant site, this phenomenon is still not understood. The process, once started, seems irreversible, but if diagnosed in time, could be prevented with non-steroid anti-inflammatory drugs that inhibit development of heterotopic ossifications. The major prerequisite for testing this possibility is to define parameters of an early diagnosis of enhanced osteogenesis. Thus, the aim of this study was to test whether serum values of some parameters related to bone regeneration could allow an early prediction of enhanced ossification following bone fracture in patients with severe head injury. Samples of sera were obtained from three groups of injured patients: fractures of long bones or large joints only (n = 6), severe head injury only (n = 8), severe head injury and fractures of long bones and large joints (n = 7) and from a group of apparently healthy volunteers (n = 10). The values for alkaline phosphatase (ALP), the bone isoenzyme, and the carboxy terminal propeptide of type I procollagen (PICP) were significantly higher (5-20 times as high) in patients with severe head injury and bone or joint fractures than in any other group. Significantly increased concentrations of PICP were already found in the 1st week after injury, and those of ALP and of the bone isoenzyme increased during the 2nd week after injury. Results show that these parameters are helpful for an early diagnosis of enhanced osteogenesis and heterotopic ossifications in patients with severe head injury and bone fractures. Further studies are necessary to verify these findings, while analysis of reasons for the specific patterns of dynamic change of these parameters could lead to better understanding of the mechanisms underlying the uncontrolled bone formation.

Adolescent↗

Serum levels of aminoterminal type III procollagen peptide in normal subjects and hepatic fibrosis.

Serum levels of aminoterminal type III procollagen peptide in normal subjects were determined by radioimmunoassay. The levels of the peptide markedly increase in infants and gradually decrease with age in childhood. The peptide increases again in prepubertal children and then decreases through adulthood. These findings suggest that increased levels of the peptide reflect accelerated collagen synthesis in the human body. Comparison of serum levels of the peptide with the degree of hepatic fibrosis revealed that the peptide increased in parallel with the progress of hepatic fibrosis.

Adolescent↗

Cryptogenic chronic active liver disease. Evaluation of serum aminoterminal peptide of type III procollagen as a marker of histological activity.

Sera from 64 patients with HBsAg-negative chronic liver disease with or without cirrhosis were investigated for aminoterminal peptide of type III procollagen (sP-III-P) as a suitable marker of hepatic fibrosis; 244 healthy control subjects were included in the study. A close correlation (p less than 0.01) between sP-III-P levels and histological activity was observed; on the contrary, no correlation was found between the same serum marker of liver fibroplasia and biochemical activity or clinical severity of the disease. We conclude that sP-III-P as a suitable marker of liver overload of collagen fibers is strongly correlated with the histological activity of the disease. Local immune reactions produce soluble substances that might stimulate fibroblastic activity. The test has a significant sensitivity and a very high specificity as a marker of chronic liver disease with histological activity.

Adult↗

A comparative study of prostate specific antigen (PSA), C-terminal propeptide of blood type I procollagen (PICP) and urine type I collagen-crosslinked N telopeptide (NTx) levels using bone scintigraphy in prostate cancer patients.

We compared the ability to diagnose skeletal metastasis between serum prostate specific antigen (PSA), C-terminal propeptide of blood type I procollagen (PICP), and urine type I collagen-crosslinked N telopeptide (NTx) in prostate cancer patients. In sixty-nine patients with prostate cancer, bone scintigraphy was performed, and serum PSA and PICP and urine NTx were measured. The median level of serum PSA in the osseous metastasis-negative group (n = 33) was 0.80 ng/ml being significantly lower as compared to the osseous metastasis-positive group (n = 36, 7.70 ng/ml) (p < 0.0001). The serum PICP and urine NTx/Cr levels appeared lower in the osseous metastasis-negative group than the osseous metastasis-positive group, but there was no significant difference. Logistic regression analysis showed that ability to diagnose skeletal metastasis of serum PSA was 68.1% and superior to those of serum PICP (56.5%) and urine NTx/Cr (53.6%). Serum PSA improved the ability to diagnose skeletal metastasis when combined with serum PICP or urine NTx/Cr. When patients were grouped according to the extent of disease grade (EOD grade) nomenclature, Spearman's correlation coefficient by rank showed that serum PSA was most significantly correlated with EOD grade (p < 0.0001). In 14 patients whose skeletal metastases progressed or regressed, the change of serum PSA more clearly separated the osseous metastasis-regression group and osseous metastasis-progression group than did serum PICP and urine NTx/Cr. Serum PSA was more reliable than bone resorption and formation markers produced by crosslinking of type I collagen.

Aged↗

Procollagen-type III peptide serum concentrations in alcoholic and non-alcoholic liver disease.

