Biochemical Discrimination of Hurler and Scheie Syndromes

1979 ◽  
Vol 57 (3) ◽  
pp. 265-272 ◽  
Author(s):  
J. J. Hopwood ◽  
Vivienne Muller
Keyword(s):  
Thermal Stability ◽  
Skin Fibroblasts ◽  
Ph Optimum ◽  
Biochemical Evidence ◽  
Cultured Skin ◽  
Thermal Stability Of ◽  
Normal Controls

1. Homogenates of cultured skin fibroblasts derived from patients with α-l-iduronidase-deficiency disorders (Hurler and Scheie syndromes) were capable of hydrolysing iduronosyl anhydro-[l-3H]mannitol 6-sulphate although at considerably reduced rates compared with normal controls. 2. The Vmax. values of α-l-iduronidase from patients with Hurler or Scheie syndromes and from normal controls were 11, 12 and 833 pmol min−1 mg−1 of protein respectively; the corresponding apparent Km values were 656, 50 and 53 μmol/l respectively. The α-l-iduronidases from normal and Scheie fibroblast homogenates were shown to exhibit pH optima at 3·6 and 4·1 and were competitively inhibited by both chloride and sulphate ions: Hurler α-l-iduronidase activity exhibited one pH optimum at 3·8 and was also inhibited by chloride and to a lesser extent by sulphate ions. 3. The thermal stability of Hurler, Scheie and normal α-l-iduronidase activities at 55°C gave half-lives of approximately 1·0, 2·5 and 1·0 h respectively. 4. These biochemical findings clearly demonstrate enzyme differences for these two clinically distinct phenotypes and provide biochemical evidence that the Hurler and Scheie syndromes result from different allelic mutations.

Isolation and Partial Characterization of Alcohol Dehydrogenase From Broad Bean (Vicia faba)

1974 ◽  
Vol 1 (4) ◽  
pp. 579 ◽  
Author(s):  
S Leblova
Keyword(s):  
Thermal Stability ◽  
Alcohol Dehydrogenase ◽  
Broad Bean ◽  
Plant Tissues ◽  
Growth Substances ◽  
Ph Optimum ◽  
Alcohol Dehydrogenase Activity ◽  
Km Value ◽  
Thermal Stability Of

Alcohol dehydrogenase isolated from broad bean was found to have a Km value of 1.0 × 1.0 -2 M, a pH optimum of 8.7 and a molecular weight of 60 000 � 5000. The enzyme lost 55 % of its activity after being heated at 55�C, and was totally inactivated at 70°C. Thermal stability of the enzyme was not enhanced by NAD+ or ethanol. The substrate specificity of the enzyme is reported. Cysteine and mercaptoethanol activated the enzyme, whilep-chloromercuribenzoate, Cu2+, Hg2+, B4O72- -, Zn2+ and EDTA inhibited it. The influence of ethanol, acetaldehyde and growth substances on alcohol dehydrogenase activity in germinating broad bean seeds and plant tissues was also studied.

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.

scholarly journals IMMOBILIZATION OF PAPAIN ON CHITOSAN

2010 ◽  
Vol 8 (3) ◽  
pp. 372-376
Author(s):  
Sari Edi Cahyaningrum ◽  
Narsito Narsito ◽  
Sri Juari Santoso ◽  
Rudiana Agustini
Keyword(s):  
Thermal Stability ◽  
Optimum Temperature ◽  
Ph Optimum ◽  
Optimum Ph ◽  
Thermal Stability Of

In this study, papain was immobilized on chitosan with Mg(II) cosslinked agent. Studies on free and immobilized papain systems for determination of optimum pH, optimum temperatur, thermal stability and reusability were carried out. The results showed that free papain had optimum pH 6.5 and optimum temperature 55 °C while the immobile papain hadoptimum pH 8 and optimum temperature 80 °C. The thermal stability of the immobilized papain, relative to that of the free papain, was markedly increased. The immobilized papain can be reused for at least six times.   Keywords: papain, immobilization, chitosan

Sulphamidase Activity in Leucocytes, Cultured Skin Fibroblasts and Amniotic Cells: Diagnosis of the Sanfilippo a Syndrome with the Use of a Radiolabelled Disaccharide Substrate

1981 ◽  
Vol 61 (6) ◽  
pp. 729-735 ◽  
Author(s):  
J. J. Hopwood ◽  
Helen Elliott
Keyword(s):  
Incubation Temperature ◽  
Reaction Rates ◽  
Skin Fibroblasts ◽  
Cultured Skin ◽  
Normal Individuals ◽  
Amniotic Cell ◽  
Routine Assay ◽  
Incubation Temperatures ◽  
Amniotic Cells ◽  
Normal Controls

