The distribution of unsulphated and sulphated glycosaminoglycans in palmar fascia from patients with Dupuytren's disease and healthy subjects.

Eighty specimens from 20 patients with Dupuytren's disease and 7 biopsies of healthy palmar fascia were analysed for their glycosaminoglycan isomer patterns with a combined enzymatic/HPLC method. The diseased portions of palmar fascia tissue were characterized by elevated total glycosaminoglycans together with a relative increase in the sulphated fractions. The macroscopic stages of nodules, bands and unaffected tissue could be classified very well by multivariate statistical analysis on the basis of their glycosaminoglycan patterns. The biochemical analysis provided evidence of the pathological process even in those specimens that did not yet show any clinical symptoms of the disease.

[A histochemical study of acid glycosaminoglycans on normal and pathological cartilage, ligaments and several other connective tissues].

Histochemical estimation of acid glycosaminoglycan was critically re-evaluated, using the meniscus, intervertebral disc, ossified yellow ligament, ganglion, Dupuytren's fascia and several other tissues. Each tissue was stained with toluidine blue, alcian blue, high iron diamine, low iron diamine, aldehyde fuchsin and dialyzed iron-ferrocyanide. Digestion techniques for GAG were used in these staining methods, and the effects of protease inhibitors (PI) on digestion were also examined. In this study, the optimal temperature for digestion with Streptomyces hyaluronidase was between 37 degrees C and 43 degrees C, which varied according to the tissue examined. The addition of PI seemed necessary because the enzymatic treatment without PI resulted in an excessive decrease of staining. Protease-free chondroitinase ABC, which did not excessively decrease staining results, was found to be more useful than chondroitinase ABC without PI.

The biomechanical effects of tight suture closure upon fascia.

The biomechanical effect of tight suture closure of fascial incisions in male rats was evaluated. The incision of rats in group 1 was closed with maximum tension on the first throw of the approximating knot. The incision of rats in group 2 was closed in a similar manner, however a 3.14 squared millimeter rod was placed in the suture loop before securing the first throw. The rod was removed after the knot was formed and fascial edges were observed to be loosely approximated. All rats were sacrificed after seven days. The abdominal fascial and muscle unit was dissected and a 1 by 6 centimeter strip containing two suture loops was excised. Scanning electron microscopic examination of tissue specimens, tensile strength measurements and energy-to-failure under tensile loading studies were performed. Specimens for hydroxyproline assay were obtained from the tissue immediately surrounding the incision and between two suture loops. The scanning electron microphotographs of group 1 showed tissue overlap from the excessive force on the initial knot throws. Wounds loosely approximated had proper alignment of the wound edges and had greater proliferative activity in the clefts of the wounds. The tensile strength and energy-to-failure studies showed loosely approximated wounds to be far stronger (p less than 0.001). No statistically significant difference (p greater than 0.05) was found between hydroxyproline assays of the two groups.

Oestrogen and progesterone hormone receptors in Dupuytren's disease.

Dupuytren's disease is associated with alcoholism and chronic liver disease, conditions frequently associated with deranged steroid hormone metabolism. The possible influence of endogenous sex steroid hormones on the development of Dupuytren's disease has therefore been investigated. An analysis of diseased palmar fascia for oestrogen and progesterone receptors was undertaken in fifteen patients. Hormone specific receptors were not found in the palmar fascia of our patients with Dupuytren's disease, thus suggesting that other mechanisms or factors contribute to the pathogenesis of this fibrotic process.

Fibrin/fibrinogen and fibrinolytic activity of the palmar fascia in Dupuytren's contracture.

Considering the proved interaction of fibrin with fibroblasts and the seemingly decisive role of structural and functional changes ("modulation") of these cells in the evolution of Dupuytren's contracture, research has been carried out in order to investigate the fibrinolytic capacity and the possible presence of fibrin/fibrinogen in the palmar fascia of subjects operated upon for Dupuytren's Disease. Fibrin/fibrinogen were detected by a direct immunofluorescence technique and fibrinolytic activity was assessed by a fibrin plate method. A remarkable decrease of fibrinolytic activity and the presence of fibrin/fibrinogen were observed in small nodules in the early stage of disease, whereas large nodules showed a high amount of plasminogen activator enzymes. Small nodules seem to form and increase by progressive adhesion of fibroblasts to the polymerizing fibrin, while high fibrinolytic activity of large nodules probably results from "modulation" of many fibroblasts into contractile myofibroblasts and could therefore be considered as a biochemical sign of the evolutionary phase of Dupuytren's contracture.

Concentrations of cefmenoxime in human tissues.

Concentration levels of cefmenoxime were studied in human bone, muscle, fascia, cerebrospinal fluid, and synovial fluid. Tissue samples were taken 30 to 50 minutes after intravenous injection of 2,000 mg of cefmenoxime just before operation in patients undergoing orthopedic surgery. Levels were compared with those in the blood serum of the same patients. Cefmenoxime levels were also compared with levels of other cephalosporins examined by the same techniques and, with one exception, were found to be higher than those produced by the other agents. Cefmenoxime does not produce undesirable side effects, as do the other cephalosporins. Assay techniques for various tissues are described.

