Normal Electrophoretic Mobility Research Articles

Electrophoretic detection of reversible chlorpromazine · HCl binding at the human erythrocyte surface

The binding of chlorpromazine · HCl at the human erythrocyte surface has been detected through its effect on cellular electrophoretic mobility. Incubation of erythrocytes (approx. 5 · 10 6/ml) in 23 μM chlorpromazine · HCl resulted in a reduction of negative electrophoretic mobility from the control value of −1.11 ± 0.01 (μm · s −1)/(V · cm −1) to −1.00 ± 0.02 (μm · s −1)/(V · cm −1) (pH 7.2, ionic strength 0.155). This mobility change was completely reversed when chlorpromazine · HCl was removed by centrifugal washing. Increasing the drug concentration to 70μM did not affect the mobility change, indicating saturation of the electrophoretically detectable drug binding sites over chlorpromazine · HCl concentration range studied here. The effect of the 23 μM chlorpromazine · HCl on electrophoretic mobility was also measured in isotonic media of reduced ionic strength. The drug-induced reduction in negative surface charge density was found to be independent of ionic strength over the range 0.155 (Debye length, 0.8 nm) to 0.00310 (Debye length, 5.7 nm). Fixation of erythrocytes with glutaraldehyde affected neither the normal electrophoretic mobility of discocytes nor the reduced electrophoretic mobility of chlorpromazine · HCl-induced stomatocytes. When these stomatocytes were first fixed with glutaraldehyde, then washed free of chlorpromazine · HCl, they retained the stomatocyte form while regaining a normal control electrophoretic mobility. Conversely, when discocytes fixed in that form were treated with chlorpromazine · HCl, they showed the same mobility change as did fixed or unfixed stomatocytes. The drug-induced mobility change is therefore independent of the shape change, but reflects a contribution to cellular surface charge density from the membrane-bound chlorpromazine · HCl molecules. From the charge reduction, it is estimated that about 10 6 chlorpromazine · HCl molecules are bound at the electrokinetic cell surface and occupy approximately 0.4% of the total surface area.

Acid Maltase Deficiency in Adults

3 adult women with distinct clinical pictures of progressive myopathy were studied. The morphological findings of biopsied skeletal muscle suggested the diagnosis of type II glycogenosis. Biochemical analysis confirmed a profound deficiency of alpha-1,4-glucosidase activity. Electrophoresis of muscle acid maltase showed the presence of one band in normal individuals. A very faint band with normal electrophoretic mobility was present in the patients' muscles. Muscle neutral maltase is composed of four bands in normal adult individuals: two of the four bands were clearly reduced in the muscles of the patients. The acid and neutral maltases were not significantly reduced in the patients' leukocytes. Acid maltase determination in urine made it possible to identify the homozygous, but not to completely segregate the heterozygous, from unaffected adult subjects.

Three glucose 6-phosphate dehydrogenase variants found in Japan.

Three Japanese glucose 6-phosphate dehydrogenase (G6PD) variants were investigated. G6PD 'Mediterranean-like' had markedly decreased activity, normal electrophoretic mobility, low Km G6P, low Km NADP, increased utilization of all three substrate analogues (2-deoxy-G6P, Gal-6P, and deamino-NADP) and slightly decreased heat stability and slightly biphasic pH curve. G6PD 'Ogori' had markedly decreased activity, but otherwise normal characteristics. G6PD 'Hofu' had moderately decreased activity, normal electrophoretic mobility, slightly reduced Km G6P, normal Km NADP, normal utilization of 2-deoxy-G6P and Gal-6P, but increased utilization of deamino-NADP and normal heat stability as well as normal pH curve.

A Special Report: Four-year Study of a Boy with Combined Immune Deficiency Maintained in Strict Reverse Isolation from Birth

