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PubMed1998

Procollagens and inflammatory cytokine concentrations in tarsal and limbal vernal keratoconjunctivitis.

To quantify the presence of inflammatory/fibrogenic cytokines and procollagens type I (PICP) and III (PIIIP) in active and non-active tarsal and limbal forms of vernal keratoconjunctivitis (VKC), tear and blood samples were collected from 27 VKC patients (20 active and 7 non-active) and 15 normal subjects. Upper tarsal conjunctival biopses were obtained from 8 controls and 8 tarsal VKC patients. From biopses of 4 tarsal VKC, fibroblasts were cultured in F12 medium with 10% FCS. TGF-beta 1, IL-1 alpha, IL-1 beta, IL-6 and TNF-alpha, PICP and PIIIP were measured in: (1) tears, (2) homogenized conjunctival tissues, (3) serum, (4) supernatants of tissue cultures at 24 hr, and fibroblast primary passage cultures. Results showed: (1) in tears, TGF-beta 1 and TNF were identified in several active VKC patients without significant differences between the tarsal and the limbal forms. IL-1 beta (27 +/- 51 pg ml-1, P = 0.03) and IL-6 (28 +/- 43 pg ml-1, P = 0.006) were significantly increased in tarsal VKC compared to controls. Both control and non-active VKC tear samples had undetectable levels of all of the above cytokines. PICP and PIIIP were significantly increased in tarsal VKC compared to both limbal VKC and controls. Non-active VKC levels were similar to controls. (2) In homogenized VKC tissues, TGF-beta 1 and IL-6 were both significantly increased compared to controls (P < 0.01) while no increases were observed in IL-1 and TNF-alpha. (3) In serum, IL-1 alpha, IL-1 beta and TNF-alpha were higher in VKC patients compared to controls. (4) In vitro fibroblasts from VKC patients showed an increased production of TGF-beta 1, IL-1 alpha, IL-1 beta, IL-6, TNF-alpha, PICP, and PIIIP over time. Increased levels of TGF-beta 1, IL-1 and IL-6 in VKC tissues and tears indicate a local production of these cytokines in active VKC. Collagen hyperproduction occurs only in active tarsal VKC and may be related to high levels of TGF-beta 1, IL-1 and IL-6. Increased serum levels of IL-1 and TNF-alpha suggests that systemic immunological changes occur in VKC. Cell culture can be used as a model to further study the pathogenesis of VKC and its characteristic local fibroblast activation.

Adolescent↗
PubMed1996

Reduced levels of alpha1(XI) procollagen mRNA in SV40-transformed cells.

We have used mRNA differential display to search for cellular genes whose expression is modulated in SV40-transformed cells. We identified a gene, the alpha1 subunit of type XI procollagen (alpha1(XI)), whose mRNA level is reduced in most F111 rat cell lines that express either wild-type SV40 T antigen or an amino-terminal fragment of T antigen (T147D). alpha1(XI) mRNA expression is also reduced in rat embryo fibroblasts transformed by SV40. This phenomenon is not unique to rat cells, because alpha1(XI) mRNA expression is repressed in C3H10T1/2 mouse cells transformed by either T147D or wild-type SV40. Interestingly, alpha1(XI) mRNA levels are not reduced in F111 cells that express only the polyomavirus large T antigen, but are reduced in F111 cells transformed by polyoma virus (which expresses all three polyomavirus T antigens) and in F111 cells that express the adenovirus E1A proteins. Thus, repression of alpha1(XI) mRNA expression may be a common feature of transformation by DNA tumor viruses.

Adenovirus E1A Proteins↗
PubMed1993

Study of serum procollagen type III peptide in patients with hepatic cirrhosis from a clinical point of view.

N-terminal procollagen-III peptide (P-III-P) has been considered a marker of fibrogenesis and inflammatory activity of the liver. We measured the P-III-P serum levels in 83 cirrhotic patients fully characterized from a clinical and laboratory point of view. Cirrhotic patients had significantly higher P-III-P serum levels than controls (P < 0.0001). Of the cirrhotic patients 73.5% had increased P-III-P. A significant negative correlation was found between P-III-P and transaminases, and patients with normal values of alanine amino-transferase had higher P-III-P serum levels than those with increased values (P = 0.03). On the other hand, no significant association was found with portal hypertension, Child classes, or alcoholic liver disease. No one independent factor appears to be responsible for the increase in P-III-P. The measurement of serum P-III-P is of little if any use in the evaluation of cirrhotic patients.

