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Biomedical subjects

L Dahlberg

Publications and source records attributed to L Dahlberg.

At least 19 recordsLinked to original sources

Delayed gadolinium-enhanced MRI of cartilage (dGEMRIC): intra- and interobserver variability in standardized drawing of regions of interest.

PURPOSE: To establish the reproducibility of a standardized region of interest (ROI) drawing procedure in delayed gadolinium-enhanced magnetic resonance imaging (MRI) of cartilage (dGEMRIC). MATERIAL AND METHODS: A large ROI in lateral and medial femoral weight-bearing cartilage was drawn in images of 12 healthy male volunteers by 6 investigators with different skills in MRI. The procedure was done twice, with a 1-week interval. Calculated T1-values were evaluated for intra- and interobserver variability. RESULTS: The mean interobserver variability for both compartments ranged between 1.3% and 2.3% for the 6 different investigators without correlation to their experience in MRI. Post-contrast intra-observer variability was low in both the lateral and the medial femoral cartilage, 2.6% and 1.5%, respectively. The larger variability in lateral than in medial cartilage was related to slightly longer and thinner ROIs. CONCLUSION: Intra-observer variability and interobserver variability are both low when a large standardized ROI is used in dGEMRIC. The experience of the investigator does not affect the variability, which further supports a clinical applicability of the method.

Adult↗

Development of methods for the determination of vitamins A, E and beta-carotene in processed foods based on supercritical fluid extraction: a collaborative study.

New methodologies based on supercritical fluid extraction (SFE) have been developed for the determination of fat-soluble vitamins in processed foods. The results obtained so far indicate that SFE is well suited to extraction of fat-soluble vitamins from food products, although validation work is required to establish accuracy and precision. The vitamins investigated were A, E and beta-carotene, and the processed foods were UHT milk, milk powder, minced meat, liver paste, infant formula, canned baby food and margarine. Extraction equipment employed analyte collection on either a solid-phase trap or in a solvent. After extraction, the samples were saponified and the vitamins determined using reversed-phase liquid chromatography with ultraviolet or fluorescence detection. Sample throughput was at least 12 samples day(-1), i.e. at least twice the number achievable with a conventional extraction methodology. The detection limits for the vitamins in different processed foods were well below 0.1 microg g(-1). Recoveries (in comparison with vitamin levels obtained using conventional solvent extraction) were close to 100% for experienced personal with access to modern automatic equipment. To reach this level, it was necessary to protect the vitamins with an antioxidant during the different steps of the analysis procedure, to add methanol or ethanol to the extraction cell to facilitate the analyte extraction from the food matrix, and when using a solid-phase trap, to employ a fractionated extraction-elution procedure to prevent breakthrough losses. The developed methods were tested in a validation exercise between five laboratories, which had taken part in the method development, and in an intercomparison between 10 laboratories including laboratories with less experience of vitamin determination. The within-laboratory RSD was generally < or = 11%. The average of the between-laboratory relative standard deviation (RSD) was about 23% in the validation, and increased to about 40% in the intercomparison. Ruggedness tests performed at different steps of the project showed that different types and models of equipment did not give large differences in recoveries. Thus, the increasing RSD can largely be ascribed to differences in experience in vitamin analysis of the participants.

Chromatography, Supercritical Fluid↗

Gd-DTPA2)-enhanced MRI of femoral knee cartilage: a dose-response study in healthy volunteers.

The negatively charged contrast agent Gd-DTPA2- distributes inversely to the cartilage fixed charged density. This enables structural cartilage examinations by contrast-enhanced MRI. In line with the development of a clinically applicable protocol for such examinations, this study describes the temporal pattern of Gd-DTPA2- distribution in femoral knee cartilage at three different doses in healthy volunteers. Nineteen volunteers (ages 21-28 years) were examined with a 1.5T MRI system. Quantitative relaxation rate measurements were made in weight-bearing central parts of femoral cartilage using sets of five turbo inversion recovery images with different inversion times. The cartilage was analyzed before and four times (1-4 h) after an intravenous injection of Gd-DTPA2- at single, double, and triple doses: 0.1, 0.2, and 0.3 mmol/kg body weight, respectively. The increase in R1 postcontrast was linearly dose-related at all times. The highest R1 values were registered at 2 and 3 h postcontrast, suggesting 2 h to be optimal in the clinical situation. The triple dose indicated a subtle compartmental difference in men, with higher contrast distribution medially than laterally. Results suggest that the triple dose is needed to detect minor cartilage matrix differences.