A 200-kd protein isolated from the fascia adherens membrane domains of chicken cardiac muscle cells is detected immunologically in fibroblast focal adhesions.

On the premise that the fascia adherens of cardiac muscle cell intercalated disk membranes is a structure that is closely homologous to the focal adhesions formed by fibroblasts, a fascia adherens preparation was isolated from chicken cardiac muscle, and was analyzed for its protein composition. A prominent 200-kilodalton (kd) protein was purified from the fascia preparation and shown to be antigenically unrelated to several previously characterized cytoskeletal proteins, including cardiac myosin and vinculin. With monospecific antibodies to the 200-kd protein, an identical or closely similar intracellular protein was shown to be associated with the focal adhesion plaques of fibroblasts.

Dupuytren's contracture: morphological and biochemical changes in palmar aponeurosis.

The palmar aponeurosis removed from ten patients with Dupuytren's contracture was studied using morphological and biochemical approaches. The histological characteristic of Dupuytren's contracture is the presence of numerous nodules among the lamellar structures of the collagen fibres. In the nodules, there are many active fibroblasts which are surrounded by immature fibres and metachromatic substances demonstrated by toluidine blue staining. Ultrastructurally, the active fibroblasts have the characteristics of myofibroblasts, as previously reported by Dr. Gabbiani. We found that some fibroblasts have intracellular collagen fibrils in the cytoplasm. When assayed by Siegel and Martin's method, lysyl oxidase activity of the palmar aponeurosis was significantly higher in Dupuytren's contracture than in normal hands. Biochemical studies such as electrophoretic analysis of mucopolysaccharides, determination of uronic acid and collagen contents were undertaken to compare the aponeurosis of Dupuytren's contracture with normal cases. The uronic acid contents were higher in Dupuytren's contracture than in the controls. However, no difference between the two groups was found in the collagen contents and in the composition of the mucopolysaccharides. These characteristic features; existence of myofibroblasts and intracellular collagen fibrils and increase in the activity of lysyl oxidase probably play a significant role in the establishment of flexion contracture of the fingers in Dupuytren's contracture.

Correlation of specific sulfated glycosaminoglycans with collagen types I, II, and III.

A qualitative and quantitative biochemical study of the glycosaminoglycans was performed in tissues constituted predominantly by one type of collagen, or in tissues containing mixtures of different types of collagen. The results obtained show the presence of dermatan sulfate, chondroitin sulfate, and heparin sulfate in tissues containing collagen types I, II, or III, respectively, suggesting a specific correlation of different glycosaminoglycans with these three types of collagen.

Dupuytren's contracture. An electron microscopic, biochemical, and clinical correlative study.

Fascial specimens were obtained from twenty-four patients treated operatively for Dupuytren's contracture. The nodules and cords were examined by electron microscopic and biochemical techniques. The clinical course and response to operative treatment were then correlated with the tissue findings. Electron microscopic analysis revealed myofibroblasts in the nodules of seven of twenty-four patients. Fibroblasts containing prominent microtubules were found in ten patients. The fascia contained type-III collagen, increased amounts of collagen per unit of dry weight, and an increase in reducible cross-links of collagen. While the nodules were noted to contain a greater increase in reducible cross-links than the cords, there was little variation in the biochemical findings from patient to patient. Clinical recurrence was not related to the age of the patient at onset, duration, or severity of disease. Recurrence was related to the electron microscopic findings of myofibroblasts in the nodules and fibroblasts containing prominent microtubules in the fascia of these patients.

[Serum and tissue concentrations after a single dose of cefaclor]. / Serum- und Gewebespiegel nach einmaliger Cefaclorgabe.

Serum and tissue concentrations of cefaclor were determined a total of 155 and 96 times respectively in 16 volunteers after a single dose of 1 g. At this dosage peak concentrations of 13.5, 14.5 and 13.4 mcg/ml were measured after 60, 90 and 120 minutes respectively. Tissues in which concentrations were measured included cortical bone, spongy bone, muscle, fascia, cutis and subcutis. By measuring blood concentrations of the tissue samples, a division could be made for purposes of calculation into intravascular and extravascular active components. Low amounts of extravascular cefaclor could be established merely in the fascia and in the cutis. The cefaclor concentrations found in spongy bone, muscles and subcutis proved to be determined to a large extent by the intravascular antibiotic. No cefaclor could be detected in cortical bone at the given dosage.

Collagen of Dupuytren's disease.

1. In contrast to collagen from the aponeurosis of normal adult subjects, the nodules, contractures and apparently unaffected aponeurosis from patients with Dupuytren's disease contained substantial amounts of type III collagen. 2. The presence of type III collagen supports the previous proposal that the initial response to injury is the synthesis of an increased proportion of this form of collagen. The increased amounts in the apparently unaffected aponeurosis indicate the disease is not strictly focal but more systemic than is usually considered.