Purification of human procollagen type I carboxyl-terminal propeptide cleaved as in vivo from procollagen and used to calibrate a radioimmunoassay of the propeptide

1994 ◽  
Vol 40 (5) ◽  
pp. 811-816 ◽  
Author(s):  
B J Pedersen ◽  
M Bonde
Keyword(s):  
Geometric Mean ◽  
Sodium Dodecyl ◽  
Gel Filtration ◽  
Type I ◽  
Procollagen Type ◽  
Amino Terminal ◽  
Fetal Fibroblasts ◽  
Procollagen Type I ◽  
Carboxyl Terminal

Abstract We purified human procollagen type I carboxyl-terminal propeptide (PICP) that had been cleaved as in vivo from procollagen. PICP in serum-free medium from cultured human fetal fibroblasts was purified by thiophilic adsorption chromatography, low-pressure gel filtration, and HPLC gel filtration. The purity and homogeneity of the protein was verified by sodium dodecyl sulfate-polyacrylamide gel electrophoresis. Amino-terminal amino acid sequencing showed that the sequences of the alpha 1 and alpha 2 chains of this PICP were identical to those of the PICP produced in vivo. The monocomponent PICP thus purified was used as calibrator in a simple equilibrium-type RIA of PICP with polyclonal antibodies raised in rabbits. The measuring range is 0.15-3.75 nmol/L, and the assay detection limit is 0.03 nmol/L. The within-run and total CVs are 2% and 4%, respectively. The reference interval for the plasma concentration of PICP in healthy women of ages > 30 years is 0.36-1.44 nmol/L (geometric mean 0.72 nmol/L, n = 154).

scholarly journals Procollagen Type I Amino-Terminal Propeptide: Pediatric Reference Data and Relationship with Procollagen Type I Carboxyl-Terminal Propeptide

2004 ◽  
Vol 50 (11) ◽  
pp. 2173-2176 ◽  
Author(s):  
Patricia M Crofton ◽  
Nancy Evans ◽  
Mervyn R H Taylor ◽  
Celia V Holland
Keyword(s):  
Reference Data ◽  
Type I ◽  
Procollagen Type ◽  
Amino Terminal ◽  
Procollagen Type I ◽  
Carboxyl Terminal

scholarly journals Longitudinal in Vivo Analysis of the Region-Specific Efficacy of Parathyroid Hormone in a Rat Cortical Defect Model

Endocrinology ◽  
2008 ◽  
Vol 150 (4) ◽  
pp. 1570-1579 ◽  
Author(s):  
David E. Komatsu ◽  
Kellie A. Brune ◽  
Hong Liu ◽  
Allen L. Schmidt ◽  
Bomie Han ◽  
...  
Keyword(s):  
Computed Tomography ◽  
Bone Healing ◽  
Quantitative Computed Tomography ◽  
Type I ◽  
Defect Model ◽  
Procollagen Type ◽  
Cortical Defect ◽  
Defect Sites ◽  
Procollagen Type I

PTH has been shown to enhance fracture repair; however, exactly when and where PTH acts in this process remains to be elucidated. Therefore, we conducted a longitudinal, region-specific analysis of bone regeneration in mature, osteopenic rats using a cortical defect model. Six-month-old rats were ovariectomized, and allowed to lose bone for 2 months, before being subjected to bilateral 2-mm circular defects in their femoral diaphyses. They were then treated for 5 wk with hPTH1–38 at doses of 0, 3, 10, or 30 μg/kg · d and scanned weekly by in vivo quantitative computed tomography. Quantitative computed tomography analyses showed temporal, dose-dependent increases in mineralization in the defects, intramedullary (IM) spaces, and whole diaphyses at the defect sites. Histomorphometry confirmed PTH stimulation of primarily woven bone in the defects and IM spaces, but not the periosteum. After necropsy, biomechanical testing identified an increase in strength at the highest PTH dose. Serum procollagen type I N-terminal propeptide concentration showed a transient increase due to drilling, but procollagen type I N-terminal propeptide also increased with PTH treatment, whereas tartrate-resistant acid phosphatase unexpectedly decreased. Analyses of lumber vertebra confirmed systemic efficacy of PTH at a nonfracture site. In summary, PTH dose dependently induced new bone formation within defects, at endocortical surfaces, and in IM spaces, resulting in faster and greater bone healing, as well as efficacy at other skeletal sites. The effects of PTH were kinetic, region specific, and most apparent at high doses that may not be entirely clinically relevant; therefore, clinical studies are necessary to clarify the therapeutic utility of PTH in bone healing.

