Søren Ribel-Madsen

Effect of Day Length on Eye Growth, Myopia Progression, and Change of Corneal Power in Myopic Children

OBJECTIVE Because of the northern location of Denmark, the length of the day over the year varies from 7 to 17.5 hours. Experimental and clinical results suggest that the development of myopia may be related to ambient light exposure. The purpose of current study was to investigate whether axial eye growth, myopia progression, or corneal power change in Danish myopic children varies with the length of the day. DESIGN Cross-sectional study. PARTICIPANTS Two hundred thirty-five children 8 to 14 years of age found to have myopia during screening for a clinical trial (ClinicalTrial.gov identifier, NCT00263471; accessed December 6, 2005). All children found to have any value of spherical equivalent that was myopic (<0 diopters [D]) at the first of 2 visits were included. METHODS Cycloplegic refraction was measured using an autorefractor, axial eye length, and corneal power using an automatic combined noncontact partial coherence interferometer and keratometer. The accumulated number of daylight hours during the measurement period was calculated for each participant using an astronomical table. MAIN OUTCOME MEASURES Change over 6 months in axial length, refraction, and corneal power. RESULTS Accumulated hours of daylight ranged from 1660 to 2804 hours. Significant correlations were found between hours of daylight and eye elongation (P = 0.00), myopia progression (P = 0.01), and corneal power change (P = 0.00). In children with an average of 2782 ± 19 hours of daylight, axial eye growth was 0.12 ± 0.09 mm, myopia progression was 0.26 ± 0.27 D, and corneal power change was 0.05 ± 0.10 D per 6 months, whereas in children with an average of 1681 ± 24 hours of daylight, axial eye growth was 0.19 ± 0.10 mm, myopia progression was 0.32 ± 0.27 D, and corneal power change was -0.04 ± 0.08 D per 6 months. CONCLUSIONS Eye elongation and myopia progression seem to decrease in periods with longer days and to increase in periods with shorter days. Children should be encouraged to spend more time outside during daytime to prevent myopia. FINANCIAL DISCLOSURE(S) The author(s) have no proprietary or commercial interest in any materials discussed in this article.

Biochemical and ultrastructural changes in rabbit sclera after treatment with 7-methylxanthine, theobromine, acetazolamide, or L-ornithine

AIMS To examine a possible effect of 7-methylxanthine, theobromine, acetazolamide, orl-ornithine on the ultrastructure and biochemical composition of rabbit sclera. METHODS Groups of pigmented rabbits, six in each group, were dosed during 10 weeks with one of the substances under investigation, and one untreated group was the control. Samples of anterior and posterior sclera were taken for determination of hydroxyproline, hydroxylysine, proline, proteoglycans, uronic acids and dermatan sulphate, chondroitin sulphate, and hyaluronic acid. Sections were examined with electron microscopy, and the diameter of the individual collagen fibrils was measured. RESULTS Treatment with theobromine produced a significant increase in the contents of hydroxylysine, hydroxyproline, and proline in both anterior and posterior sclera, while 7-methylxanthine increased the contents of hydroxyproline and proline selectively in posterior sclera. Acetazolamide, on the other hand, significantly decreased the contents of hydroxyproline and proline in samples from anterior sclera. Uronic acids in both anterior and posterior sclera were significantly reduced by treatment with 7-methylxanthine, and l-ornithine significantly reduced uronic acids in posterior sclera. An inverse correlation between contents of hydroxyproline and uronic acids was found. The mean diameter of collagen fibrils was significantly higher in the posterior sclera from rabbits treated with 7-methylxanthine or theobromine, and significantly lower in rabbits treated with acetazolamide or l-ornithine compared with controls. In the anterior sclera, fibril diameter was significantly reduced in all treatment groups compared with controls. A positive, significant correlation between fibril diameter and content of hydroxyproline and proline was found in posterior sclera. CONCLUSION 7-Methylxanthine, a metabolite of caffeine, increases collagen concentration and the diameter of collagen fibrils in the posterior sclera, and may be useful for treatment or prevention of conditions associated with low level and/or inferior quality of scleral collagen, such as axial myopia, chronic open angle glaucoma, and possibly neovascular age related macular degeneration. The apparent loss of collagen induced by chronic treatment with acetazolamide should be taken into consideration as a potentially harmful side effect. These results may indicate that scleral biochemistry and ultrastructure are influenced by the retinal pigment epithelium. One possible explanation is that the scleral fibroblasts which produce the collagen are sensitive to changes in the physiological electric field created by the retinal pigment epithelium.

Cytokine measurements and possible interference from heterophilic antibodies--problems and solutions experienced with rheumatoid factor.

