Gene/Protein Disease Symptom Drug Enzyme Compound
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Previous studies of the collagen synthesis markers hyaluronan (hyaluronic acid) (HA) and procollagen type III aminoterminal peptide (PIIINP) in pulmonary fibrosis have reported elevated levels in bronchoalveolar lavage fluid (BALF) and suggested an association with disease activity. The objective of the present study was to evaluate whether HA and PIIINP in BALF and serum (S) correlated with paraclinical markers of disease activity (chest X-ray profusion score, pulmonary function tests (FEV1, FVC, TLC, DLCO)) in patients with pulmonary fibrosis. The material comprised 27 patients with biopsy-proven pulmonary fibrosis (12 cryptogenic and 15 due to connective tissue diseases) and 24 control subjects with normal lung function. BAL was performed in the right middle lobe with 250 ml saline. HA and PIIINP were measured in the cell-free BALF supernatant and in serum. Patients had higher BALF-HA (mean 86 +/- 17 (SEM) micrograms/l) than controls (39 +/- 2 micrograms/l (p < 0.01)), higher BALF-albumin (124 +/- 24 mg/l) than controls (58 +/- 4 mg/l (p < 0.01)) and higher BALF/S-HA ratio (2.4 +/- 0.6) than controls (1.2 +/- 0.6 (p < 0.05)). There were no significant differences respecting BALF-PIIINP, S-HA, or S-PIIINP. Patients (n = 14) with progressive disease had higher BALF-HA, higher BALF-albumin, higher S-PIIINP, and higher S-immunoglobulins than those with stable disease, but the differences did not reach statistical significance. Smokers (n = 18) had lower BALF-HA, lower S-HA, lower S-PIIINP, lower S-immunoglobulins, and higher lung function tests than non-smokers, but the differences did not reach statistical significance. In patients, significantly positive correlations were found between BALF-HA and BALF-albumin, between BALF-PIIINP and BALF-albumin, and a significantly negative correlation between S-PIIINP and TLC. None of the BALF or serum markers correlated with chest X-ray profusion score or any of the other lung function measurements. It is concluded that disease activity may be associated with elevated HA and PIIINP levels. Smoking may influence the immunological processes in pulmonary fibrosis and may be a confounder in studies of these patients.
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PMID:Hyaluronan and procollagen type III aminoterminal peptide in serum and bronchoalveolar lavage fluid from patients with pulmonary fibrosis. 853 35

Postoperative adhesions account for a significant morbidity after abdominal, gynecological, or cardiac surgery. A large number of compounds have been suggested to prevent such adhesions, but none is generally accepted. We have compared eight different substances that could be beneficial for the prevention of postoperative adhesions in a new standardized rabbit model with measurement of the areas of adhesion. In 10 groups of 20 rabbits an area of abrasion of the serosa of the ileum, the appendix, and the abdominal wall measuring 10,000 mm2 was created by an emery piston during celiotomy. The controls received no medication. The treatment groups received a single intraperitoneal administration of 1 ml per 100 g body wt of normal saline (NaCl), 5 mg taurolidine (T), 0.5 U plasmin/300 U DNase (PD), 2000 IU streptokinase/500 IU streptodornase (SS), 7 mg phosphatidylcholine (PC), 4 mg hyaluronic acid (HA), 7 mg sphingolipid (SL), 7 mg galactolipid (GL), or 0.5 ml tetrachlorodecaoxide (TCDO), respectively. Ten days later the extent of adhesions was quantified by morphometry. The total area of adhesions (+/- SEM) was found to be 1998 +/- 124 mm2 in controls. The application of NaCl reduced the adhesions to 1368 +/- 58 mm2, of T to 1012 +/- 48 mm2, of PD to 673 +/- 33 mm2, of SS to 360 +/- 44 mm2, of PC to 335 +/- 84 mm2, of HA to 328 +/- 76 mm2, of SL to 278 +/- 80 mm2, of GL to 261 +/- 67 mm2, and of TCDO to 240 +/- 45 mm2. The effects of PD, SS, PC, HA, SL, GL, and TCDO were significant in comparison to controls and NaCl. Our experimental data suggest that the two new lipid substances, SL and GL, are the most likely candidates for routine clinical use in the prevention of postsurgical adhesions.
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PMID:Prevention of postoperative adhesions by single intraperitoneal medication. 853 79