The type III procollagen aminopeptide (sPIIIP) serum levels were measured in 197 patients with liver disease and were correlated with morphological and serological alterations and with alcohol drinking habits. The sPIIIP levels resulted significantly increased in 51% of 43 patients with untreated chronic active hepatitis (CAH), in 61% of 36 patients with CAH plus cirrhosis, in 69% of 26 patients with inactive cirrhosis, in 4 out of 8 patients with alcoholic steatosis and fibrillogenesis, but remained unchanged in 38 cases of alcoholic steatosis plus siderosis and in 13 cases of chronic persistent hepatitis. A correlation between sPIIIP levels and the histological pattern of fibrosis could not be demonstrated in a single type of fibrotic liver disease and no differences were found between alcoholic and non-alcoholic patients. We agree upon the opinion that high sPIIIP levels may identify liver fibrogenic activity, but this test needs further technical improvements before it could be widely used in the clinical practice.

Adult↗

Predictive role of serum procollagen III peptide and Knodell's index in survival prognosis of patients with hepatitis B virus liver cirrhosis.

OBJECTIVE: The objective of the study was to improve the accuracy of survival prognosis in patients with liver cirrhosis using procollagen III peptide (PIIIP), as a marker of inflammation and fibrogenesis, and Knodell's histologic activity index (KI) in addition to previously used prognostic factors. PATIENTS AND METHODS: Five-year survival was followed in a group of 75 patients with hepatitis B virus (HBV) liver cirrhosis (patients testing anti-HBe positive and HBV-DNA negative). There were 31 patients with compensated cirrhosis and 44 with decompensated cirrhosis. The diagnostic procedure included clinical, laboratory, ultrasound and pathohistologic examination. We combined PIIIP and KI with other significant variables to achieve the highest possible sensitivity, specificity and accuracy for survival prognosis in HBV liver cirrhosis. The models were compared using ROC analysis. RESULTS: At the end of the five-year period of survival follow-up, there were 39 survivors and 36 patients had died (only three died from an extrahepatic cause). In the quantitative model, the discriminant canonical function (DCF) identified PIIIP, bilirubin, prothrombin time, ascites and KI as statistically significant parameters in the prognosis of five-year survival. Calculation of the score based on DCF yielded an accuracy of 89.3%. In the semiquantitative model, the analysis of variance identified PIIIP, bilirubin, albumin, pro-thrombin time, alkaline phosphatase, ascites and KI as significant variables. When PIIIP was added to the clinicohistologic diagnosis, Child-Pugh score and KI, the level of accuracy improved by 12% (from 78% to 90%), 11% (from 79% to 90%) and 10.6% (from 80% to 90.6%), respectively. When calculated with the three biochemical parameters (alkaline phosphatase, PIIIP and bilirubin) and KI identified by DCF, the accuracy was 90.6%. CONCLUSION: Combining PIIIP and KI with other prognostic parameters is useful in achieving a better precision of survival prognosis in patients with HBV liver cirrhosis.

Adult↗

Long-term monitoring of rec-GH treatment by serial determination of serum aminoterminal propeptide of type III procollagen in children and adults with GH deficiency.

Serum aminoterminal propeptide of type III procollagen (PIIINP) levels, a reliable marker of collagen formation, were evaluated in children (C=7) and adults with childhood-onset (CO=10) and acquired (A=18) GH deficiency (GHD) before, during and after withdrawal of rec-GH therapy (C=0.6 IU/kg/week, CO=0.5 IU/kg/week, A=0.25 IU/kg/week). The duration of treatment was 12 months for C and A and 6 months for CO; investigations were carried out before and at 3, 6, 9 and 12 months (for C and A) and at 3 and 6 months (for CO) of GH treatment and 6 months after the withdrawal of therapy (for A and CO). Data obtained from patients were compared with those recorded in two age- and sex-matched control groups. Before treatment, serum PIIINP levels were significantly lower (p<0.001) in C with GHD (mean+/-SE: 2.9+/-0.4 ng/ml) than in controls (6.1+/-0.4 ng/ml), while no significant differences were recorded between adults with CO/A-GHD (3.7+/-0.5 ng/ml and 3.4+/-0.2 ng/ml) and controls (3.2+/-0.2 ng/ml). GH treatment caused a significant increase (p<0.0001) of PIIINP levels both in C (3rd month: 4.4+/-0.2 ng/ml, 6th month: 5.1+/-0.4 ng/ml, 12th month: 5.1+/-0.5 ng/ml), CO-GHD (3rd month: 12.7+/-1.2 ng/ml; 6th month: 10.2+/-0.6 ng/ml) and A-GHD (3rd month: 10.0+/-1.0 ng/ml; 6th month: 8.4+/-0.6 ng/ml; 12th month: 7.0+/-0.7 ng/ml), the increase being dose-dependent (more marked and sustained in adults with CO-GHD). The maximal stimulation of collagen synthesis occurred after 3 months of GH treatment in adults with GHD, while a more gradual and less relevant increase was observed in C with GHD. Six months after the withdrawal of GH therapy, serum PIIINP levels of adults with CO-GHD (3.6+/-0.3 ng/ml) were similar to those recorded before treatment, while in adults with A-GHD serum PIIINP levels (2.6+/-0.2 ng/ml) were significantly lower (p<0.01) than in basal condition. In conclusion, our study shows that: a) GHD is associated with a reduction of soft tissue formation in children, while it seems to exert no relevant effects in adults with GHD; b) GH therapy causes a rapid stimulation of collagen turnover, which shows a different pattern in children and adults; c) the GH-induced stimulation of collagen synthesis is rapidly removed after the withdrawal of GH treatment. For these reasons, the determination of peripheral markers of GH effects appears useful for the monitoring of GH therapy and can contribute to assess the "tailored" substitutive dose for the individual patient.