Adult↗

Determination of food constituents based on SFE: applications to vitamins A and E in meat and milk.

A method has been developed for the determination of vitamins A and E in food using supercritical fluid extraction (SFE), applying liquid or solid trapping, with an accuracy equal to conventional solvent extraction methods. Under optimal conditions, using methanol modified carbon dioxide as a supercritical fluid, Hydromatrix as a water adsorbent, and with a small amount of ascorbic acid and methanol added to the sample, the extraction time is reduced to 80 min. This time is considerably shorter than in conventional methods. Other advantages are the reduction of manual manipulations leading to lower labour costs and reduced consumption of organic solvents in the sample preparation step.

Animals↗

Comparison of the degradation of type II collagen and proteoglycan in nasal and articular cartilages induced by interleukin-1 and the selective inhibition of type II collagen cleavage by collagenase.

OBJECTIVE: To compare interleukin-1alpha (IL-1alpha)-induced degradation of nasal and articular cartilages in terms of proteoglycan loss and type II collagen cleavage, denaturation, and release; to examine the temporal relationship of these changes; and to investigate the effects of an inhibitor of collagenase 2 and collagenase 3 on these catabolic processes. METHODS: Discs of mature bovine nasal and articular cartilages were cultured with or without human IL-1alpha (5 ng/ml) with or without RS102,481, a selective synthetic inhibitor of collagenase 2 and collagenase 3 (matrix metalloproteinase 8 [MMP-8] and MMP-13, respectively) but not of collagenase 1 (MMP-1). Immunoassays were used to measure collagenase-generated type II collagen cleavage neoepitope (antibody COL2-3/4C(short)) and denaturation (antibody COL2-3/4m), as well as total type II collagen content (antibody COL2-3/4m) in articular cartilage and culture media. A colorimetric assay was used to measure total proteoglycan concentration (principally of aggrecan) as sulfated glycosaminoglycans (sGAG). RESULTS: IL-1alpha initially induced a decrease in tissue proteoglycan content in nasal cartilage. A progressive loss of proteoglycan was noted during culture in articular cartilages, irrespective of the presence of IL-1alpha. In both cartilages, proteoglycan loss was followed by IL-1alpha-induced cleavage of type II collagen by collagenase, which was often reflected by increased denaturation. The inhibitor RS102,481 had no clear effect on the reduction in proteoglycan content (measured by sGAG) and collagen denaturation in either cartilage, but at 10 nM it inhibited the enhanced cleavage of type II collagen, partially in nasal cartilage and completely in articular cartilage. CONCLUSION: IL-1alpha-induced cleavage and denaturation of type II collagen is observed in both hyaline cartilages and is secondary to proteoglycan loss. It probably involves different collagenases, since there is no evidence of a rate-limiting role for collagenase 1 in articular cartilage, unlike the case for nasal cartilage. Inhibitors of this kind may be of value in the treatment of cartilage damage in arthritis. Also, the ability to detect the release of type II collagen collagenase-generated fragments from degraded cartilage offers the potential to monitor cartilage collagen damage and its control in vivo.

Animals↗

Selective enhancement of collagenase-mediated cleavage of resident type II collagen in cultured osteoarthritic cartilage and arrest with a synthetic inhibitor that spares collagenase 1 (matrix metalloproteinase 1).