Adult↗
PubMed1995

Non-radioactive multiplex-SSCP analysis: detection of a new type II procollagen gene (COL2A1) mutation.

We have developed a protocol that allows fast and efficient mutation screening of the 54 exons from the type II procollagen (COL2A1) gene. The protocol is based on the multiple non-radioactive hybridization of blotted single-strand conformation polymorphism gels. Using this screening procedure we have been able to identify a new (Gly895 to Ser) mutation in the COL2A1 gene of a patient with a mild form of spondyloepiphyseal dysplasia congenita.

Amino Acid Sequence↗
PubMed1993

Deletion of 19 base pairs in intron 13 of the gene for the pro alpha 2(I) chain of type-I procollagen (COL1A2) causes exon skipping in a proband with type-I osteogenesis imperfecta.

Skin fibroblasts from a proband with mild osteogenesis imperfecta (type I) synthesized normal pro alpha 2(I) chains and shortened pro alpha 2(I) chains of type-I procollagen. The type-I collagen that contained the shortened alpha 2(I) chains was thermally unstable in that it was cleaved at 30 degrees C by a mixture of trypsin and chymotrypsin. The mutation generating the shortened pro alpha 2(I) chains was shown to be a deletion of 19 base pairs from +4 to +22 of intron 13 of the COL1A2 gene by sequencing of genomic DNA and allele-specific oligonucleotide hybridization. The same mutation was found in the proband's affected father. Probe-protection experiments with S1 nuclease demonstrated that about 88% of the RNA transcripts from the mutated allele were spliced by exon skipping from exon 12 to exon 14 and that about 12% of the RNA transcripts were normally spliced. There was no evidence for use of cryptic splice sites, even though two cryptic splice sites had more favorable statistical scores and delta G degree 37 values than the new site that was created by the mutation and that was used for splicing of 12% of the transcripts into a normal mRNA. Comparison of the results with observations on 17 previously reported mutations that produced in-frame deletions of amino acids from the triple-helical domain of type-I collagen indicated that deletions in the N-terminal half of the alpha 2(I) chain tended to produce milder phenotypes than similar deletions elsewhere in the alpha 1(I) or alpha 2(I) chains.

Adolescent↗
PubMed1986

The single copy gene coding for human alpha 1 (IV) procollagen is located at the terminal end of the long arm of chromosome 13.

Using dual-laser sorted chromosomes and spot-blot analysis, we have previously assigned genomic DNA sequences coding for human alpha 1 (IV) procollagen to chromosome 13 (Pihlajaniemi et al. 1985). By in situ hybridization to normal chromosomes and chromosomes with 13q deletions, we now report the localization of this gene to the terminal end of the long arm of chromosome 13. In addition, Southern and slot blot hybridization analysis clearly show that these genomic sequences are present only once per haploid genome.

Base Sequence↗
PubMed1994

Carboxyterminal propeptide of type I procollagen in osteomalacia.

Carboxyterminal propeptide of type I procollagen (PICP) was measured in sera from 25 patients with osteomalacia due to privational vitamin D deficiency. The mean value of serum PICP was significantly raised in patients with osteomalacia compared with 40 normal subjects (mean +/- SD, 161 +/- 82 ng/ml and 113 +/- 31 ng/ml, respectively; P = 0.01). The raised serum PICP reflected the accelerated bone turnover in this condition as did the elevated serum total alkaline phosphatase (ALP), but the variation in values of serum PICP noted in individual patients may reflect the different stages of osteomalacia. The normalization of serum PICP and ALP, indicating a healing process of the bone disorder, needed longer time of vitamin D and calcium therapy. In contrast, adjusted serum calcium and inorganic phosphate (IP) responded and normalized rapidly. Serum PICP and total ALP appeared to behave differently in the disease and in its response to vitamin D therapy suggesting that these two markers may represent different functions of osteoblasts in osteomalacia.

Adult↗
PubMed1993

Procollagen type III N-peptide and type IV collagen 7S-domain in the sera of breast cancer patients.

We investigated the usefulness of serum levels of procollagen type III N-peptide (P III P) and the type IV collagen, 7S-domain, (IV-C) as markers of recurrent and metastatic breast cancer. Serum P III P and IV-C levels were found to be significantly higher in patients who showed postoperative recurrence, with 76.9% of P III P-positive cancer bearing patients and 68.0% of IV-C-positive cancer bearing patients having metastases in the bone and/or liver. Furthermore, 66.7% of all patients with metastases in the bone and/or liver were P III P-positive and 75.6% were IV-C-positive. Patients with liver metastases were either P III P- or IV-C-positive, and the levels were higher than those in patients with bone metastases. Thus, a positive correlation of serum P III P and IV-C levels was observed. These findings suggest that both serum P III P and IV-C levels are useful markers of recurrent and metastatic breast cancer, especially of disease in the bone and/or liver.