The uptake of hyaluronic acid (HA) was used to assess preservation damage to sinusoidal endothelial cells (SEC) during cold storage and subsequent normothermic reperfusion of rat livers. After 8, 16, 24, and 48 h storage in University of Wisconsin (UW) solution, livers were gravity-flushed via the portal vein with a standard volume of cold UW solution containing 50 micrograms/l HA. The effluent was collected for analysis of HA, aspartate aminotransferase (AST), and lactate dehydrogenase (LDH). The mean uptake of HA at 0 h was 59.1% +/- 4.6% (mean +/- SEM). After 8 h of storage, HA uptake was similar (55.5% +/- 7.3%), whereas after 16 h of storage it was reduced to 34.7% +/- 5.8%. At 24 and 48 h of storage, no uptake of HA was found. In a second series of experiments, livers were stored in UW solution and subsequently reperfused for 90 min with a Krebs-Henseleit solution (37 degrees C) in a recirculating system containing 150 micrograms/l HA. Following 8 h of storage, 34.6% +/- 8.0% of the initial HA concentration was taken up from the perfusate. After 16 and 24 h of storage, no uptake of HA was found. The results of this study indicate that damage to SEC occurs progressively during storage, leading to zero uptake of HA by the rat livers at 24 h of cold ischemia time. Additional reperfusion injury to the SEC was demonstrated by the reduced ability of the SEC to take up HA following normothermic reperfusion. The uptake of exogenous HA in preserved livers, used as a tool to assess SEC injury, enables the detection of early preservation damage.
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PMID:Hyaluronic acid uptake in the assessment of sinusoidal endothelial cell damage after cold storage and normothermic reperfusion of rat livers. 887 86

1. Orbital accumulation of hydrophilic, interstitial glycosaminoglycans (GAGs) and subsequent expansion of retrobulbar tissue lead to the clinical manifestation of exophthalmos in patients with Graves' eye disease. 2. A highly specific method to determine the concentration and biochemical composition of different GAGs was developed in order to obtain a sensitive test system for the activity of the disease. By means of this method, GAG excretion in 24 h urine collections of 56 patients and 21 controls was analysed by precipitation with cetylpyridinium chloride and potassium acetate in ethanol, followed by sequential enzymic hydrolysis with chondroitin AC lyase, chondroitin ABC lyase and hyaluronate lyase, with HPLC analysis of the resulting alpha, beta-unsaturated disaccharides by anion-exchange chromatography. 3. Concentrations of GAG, chondroitin sulphate A (CA), dermatan sulphate (DS) and hyaluronic acid (HA) were determined in patients and controls, with high recovery rates [72.2 +/- 5.3%, mean +/- SEM; detection limit, 4.2 micrograms/l (0.01 mumol/l)], revealing marked differences in urinary concentrations of total GAG and HA, as well as an elevation of CA in patients compared with controls. 4. Method sensitivity was 0.86 for patients with active Graves' eye disease, and 0.87 for patients with untreated ophthalmopathy, whereas specificity was 1.0 for patients with inactive disease. Patients with increased GAG concentration responded well to steroids and/or orbital irradiation (before therapy: GAG, 111.49 +/- 40.32; CA, 59.58 +/- 21.34; DS, 25.05 +/- 8.12; HA, 26.88 +/- 11.63 mg/24 h; during therapy: GAG, 54.22 +/- 10.94; CA, 20.52 +/- 4.58; DS, 17.65 +/- 3.46; HA, 16.05 +/- 3.69 mg/24 h), whereas GAG excretion increased markedly 2-3 months after stopping prednisone therapy in patients with still active eye disease (GAG, 109.9 +/- 10.51; CA, 63.8 +/- 7.34; DS, 24.1 +/- 5.07; HA, 22.0 +/- 6.28 mg/24 h). 5. This sensitive method determines the nature of renally excreted GAGs, reflecting the aberrant synthesis pattern of fibroblasts in patients with Graves' disease.
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PMID:HPLC glycosaminoglycan analysis in patients with Graves' disease. 917 26