A 4-year study of a boy with combined immune deficiency is presented, and the impact of this disease on various aspects of his growth and development is examined. There is no evidence of immune deficiency in either parent or in the genetic background on the maternal side. Three children of a brother of the mother's father may have had immune deficiencies but two have grown to be teenagers with no problems. Another died. At autopsy, however, lymph nodes appeared normal. The deceased older brother had severe combined immune deficiency (SCID). The autopsy findings showed Pneumocystis carinii pneumonia to be the direct cause of death and these findings contributed to the diagnosis of SCID. After a successful germ-free birth, the male infant (DV) was placed in the isolator. Laboratory tests were normal except that the x-rays showed no thymic shadow, his absolute lymphocyte count ranged from 399-440/mm and the lymphocytes showed no proliferative response to phytohemagglutinin (PHA). Specific tests showed the antibody-producing immune system and the cell-mediated immune system to be severely defective. The patient's lymphocytes elicited positive responses by lymphocytes from father, mother, and sister. Subsequent search in national and international tissue-typing laboratories has shown four HLA matches but none has been nonreactive in mixed lymphocyte culture (MLC). therefore, this patient has remained in isolation to the present; now he is 4 years old. Approximately 35 species of microorganisms, mostly transient contaminants, have been isolated, taking into account that the same organism may have been identified under different names in different laboratories. Those isolated frequently and in sufficiently high concentration to indicate colonization have been speciated as follows: anaerobes-Propionibacterium acnes, Lactobacillus catenaforme (disappeared spontaneously), Bacteroides oralis ss. elongatus, Clostridium (perenne, hastiforme, bifermentans), Bacteroides clostridiiformis ss. clostridiiformis; aerobes-Alcaligenes faecalis (eradicated by antibiotics), Staphylococcus epidermidis, Enterobacter agglomerans, Microcossu sp. subgroup 1, Bacillus pulvifaciens (disappeared spontaneously); yeasts-Candida (tropicalis, parapsilosis). Seven are considered to be probable components of the current autoflora: P. acnes, C. bifermentans, B. clostridiiformis ss. clostridiiformis, S. epidermidis, Micrococcus sp. subgroup 1, E. agglomerans, C. parapsilosis. No viruses or protozoa have been isolated. At age 3 years, the mean quantitation of anaerobic cells was 7.9 X 10(7) viable cells/g feces; this falls short of the mean anaerobic load from normal children. The mean aerobic concentration was 1.2 X 10(8) viable cells/g feces, not unlike normal children. Qualitatively his flora has abnormally few species and lacks those most common in normal subjects. This child has had no evidence of infection and has always been in excellent health even though some organisms which could be pathogenic under some circumstances have been isolated...

Studies on variant glucose-6-phosphate dehydrogenases: G6PD fort worth

A new variant (G6PD Fort Worth) of glucose-6-phosphate dehydrogenase has been discovered in a young man of English-Irish descent. Although the enzyme deficiency is severe, it is not associated with chronic hemolytic anemia. The variant has normal electrophoretic mobility and normal stability, but altered kinetic characteristics and altered utilization of substrate analogs. A second variant G6PD has been found in a young man of Irish-Scotch descent. It is associated with chronic hemolytic anemia and is very unstable. The requirement of this variant for high concentrations of NADP for stability at 2–4° suggests that it is either identical to G6PD Torrance or closely related to it. Inhibition studies with NADPH and other kinetic studies have been carried out to evaluate the possible in vivo function of these two variants and also that of G6PD Kilgore.

G6PD hillbrow: a new variant of glucose-6-phosphate dehydrogenase associated with drug-induced haemolytic anaemia.

A new genetic variant of the red cell enzyme glucose-6-phosphate dehydrogenase is described. It was observed in a patient presenting with severe haemolytic anaemia and renal failure following ingestion of an overdose of Beserol (paracetamol and chlormezanone). The enzyme in the red cell had 12% of the activity of a normal B+ control, but only slightly lower activity in the kidney compared with a normal control. The red cell enzyme showed normal electrophoretic mobility and thermostability, a biphasic pH optimum curve, higher than normal utilization of the substrate analogues 2-deoxy-glucose-6-phosphate and deamino-NADP, and lower than normal Michaelis constants for both substrates, glucose-6-phosphate and NADP. The enzyme was strongly inhibited in vitro by high concentrations of paracetamol and chlormezanone. The extent of inhibition was similar to that for the enzyme from a normal B+ individual.

Electrophoretic characterization of human phosphoribosylpyrophosphate synthetase

A method has been developed for the electrophoretic separation of 5-phosphoribosyl-1-pyrophosphate synthetase on cellulose acetate. The human enzyme in extracts of various tissues appears as a narrow symmetrical peak, indicating a single genotype. Similar results were obtained with rabbit and rat tissues. A mutant erythrocyte 5-phosphoribosyl-1-pyrophosphate synthetase from two siblings having gout associated with excessive purine production was found to have normal electrophoretic mobility.

