Search PubMed⌕ Search

Biomedical subjects

R G Davidson

Publications and source records attributed to R G Davidson.

At least 37 records · Page 2Linked to original sources
PubMed1985

A survey of injuries to the anterior cruciate ligament of the knee in female basketball players.

This study surveyed 76 female basketball-related injuries that occurred during a 30-month period at the B.C. Sports Medicine Clinic. The knee was the most common site of injury (72%), and anterior cruciate ligament (ACL) rupture accounted for 25% of all basketball injuries seen. A total of 19 ACL ruptures in females were seen as compared to only 4 ACL ruptures in male basketball players during the same time period. During this time period a total of 151 males and 76 female basketball players were seen. Each patient was assessed as to age, height, weight, and alignment and questioned as to mechanism of injury, playing position, experience, and training plus previous injuries. Possible etiological factors postulated included player position, joint laxity, weak quadriceps mechanism, and a possible hormonal basis.

Adolescent↗
PubMed1984

Diagnosis of pseudo-arylsulfatase A deficiency with electrophoretic techniques.

Deficient arylsulfatase A activity in man has long been associated with the neurodegenerative disease, metachromatic leukodystrophy. However, similar deficiency has been noted in clinically normal individuals, and is referred to as the pseudoarylsulfatase A deficiency condition. Although direct quantitative analysis of arylsulfatase A activity failed to differentiate between these two conditions, analysis of residual arylsulfatase A activity with either Cellogel electrophoresis or isoelectric focusing in polyacrylamide gels now has been shown to distinguish between them unequivocally. With both techniques, cultured fibroblasts from patients with pseudo-arylsulfatase A deficiency showed faint but clear bands of arylsulfatase A activity. Under identical conditions, fibroblasts from patients with metachromatic leukodystrophy showed no trace of activity. These methods can be adapted easily for general laboratory analysis in cases when results from quantitative arylsulfatase A assays are noninformative.

Cerebroside-Sulfatase↗
PubMed1984

Post-natal development of interscapular (brown) adipose tissue in the guinea pig: effect of environmental temperature.

The post-natal development of interscapular brown adipose tissue (BAT) was studied in the guinea pig by monitoring changes in DNA, triglycerides and mitochondrial protein as well as the activity of cytochrome oxidase and atractyloside insensitive GDP-binding. The results demonstrate that neonatal brown fat develops into a tissue with the gross characteristics of white adipose tissue. In this respect the post-natal development of guinea pig BAT is analogous to that of the tissue in man. Furthermore the apparent transformation is not irreversible since cold exposure (+4 degrees C) of adult guinea pigs for six weeks resulted in restoration of cytochrome oxidase, GDP-binding, etc, to levels characteristic of neonatal BAT. However, the interscapular fat pad temperature response to noradrenaline and the presence of 32 000 mol. wt mitochondrial inner membrane protein indicate that the tissue retains thermogenic activity even in warm acclimated adult guinea pigs.

Adipose Tissue, Brown↗
PubMed1983

Biochemical variability of arylsulphatases -A, -B and -C in cultured fibroblasts from patients with multiple sulphatase deficiency.

Multiple sulphatase deficiency (MSD) in man is inherited as an autosomal recessive trait and associated with deficient activities of various sulphohydrolases. Cultured fibroblasts from seven different patients were assayed for arylsulphatases-A, -B and -C activities. On the basis of the results, they may be classified into three groups: I, deficient in all three arylsulphatases; II, deficient only in arylsulphatases-A and -C with half or near-normal arylsulphatase-B; electrophoretically, arylsulphatase-A activity bands are undetectable as in metachromatic leukodystrophy; III, same as in II except electrophoretically, the residual arylsulphatase-A is detectable as faint activity bands similar to those in pseudo arylsulphatase-A deficiency. In addition to the variability among different strains, within the same strain of MSD or normal cells, each enzyme activity increased several fold with increasing time in culture. These sources of biochemical variability among and within different cell strains have not been recognized before in the study of this apparently monogenic trait with multiple enzyme deficiencies. They may account for some of the discrepancies reported in the literature on arylsulphatase activities among cultured cells from different multiple sulphatase deficient patients.