scholarly journals Effect of Active Acromegaly and Its Treatment on Parathyroid Circadian Rhythmicity and Parathyroid Target-Organ Sensitivity

2006 ◽  
Vol 91 (3) ◽  
pp. 913-919 ◽  
Author(s):  
H. D. White ◽  
A. M. Ahmad ◽  
B. H. Durham ◽  
S. Chandran ◽  
A. Patwala ◽  
...  
Keyword(s):  
Reference Range ◽  
Target Organ ◽  
Oral Glucose ◽  
Type I ◽  
Procollagen Type ◽  
Amino Terminal ◽  
Igf I ◽  
Procollagen Type I ◽  
Biochemical Cure ◽  
Active Acromegaly

Abstract Context: Patients with active acromegaly have increased bone turnover and skeletal abnormalities. Biochemical cure of acromegaly may represent a functional GH-deficient state and result in cortical bone loss. Reduced PTH target-organ sensitivity occurs in adult GH deficiency and may underlie the associated development of osteoporosis. Objective: We examined the effect of active and treated acromegaly on PTH concentration and target-organ sensitivity. Patients: Ten active acromegalic subjects (GH nadir > 0.3 μg/liter after 75-g oral glucose load and IGF-I above age-related reference range) and 10 matched controls participated in the study. Design: Half-hourly blood and 3-h urine samples were collected on patients and controls for 24 h. Samples were analyzed for PTH, calcium (Ca), nephrogenous cAMP (NcAMP, a marker of PTH renal activity), β C-telopeptide (bone resorption marker), and procollagen type-I amino-terminal propeptide (bone formation marker). Serum calcium was adjusted for albumin (ACa). Eight acromegalic subjects who achieved biochemical cure (GH nadir < 0.3 μg/liter after 75-g oral glucose load and IGF-I within reference range) after standard surgical and/or medical treatment reattended and the protocol repeated. Results: Active acromegalic subjects had higher 24-h mean PTH, NcAMP, ACa, urine Ca, β C-telopeptide, and procollagen type I amino-terminal propeptide (P < 0.05), compared with controls. Twenty-four-hour mean PTH increased (P < 0.001) in the acromegalic subjects after treatment, whereas NcAMP and ACa decreased (P < 0.05). Conclusion: Increased bone turnover associated with active acromegaly may result from increased PTH concentration and action. Biochemical cure of acromegaly results in reduced PTH target-organ sensitivity indicated by increased PTH with decreased NcAMP and ACa concentrations. PTH target-organ sensitivity does not appear to return to normal after successful treatment of acromegaly in the short term and may reflect functional GH deficiency.

Bone Regeneration and Neovascularization Processes in a Pellet Culture System for Periosteal Cells

2009 ◽  
Vol 18 (4) ◽  
pp. 443-452 ◽  
Author(s):  
Mari Akiyama ◽  
Masaaki Nakamura
Keyword(s):  
Ascorbic Acid ◽  
Bone Regeneration ◽  
Nude Mice ◽  
Vascular Endothelial Cells ◽  
Type I ◽  
Vascular Endothelial ◽  
Procollagen Type ◽  
Procollagen Type I ◽  
Periosteal Cells

Reliable bone regeneration can be achieved with a pellet culture system using bovine periosteal cells. However, bone regeneration and neovascularization processes in this system have remained unclear. The present study aimed to clarify the extracellular environment and neovascularization process. To detect components of the extracellular matrix secreted by cells and to identify the conditions necessary for bone regeneration in the body, Western blotting and in vivo tests in nude mice were performed. Cells were cultured with or without ascorbic acid and culture supernatant was precipitated. Western blotting showed that culture supernatant contained collagen type I, procollagen type I, and procollagen type I C-terminus when cells were cultured with ascorbic acid. Cells cultured with ascorbic acid formed partial bony tissues at 2 weeks after grafting to nude mice, while bone formation was missing without ascorbic acid. Immunostaining was performed using species-specific vascular endothelial cell markers to ascertain whether vascular endothelial cells were bovine or murine (nude mouse). Immunohistological methods showed vascular endothelial cells in osseous tissue formed in the subcutaneous tissue of nude mice were murine. Extracellular matrix synthesis in vitro and host blood flow in vivo are essential for bone regeneration.

