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Biomedical subjects

H J Møller

Publications and source records attributed to H J Møller.

At least 19 recordsLinked to original sources
PubMed2006

Differential expression of monocyte CD163 in single- and dual-asthmatic responders during allergen-induced bronchoconstriction.

BACKGROUND: Mononuclear phagocytes play an important role in modulating inflammatory reactions in response to antigen challenge. OBJECTIVE: To investigate the regulation of CD163, a marker of anti-inflammatory macrophages, during Dermatophagoides pteronyssinus (Dp)-induced bronchoconstriction. METHODS: Among 110 Dp-sensitive patients who underwent bronchial challenge with Dp, there were 51 dual responders (DR), 32 single responders (SR) and 27 non-responders (NR). Monocyte expression of CD14 and CD163 was evaluated by flow cytometry. Exhaled NO (eNO) concentration was determined on-line using a chemiluminescence analyser. In 21 DR, nine SR and 13 NR-soluble CD163 in plasma was measured by ELISA before, 1, 8 and 24 h after the challenge. RESULTS: In DR, the baseline expression of monocyte CD163 and eNO were significantly greater than in SR and NR. A pattern of (1) decreased monocyte CD163 expression, (2) unchanged sCD163 and (3) increased eNO in DR was contrasted by a pattern of (1) increased CD163 expression, (2) increased sCD163 and (3) unchanged eNO in SR. During allergen challenge, the changes in monocyte CD163 expression and sCD163 inversely correlated with the changes in eNO. CONCLUSION: Both pro-inflammatory and anti-inflammatory responses to allergen challenge are uniquely expressed among SR and DR.

Adolescent↗
PubMed2005

Predictive value of soluble haemoglobin scavenger receptor CD163 serum levels for survival in verified tuberculosis patients.

Pre-treatment serum levels of sCD163 were measured in a cohort of 236 suspected tuberculosis (TB) cases from Guinea-Bissau, with a median follow-up period of 3.3 years (range 0-6.4 years). In 113 cases, the diagnosis of TB was verified by positive sputum microscopy and/or culture. Among the verified TB cases, a decreased survival rate was found in 27 patients with sCD163 levels above the upper reference limit (3.95 microg/mL). The difference in survival was significant during TB treatment (log rank, p<0.02) and after long-term follow-up (log rank, p<0.001). The decrease in survival rate during TB treatment remained significant in a multivariate Cox model controlling for human immunodeficiency virus (HIV) status, age and gender, with a mortality increase of 1.19 (95% CI, 1.04-1.36) per microg of sCD163, and a hazard ratio (HR) for sCD163 levels above the upper reference limit of 4.18 (95% CI, 1.06-16.4). The difference was not significant after excluding patients with concomitant HIV-1 and HIV-2 infection in Kaplan-Meier analyses (log rank, p 0.11). In contrast, the difference in survival remained significant in Kaplan-Meier analyses after long-term follow-up, even after excluding patients with concomitant HIV-1 and HIV-2 infection (log rank, p 0.002). In the Cox model, the mortality increase per microg of sCD163 was 1.27 (95% CI, 1.14-1.40), with an HR for elevated sCD163 levels of 2.85 (95% CI, 1.44-5.63). The HRs for concomitant HIV-1 and HIV-2 infection were 6.92 (95% CI, 3.28-14.58) and 2.48 (95% CI, 1.09-5.67), respectively. Thus, sCD163 levels appeared to be an independent predictor of survival in verified TB patients.

Adult↗
PubMed2003

Biological variation of soluble CD163.

A soluble plasma form of CD163 (sCD163) was recently identified. The protein has anti-inflammatory effects in vitro and is elevated in patients with myelo-monocytic leukaemia and infection. For rational use and evaluation of this potential new quantity it is important to have knowledge of its biological variability and to use a methodology that has a sufficiently analytical quality. The day-to-day and diurnal biological variabilities of sCD163 were studied in 12 healthy people using a sandwich ELISA. The within-run-, between run- and total analytical coefficients of variation were estimated to 3.6%, 4.8% and 6.0%, respectively. The day-to-day within-subject biological variation was estimated to 9.0%, and the between-subject biological variation to 35.9%. A diurnal variation in sCD163 concentrations with 14% lower values in the night (supine position) was observed. The ratio between total analytical variation and within-subject biological variation was 0.67. The index of individuality, calculated as the ratio between within-subject biological variation and between-subject biological variation, was low and similar to complement factors and immunoglobulins. A low index of individuality is important in a monitoring situation where small changes from the set point of the individual can be detected in serial measurements. For this purpose, the critical difference for a series of results in the same patient (significant at p < 0.05) was calculated to 30% for samples taken on different days and measured in separate runs.