Biomarkers, Tumor↗
PubMed1996

Assessment of hepatic graft injury by graft effluent in rodents: N-acetyl-beta-glucosaminidase and type III procollagen peptide.

We studied the significance of N-acetyl-beta-glucosaminidase (beta-NAG) and type III procollagen peptide (P-III-P) in the effluent of rodent hepatic grafts. After total hepatectomy, the livers were preserved in chilled, lactated Ringer's solution and then divided into five groups (n = 10 each): group 1, 4 h preservation only; group 2, 4 h preservation and rewarming; group 3, 6 h preservation only; group 4, 6 h preservation and rewarming; and group 5, minimal preservation only. The beta-NAG of groups 2 and 4 was significantly higher than that of groups 1 and 3 (0.98 +/- 0.5 U/l vs 0.21 +/- 0.12 U/l; P < or = 0.01 and 1.76 +/- 0.67 U/l vs 0.38 +/- 0.25 U/l, respectively; P < or = 0.01), while that of group 4 was significantly higher than that of group 2 (1.76 +/- 0.67 U/l vs 0.98 +/- 0.50 U/l; P < or = 0.05). The P-III-P of group 4 was significantly higher than that of group 2 (0.133 +/- 0.008 U/ml vs 0.110 +/- 0.015 U/ml; P < 0.01). We conclude that beta-NAG is a novel parameter of parenchymal and nonparenchymal cells, while P-III-P reflects the integrity of the hepatic sinusoidal extracellular matrix.

Acetylglucosaminidase↗
PubMed1994

Serum concentration of procollagen type I carboxyterminal propeptide in systemic sclerosis.

The serum level of procollagen type I carboxyterminal propeptide (P1CP), which has been used as an index of collagen synthesis in patients with various fibrotic diseases during the active stage, was measured using enzyme-linked immunosorbent assay in 61 patients with systemic sclerosis (SSc) and in 21 control subjects. The mean P1CP level in the SSc patients was significantly higher than in the normal controls (mean +/- SD, 326 +/- 319 vs 128 +/- 87 ng/ml; p < 0.005). In 36% of the SSc patients, the serum P1CP level was significantly elevated more than two standard deviations above the mean control value. The mean serum P1CP level in patients with diffuse SSc was significantly higher than in those with limited SSc (411 +/- 373 vs 255 +/- 199 ng/ml; p < 0.05). In addition, the SSc patients with elevated serum P1CP levels showed a significantly greater incidence of lung fibrosis and joint involvement than those with normal P1CP levels (p < 0.005 and p < 0.05, respectively). These results suggest that the serum P1CP level is a useful indicator of the severity of disease in SSc patients.

Adolescent↗
PubMed1994

Respiratory effects and serum type III procollagen in potato sorters exposed to diatomaceous earth.

Exposure to diatomaceous earth with low crystalline silica content (< 1%) is rarely reported to cause pneumoconiotic disease, whereas airway obstruction and bronchitis are more frequently reported. We investigated the occurrence of pneumoconiosis and airflow limitation in 172 male workers from 5 potato sorting plants (55 controls, 29 salesmen, 72 currently exposed, and 16 retired exposed) exposed to inorganic dust from former sea terraces (7.7-15.4 mg/m3), high in diatomaceous earth. The presence of fibrosis was evaluated by chest radiographs (exposed only) and serum levels of type III procollagen (P-III-P) were measured as an estimate of fibrogenetic activity. Lung function was assessed by flow volume curves and impedance measurements. A validated questionnaire was used to record respiratory symptoms. No pneumoconiotic abnormalities were demonstrated by chest radiographs. In line with this finding, serum P-III-P levels were not elevated in exposed workers as compared to controls, suggesting no differences in fibrogenetic activity. In fact, serum P-III-P levels decreased significantly (P < 0.03) with increasing cumulative exposure. Flow volume parameters indicated airflow obstruction, dose-related to (cumulative) dust exposure; the annual decline in forced expiratory flow volume (FEV1) was estimated at 10.5 ml/year (P < 0.05). Airway obstruction was confirmed by impedance analysis: In the retired group impedance changes were compatible with airway obstruction extending into the peripheral airways. We conclude that this exposure to quartz during potato sorting does not result in an increased risk for pneumoconiosis, but that (prolonged) surveillance in this group is desirable in order to detect early indications of airflow obstruction.