1. Sulphamidase activity was assayed by incubation of the radiolabelled disaccharide O-(α-2-sulphamino-2-deoxy-d-glucopyranosyl)-(1 → 3)-l-[6-3H]idonic acid with homogenates of leucocytes and cultured skin fibroblasts and concentrates of urine derived from normal individuals, patients affected with sulphamidase deficiency disorder [mucopolysaccharidosis type IIIA (MPS IIIA): the Sanfilippo A syndrome], parents of such patients and patients affected with other mucopolysaccharidoses and lysosomal enzyme deficiencies. 2. the assay clearly distinguished affected homozygotes from normal controls, heterozygotes and other mucopolysaccharidoses types. 3. Sulphamidase displayed remarkable thermal stability; reaction rates were constant for at least 24 h at 60°C for leucocyte and 20 h at 37°C for cultured fibroblast preparations. Apparent Km values for fibroblast sulphamidase were 71 μmol/l at 37°C and 100 μmol/l at 50°C; the corresponding Vmax. values were 21 and 71 pmol min−1 mg−1 of protein respectively. an incubation temperature of 60h°C was used for the routine assay of sulphamidase activity in leucocytes, urine and amniotic supernatant preparations. the specific activities of fibroblast and amniotic cell sulphamidase, assessed at incubation temperatures of 37°C, were more than 10-fold the leucocyte enzyme activity at 60°C. 4. We recommend the use of radiolabelled disaccharide substrate for the assay of sulphamidase in leucocytes, skin fibroblasts and urine, for the routine enzymic detection of the sulphamidase deficiency disorder of the Sanfilippo A syndrome.

The Ordered Phase Formation in Ti-Al-Ga Alloys

1970 ◽  
Vol 28 ◽  
pp. 440-441
Author(s):  
Shiro Fujishiro ◽  
Harold L. Gegel
Keyword(s):  
Thermal Stability ◽  
High Temperature ◽  
Titanium Alloys ◽  
Growth Kinetics ◽  
Stoichiometric Composition ◽  
Good Potential ◽  
Alpha Titanium ◽  
Cold Rolled ◽  
Kinetics Of ◽  
Thermal Stability Of

Ordered-alpha titanium alloys having a DO19 type structure have good potential for high temperature (600°C) applications, due to the thermal stability of the ordered phase and the inherent resistance to recrystallization of these alloys. Five different Ti-Al-Ga alloys consisting of equal atomic percents of aluminum and gallium solute additions up to the stoichiometric composition, Ti3(Al, Ga), were used to study the growth kinetics of the ordered phase and the nature of its interface.The alloys were homogenized in the beta region in a vacuum of about 5×10-7 torr, furnace cooled; reheated in air to 50°C below the alpha transus for hot working. The alloys were subsequently acid cleaned, annealed in vacuo, and cold rolled to about. 050 inch prior to additional homogenization

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.

Microstructure studies of tungsten silicide schottky contacts on Gaas

1987 ◽  
Vol 45 ◽  
pp. 328-329
Author(s):  
Yih-Cheng Shih ◽  
E. L. Wilkie
Keyword(s):  
Thermal Stability ◽  
Field Effect Transistors ◽  
Schottky Contact ◽  
Schottky Contacts ◽  
Excellent Thermal Stability ◽  
Barrier Heights ◽  
Gaas Substrates ◽  
Film Adhesion ◽  
Threshold Voltages ◽  
Thermal Stability Of

Tungsten silicides (WSix) have been successfully used as the gate materials in self-aligned GaAs metal-semiconductor-field- effect transistors (MESFET). Thermal stability of the WSix/GaAs Schottky contact is of major concern since the n+ implanted source/drain regions must be annealed at high temperatures (∼ 800°C). WSi0.6 was considered the best composition to achieve good device performance due to its low stress and excellent thermal stability of the WSix/GaAs interface. The film adhesion and the uniformity in barrier heights and ideality factors of the WSi0.6 films have been improved by depositing a thin layer of pure W as the first layer on GaAs prior to WSi0.6 deposition. Recently WSi0.1 has been used successfully as the gate material in 1x10 μm GaAs FET's on the GaAs substrates which were sputter-cleaned prior to deposition. These GaAs FET's exhibited uniform threshold voltages across a 51 mm wafer with good film adhesion after annealing at 800°C for 10 min.

Thermal stability of Al-Cu-Fe icosahedral alloys

1991 ◽  
Vol 1 (12) ◽  
pp. 1823-1836 ◽  
Author(s):  
M. Bessière ◽  
A. Quivy ◽  
S. Lefebvre ◽  
J. Devaud-Rzepski ◽  
Y. Calvayrac
Keyword(s):  
Thermal Stability ◽  
Thermal Stability Of
Sign in / Sign up

Export Citation Format

Share Document