Aged↗
PubMed2002

Characterization of an enzyme-linked immunosorbent assay for soluble CD163.

We have recently identified a soluble plasma form of CD163 sCD163, the macrophage receptor for clearance of haptoglobin-haemoglobin complexes, and we have observed highly elevated levels of sCD163 in subgroups of haematological patients. In the present study, we describe the optimization and characterization of a sandwich ELISA for the determination of the concentration of sCD163 in plasma and serum. The optimal concentrations of antibodies were determined systematically and the assay was calibrated by CD163 purified from human spleen membranes. The minimum detection limit was below 6.25 microg/L. Recovery of CD163 added to plasma samples was 106%. The concentration of sCD163 in paired serum and plasma samples correlated well (r2=0.99); however, serum levels were 1.1 times higher than the plasma levels. The addition of haptoglobin-haemoglobin complexes did not influence the assay. A very high stability of sCD163 was measured in whole blood and in plasma subjected to different temperatures and after repeated freezing and thawing.

Antibodies↗
PubMed2002

Soluble CD163: a marker molecule for monocyte/macrophage activity in disease.

By immunoprecipitation we have identified a soluble plasma form of CD163 (sCD163), the IL-6 inducible macrophage-receptor for clearing haptoglobin-haemoglobin complexes. A sandwich ELISA for measuring sCD163 was established and used to determine the sCD163 levels in normal subjects and patients with inflammatory and myeloproliferative diseases. In normal subjects, the concentration of sCD163 was high (median 1.9 mg/l) with low intra-individual variation. Highly increased levels were seen in patients with sepsis, myeloid leukaemia and in patients with Gaucher disease characterized by accumulation of tissue macrophages. Although the physiological role of sCD163 remains unknown, our present data suggest that sCD163 might prove to be a valuable marker molecule in infectious and myeloproliferative diseases.

Antigens, CD↗
PubMed2001

[Biochemical changes in fibromyalgia. Can serum hyaluronic acid be used diagnostically?].

AIM: To assess the levels of hyaluronic acid (HA) in Danish patients with fibromyalgia (FM). METHODS: Serum levels of HA were determined in 53 patients with established FM and 55 control samples with a radiometric assay (Pharmacia). Values were correlated to clinical parameters of disease severity (duration of disease, tender point scales, visual analogue scales). RESULTS: There were no differences in the HA levels of patients and controls. In all the patients, except one, values were within the reference intervals. Nor was there an association between HA levels and clinical findings. CONCLUSIONS: Patients with FM do not generally have increased serum levels of HA, and other serum measurements have not been helpful in the diagnosis of FM. Some biochemical changes have been described in FM, however, and these have mainly been observed in the spinal fluid.

Adult↗
PubMed2001

Evaluation of potential organ culture media for eye banking using human donor corneas.

AIM: To evaluate the ability of different commercially available cell culture solutions to preserve human donor corneas during 3 weeks of "closed system" organ culture at physiological temperature. This screening was performed in an attempt to establish a rational basis for the development of a serum-free organ culture medium for eye banking. METHODS: 72 normal human donor corneas were organ cultured for 21 days at 31 degrees C in eight different test media (nine corneas in each group). The basic culture solutions included: minimal essential medium (MEM), MEM with stabilised L-glutamine, M199, DIF-1000, SFM, F99, and F99 with ascorbic acid, insulin, bFGF, transferrin, selenium, and lipids (termed F99-Sr). All media were supplemented with 2% fetal calf serum (FCS), except for MEM, which was also studied at 8% FCS. The evaluation parameters included: (1) the endothelial cell loss as evaluated using trypan blue staining; (2) the ability of keratocytes and endothelial cells to incorporate tritiated uridine into RNA as evaluated using autoradiography and digital image analysis; (3) the leakage of immunogenic keratan sulphate as assessed using ELISA; and (4) changes in storage medium pH, glucose, and lactate content. RESULTS: SFM induced the lowest endothelial cell loss of 14% (SD 2%) and the highest RNA synthesis rates of all test solutions supplemented with 2% FCS. Corneas stored in SFM also showed the least leakage of keratan sulphate and the highest glucose consumption and lactate production. In five media (MEM with 2% FCS, MEM with stabilised L-glutamine, M199, F99, and F99-Sr), comparable and intermediate potentials for organ culture were observed with endothelial cell loss of 16-19%. By contrast, 29% (4%) of the endothelium was lost after storage in DIF-1000. Interestingly, the use of 8% FCS (in MEM) had a marked protective effect on the endothelium, which showed the highest RNA synthetic activity combined with a cell loss of only 11% (4%), compared with 19% (6%) at 2% FCS (p<0.05). CONCLUSION: Among the present test solutions, SFM appears to be the most prominent candidate for a new corneal organ culture medium and should be further tested and possibly refined to effectively substitute serum addition.