OBJECTIVE: To examine whether type II collagen cleavage by collagenase and loss of proteoglycan are excessive in human osteoarthritic (OA) articular cartilage compared with nonarthritic articular cartilage, and whether this can be inhibited by a selective synthetic inhibitor that spares collagenase 1 (matrix metalloproteinase 1 [MMP-1]). METHODS: Articular cartilage samples were obtained during surgery from 11 patients with OA and at autopsy from 5 adults without arthritis. The articular cartilage samples were cultured in serum-free medium. A collagenase-generated neoepitope, which reflects cleavage of type II collagen, and proteoglycan glycosaminoglycan (GAG), which predominantly reflects aggrecan release, were assayed in culture media. In addition, cultures were performed using either of 2 synthetic MMP inhibitors, both of which inhibited collagenase 2 (MMP-8) and collagenase 3 (MMP-13), but one of which spared collagenase 1. Cultures were also biolabeled with 3H-proline in the presence and absence of these inhibitors to measure collagen synthesis (as tritiated hydroxyproline) and incorporation in articular cartilage. RESULTS: As a group, cleavage of type II collagen by collagenase was significantly increased in OA cartilage samples. In contrast, proteoglycan (GAG) release was not increased. This release of a collagenase-generated epitope was inhibited by both MMP inhibitors in 2 of 5 nonarthritic samples and in 9 of 11 OA cartilage samples. The inhibitor that spared collagenase 1 was generally more effective and inhibited release from 4 of 5 nonarthritic cartilage samples and the same OA cartilage samples. Group analyses revealed that the inhibition of collagenase neoepitope release by both inhibitors was significant in the OA patient cartilage, but not in the nonarthritic cartilage. Proteoglycan loss was unaffected by either inhibitor. Newly synthesized collagen (predominantly, type II) exhibited increased incorporation in OA cartilage, but only in the presence of the inhibitor that arrested collagenase 1 activity. CONCLUSION: These results further indicate that the digestion of type II collagen by collagenase is selectively increased in OA cartilage, and that this can be inhibited in the majority of cases by a synthetic inhibitor that can inhibit collagenases 2 and 3, but not collagenase 1. The results also suggest that in OA, newly synthesized collagen is digested, but in a different manner than that of resident molecules. Proteoglycan release was not increased in OA cartilage and was unaffected by these inhibitors. Inhibitors of this kind may be of value in preventing damage to type II collagen in human arthritic articular cartilage.

Adult↗

Genetic and behavioral risk factors for self-reported joint pain among a population-based sample of Swedish twins.

Self-reported joint pain, a typical manifestation of osteoarthritis, was examined using 335 twin pairs from the Swedish Adoption/Twin Study of Aging to estimate relative genetic and environmental influences on self-reported joint pain and to examine the relationships between joint pain, health behavior, and psychological variables. Findings suggest that family resemblance for self-reported joint pain represents similar environments more than genetic similarity. Data from the early 1970s, including exercise, physical activity at work, obesity, and neuroticism, were used to predict joint pain in 1993. For men, moderate amounts of exercise decreased the likelihood of joint pain, but strenuous amounts of physical activity in the workplace had the opposite effect. For women, exercise and physical activity were not significant predictors, but past obesity and higher levels of neuroticism increased the likelihood of reporting joint pain in 1993.

Aged↗

High prevalence and incidence of sexually transmitted diseases in urban adolescent females despite moderate risk behaviors.

To better understand the prevalence, incidence, and risk factors for sexually transmitted diseases (STDs) among female adolescents, a prospective 6-month cohort study was conducted at four teen clinics in a southeastern city. At enrollment, 260 (40%) of 650 sexually active females ages 14-19 years had an STD: chlamydia, 27%; herpes simplex virus type 2 (HSV-2), 14%; gonorrhea, 6%; trichomoniasis, 3%; and hepatitis B, 2%. At follow-up, 112 (23%) of 501 participants had an incident infection: chlamydia, 18%; HSV-2, 4%; gonorrhea, 4%; and trichomoniasis, 3%. At either enrollment or follow-up, 53% had >/=1 STD; of those with 1 lifetime partner, 30% had an STD. Having a new partner (odds ratio [OR], 2.2; 95% confidence interval [CI], 1. 1-4.2) or friends who sell cocaine (OR, 1.6; CI, 1.0-2.6) was independently associated with incident infection. STD incidence and prevalence were extremely high in this population, even in teenagers with only 1 lifetime partner. Individual risk behaviors appeared less important for STD risk than population factors.

Adolescent↗

Causes of strain affecting relatives of Swedish oldest elderly. A population-based study.