Adolescent↗

1,25 dihydroxyvitamin-D3 attenuates the confluence-dependent differences in the osteoblast characteristic proteins alkaline phosphatase, procollagen I peptide, and osteocalcin.

In the present study a cell culture model of primary human osteoblasts based on degrees of confluence was investigated by measuring basal and 1,25(OH)2D3stimulated levels of the osteoblast characteristic proteins alkaline phosphatase (AP), procollagen I-peptide (PICP), and osteocalcin (OC), as well as the corresponding gene expression. Primary osteoblast-like cell cultures from seven donors were treated in the second passage with 1,25(OH)2D3 (5 x 10(-8) M for 48 hours) and investigated at four stages of confluence (stage I 50%, stage II 75%, stage III 100%, and stage IV 7 days postconfluence). In untreated cultures passing through the different stages of confluence, we saw a 1.8-fold increase of AP activity, a 2. 3-fold increase of OC secretion, but a decrease of PICP levels to 0. 36-fold. Gene expression showed only minor variation between the different confluence stages. 1,25(OH)2D3 did not significantly affect PICP production. Alkaline phosphatase protein was stimulated during proliferation until confluence, with no effect thereafter. Surprisingly, OC secretion and mRNA expression were stimulated in all four stages to the same absolute level independent of basal values. We conclude that our results correspond to other studies showing differentiation-stage dependent changes of basal levels of osteoblast-specific proteins. However, 1,25(OH)2D3 stimulation decreased the confluence-dependent difference for AP and abolished it for osteocalcin, thus leading to a more differentiated phenotype of the osteoblast. Therefore, 1,25(OH)2D3 stimulation might improve the reproducibility of results obtained at different confluence stages from cultures of clinical samples.

Alkaline Phosphatase↗

Serum tartrate-resistant acid phosphatase 5b in monitoring bisphosphonate treatment with clodronate: a comparison with urinary N-terminal telopeptide of type I collagen and serum type I procollagen amino-terminal propeptide.

Osteoclastic tartrate-resistant acid phosphatase activity in serum (S-TRACP 5b) was measured in postmenopausal women ( n =59, mean age 56.1 years) with vertebral osteopenia before and during 2-year treatment with an 800-mg daily dose of clodronate, with a non-amino bisphosphonate. Changes in TRACP 5b were compared with those in urinary excretion of type I collagen amino-terminal telopeptide (U-NTX), corrected for creatinine excretion, a well-established marker of bone resorption, and to serum type I procollagen amino-terminal propeptide (S-PINP), a marker of bone formation. Marker changes 1 year after start of treatment were correlated with changes in bone mineral density (BMD). The least significant change (LSC) for each marker and BMD was calculated from values for subjects receiving placebo. Responders to treatment were those exhibiting a change larger than LSC. In response to clodronate treatment S-TRACP 5b (mean change up to -18%) decreased less than did U-NTX (up to -51%) or S-PINP (up to -46%). Marker changes correlated with changes in lumbar spine and trochanter BMD. The most efficient marker for finding responders to treatment was S-PINP, which changed more than the LSC (32%) in 72% of the subjects at the 1-year time point and in 79% at the 2-year time point. S-TRACP 5b change exceeded the LSC (27%) in 40% and 34% of the subjects at each time point, while U-NTX change exceeded the LSC (55%) in 55% and 40%, respectively. We conclude that, in terms of the proportion of subjects exhibiting any change exceeding the LSC, S-TRACP 5b did not appear to be superior to U-NTX and S-PINP in the follow-up of clodronate treatment. The reason may lie in the mechanism of action of clodronate, which rather than reducing the number of TRACP 5b-secreting osteoclasts, reduces the activity of bone proteolytic enzymes and thus the rate of bone organic matrix degradation. This is seen in decreased amounts of type I collagen breakdown products (U-NTX), and through coupling of bone resorption with bone formation, in a decrease in circulating levels of the marker that reflects new collagen formation (S-PINP).

Acid Phosphatase↗