The half-life of [3H]hyaluronic acid in rabbit knee joints was estimated using two methods: (i) by following the [3H]hyaluronan content of the synovial fluid after intra-articular injection and (ii) by following the 3H2O radioactivity of plasma after intra-articular injection of [3H]hyaluronan. For normal rabbits we obtained a half-life of 15.8 hours (method I) and 17.5 +/- 1.0 hours (mean +/- SEM, method II), respectively. The second method was used to estimate the kinetics of the hyaluronan elimination from normal, sham-operated, as well as from osteoarthritic rabbit knee joints (Colombo model of osteoarthritis). Four weeks after injury, during the developing phase of osteoarthritis, the half-life of hyaluronan rose significantly to 23.5 +/- 2.1 hours and returned to normal levels (17.4 +/- 2.7 hours) 12 weeks after the operation (osteoarthritis developed). At the stage of developed osteoarthritis, the clearance rates were considerably higher than in normal rabbits.
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PMID:Elimination of tritium-labelled hyaluronic acid from normal and osteoarthritic rabbit knee joints. 918 39

The aim of this work was to evaluate the cytosolic contents of hyaluronic acid (HA) and cathepsin D (CatD) in gastric carcinomas and their possible relationships with the clinicopathological parameters of the tumors. Our study demonstrated a wide variability in the cytosolic levels of HA (mean +/- SEM: 3748 +/- 411 ng/mg protein) and cathepsin D (52 +/- 4 pmol/mg protein) in the tumors of 78 gastric cancer patients. In addition, the tumoral contents of HA and CatD were significantly higher (p<0.005) in diffuse type (HA: 6027 +/- 1099 ng/mg protein; CatD: 75 +/- 13 pmol/mg protein) than in intestinal type (HA: 2735 +/- 242 ng/mg protein; CatD: 42 +/- 3 pmol/mg protein) carcinomas. These data suggest that both markers may contribute to the biological characterization of gastric carcinomas.
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PMID:Relationship of tumoral hyaluronic acid and cathepsin D contents with histological type of gastric carcinoma. 1101 96

A novel, biomimetic, interpenetrating polymer network (IPN) between poly-L-lysine (PLL) and ultra high molecular weight polyethylene (UHMWPE) has been synthesized in an attempt to decrease wear in joint prostheses. A biomaterial with a gradient IPN of cationic PLL and UHMWPE has been synthesized, in the surface of bulk UHMWPE, to recruit the poly-anion, hyaluronic acid, from the synovial fluid. It is hypothesized that the hyaluronic acid molecules and their associated hydration layer will improve lubrication between the articulating surfaces, thus lowering both friction and wear. The synthesis involves four steps. Silylation of the PLL-HBr to PLL-SiMe3 utilizing bis(trimethylsiyl)acetamide (BSA). Swelling of the UHMWPE in a solution of PLL-SiMe3/xylenes at 60 degrees C with ultrasonics. Crosslinking of the PLL-SiMe3 within the UHMWPE with 1,8-diisocyanatooctane (a.k.a. OMDI). Finally, de-swelling and drying of the IPN under vacuum at 50 C. Visual observations show an adhered film on the IPN surface. Reflective FTIR spectra contain the characteristic peaks associated with UHMWPE. Two additional peaks, at 3410 and 1690 cm-1, are associated with PLL. SEM shows a morphology dominated by PLL spheres with diameters ranging from < 1 micron up to 3 micron. This shows that the PLL-SiMe3 has been crosslinked by the OMDI and was not rinsed away by either xylenes or sonicated water rinses. High contact angle of the PLL in contact with the UHMWPE demonstrate that the PLL has been de-silylated and returned to its hydrophilic nature. The spheres attached to the surface of the UHMWPE indicate that PLL has infiltrated the UHMWPE physical network and is entangled there. XPS confirms the presence of nitrogen by a 3-5 atomic percent signal in the outer 100 A of the IPN surface.
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PMID:Synthesis and characterization of a novel UHMWPE interpenetrating polymer network. 1114 88

