scholarly journals The disorder of hyaluronic acid metabolism in cultured skin fibroblasts derived from a patient with the Hurler syndrome

1973 ◽  
Vol 132 (3) ◽  
pp. 403-408 ◽  
Author(s):  
Ralph J. Germinario ◽  
Arthur Kahlenberg ◽  
Leonard Pinsky
Keyword(s):  
Hyaluronic Acid ◽  
Acid Content ◽  
Specific Radioactivity ◽  
Skin Fibroblasts ◽  
Hurler Syndrome ◽  
Acid Fraction ◽  
Normal Cells ◽  
Cultured Fibroblasts ◽  
Cultured Skin ◽  
Corrective Factor

The metabolism of hyaluronic acid in cultured skin fibroblasts derived from a patient with the Hurler syndrome and from a normal subject was examined. 1. An increased net incorporation of [3H]glucose into the hyaluronic acid fraction of the Hurler-syndrome cells occurred when compared with normal cells. 2. During a `chase' period, approx. 35% of the radioactivity derived from glucose was lost from the hyaluronic acid fraction of the Hurler-syndrome cells, whereas the normal cells retained all their radioactivity. 3. Although the Hurler-syndrome cells contained a ninefold greater amount of hyaluronic acid than normal cells, simultaneous determination of the specific radioactivity derived from the label revealed a value for the Hurler-syndrome cells one-half that of normal cells. These results are taken to indicate that the Hurler cells synthesize hyaluronic acid de novo at a higher rate than do normal cells. 4. Exposure of Hurler-syndrome cultured fibroblasts to a crude urine corrective-factor preparation (Neufeld & Cantz, 1971), now known to contain α-l-iduronidase, the specific Hurler-syndrome corrective factor (Bach et al., 1972), decreased the hyaluronic acid content to near-normal values before any effect was observed on [3H]glucose incorporation into the hyaluronic acid fraction. 5. In addition, the hyaluronic acid content of the normal cells decreased after exposure to the corrective factor of urine. 6. The mobilization of hyaluronic acid in Hurler-syndrome and normal cells exposed to the crude corrective-factor preparation of urine caused a decrease in specific radioactivity in the `corrected' Hurler-syndrome cells and an increase in specific radioactivity in the `corrected' normal cells.

scholarly journals Biochemical studies on the sulphated glycosaminoglycan fraction of skin fibroblasts cultured from a patient with the Hurler syndrome

1973 ◽  
Vol 132 (3) ◽  
pp. 395-402 ◽  
Author(s):  
Ralph J. Germinario ◽  
Arthur Kahlenberg ◽  
Leonard Pinsky
Keyword(s):  
Specific Radioactivity ◽  
Cell Types ◽  
Skin Fibroblasts ◽  
Hurler Syndrome ◽  
Normal Cells ◽  
Relative Retention ◽  
Cultured Skin ◽  
Corrective Factor ◽  
Sulphated Glycosaminoglycans ◽  
Sulphated Glycosaminoglycan

1. The metabolism of the sulphated glycosaminoglycan fraction in cultured skin fibroblasts derived from a patient with the Hurler syndrome and from a normal subject was studied. Two labelled precursors, Na235SO4 and d-[2-3H]glucose, were used and their intracellular fates during uptake and `chase' periods were assessed after separation of sulphated glycosaminoglycans from hyaluronic acid. After 4 or 8h of exposure to culture medium containing both labels, [35S]sulphate incorporation into the sulphated glycosaminoglycan fraction was twofold greater in Hurler-syndrome cells than in normal cells. At the same time, the rate of incorporation of [3H]glucose into the sulphated glycosaminoglycan fraction was approximately the same for both cell types. Consequently, an increased 35S/3H ratio (nmol of [35S]sulphate incorporated/nmol of [3H]glucose incorporated) was observed for Hurler-syndrome cells compared with normal cells. 2. The results of `chase' experiments revealed that although the expected loss and relative retention of labelled sulphate occurred in the sulphated glycosaminoglycan fraction of normal and Hurler-syndrome cells, both cell types retained all of their radioactivity derived from [3H]glucose. 3. After 34h exposure to a `corrective-factor' preparation from urine, the sulphated glycosaminoglycan content (as hexosamine and [35S]sulphate) of the Hurler-syndrome cells approached normal values. At the same time, there was an increase in specific radioactivity of `corrected' Hurler-syndrome cells.

scholarly journals Characterization of lysosomes and lysosomal enzymes from Chediak-Higashi-syndrome cultured fibroblasts

