Thorkil Ammitzbøll

CHANGES IN THE CONTENT AND COMPOSITION OF COLLAGEN IN THE GLAUCOMATOUS EYE - BASIS FOR A NEW HYPOTHESIS FOR THE GENESIS OF CHRONIC OPEN ANGLE GLAUCOMA - A Preliminary Report

The pressure theory is still predominant in explaining the pathophysiology of the chronic open angle glaucoma. An insufficient drainage system resulting in an increased intraocular pressure is the hasis for this theory. The pressure will exert an effect upon the optic disc which either directly on the nerve fibres or indirectly via the vascular system will result in a characteristic optic atrophy. The collagen fibres, both in the trabeular meshwork of the anterior chamber and in the lamina cribrosa of the optic disc, form a mesh through which the aqueous humour and the nerve fibres, respectively, pass through the wall of the eye. A hypothesis explaining the pathophysiology of this disease, and based on the assumption that there is a primary change in the collagen molecules, resulting in a weaker structure than normal both in the trabeculae and in the laminae, is forwarded. The structures analysed for the content of hydroxypro‐line, hydroxylysine and proline were the trabecular meshwork, the sclera and the lamina cribrosa. Three categories of autopsy eyes were studied, i.e. normal eyes, glaucomatous eyes, and eyes under a suspicion of glaucoma. In the normal eyes, the collagen composition in the trabecular meshwork was different from that in the sclera and the lamina cribrosa. There is also a difference in the composition between the sclera and the lamina cribrosa. In glaucoma, the content and/Or the composition of the collagen molecules in the lamina were significantly changed. In the eyes under suspicion of glaucomathe same changes as in the glaucomatous eyes could be demonstrated. However, 5 of the 7 eyes in this category had no demonstrable nerve atrophy. The findings suggest that the change in collagen pattern is primary. This study has not demonstrated which types of collagen are present or the physical properties of this collagen. Further tests to demonstrate the different types of collagen and their rigidity are planned.

Regional glycosaminoglycans composition of the human sclera

Abstract 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 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.

Electrically-induced collagen calcification in pig skin. A histopathologic and histochemical study

Deposition of calcium salts on collagen fibres in skin of fully anaesthetized pigs was induced by exposure to direct current (d.c.). In biopsies obtained from cathode areas successively from day 1 to day 7 after exposure the histopathologic and histochemical changes before and after the initial deposition of calcium salts have been examined. For comparison skin sites with intradermal injected calcium hydroxyapatite crystals were studied in addition. Small areas of calcified collagen and elastic fibres were noted in viable tissue 2 days after d.c. exposure. In succeeding days the calcified areas enlarged with new deposits always more superficial and closer to the epidermis than the original calcium deposits. Preconditions for calcification appear to be (1) a pH change in basic direction and/or the electrochemical processes specific to the cathode area and (2) a viable tissue. Elastic fibres appear to have a lower calcification threshold than collagen fibres. A positive staining for glycoproteins (PAS) and glycosaminoglycans (alcian blue pH 2.5) was noted in the calcified collagen fibres simultaneously with the calcification. In succeeding days the intensity of the staining reactions increased. Whether changes in the glycoproteins, collagen and its intimately bound glycosaminoglycans precede the calcification or the staining reactions develop secondarily to this deposition is not known. However, seven days after intradermal injections of Ca-apatite crystals in pig skin small and large crystals were observed ultrastructurally without any relation to collagen fibrils, but the calcified tissue presented a positive PAS and alcian blue reaction from day 2. Thus the PAS and alcian blue stainings in this model develop secondary to the deposition of calcium salts.

A study of cell proliferation kinetics in the small intestinal epithelium of psoriasis patients

Small intestinal biopsies from psoriasis patients and from normal controls were obtained with a multipurpose suction tube. The tissue was incubated with tritiated thymidine and processed for autoradiography.

Bioavailability of D-penicillamine in relation to gastrointestinal involvement of generalized scleroderma.

The bioavailability of D-penicillamine was measured in 24 patients with generalized scleroderma (Progressive Systemic Sclerosis, PSS). Esophageal changes characteristic of generalized scleroderma were present in 15 of the patients, and 3 of those patients had duodenal involvements as well. The plasma concentrations of D-penicillamine were measured at 0 h, 1 h, 2 h, and 4 h after an oral dose of 300 mg D-penicillamine. Patients with duodenal and/or esophageal changes specific for scleroderma had significantly lower bioavailability of D-penicillamine than scleroderma patients without gastrointestinal manifestations. The decreased plasma D-penicillamine in scleroderma patients with involvement of the gastrointestinal tract may be due to an increased degradation of D-penicillamine in the gastrointestinal tract and/or an impaired absorption of the drug. Since the plasma level of D-penicillamine is so sensitive to pathological changes of the gastrointestinal tract, it may be advisable to adjust the dose of D-penicillamine on the basis of measurements of the plasma concentration of D-penicillamine.

Glycosaminoglycans in Localized Scleroderma (Morphoea)

The composition of glycosaminoglycans (GAGs) was analyzed in skin samples of eight patients suffering from localized scleroderma, i.e., three having generalized morphoea and five localized morphoea plaques. From each patients, biopsies were obtained from sclerotic and perilesional areas, and from clinically uninvolved skin of the same region. In the perilesional areas hyaluronic acid concentration was increased (p less than 0.05), while it was decreased (p less than 0.01) in sclerotic areas. Dermatan sulfate concentration was increased in the perilesional (p less than 0.05) as well as in the sclerotic (p less than 0.01) areas. Chondroitin 4/6 sulfate was increased in the sclerotic areas (p less than 0.05). Heparan sulfate showed no changes. No major differences were found in the total concentrations of uronic acid and hexosamine. This study demonstrates that changes in GAG composition in localized scleroderma follow previously described sequences of events of inflammation and fibrosis of connective tissue.

Collagen and uronic acid distribution in the human sclera

Abstract In the normal adult eye the content of uronic acid, hydroxyproline, hydroxylyxine 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.

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.

Bioavailability of D-Penicillamine in a Patient with Gastrointestinal Progressive Systemic Sclerosis

D-penicillamine pharmacokinetics were studied in a patient with gastrointestinal progressive systemic sclerosis possibly complicated by malabsorption. D-penicillamine bioavailability was examined after oral, duodenal, intravenous and rectal administration. No D-penicillamine was detectable in plasma after administration to the gastrointestinal tract. The pharmacokinetics after intravenous administration agreed closely with the corresponding situation in healthy volunteers.