The blood-contacting properties and the effect on bacterial adhesion of a material based on polyurethane and poly(amido-amine) (PUPA), both in its native form and with the anticoagulant molecules heparin or sulphated hyaluronic acid (HyalS3.5) electrostatically bonded to its surface, were evaluated and compared in vitro. The presence of the biological molecules on the surface was revealed by a dye test and ATR/FTIR analysis. Bound heparin was found to maintain its physiological action, in terms of thrombin inactivation, as well as did free heparin. Moreover, it reduced the degree of platelet adhesion. On the contrary, bound HyalS3.5 lost its anticoagulant activity, though it reduced platelet adhesion. The number of platelets on both modified surfaces was low. Their shape distribution, as determined by SEM, did not differ significantly on the two modified surfaces or with respect to the bare PUPA surface. HyalS3.5 and heparin also inhibited adhesion of Staphylococcus epidermidis to the material. A possible relationship between the platelet and bacterial adhesion is ascribed to the mediating role of plasma proteins.
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PMID:In vitro study of blood-contacting properties and effect on bacterial adhesion of a polymeric surface with immobilized heparin and sulphated hyaluronic acid. 1121 Oct 93

The uppermost superficial surface layer of articular cartilage, the 'lamina splendens' which provides a very low friction lubrication surface in articular joints, was investigated using atomic force microscopy (AFM). Complementary specimens were also observed under SEM at -10 degrees C without dehydration or sputter ion coating. Fresh adult pig osteochondral specimens were prepared from the patellas of pig knee joints and digested with the enzymes, hyaluronidase, chondroitinase ABC and alkaline protease. Friction coefficients between a pyrex glass plate and the osteochondral specimens digested by enzymes as well as natural (undigested) specimens were measured, using a thrust collar apparatus. Normal saline, hyaluronic acid (HA) and a mixture of albumin, globulin, HA (AGH) were used as lubrication media. The surface irregularities usually observed in SEM studies were not apparent under AFM. The articular cartilage surface was resistant to hyaluronidase and also to chondroitinase ABC, but a fibrous structure was exhibited in alkaline protease enzymes-digested specimens. AFM analysis revealed that the thickness of the uppermost superficial surface layer of articular cartilage was between 800 nm and 2 microm in adult pig articular cartilage. The coefficient of friction (c.f.) was significantly higher in chondroitinase ABC and alkaline protease enzymes digested specimens. Generally, in normal saline lubrication medium, c.f. was higher in comparison to HA and AGH lubrication media. The role of the uppermost, superficial surface layer of articular cartilage in the lubrication mechanism of joints is discussed.
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PMID:Role of uppermost superficial surface layer of articular cartilage in the lubrication mechanism of joints. 1155 3

The skin consists of epidermis and dermis. In order to develop a biomimetic material for more realistic tissue regeneration, we fabricated a double layer gelatin sponge with different pore sizes by freeze-drying at different temperatures -20 [symbol: see text] and -196 [symbol: see text], respectively. A pseudo microporous structure, which provided the paracrine crosstalk in epithelial-mesenchymal interactions similar to the basement membrane in epidermal-dermal junction was formed between this two compartment. Chrondroitin-6-Sulphate (C-6-S) and hyaluronic acid (HA) were incorporated within the gelatin scaffold to create an appropriate microenvironment for cell proliferation and migration. The mechanical strength of gelatin sponges was improved by cross-linking of gelatin with carbodiimide in 95% ethanol solution two times. The lattice structure and pore sizes were evaluated by SEM to confirm that the interconnected porous structures were retained. We chose a dynamic spinner flask seeding method for more even distribution followed by a culture system in the air-liquid interface cultured for 21 days. Differentiation and phenotypic expression of keratinocytes were investigated by histological analysis and immunohistochemical identification. In this study, we found multiple epidermis-like layers constructed by cultured keratinocytes. It is suggested that the bi-layer scaffolds have the potential to be used as skin equivalents for application in burn wounds. In the future, the qualitative of chemical reagents and in vivo animal model tests will be evaluated.
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PMID:Organotypic keratinocyte-fibroblast cocultures on a bilayer gelatin scaffold as a model of skin equivalent. 1272 46


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