Cytokines are important in the understanding of the immune process in health and disease and are valuable indicators in diagnostics. Measurements of cytokines are based on immunometric methods, and it is important to understand possible pitfalls in these methods to produce reliable cytokine data. This paper focuses on obtaining optimal measurements when applying enzyme-linked immunosorbent assay (ELISA) or multiplex immunoassays (MIA). Cytokines are measured in serum or plasma, as well as in various other body fluids, all containing a series of antibodies and the possibility of interference from these. Some antibodies, such as heterophilic and human anti-animal antibodies, are able to interfere with all immunoassays, but the immunometric techniques are most prone to serious interference from this source. Another type, rheumatoid factor (RF) is a composite of different autoimmune antibodies which can be present in both blood and synovial fluid. RF is present in some arthritic diseases as well as in some other medical conditions. When present, especially RF IgM is known to interfere with the immunometric measurements. A possible and affordable solution to diminish this interference is PEG precipitation, but other efficient, but more expensive, methods, such as precipitation using Protein L or commercially available blocking agents, are also available. Interference of RF is at present not tested in all cytokine assays, but degree of interference from RF, human anti-animal and heterophilic antibodies, as well as from other possible disease-specific antibodies, must always be considered when developing and applying new assays for cytokine measurements.

A Synoviocyte Model for Osteoarthritis and Rheumatoid Arthritis: Response to Ibuprofen, Betamethasone, and Ginger Extract—A Cross-Sectional In Vitro Study

This study aimed at determining if synovial cell cultures from rheumatoid arthritis (RA), osteoarthritis (OA), and healthy controls (HC) differ and are suitable disease models in pharmacological studies, and tested their response to some anti-inflammatory drugs. Synovial cells were isolated from synovial membrane or joint fluid. Cells were cultivated and exposed to no or TNF-α stimulation without, or in the presence of, betamethasone, ibuprofen, or a standardized ginger extract. Concentrations of a panel of cytokines, growth factors, and chemokines were mapped for each culture and condition. Our cells secreted an increased amount of the cytokines IL-1β, IL-6, and IL-8 in response to TNF-α stimulation in all conditions. OA cells showed a higher IL-6 and IL-8 and a lower IL-1β production, when not stimulated, than RA and HC cells, which were similar. TNF-α stimulation caused similar IL-1β, IL-6, and IL-8 release in all groups. Ibuprofen showed no effect on cytokine production, while ginger extract was similar to betamethasone. Ginger extract was as effective an anti-inflammatory agent as betamethasone in this in vitro model. Cultured fibroblast-like synoviocytes from OA and RA subjects promise to be a useful pharmacological disease model, but further studies, to support results from such a model are needed.

Rheumatoid Factor and Its Interference with Cytokine Measurements: Problems and Solutions

Use of cytokines as biomarkers for disease is getting more widespread. Cytokines are conveniently determined by immunoassay, but interference from present antibodies is known to cause problems. In rheumatoid arthritis (RA), interference of rheumatoid factor (RF) may be problematic. RF covers a group of autoantibodies from immunoglobulin subclasses and is present in 65–80% of RA patients. Partly removal of RF is possible by precipitation. This study aims at determining the effects of presence of RF in blood and synovial fluid on cytokine measurements in samples from RA patients and finding possible solutions for recognized problems. IL-1β, IL-4, IL-6, and IL-8 were determined with multiplex immunoassays (MIA) in samples from RA patients prior to and after polyethylene glycol (PEG 6000) precipitation. Presence of RF does interfere with MIA. PEG 6000 precipitation abolishes this RF interference. We recommend PEG precipitation for all immunoassay measurements of plasma samples from RA patients.

Expression of full-length and splice forms of FoxP3 in rheumatoid arthritis

Objective: The aim of our study was to compare the presence of full-length and alternative splice forms of FoxP3 mRNA in CD4 cells from rheumatoid arthritis (RA) patients and healthy controls. Methods: A quantitative real-time polymerase chain reaction (QRT-PCR) method was used to measure the amount of FoxP3 mRNA full-length and splice forms. CD4-positive T cells were isolated from peripheral blood from 50 RA patients by immunomagnetic separation, and the FoxP3 mRNA expression was compared with the results from 10 healthy controls. Results: We observed an increased expression of full-length FoxP3 mRNA in RA patients when compared to healthy controls, as well as an increase in CD25 mRNA expression, but no corresponding increase in CTLA-4 mRNA expression. The presence of an alternative splice form of FoxP3 lacking exon 2 was confirmed in both RA patients and healthy controls, but with no significant difference in expression between the two groups. There was a positive correlation between the amount of FoxP3 mRNA and the clinical inflammation parameters C-reactive protein (CRP) and erythrocyte sedimentation rate (ESR), and a negative correlation between FoxP3 mRNA and the dose of methotrexate (MTX) given to the patients. Conclusion: RA patients express more full-length FoxP3 than healthy controls in peripheral blood CD4-positive cells, suggesting an increased number of regulatory T cells (Tregs). However, no concomitant increase in CTLA-4 expression was seen. We therefore propose that the Tregs are left unable to suppress the ongoing inflammation due to a deficiency in CTLA-4 needed for cell contact-dependent suppression.