We examined determinants of carer strain in a random sample of 170 relative of dependent elderly persons in Sweden aged 80 years or more. These persons were interviewed at home by means of the Sickness Impact Profile (SIP). Carer strain was assessed in interviews with carers who bore the main responsibility for care of an elderly relative. The response rate was 92%. Multiple logistic regression showed that cohabitation of carer and recipient was the sociodemographic variable most significantly associated with various stress factors and consequences or manifestations of stress; strain connected with ADL assistance given to the elderly person, strain arising from the elderly person's uncooperative or recalcitrant attitude or behaviour, worsening of the elderly-carer relationship, and encroachment on the carer's social life. Correlations between strain and various categories of physical and mental impairment were much lower, and in some respects were even negative. We conclude that support programs for relatives caring for dependent elderly persons should aim primarily at creating continuous relief for the carers and improving their opportunities for contact with other people generally.

Aged↗

Evidence for altered synthesis of type II collagen in patients with osteoarthritis.

There is evidence to suggest that the synthesis of type II collagen is increased in osteoarthritis (OA). Using an immunoassay, we show that the content of the C-propeptide of type II procollagen (CPII), released extracellularly from the newly synthesized molecule, is directly related to the synthesis of this molecule in healthy and osteoarthritic articular cartilages. In OA cartilage, CPII content is often markedly elevated (mean 7.6-fold), particularly in the mid and deep zones, reaching 29.6% of the content in newborn. Synthesis is also directly related to total collagen II content in OA, suggesting its importance in maintaining collagen content and cartilage structure. The release of CPII from cartilage is correlated directly with cartilage content. However, the increase in CPII in OA cartilage is not reflected in serum, where a significant reduction is observed. Together these studies provide evidence for alterations in procollagen II synthesis in vivo in patients with OA.

Adult↗

Enzymatic specificity and hydrolysis pattern of the catalytic domain of the xylanase Xynl from Rhodothermus marinus.

The catalytic domain of a xylanase from Rhodothermus marinus was produced in Escherichia coli. The catalytic domain belongs to glycosyl hydrolase family 10. The produced protein has a 22-amino acid leader peptide followed by a 411-amino acid truncated xylanase. The molecular mass was 48 kDa and the recombinant xylanase had a pI of 4.9. The pH and temperature optima for activity were determined to be 7.5 and 80 degrees C, respectively. At that temperature the enzyme had a half-life of 1 h 40 min. An addition of 1 mM calcium stabilized the activity of the enzyme at 80 degrees C. The xylanase had its highest specific activity on oat spelt xylan but was active also on other xylans and to a limited extent on some other polysaccharides (soluble glucans). No exo- or endo-cellulase activity was observed. Hydrolysis of xylo-oligomers and oat spelt xylan was studied and the predominant products of hydrolysis were xylobiose and xylotriose. The enzyme was inactive on xylobiose, xylotriose and on the soluble fraction from oat spelt xylan. The R. marinus xylanase is shown to have a strong preference for internal linkages and is therefore classified as an endo-xylanase.

Amino Acid Sequence↗

Longitudinal and cross-sectional variability in markers of joint metabolism in patients with knee pain and articular cartilage abnormalities.

OBJECTIVE: To determine the within- and between-patient variability in the concentrations of synovial fluid, serum and urine markers of joint tissue metabolism in a cohort of patients with knee pain and cartilage changes consistent with early-stage knee osteoarthritis. DESIGN: Samples of synovial fluid, serum, and urine were obtained from 52 patients on eight different occasions during 1 year, as part of a clinical trial in patients with cartilage abnormalities and knee pain. In joint fluid, aggrecan fragments were quantified by dye precipitation and enzyme-linked immunosorbent assay (ELISA), and matrix metalloproteinases-1 and -3, and tissue inhibitor of metalloproteinases-1 by sandwich ELISAs. In serum, keratan sulfate was quantified by ELISA. Type I collagen N-telopeptide cross-links in urine were determined by ELISA. RESULTS: The degree of cross-sectional variability in marker concentrations did not vary between the different sampling occasions, and did not differ between the periods of weeks 0 (baseline), 1-4 (treatment) and 13-26 (follow-up). Both between-patient and within-patient coefficients of variation varied for markers in different body fluid compartments, with the lowest variability for serum keratan sulfate, followed by urine type I collagen N-telopeptide crosslinks, and the highest for synovial fluid markers. For synovial fluid, aggrecan fragments showed the least variability, and matrix metalloproteinases the highest. One patient with septic arthritis showed a fivefold peak increase in joint fluid aggrecan fragment concentrations, while the concentration of matrix metalloproteinase-3 increased 100-fold. CONCLUSIONS: Molecular markers of joint tissue metabolism have been suggested as, for example, outcome measures for clinical trials of disease-modifying drugs in osteoarthritis. This report is the first to present data on between- and within-patient variability for such molecular markers in three different body fluid compartments in stable cohort of patients. The availability of such data enables calculations to determine the number of patients needed in prospective studies using these markers as outcome measures.