Adult↗
PubMed2000

Patients with fibromyalgia have normal serum levels of hyaluronic acid.

OBJECTIVE: To investigate the levels of hyaluronic acid (HA) in Danish patients with fibromyalgia (FM). METHODS: Serum levels of HA were determined in 53 patients with established FM and 55 control samples using a radiometric assay. Values were correlated to clinical disease severity variables (duration of disease, tender point scales, visual analog scales). RESULTS: There were no differences in HA levels between patients and controls. HA levels of all patients except one were within the reference ranges. There was no association between HA levels and clinical findings. CONCLUSION: Patients with FM do not generally have increased serum levels of HA.

Aging↗
PubMed1998

Connective tissue markers of rheumatoid arthritis.

Rheumatoid arthritis (RA) is a common systemic autoimmune disorder of unknown aetiology. The most common outcome of RA is a progressive development of joint destruction and deformity. Early introduction of disease-modifying antirheumatic drugs seems important for prevention of the long term injuries of articular cartilage and bone. Early diagnosis and selection of patients with rapidly progressive disease therefore is of clinical significance. Routine laboratory tests are valuable in monitoring for renal, hepatic and haematological side effects of medical treatment. Determination of rheumatoid factor contributes to the classification of arthritis as RA, and acute phase reactants are useful for quantifying and comparing the level of inflammatory activity in the course of a given patient. There is, however, a lack of sensitive and specific biochemical markers for RA, and frontline biochemical research is devoted to characterizing molecules which are of diagnostic and prognostic value, as well as molecules which are indicators of the degree of joint cartilage and bone destruction. The present survey summarizes current knowledge concerning possible tissue-specific marker molecules of RA.

Aggrecans↗
PubMed1996

Viability of human corneal keratocytes during organ culture.

The viability of human corneal keratocytes was assessed during four weeks of 'closed system' organ culture at 31 degrees C. After 28 days of culturing, the entire keratocyte population was still alive and viable because all cells incorporated uridine; a parameter for RNA-synthesis. During the first 14 days, mitoses were found in the anterior half of the stroma (0.23% mitoses per 48 h), while only few keratocytes were able to divide at day 28 (0.01% mitoses per 48 h). Metabolic parameters revealed a progressing acidosis in the medium with oxygen and glucose depletion. Immunological measurements of keratan sulphate proteoglycan suggested that approximately 1% of the total content was lost during the period. In conclusion, our current organ culture technique can maintain a viable keratocyte population for four weeks; a viable stroma can be grafted within this period.

Aged↗
PubMed1995

Demonstration of immunogenic keratan sulphate in commercial chondroitin 6-sulphate from shark cartilage. Implications for ELISA assays.

The prototype monoclonal keratan sulphate (KS) antibody 5D4 that is widely used for detection of KS in tissues and biological fluids reacts strongly with commercial low grade shark cartilage chondroitin 6-sulphate. Characterization of the immunogenic material by chondroitinase ABC digestion, ELISA inhibition studies, immunoblotting and HPLC analyses confirmed the presence of substantial amounts of KS, probably as a large proteoglycan (> 120 kDa). Commercial and heterogenic glycosaminoglycan preparations therefore must be used with great caution in immunological analyses. On the other hand the shark cartilage chondroitin 6-sulphate is an easy accessible source of immunogenic KS that can be used as a reference standard and as coating antigen in KS-ELISAs. The concentration of immunogenic KS in synovial fluid measured with an ELISA based solely on reagents of shark cartilage chondroitin 6-sulphate correlated well (r = 0.90) with the concentrations obtained with a traditional KS-ELISA that uses purified aggrecan as standard and coating antigen, and KS in both serum and synovial fluid could be measured with sufficient linearity.

Animals↗
PubMed1995

Improved method for silver staining of glycoproteins in thin sodium dodecyl sulfate polyacrylamide gels.

