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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, or L-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.

Biochemical changes in rabbit sclera following destruction of pigment epithelium.

The long-term effect of destruction of the pigment epithelium by sodium iodate on the biochemistry of rabbit sclera was studied in one group with intravenous injection of sodium iodate, and in a second group with injection of sodium iodate into the right eye. Intravenous treatment produced a non-significant increase in the uronic acid concentration. In the second group the untreated fellow eye was microscopically intact, but was shown (as the treated eye) to alter the concentration of uronic acid in different parts of the sclera. All eyes from treated animals exhibited changes in the relative content of the various glycosaminoglycans and in the content of hydroxyproline, hydroxylysine and proline. This work indicates that the pigment epithelium may play a key role in the control mechanism of the scleral connective tissue, and this again has major implications in terms of a possible medical treatment of axial myopia.

The effect of hyaluronic acid on cartilage in the immobilized rabbit knee.

Out of 30 adult rabbits, 20 had one knee immobilized with a plaster of Paris cast for 6 or 12 weeks, and 10 rabbits were used as untreated controls. Prior to immobilization, 10 knees were injected with high-molecular weight hyaluronic acid. The articular cartilage of the femoral condyles was studied by light microscopy, whereas that of the patella and tibia was analyzed biochemically. Degenerative changes of the articular cartilage similar to those seen in arthrosis were observed after 6 weeks. The intraarticular injection of hyaluronic acid did not prevent these changes; instead, the reparative processes seemed inhibited.

Facet tropism: comparison of plain film and computed tomography examinations.

This study compares the findings of plain film X-ray and computed CT examination in the diagnosis of facet orientation and the presence of tropism. Twenty consecutive patients having lumbar disc disease with sciatica were studied utilizing plain X-ray as well as CT scanning. A chiropractic radiologist read the films to determine if facet facings were sagittally, semi-sagittally or coronally oriented on both CT and plain X-ray study. CT was accepted as the most accurate method to determine the true facet orientation, and plain X-ray interpretation of facet orientation was compared to the CT reading. There was a statistically significant relationship in diagnosing tropism between plain film X-ray and CT readings, with a predictive accuracy that ranged from 58-84% across the three segmental levels. However, the exact concordance of plain film X-ray and CT readings for right and left facet facings was very low. This raises the question of how the profession defines diagnostic accuracy.

Collagen and uronic acid distribution in the human sclera.

In the normal adult eye the content of uronic acid, hydroxyproline, hydroxylysine and proline from different parts of sclera was examined. Scleral samples from the posterior eyeball showed a higher content of uronic acid than the scleral samples from papilla, equator and limbus. Apart from a lower concentration of hydroxyproline in sclera from equator, no major differences in concentration of hydroxyproline, hydroxylysine or proline was found in sclera samples.

Regional glycosaminoglycans composition of the human sclera.

Human sclera from different parts of the normal adult eye was analyzed for content of uronic acid and the distribution of specific glycosaminoglycan types. Uronic acid is widely determined as representative of glycosaminoglycans in biological substances and variations of about 2-fold were found. Differences of up to 50% were detected in the relative abundance of hyaluronic acid and of dermatan sulphate, depending on the site on the globe from which the sample was taken. Sclera from around the papilla was found to be rich in content of dermatan sulphate. Sclera from the posterior eyeball showed a higher percentage of chondroitin sulphate as compared with sclera from equator and limbus. Equatorial sclera was richer in hyaluronic acid than posterior sclera. Sclera from the posterior eyeball showed a higher content of uronic acid as compared with sclera from equator and limbus. This work is an addition to the understanding of the scleral biochemistry and this finding may have some relevance in connection with ocular disease, as the proteoglycans and their content of glycosaminoglycans play a key role in the regulation of collagen fibril assembly and in the biomechanical strength of collagen fibrils.

Glycosaminoglycans in patellar cartilage.

We obtained samples of patellar cartilage at the time of surgery from 20 patients: 11 with normal and 9 with macroscopic cartilaginous degeneration. The glycosaminoglycans were separated by cellulose acetate electrophoresis, and the relative content of major glycosaminoglycans was measured by optic scanning. The total content of glycosaminoglycans was estimated by uronic acid analysis. We found that degenerated cartilage contained an increase in dermatan sulfate and hyaluronic acid and a decrease in chondroitin 6-sulfate and total uronic acid concentration, with only slight changes in chondroitin 4-sulfate and keratan sulfate compared with normal cartilage.

Glycosaminoglycans in human breast cancer.

Abnormal concentrations of glycosaminoglycans (GAG) have been reported for various types of tumors, suggesting that they may play a role in neoplasia. The correlation between the content of individual GAGs was studied in breast tumor tissues. The total content of GAG was estimated by uronic acid analysis. The relative distributions of dermatan sulphate, heparan sulphate, hyaluronic acid and chondroitin sulphate were measured after cellulose acetate electrophoresis. Mammary tissue samples were obtained at the time of surgery from 11 women, 6 with fibroadenomata and 5 with carcinoma. From each patient, biopsies were obtained centrally in the tumor and perilesional areas adjacent to the tumor, and also from clinically uninvolved tissue in the same region. In the central areas, it was found that carcinoma had a significant increase in chondroitin sulphate and uronic acid content, and a significant decrease in dermatan sulphate content, as compared with fibroadenoma. The chondroitin sulphate content in perilesional carcinomatous tissue was significantly greater than in clinically uninvolved tissue.

Short leg correction: a clinical trial of radiographic vs. non-radiographic procedures.

Visual leg length insufficiency detection and correction is compared with established radiographic procedures on 41 consecutive patients presenting to a chiropractic clinic with low back pain. It is commonly accepted that the most accurate procedure of short leg demonstration is the standing X ray. Visual correction, as described by Rene Cailliet, uses three anatomical points of reference: a) iliac crest levelness, b) vertical appraisal of the spine from the sacral base (the spine should be perpendicular to the sacral base) and c) levelness of the posterosuperior iliac spine (PSIS) dimples. Lifts of varying thickness were placed under the foot of the short leg in both leg length corrective procedures. This study found that the visual method of measurement did not differ significantly from the X-ray method of measurement for leg length insufficiency. Further, it was found that when comparing those in which the visual measure was less than the X-ray measure and those in which the visual measure was greater, there was a significant relationship between visual and X-ray measures. Eta (eta 2) demonstrates that there is a very strong relationship between visual and X-ray methods of measurement. A review of the literature is presented regarding the correlation of leg length insufficiency and musculoskeletal disorders, as well as the discrepancy required to alter biomechanical properties of the trunk and lower extremity.

Exercise and smoking habits in patients with and without low back and leg pain.

The exercise and smoking habits of low back or leg pain sufferers vs. persons not having low back or leg pain are compared. The type, frequency and length of exercise is determined from a study of 576 low back or leg pain sufferers compared to 50 persons who state they are symptomatic. The same was done for smoking habits. Thirty-three percent of low back or leg pain sufferers smoked as compared to 14% of those without pain. Forty-seven percent of low back or leg pain sufferers as compared to 86% of non-sufferers exercised regularly. The level of physical activity and general exercise have been found to improve strength, mobility and endurance; this might prevent future back injury. Our study is to determine differences in the exercise habits of persons with low back and/or leg pain vs. those who do not have pain, with the intention being to see if pain sufferers exercise less.