Characterization of glucose-6-phosphate dehydrogenase in Thailand

Characterization of partially purified eryrhrocyte G-6-PD from 50 enzymedeficient males in 45 unrelated Thai families revealed 6 enzyme variants. Thirty-five subjects in 31 families had G-6-PD variant with normal electrophoretic mobility, slightly low Km G-6-P, normal substrate-analog utilization, normal pH-optimum curve, and slightly increased heat stability. This enzyme variant is called G-6-PD Mahidol.

Surface charge on Plasmodium knowlesi and P. coatneyi-infected red cells of Macaca mulatta

Red cells infected by Plasmodium falciparum, P. coatneyi, and P. knowlesi adhere to venous endothelium. The possible role of red cell surface charge in the adhesiveness was investigated. The electrophoretic mobility of red cells from monkeys infected by P. knowlesi and P. coatneyi was not significantly different from mobilities determined before infection. When the infected red cells were concentrated by density gradient, they still showed normal electrophoretic mobility. Surface charge distribution was studied by binding positively charged ferric oxide hydrosols to glutaraldehyde-fixed red cells. Colloid binding was similar between the red cells infected with P. knowlesi and P. coatneyi and the uninfected red cells. Furthermore, knoblike membrane protrusions on red cells infected by P. coatneyi showed similar colloid binding as adjacent membranes. Therefore, neither the net surface charge nor the charge distribution on the red cell surface was altered by these infections. The knoblike protrusions in P. coatneyi-infected red cells have radii of curvature of 28–98 nm. Since these low radii would theoretically allow two similarly curved surfaces to approach within 0.5–1.0 nm of each other, knoblike protrusions may form the basis of adhesion. Knoblike protrusions of P. falciparum-infected red cells have similar dimensions and are the apparent site of attachment to venous endothelium.

Plasmalipoproteine bei Patienten mit Hyperthyreose: Isolierung und Charakterisierung eines abnormen High-Density-Lipoproteins

Article Plasmalipoproteine bei Patienten mit Hyperthyreose: Isolierung und Charakterisierung eines abnormen High-Density-Lipoproteins was published on January 1, 1972 in the journal Clinical Chemistry and Laboratory Medicine (CCLM) (volume 10, issue 7).

Platelet electrophoresis: effect of defibrination by ancrod (Arvin).

The changes in platelet electrophoretic mobility induced by adenosine diphosphate (ADP) and noradrenaline in normal subjects, in patients with acute illnesses, and in patients with chronic vascular disease described by Hampton and Mitchell (1966 a, b, c) are confirmed. The platelet electrophoretic mobility dose response curves in nine patients undergoing therapeutic defibrination by ancrod have been studied. Pretreatment platelet sensitivity to ADP was increased, during treatment at 48 hr and at 96 hr was normal, and one week after therapy was stopped reverted to the abnormal sensitivity pattern. During ancrod therapy the normal electrophoretic mobility response to ADP was associated with low levels of fibrinogen; one possible explanation of the altered platelet sensitivity found before treatment might be an abnormality of the plasma fibrinogen in disease states.

G-6-PD Bangkok: A New Variant Found in Congenital Nonspherocytic Hemolytic Disease (CNHD)

Abstract A new variant of G-6-PD was found in a Thai boy who had typical clinical and hematologic findings of congenital nonspherocytic hemolytic anemia. This mutant has normal electrophoretic mobility, normal affinity for G-6P and NADP, and is very labile to heat. It has an activity of about 5 per cent of normal and shows increased utilization of 2-deoxyglucose-6-phosphate and NAD and decreased utilization of deamino NADP. It is different from previously described mutants, and is named G-6-PD Bangkok.

A study of the thyroglobulin, thyroidal protease and iodoproteins in two congenital goitrous cretins

Detailed studies of thyroid functions were carried out in two adult goitrous cretins who were siblings. Both patients had high levels of serum protein-bound iodine, only a fraction of which could be accounted for as iodothyronines. The major circulating iodoprotein possessed many physical characteristics resembling those of serum albumin. Data are presented to indicate that this iodoprotein was not related to thyroglobulin as a precursor or as a degradation product but was rather the product of a collateral pathway of iodine metabolism in the thyroid. Thyroglobulin isolated from these patients showed normal electrophoretic mobility, sedimentation and solubility. The protease from their thyroid glands was able to hydrolyze the thyroglobulin from those glands in an apparently normal manner. The etiology of this disorder may be related to a reduced level of an enzyme in thyroglobulin synthesis with consequent glandular hypertrophy and increased synthesis of other thyroidal iodoproteins.