A method for detection of glycoproteins in thin sodium dodecyl sulfate polyacrylamide gels was developed by a combination of (i) initial periodic acid oxidation/Alcian blue staining and (ii) subsequent staining with silver nitrate. The procedure allowed detection of as little as 1.6 ng of alpha 1-acid glycoprotein and 8-40 ng of a polydisperse mucin sample, which is at least 10 times more sensitive than previously published methods. The method should be very useful for assessment of sample purity and detection of glycoproteins in dilute mixed samples.

Alcian Blue↗
PubMed1994

Effect of flunarizine and calcium on serotonin uptake in human and rat blood platelets and rat synaptosomes.

Calcium and the calcium overload blocker flunarizine exert profound effects on mood. We therefore studied the effect of calcium and flunarizine on serotonin uptake in human and rat blood platelets and in rat synaptosomes. Calcium (1.3 mmol/L) had a weak inhibiting effect on serotonin uptake in blood platelets, whereas no effect was observed in synaptosomes. Flunarizine inhibited serotonin uptake in a concentration dependent manner with an IC50 value of 1 mumol/L in blood platelets and 5 mumol/L in synaptosomes. The inhibition did not depend on the presence of extracellular calcium indicating that the effect is not coupled to a blockade of cellular calcium influx. In human blood platelets, the inhibition was of the noncompetitive type. These results indicate that flunarizine interacts directly with the 5-HT uptake site. The relatively high concentration of flunarizine required to inhibit 5-HT uptake may question the clinical importance of this effect.

Animals↗
PubMed1994

ELISA for the core protein of the cartilage large aggregating proteoglycan, aggrecan: comparison with the concentrations of immunogenic keratan sulphate in synovial fluid, serum and urine.

Immunological assays for fragments of the cartilage large aggregating proteoglycan, aggrecan, have been widely used to monitor cartilage turnover. These assays have commonly employed the monoclonal keratan sulphate antibody, 5D4. Keratan sulphate, however, is present in many tissues and 5D4 affinity is critically dependent on antigen structure. We have therefore raised and characterized a monoclonal antibody (1-F21) that reacts with the core protein of aggrecan without interference from the glycosaminoglycan side chains and, using this antibody, we have optimized a sensitive, competitive ELISA. The within-assay and between-assay coefficients of variation were 4.9-8.9% and 11.1-13.0%, respectively. The mean concentrations of core protein in synovial fluid, serum and urine were 76.4 micrograms/ml, 104.0 ng/ml and 81.0 ng/ml, respectively. In synovial fluids the concentrations were closely correlated with the concentrations of keratan sulphate as determined by 5D4 (r = 0.94), whereas in serum and urine there was no obvious correlation between the determinations. These findings show that measurement of both core protein and keratan sulphate results in a more precise description of aggrecan turnover.

Aggrecans↗
PubMed1994

The epidermal growth factor-like domain of the human cartilage large aggregating proteoglycan, aggrecan: increased serum concentration in rheumatoid arthritis.

The large aggregating proteoglycan from human cartilage, aggrecan, has recently been shown to possess an immunologically detectable domain with close homology to epidermal growth factor (EGF), that is variably expressed by alternative mRNA splicing. Using a competitive ELISA we detected this domain in sera from both patients with RA and normal controls. The EGF-like domain could only be detected after digestion of sera with chondroitinase ABC, which demonstrates its proteoglycan origin. The concentration of the aggrecan EGF-like domain was considerably elevated in sera from patients with RA as compared to the control sera (P < 0.001), and the concentration of the domain increased with age in both the patient (r = 0.58, P < 0.05) and the control (r = 0.66, P < 0.01) group. These results demonstrate that the EGF-like domain of aggrecan could be of value as a marker of RA.

Adult↗
PubMed1993

Combined alcian blue and silver staining of subnanogram quantities of proteoglycans and glycosaminoglycans in sodium dodecyl sulfate-polyacrylamide gels.

Proteoglycans stain weakly in polyacrylamide gels by traditional protein stains such as coomassie brilliant blue or silver. In the present work preparations of large aggregating proteoglycan from human articular cartilage were used to evaluate a convenient staining method based on successive staining with alcian blue and neutral silver. The method is developed for sodium dodecyl sulfate-polyacrylamide gel electrophoresis; with ultrathin minigels in a semiautomated electrophoresis system, takes 1 1/2 h, and uses stable reagents. Preparations, electrophoresis, and staining of up to 24 samples can be completed within 2 1/2 h. The method has a detection limit of 0.04-1 ng proteoglycan and less than 0.5 ng of glycosaminoglycan. In addition the method can be adjusted for selective staining of proteoglycans and glycosaminoglycans.

Acrylic Resins↗