Adult↗
PubMed1983

The significance of the aminoterminal propeptide of type III procollagen in paroxysmal nocturnal haemoglobinuria and myelofibrosis.

The level of the aminoterminal propeptide Col 1-3 of type III procollagen (PC-III) was determined in patients with paroxysmal nocturnal haemoglobinuria (PNH) and primary myelofibrosis (PMF), to study whether PC-III can be used as a parameter for the rate and/or degree of bone marrow replacement with collagen. Normal PC-III levels were found in PNH (6.6 +/- 1.1 micrograms/l; N: 8.6 +/- 1.8 micrograms/l), while significantly increased levels were found in PMF (24.8 +/- 2.2 micrograms/l). During a follow-up of 1 year, a slight increase of 2 micrograms/l occurred in three patients with a stable fibrosis, while one patient with more active disease demonstrated an increase of 25 micrograms/l. Treatment with acetylsalicylic acid led to a decline of PC-III as well as beta-thromboglobulin level, although normalization did not occur. It was demonstrated by means of gel filtration that the antigens related to the PC-III peptide were heterogenous, and that in PMF at least two main peaks were present, with molecular masses equal to and smaller than PC-III peptide. These data demonstrate that the radioimmunoassay cannot be used for the quantitative determination of PC-III; nevertheless it gives some insight in the process of bone marrow fibrosis.

Aged↗
PubMed1988

Altered expression of collagen type VI in brain vessels of patients with chronic hypertension. A comparison with the distribution of collagen IV and procollagen III.

The vascular extracellular matrix (ECM) plays an important role in the histopathology of cerebral microcirculation, but its characterization is still incomplete. For that reason we investigated paraffin-embedded and cryostat sections of intracerebral and meningeal vessels from eight normotensive and six hypertensive humans using monospecific affinity-purified polyclonal antibodies against human/monkey amino-terminal procollagen I + III peptide (P I P, P III P), collagen IV (7-S and NC1 domains), VI, and laminin (P 1 fragment) by applying peroxidase-antiperoxidase- and alkaline phosphatase-antialkaline phosphatase techniques. In normotensives, laminin and collagen IV were codistributed in the basal lamina of meningeal and intraparenchymal vessels. Collagen VI was only present in the adventitia of meningeal vessels and larger intraparenchymal arteries and veins, whereas it was absent from cortical vessels including capillaries. Intensive staining for collagen VI was observed in the choroid plexus, the superficial glia and sheath of cranial nerves. In hypertensives, the basement membrane constituents laminin and collagen IV appeared ubiquitously increased. Here, collagen VI was also deposited in the broadened vascular intima and media of larger arteries and in cortical vessels. In both groups collagen VI and P III P appeared to be codistributed. Our results indicate that significant qualitative change sin ECM of cerebral blood vessels are taking place during the development of hypertension, such as (1) an atypical deposition or an increase of collagen VI which by interconnecting collagen fibrils (I and III) might exert a stabilizing (sclerosing) function in the ECM, and (2) a thickening of vascular basement membranes caused by an accumulation of its major components laminin and collagen IV.

Aged↗
PubMed1993

Impact of chemotherapy on collagen metabolism: a study of serum PIIINP (aminoterminal propeptide of type III procollagen) in advanced sarcomas.

We have previously shown that the serum aminoterminal propeptide of type III procollagen (PIIINP) is a prognostic factor for survival in localised soft-tissue sarcomas, and that elevated values are frequent in metastatic disease. In the present study PIINP is analysed during chemotherapy in 26 patients with advanced sarcomas. Non-responders had a significantly higher pretreatment level of PIIINP than responders (P = 0.05), when only patients with no recent therapeutic interventions were studied. However, during chemotherapy PIIINP followed the clinical course of the malignant disease in only a minority of patients. Patients with recent surgery or recently completed chemotherapy had an increased pretreatment PIIINP value (P = 0.03). In these patients PIIINP declined during chemotherapy irrespective of tumour response. A pretreatment PIIINP level within the reference range tended to increase with time irrespective of response. Moreover, the values taken during a chemotherapy infusion were significantly higher than those immediately preceding the corresponding cycle (P = 0.001). Our results suggest that pretreatment PIIINP is of value as a prognostic factor for chemotherapy response in patients with advanced sarcomas. During chemotherapy PIIINP is of minor importance in monitoring response because of the influence of chemotherapy and other therapeutic interventions on the level of PIIINP.