Adult↗

Enhanced cleavage of type II collagen by collagenases in osteoarthritic articular cartilage.

We demonstrate the direct involvement of increased collagenase activity in the cleavage of type II collagen in osteoarthritic human femoral condylar cartilage by developing and using antibodies reactive to carboxy-terminal (COL2-3/4C(short)) and amino-terminal (COL2-1/4N1) neoepitopes generated by cleavage of native human type II collagen by collagenase matrix metalloproteinase (MMP)-1 (collagenase-1), MMP-8 (collagenase-2), and MMP-13 (collagenase-3). A secondary cleavage followed the initial cleavage produced by these recombinant collagenases. This generated neoepitope COL2-1/4N2. There was significantly more COL2-3/4C(short) neoepitope in osteoarthritis (OA) compared to adult nonarthritic cartilages as determined by immunoassay of cartilage extracts. A synthetic preferential inhibitor of MMP-13 significantly reduced the unstimulated release in culture of neoepitope COL2-3/4C(short) from human osteoarthritic cartilage explants. These data suggest that collagenase(s) produced by chondrocytes is (are) involved in the cleavage and denaturation of type II collagen in articular cartilage, that this is increased in OA, and that MMP-13 may play a significant role in this process.

Adult↗

Osteoarthritis of the knee after injury to the anterior cruciate ligament or meniscus: the influence of time and age.

The degree of cartilage changes by arthroscopic and radiologic examination was evaluated in a retrospective, cross-sectional study of 1012 patients with knee complaints at different times after knee injury. Patients were classified by age, sex, trauma date and type of injury. The condition of the cartilage was graded on a 1-10 scale based on findings at arthroscopy and on weight-bearing radiographs. Patients with injury to the anterior cruciate ligament (isolated or combined with injury to meniscus or collateral ligaments) showed the first radiologic signs (joint space narrowing) of osteoarthritis (OA) at an average age of about 40 years, while patients with isolated meniscus injury had the same stage of disease at an average age of about 50 years. Both study groups displayed the first radiologic signs of OA on average about 10 years after the injury and showed increasingly serious arthroscopic and radiologic signs of joint damage with increased time between injury and examination. For patients who sustained an isolated meniscus injury between the ages of 17 and 30, the average time until development of radiologic signs of OA was about 15 years, while for those who had the same injury over the age of 30, the corresponding time interval was only about 5 years. We conclude from this that knee OA becomes increasingly severe with increased time between joint injury and examination. OA changes appear sooner in older patients with knee injury than in the young.

Adolescent↗

Markers of cartilage matrix metabolism in human joint fluid and serum: the effect of exercise.

The concentrations of cartilage proteoglycan (aggrecan), stromelysin-1, tissue inhibitor of metalloproteinases-1 (TIMP-1) and procollagen II C-propeptide in knee joint fluid and the levels of aggrecan, hyaluronan and keratan sulfate in serum were measured before and after exercise in 33 healthy athletes. The samples before exercise were obtained after 24 h rest from running or soccer and the samples after exercise were obtained 30-60 min after the exercise. Nine athletes ran on a treadmill for 60 min, 16 ran on road for 80 min and 8 played one soccer game (90 min). A reference group of 28 patients with knee pain but not evidence of joint pathology or injury was used for comparison. In joint fluid no single marker from the degradative processes in cartilage matrix changed significantly with exercise but all showed a rising trend. All markers except stromelysin showed lower concentrations in athletes at rest compared to the reference group. In serum from runners before exercise the concentration of keratan sulfate was significantly higher than in both the soccer and reference groups and further increased after exercise. The increase in markers after exercise may reflect an effect of mechanical loading in combination with a possible high turnover rate of body cartilage matrix in these individuals.

Adult↗