scholarly journals A Monoclonal Antibody Specific for the Amino Terminal Cleavage Site of Procollagen Type I

1983 ◽  
Vol 134 (1) ◽  
pp. 183-189 ◽  
Author(s):  
Harald G. FOELLMER ◽  
Kazuko KAWAHARA ◽  
Joseph A. MADRI ◽  
Heinz FURTHMAYR ◽  
Rupert TIMPL ◽  
...  
Keyword(s):  
Monoclonal Antibody ◽  
Cleavage Site ◽  
Type I ◽  
Procollagen Type ◽  
Amino Terminal ◽  
Procollagen Type I

scholarly journals Serum carboxyl-terminal propeptide of procollagen type I in exercise-induced left ventricular hypertrophy

2004 ◽  
Vol 27 (8) ◽  
pp. 471-474 ◽  
Author(s):  
Eduardo M. Escudero ◽  
Ana L. Tufare ◽  
Oscar Rebolledo ◽  
Laura Pellegrini ◽  
Carlos Lobrutto
Keyword(s):  
Left Ventricular Hypertrophy ◽  
Ventricular Hypertrophy ◽  
Left Ventricular ◽  
Type I ◽  
Procollagen Type ◽  
Procollagen Type I ◽  
Carboxyl Terminal ◽  
Exercise Induced

scholarly journals Serum concentrations of carboxyl-terminal propeptide of type I procollagen, amino-terminal propeptide of type III procollagen, cross-linked carboxyl-terminal telopeptide of type I collagen, and their interrelationships in schoolchildren

1997 ◽  
Vol 43 (9) ◽  
pp. 1577-1581 ◽  
Author(s):  
Patricia M Crofton ◽  
Jean C Wade ◽  
Mervyn R H Taylor ◽  
Celia V Holland
Keyword(s):  
Type I Collagen ◽  
Reference Intervals ◽  
Type I ◽  
High Turnover ◽  
Childhood Growth ◽  
Type Iii ◽  
Procollagen Type ◽  
Amino Terminal ◽  
Type I Procollagen ◽  
Carboxyl Terminal

Abstract We report pediatric age- and sex-specific 95% reference intervals for procollagen type I C-terminal propeptide (PICP), the cross-linked C-terminal telopeptide of type I collagen (ICTP), and procollagen type III N-terminal propeptide (P3NP), measured in plasma from 302 schoolchildren (156 boys, 146 girls) ages 4–19 years. All three markers displayed a significant variation with age (ANOVA P ≤0.0015). PICP showed no detectable increase during adolescence for either sex, but decreased towards adult concentrations after the age of puberty, with an earlier decrease for girls than for boys (P <0.01). ICTP and P3NP both increased in pubertal-aged children (P <0.05), with an earlier increase in girls than in boys (P <0.05), before decreasing towards adult concentrations (P <0.01). All three collagen markers were highly correlated with one another (P <0.001). The patterns observed mirrored the childhood growth curve and reflected the high turnover of bone and soft tissue during childhood growth.

scholarly journals Osteokines and Bone Markers at Rest and following Plyometric Exercise in Pre- and Postmenopausal Women

2020 ◽  
Vol 2020 ◽  
pp. 1-10
Author(s):  
Katlynne Nelson ◽  
Rozalia Kouvelioti ◽  
Alexandros Theocharidis ◽  
Bareket Falk ◽  
Peter Tiidus ◽  
...  
Keyword(s):  
Postmenopausal Women ◽  
Type I Collagen ◽  
Premenopausal Women ◽  
Type I ◽  
Bone Homeostasis ◽  
Younger Women ◽  
Procollagen Type ◽  
Amino Terminal ◽  
Plyometric Exercise ◽  
Procollagen Type I