1986 ◽  
Vol 238 (2) ◽  
pp. 589-595 ◽  
Author(s):  
A L Miller ◽  
R Stein ◽  
M Sundsmo ◽  
R Y Yeh
Keyword(s):  
Lysosomal Enzymes ◽  
Specific Activity ◽  
Skin Fibroblasts ◽  
Cultured Fibroblasts ◽  
Cultured Skin ◽  
Lysosomal Dysfunction ◽  
Chediak Higashi Syndrome ◽  
Normal Controls

Chediak-Higashi-syndrome cultured skin fibroblasts were used to study the possible involvement of lysosomal enzymes and lysosomal dysfunction in this disorder. Our evidence indicated that Chediak-Higashi fibroblasts displayed a significant decrease in the specific activity of the acidic alpha-D-mannosidase (pH 4.2) compared with normal controls. Additional studies revealed a small, but significant, decrease in the rate of degradation of 125I-labelled beta-D-glucosidase that had been endocytosed into Chediak-Higashi cells.

METACHROMASIA AND ASSAY FOR LYSOSOMAL ENZYMES IN SKIN FIBROBLASTS CULTURED FROM PATIENTS WITH CYSTIC FIBROSIS AND CONTROLS

PEDIATRICS ◽  
1971 ◽  
Vol 47 (6) ◽  
pp. 1010-1018
Author(s):  
Ilana Kraus ◽  
Irena Antonowicz ◽  
Harnish Shah ◽  
Herbert Lazarus ◽  
Harry Shwachman
Keyword(s):  
Cystic Fibrosis ◽  
Lysosomal Enzymes ◽  
Skin Fibroblasts ◽  
Control Group ◽  
Staining Reaction ◽  
Cultured Fibroblasts ◽  
Cultured Skin ◽  
Cell Strains ◽  
Absolute Correlation ◽  
Metachromatic Granules

The ability of Toluidine blue 0 to stain certain cytoplasmic elements metachromatically in cultured skin fibroblasts derived from patients with cystic fibrosis (CF) has been investigated. Fifteen of 18 (83.3%) cell strains from patients with cystic fibrosis were positive, two (11.1%) borderline, and one negative (5.5%). In a control group, 8 of 18 (44.4%) were positive, three borderline (16.6%), and seven negative (38.8%). In addition to the lack of an absolute correlation between metachromasia and CF, inconsistencies in staining reaction were continually noted. Several parameters which account for these variations have been examined. Since metachromatic granules are felt to represent lysosomal structures, a study of four lysosomal enzymes in the skin cultured fibroblasts was carried out in material from 10 patients with CF and 11 controls. No differences were noted in the activity of β-galactosidase, β-glucuronidase, aryl sulfatase, and acid phosphatase. While these studies do not support the possibility that CF is due to a lysosomal hydrolase deficiency in the metabolism of mucopolysaccharides, they do not exclude it either.

scholarly journals Glutathione reverses the growth abnormalities of skin fibroblasts from insulin-dependent diabetic patients with nephropathy.

1998 ◽  
Vol 9 (6) ◽  
pp. 1060-1066
Author(s):  
A Morocutti ◽  
M Sethi ◽  
A Hayward ◽  
A Lee ◽  
G Viberti
Keyword(s):  
Oxidative Stress ◽  
Superoxide Dismutase ◽  
Fold Increase ◽  
Skin Fibroblasts ◽  
Diabetic Patients ◽  
Cultured Fibroblasts ◽  
Cultured Skin ◽  
Insulin Dependent ◽  
Insulin Dependent Diabetic ◽  
Growth Abnormalities