Arylsulfatases↗
PubMed1983

Gene dosage effects in human diploid and tetraploid fibroblasts.

The effects of cell ploidy on the biochemical characteristics of cultured cells were compared using human diploid vs tetraploid fibroblasts isolated with a non-selective method. Their DNA replication was compared by thymidine incorporation, and DNA content by Feulgen staining and quantitative analysis. Their RNA and protein content, cell sizes and the specific activities of glucose-6-phosphate dehydrogenase (G-6-PD) and 6-phosphogluconate dehydrogenase (6-PGD) were assayed quantitatively. With the exception of RNA content, all other parameters demonstrated a 2-fold increase reflecting the increase in cell ploidy. These direct gene dosage effects on the genetic material and functional expression of the human genome were in contrast to previous observations in other species and validate the use of human intraspecific euploid hybrids for biochemical and genetic studies.

Adult↗
PubMed1983

Pseudo arylsulfatase-A deficiency in healthy individuals: genetic and biochemical relationship to metachromatic leukodystrophy.

Metachromatic leukodystrophy is a hereditary neurodegenerative disorder in man associated with deficient arylsulfatase-A activity (aryl-sulfate sulfohydrolase, EC 3.1.6.1). The same enzyme deficiency has been noted in clinically normal individuals, a condition known as pseudo arylsulfatase-A deficiency. With a nonselective method, somatic cell hybrids were obtained from cultured fibroblasts of these two types of individuals; the hybrids showed no restoration of arylsulfatase-A activity. Thus, metachromatic leukodystrophy and pseudo arylsulfatase-A deficiency are allelic conditions. Although these conditions cannot be distinguished by simple quantitative arylsulfatase-A activity assays, they can be differentiated with sucrose density gradient centrifugation, Cellogel electrophoresis, or isoelectric focusing in polyacrylamide gels. In each case, a small amount of activity with characteristics of arylsulfatase-A was found only from fibroblasts of pseudo arylsulfatase-A-deficient individuals and not from those of metachromatic leukodystrophy patients. This residual enzyme has the same pH optimum, heat stability, inhibitor sensitivity, and Km as the normal enzyme but slightly different isoelectric points. In conclusion, although pseudo arylsulfatase-A deficiency and metachromatic leukodystrophy have very different clinical outcomes, they are due to mutations of the same structural gene, coding for arylsulfatase-A. These two conditions can be differentiated now by simple electrophoretic analysis of the residual arylsulfatase-A activity.

Arylsulfatases↗
PubMed1982

Somatic cell hybridization studies on the genetic regulation and allelic mutations in metachromatic leukodystrophy.

Metachromatic leukodystrophy is a hereditary neurodegenerative disease associated with deficient arylsulfatase A activity. Clinical variants differ in onset times and severity of the disease but each breeds true within families. Somatic cell hybridization techniques were used to clarify the genetic relationship among these mutants. Hybrid clones isolated with a nonselective method from fusing fibroblasts of an infantile and a juvenile variant did not show complementation of arylsulfatase A activity. Hence, these clinical variants are allelic mutants. Previous somatic cell hybridization studies suggested that "arylsulfatase A-deficiency" is a dominant phenotype, in contrast to its apparent recessive mode of inheritance. To resolve this discrepancy, hybrid clones from fusing normal and arylsulfatase A-deficient fibroblasts were isolated nonselectively. They continued to express arylsulfatase A activity. Hence, even in vitro, "arylsulfatase A-deficiency" remains as a recessive phenotype.

Adult↗
PubMed1980

Complementation of arylsulfatase A in somatic hybrids of metachromatic leukodystrophy and multiple sulfatase deficiency disorder fibroblasts.