The effect of plyometric exercise on bone biomarkers has been studied in pediatric and young adult populations in order to better understand how exercise influences bone homeostasis. However, there are no such data in postmenopausal women, a group characterized by an uncoupling of the bone resorption-formation cycle. This study examined the serum concentrations of sclerostin, dickkopf-1 (DKK1), c-terminal crosslinking telopeptides of type I collagen (CTXI), and procollagen type I amino-terminal propeptide (PINP) at rest and following a single bout of plyometric exercise in 20 premenopausal ( 23.1 ± 2.3 years) and 20 postmenopausal women ( 57.9 ± 4.3 years). The exercise consisted of 128 jumps, organized into 5 circuit stations. Blood samples were obtained prior to and 5 min, 1 h, and 24 h postexercise. At rest, postmenopausal women had significantly higher sclerostin and CTXI, but lower DKK1 than premenopausal women. Sclerostin increased 5 min postexercise only in the premenopausal group. DKK1 decreased 24 h postexercise in the premenopausal women while it decreased 1 h postexercise in the postmenopausal women. In both groups, CTXI did not change across time and PINP decreased 5 min and 1 h postexercise ( p < 0.05 ). The PINP/CTXI ratio decreased 5 min and 1 h postexercise then significantly increased 24 h postexercise only in premenopausal women. These results indicate that although plyometric exercise is effective in eliciting osteoanabolic effects in younger women; such an effect is not evident in postmenopausal women.

Abstract 74: The Inflammatory Phenotype Of Mesenchymal Fibroblasts And Its Role In Aging Dependent Cardiac Fibrosis- A Target For Statins?

2014 ◽  
Vol 115 (suppl_1) ◽  
Author(s):  
Katarzyna A Cieslik ◽  
JoAnn Trial ◽  
Mark L Entman
Keyword(s):  
Endothelial Cells ◽  
Myeloid Cell ◽  
Transendothelial Migration ◽  
Mouse Heart ◽  
Type I ◽  
Aged Mouse ◽  
Vitro Assay ◽  
Procollagen Type ◽  
Procollagen Type I ◽  
Inflammatory Phenotype

In the aging mouse (C57BL/6) myocardium fibrosis steadily increases after 14 months of age and is accompanied by elevated numbers of myeloid derived fibroblasts. Recently, we proposed a mechanism by which inflammatory mesenchymal fibroblasts (IMF) derived from mesenchymal stem cells secrete monocyte chemoattractant protein-1 (MCP-1) necessary for myeloid fibroblast induction in the aging heart. The current study extends the characterization of this inflammatory phenotype by describing elevated interleukin-6 (IL-6) secretion and increased expression of IL-6 receptor (IL-6R) in IMF. Since IL-6R lacks an intracellular domain it requires a co-receptor gp130 (generally expressed) to induce an intracellular signal. Thus, generation of an IL-6R soluble receptor allows IL-6 signaling on cells that do not express IL-6R (or expression is low), such as endothelial cells. We investigate the function of IL-6 and IL-6R in the promotion of transendothelial migration of monocytes through cardiac endothelium and their maturation into myeloid fibroblasts in in vitro assay. Treatments with IL-6 and more extensively IL-6+IL-6R resulted in a 3-5 fold increase (above the control level) in myeloid cell migration and maturation into myeloid fibroblasts. Thus IMF can contribute both IL-6 and IL-6R to endothelial cells and facilitate myeloid cell transendothelial migration. In agreement with these data, analysis of the aged mouse heart revealed the presence of fibroblasts expressing IL-6 (procollagen type I + IL-6 + cells), M1 macrophages (CD86 + cells) and M2 macrophages (CD301 + procollagen type I + cells) that were absent in hearts from young mice. The mechanisms by which expression of these factors is upregulated in IMF are being investigated; our data suggest that MCP-1 and IL-6 expression are controlled by the farnesyltransferase (FTase)-Ras-Erk1/2 pathway. Interestingly, since atorvastatin interferes with farnesyl synthesis it also reduced MCP-1 and IL-6 expression in IMF. These data may introduce a new use of this class of drugs in the prevention of the age-related fibrosis.

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