FoxP3 mRNA splice forms in synovial CD4+ T cells in rheumatoid arthritis and psoriatic arthritis.

Ryder LR, Bartels EM, Woetmann A, Madsen HO, Ødum N, Bliddal H, Danneskiold‐Samsøe B, Ribel‐Madsen S, Ryder LP. FoxP3 mRNA splice forms in synovial CD4+ T cells in rheumatoid arthritis and psoriatic arthritis. APMIS 2012; 120: 387–96.

Urinary markers of altered collagen metabolism in fibromyalgia patients

Objective: To assess the metabolism of collagen in fibromyalgia (FM) patients, and to compare the occurrence of collagen metabolism markers to the severity of FM symptoms. Methods: Morning urine was collected from 27 FM women fulfilling the American College of Rheumatology (ACR) criteria for FM, and from seven controls. FM patients completed the Fibromyalgia Impact Questionnaire (FIQ). Bone mineral density (BMD), isokinetic muscle strength in knee and elbow, and hand‐grip strength were measured. Urinary concentrations of collagen type I cross‐linked C‐telopeptide (CTX‐I) and collagen type II cross‐linked C‐telopeptide (CTX‐II) were determined by enzyme‐linked immunosorbent assay (ELISA). Pyridinoline (Pyd) and deoxypyridinoline (Dpd) were determined by liquid chromatography, and hydroxyproline (Hyp) by spectrophotometry. All concentration data were normalized to creatinine. Results: Mean values in the FM group and the control group, respectively, were: urinary CTX‐I 246.8 and 337.5 µg/mmol (p = 0.060); CTX‐II 110.4 and 185.1 ng/mmol (p = 0.035); Pyd 56.1 and 52.3 nmol/mmol (NS); Dpd 15.1 and 14.0 nmol/mmol (NS); Pyd : Dpd ratio 4.05 and 3.96 (NS); Hyp 26.1 and 21.1 µmol/mmol (NS). Significant inverse correlations were seen between CTX‐I and the intensity of fatigue, and between CTX‐II and anxiety. An inverse correlation between CTX‐I and muscle strength was apparent, but relied on extreme values from one patient, and no significant correlation was found between CTX‐I or CTX‐II and tender points or BMD in the FM group. Conclusions: Low urinary concentrations of CTX‐II and CTX‐I and normal levels of Pyd and Dpd were found in FM, but their relationship to the intensity of FM symptoms was unclear.

Clinical Performance of the ASR and ReCap Resurfacing Implants—7 Years Follow-Up

We perform a non-randomized, consecutive pilot study on the ASR and ReCap resurfacing hip implants and have completed 7 years follow-up. Forty-six non-osteoporotic patients with hip osteoarthritis and anatomical conditions suitable for resurfacing were divided into 2 equal groups and operated sequentially, starting with the ASR implants. Sixteen patients operated with ASR and 19 patients with ReCap have been followed-up. There were no significant differences between the two groups preoperatively as to physical function, pain, or femoral BMD. The serum concentrations of cobalt and chromium were higher in the ASR group from 1/2 to 7 years postoperatively. Five of 16 ASR implants have been revised, and none of the ReCap implants. BMD below the femoral component increased in both groups.

Latanoprost eye drops increase concentration of glycosaminoglycans in posterior rabbit sclera.

PURPOSE Glycosaminoglycans are important components of ocular tissues such as the sclera. The pressure reducing effect of a new antiglaucoma drug, latanoprost, is based on an increase in the uveo-scleral outflow by way of modulation of the intracellular matrix of the ciliary body. The purpose of the study was to test the effect of latanoprost on the content of glycosaminoglycans in rabbit cornea and sclera. METHODS Twelve rabbits were studied. Six rabbits were treated for 12 weeks with latanoprost eye drops and 6 with hydroxypropylmethylcellulose, dextran 70 eye drops for control. Samples were taken from cornea and anterior, lateral, and posterior sclera. Glycosaminoglycans were determined quantitatively by spectrophotometry (uronic acids). RESULTS A significant increase in the concentration of uronic acids was found in all three localisations of sclera from latanoprost-treated animals. The increase was 26%, 24%, and 20% in anterior, lateral, and posterior sclera, respectively. CONCLUSIONS Long-term treatment with latanoprost induces biochemical changes in sclera. The results indicate that topically applied latanoprost reaches the posterior parts of the rabbit eye.