Metachromatic leukodystrophy and multiple sulfatase deficiency disorder are severe neurodegenerative diseases inherited as separate autosomal recessive traits. Arylsulfatase A (aryl-sulfate sulfohydrolase, EC 3.1.6.1) activity is deficient in both diseases but in multiple sulfatase deficiency disorder, activities of arylsulfatases B and C and other sulfatases are also reported to be reduced. Somatic hybrid cell clones produced by fusing cultured fibroblasts from patients with these diseases were isolated by a nonselective technique based on unit-gravity sedimentation. Arylsulfatase A activity was restored in these hybrids. The complemented enzyme resembled the normal arylsulfatase A in heat stability, pH optimum, Km, electrophoretic mobility, and immunologic reactivity. Because a structurally normal enzyme can be restored in a hybrid only though intergenic complementation, these results indicate that the mutations responsible for the deficiency of arylsulfatase A activity in metachromatic leukodystrophy and multiple sulfatase deficiency disorder are nonallelic and that at least two genetic loci control the expression of arylsulfatase A activity in the human genome. Furthermore, arylsulfatase C activity was also restored to normal in the hybrids, indicating that a common sulfatase inhibitor is not the cause of the multiple sulfatse deficiency.

Cells, Cultured↗
PubMed1980

Gaucher's disease II. Studies on the kinetics of beta-glucosidase and the effects of sodium taurocholate in normal and Gaucher tissues.

beta-Glucosidase activity from normal human cultured fibroblasts was not affected by the presence of up to 0.1% (w/v) (1.86 mM) purified sodium taurocholate. At concentrations greater than 0.1%, there was a gradual decrease in activity. Conversely, beta-glucosidase activity from fibroblasts of three patients with juvenile onset Gaucher's disease was preferentially inhibited by the detergent at concentrations as low as 0.025% (0.46 mM). A 40% decrease in activity was observed at 0.1%. Crude sodium taurocholate was more potent in inhibiting beta-glucosidase activity from both the normal and Gaucher fibroblasts. However, very high background fluorescence and inconsistent results were observed when different batches of the crude taurocholate from the same or different sources were used. Similarly, beta-glucosidase activity from Gaucher splenic tissue homogenates, supernatant fluids (40,000 x g) and residue pellets was preferentially inhibited by purified sodium taurocholate. These findings indicate that the reliability and sensitivity of the enzyme assay for Gaucher's disease can be enhanced by determining beta-glucosidase activity in both the absence and presence of purified sodium taurocholate, particularly when variant cases with relatively high residual enzyme activity are encountered. In contrast to the enzyme from fibroblasts and spleens, beta-glucosidase activity from human placenta was markedly activated (greater than 300%) by the presence of 0.08% (1.49 mM) purified sodium taurocholate or 0.1 mM phosphatidyl serine, suggesting the presence of a predominate form of beta-glucosidase, possibly glucocerebroside beta-glucosidase, which is activated by the detergent. The apparent Michaelis constant (Km) for both the soluble and membrane-bound enzyme from normal fibroblasts was 1.6 +/- 0.1 mM. Kms from a patient with severe juvenile Gaucher's disease and two other patients with milder manifestations were 0.8 +/- 0.2 and 3.3 +/- 0.3 mM, respectively.

Adolescent↗
PubMed1980

Gaucher disease. III. Substrate specificity of glucocerebrosidase and the use of nonlabeled natural substrates for the investigation of patients.

A reproducible and convenient method for assaying glucocerebrosidase activity using the natural substrates has been developed. From the insoluble pellet fraction of cultured skin fibroblast homogenates, released glucose was measured enzymically using hexokinase coupled with the glucose-6-phosphate dehydrogenase (G6PD) and nicotinamide adenine dinucleotide phosphate (NADP) system. Optimal enzyme assay conditions required both Triton X-100 and sodium taurocholate, pH 4.8. Glucocerebrosidase activities from three patients with type 1 Gaucher disease were 17.5%, 15.8%, and 11.2% of normal (normal = 198 +/- 14 nmol/hr per mg protein, n = 3). The first patient had normal beta-glucosidase activity with the artificial fluorogenic umbelliferone substrate. Interference with the accuracy of the glucose-dependent assay system by either glycolytic or gluconeogenic enzyme activites was not detected under these experimental conditions, and when substrates with long fatty-acid chain lengths (C = 22) were used, markedly decreased glucocerebrosidase activity occurred in both normal individuals and patients. The apparent Km's for the natural substrates were 0.56 +/- 0.05 mM for controls and 2.2-3.3 mM for Gaucher fibroblasts. These data further support the hypothesis that a structurally altered and catalytically deficient enzyme is synthesized in patients with type 1 Gaucher disease and illustrate the value of the natural substrate in investigating patients.

Cells, Cultured↗