Antineoplastic Combined Chemotherapy Protocols↗
PubMed1987

Is determination of serum N-terminal procollagen type III peptide (sPIIIP) a marker of hepatic fibrosis?

Serum N-terminal procollagen type III peptide (sPIIIP) levels were evaluated in 58 patients affected by chronic liver disease, in order to assess the usefulness of sPIIIP as a marker of hepatic fibrosis. In 45 patients sPIIIP was also correlated to liver histology; biopsies were scored by two of the authors, without knowledge of diagnosis. Compared to normal controls, sPIIIP concentration was found to be significantly elevated in chronic active hepatitis (CAH) and in cirrhosis, but not in fatty liver. Patients affected by chronic persistent hepatitis (CPH) had values of sPIIIP higher than normal in four of 11 cases considered. A close correlation was found between sPIIIP values and histological parameters of inflammation, necrosis, and degeneration, while the relationship between sPIIIP levels and fibrosis was weaker. These data suggest that sPIIIP determination may reflect the extent of inflammatory changes in the liver; but it cannot be considered a reliable index of hepatic fibrosis.

Adult↗
PubMed1979

Activity of procollagen-prolyl-hydroxylase and N-acetyl-beta-glucosaminidase in liver biopsies from patients with chronic liver diseases.

In 5 mg of liver tissue obtained by needle biopsy from patients with chronic liver diseases, activity of both Procollagen-prolyl-hydroxylase and N-Acetyl-beta-glucosaminidase was measured. The activity of both enzymes increased due to the activity of fibrosis, but prolylhydroxylase increased more. The relation of these two enzyme activities in the development of liver fibrosis is discussed.

Acetylglucosaminidase↗
PubMed1991

Differential patterns of PMN-elastase and type III procollagen peptide in knee joint effusions due to acute and chronic sports injuries.

In 38 traumatic knee joint effusions the proteolytic enzyme PMN-elastase (PMN-E) and the repair marker procollagen III aminoterminal peptide (PIIINP) were determined. According to the period between trauma and first aspiration of the effusion, the patients were divided into 3 groups. Group I (17 patients; period between trauma and first aspiration not longer than 72 hours) showed high concentrations of PMN-E (up to 5400 ng/ml) and low concentrations of PIIINP (less than 13 U/ml). Group II (11 patients; aspiration within 4 to 14 days) had mean PMN-E and PIIINP concentrations of 125.6 ng/ml and 52.1 U/ml, respectively. In group III (10 patients, aspiration after 14 days) mean PMN-E concentration was 123.8 ng/ml and mean PIIINP concentration was 63.4 U/ml. Graphic depiction of PMN-E and PIIINP levels in each individual sample as a function of time between trauma and fluid collection revealed highly increasing PMN-E levels during the first 24 posttraumatic hours, followed by rapidly decreasing levels within 72 hours post trauma, and no change after the 4th posttraumatic day. In contrast, PIIINP increased continuously up to the first posttraumatic week and stayed at high levels up to 90 days (end of the observation period). The differential patterns of PMN-E and PIIINP concentration in knee joint effusions may be useful in estimating the period between trauma and first treatment (aspiration of effusion) and should, therefore, be helpful in detecting degenerative lesions, which seem to be characterized by low PMN-E concomitantly with high PIIINP levels.

Adolescent↗
PubMed1992

Transpulmonary gradient of type III procollagen peptides: acute effects of cardio-pulmonary bypass.

Type III procollagen N-peptides (PIIINPs) are believed to be released in stoichiometric amounts as type III collagen molecules are secreted from cells. We hypothesized that if the human lung actively produces type III collagen a detectable transpulmonary gradient in PIIINPs would exist in normal individuals that might be altered following a pulmonary insult. PIIINPs were therefore measured by radioimmunoassay in serum taken simultaneously from the pulmonary artery (PA) and left ventricle/aorta (LV) in 11 patients undergoing routine cardiac catheterisation. Mean PIIINP levels +/- SEM in LV were 66.8 +/- 5.4 micrograms.ml-1 and 59.9 +/- 4.1 micrograms.ml-1 in PA (p less than 0.04). In 6 patients, repeat measurements taken 4 h after cardiopulmonary bypass revealed a significant fall in PA values to 43.8 +/- 2.6 micrograms.ml-1 (p less than 0.001) and abolition of the transpulmonary gradient. These results suggest the adult human lung actively synthesis type III collagen and that, in the short term, cardiopulmonary bypass inhibits this process.

Cardiac Catheterization↗