Oxidative stress has been proposed as a possible pathogenic factor for diabetic complications. It is relevant in determining cell replicative capacity and life span, and in vitro antioxidant treatment is able to reverse the impaired proliferative activity of different cell types. It was recently demonstrated that cultured skin fibroblasts from insulin-dependent diabetic patients with nephropathy age prematurely and have a shorter life cell cycle. To test whether the growth phenotype of cells from patients with diabetic nephropathy was related to a lack of protection from oxidative stress, the effect of reduced glutathione (GSH) on cultured skin fibroblasts from 13 insulin-dependent diabetes mellitus (IDDM) patients with nephropathy (DN), 10 IDDM patients without kidney disease (D), and 10 nondiabetic control subjects (C), in normal (5 mM) glucose (NG) and high (22 mM) glucose (HG) medium was studied. After 6 to 8 passages, fibroblasts from DN showed impaired growth both in NG (mean +/- SD fold increase over baseline counts in DN 1.17 +/- 0.6 versus D 1.7 +/- 0.5 versus C 1.95 +/- 0.8; P = 0.04 by ANOVA) and in HG (mean +/- SD fold increase over baseline counts DN 1.16 +/- 0.41 versus D 1.89 +/- 0.66 versus C 2.24 +/- 0.9; P = 0.003 by ANOVA). GSH prevented the growth abnormalities of cells from DN restoring it to values similar to that of the other two groups (mean +/- SD fold increase over baseline counts NG +/- GSH: DN 1.68 +/- 0.9 versus D 1.78 +/- 0.49 versus C 1.99 +/- 0.7, P = 0.6; and in HG + GSH: DN 1.66 +/- 0.69 versus D 1.87 +/- 0.75 versus C 2.2 +/- 0.9, P = 0.3). Growth rates were not affected by the addition of GSH in fibroblasts from D and C. The treatment of fibroblasts from D and C with the inhibitor of the gamma-glutamylcysteine synthetase activity, L-buthionine-S,R-sulfoximine, resulted in growth impairment, and the addition to the culture medium of another antioxidant, superoxide dismutase, corrected the growth abnormalities in fibroblasts from DN. The impaired growth of cultured fibroblasts from IDDM patients with nephropathy is prevented by GSH and superoxide dismutase and is independent of prevailing glucose concentrations. This suggests that oxidative stress is an important mechanism of intrinsic cell dysfunction in these patients.

Electron Microscopy of Fibroblasts from Myotonic Muscular Dystrophy Patients

1981 ◽  
Vol 39 ◽  
pp. 498-499
Author(s):  
S. E. Miller ◽  
G. B. Hartwig ◽  
R. A. Nielsen ◽  
A. P. Frost ◽  
A. D. Roses
Keyword(s):  
Electron Microscopy ◽  
Muscular Dystrophy ◽  
Genetic Diseases ◽  
Skin Fibroblasts ◽  
Inborn Errors ◽  
Cytoplasmic Inclusions ◽  
Cultured Skin ◽  
Myotonic Muscular Dystrophy ◽  
Glycosaminoglycan Metabolism

Many genetic diseases can be demonstrated in skin cells cultured in vitro from patients with inborn errors of metabolism. Since myotonic muscular dystrophy (MMD) affects many organs other than muscle, it seems likely that this defect also might be expressed in fibroblasts. Detection of an alteration in cultured skin fibroblasts from patients would provide a valuable tool in the study of the disease as it would present a readily accessible and controllable system for examination. Furthermore, fibroblast expression would allow diagnosis of fetal and presumptomatic cases. An unusual staining pattern of MMD cultured skin fibroblasts as seen by light microscopy, namely, an increase in alcianophilia and metachromasia, has been reported; both these techniques suggest an altered glycosaminoglycan metabolism An altered growth pattern has also been described. One reference on cultured skin fibroblasts from a different dystrophy (Duchenne Muscular Dystrophy) reports increased cytoplasmic inclusions seen by electron microscopy. Also, ultrastructural alterations have been reported in muscle and thalamus biopsies from MMD patients, but no electron microscopical data is available on MMD cultured skin fibroblasts.

scholarly journals Glycosaminoglycan synthesis by cultured skin fibroblasts from a patient with Lowe's syndrome.

1981 ◽  
Vol 256 (20) ◽  
pp. 10313-10318
Author(s):  
S. Fukui ◽  
H. Yoshida ◽  
T. Tanaka ◽  
T. Sakano ◽  
T. Usui ◽  
...  
Keyword(s):  
Skin Fibroblasts ◽  
Glycosaminoglycan Synthesis ◽  
Cultured Skin

scholarly journals Elevated Cystine Levels in Cultured Skin Fibroblasts From Patients With I-Cell Disease

1979 ◽  
Vol 13 (12) ◽  
pp. 1350-1355 ◽  
Author(s):  
Frank Tietze ◽  
Jean DeBrohun Butler
Keyword(s):  
Skin Fibroblasts ◽  
Cell Disease ◽  
Cultured Skin

Heme content of normal and porphyric cultured skin fibroblasts

1977 ◽  
Vol 15 (11-12) ◽  
pp. 1061-1070 ◽  
Author(s):  
David A. Brenner ◽  
Joseph R. Bloomer
Keyword(s):  
Skin Fibroblasts ◽  
Cultured Skin
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