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J Elion

Publications and source records attributed to J Elion.

At least 145 records · Page 8Linked to original sources
PubMed1990

HLA-DQB 1 codon 57 and genetic susceptibility to type 1 (insulin-dependent) diabetes mellitus in French children.

Variations in the incidence rate of Type 1 (insulin-dependent) diabetes mellitus might relate to ethnic-specific genetic backgrounds. HLA-DQB 1 alleles were typed in 75 French diabetic children and 85 matched control children. Enzymatically-amplified DQB 1 exon 2 was hybridized with oligoprobes specific for the six most common alleles. Alleles coding for an Asp residue at position 57 in the DQ beta chain are strongly negatively associated with Type 1 diabetes in the French population. Nevertheless, one of the diabetic children was an Asp 57 homozygote (DQB 1.2/3.1). Among alleles coding for a residue other than Asp at position 57, alleles 3.2 and 2 (Ala 57) are positively associated with diabetes but not allele 1.1 (Val 57) which is less frequent in diabetic children than in control children. Heterozygosity for 2/3.2 alleles is the genotype most strongly associated with diabetes (Odds ratio = 52.9). Large comparative population studies will be necessary to determine whether the frequency of DQB 1 alleles positively associated with Type 1 diabetes (2 and 3.2) in a given ethnicity is related to its incidence rate in the same population.

Alanine↗
PubMed1989

Activation of the human epsilon- and beta-globin promoters by SV40 T antigen.

We have studied the effect of the SV40 T antigen on expression from human globin promoters fused to the bacterial chloramphenicol acetyltransferase (CAT) gene and compared its effect with the SV40 enhancer and the adenovirus E1A protein. We have observed that expression of p epsilon GLCAT and p beta GLCAT (the epsilon-globin or beta-globin promoter linked to the CAT gene) was significantly stimulated when cotransfected with a cloned T antigen plasmid into CV-1 cells, indicating that trans-activation of the globin promoters was mediated by SV40 T antigen. Transfection of the p beta GLCAT-SV (p beta GLCAT containing the SV40 enhancer element) into CV-1 cells resulted in a 50-60-fold increase in CAT activity as compared to p beta GLCAT (no enhancer). However, cotransfection of the p beta GLCAT-SV with the cloned T antigen resulted in an additional increase of CAT expression, which suggests that T antigen and the SV40 enhancer activate globin gene expression independently. We found that T antigen but not E1A could further stimulate the expression of an enhancer-containing plasmid in CV-1 cells; whereas E1A but not T antigen could further stimulate p epsilon GLCAT expression in COS-1 cells which constitutively express the SV40 T antigen. These results suggest that T antigen and E1A also act independently. Deletion analysis showed that the minimum sequence required for a detectable level of stimulation of the epsilon-globin promoter by T antigen is 177 bp 5' to the cap site, suggesting that the target sequences for response to T antigen do not reside in the canonical 100 bp promoter region, but rather reside in sequences further upstream, and therefore the cellular factors interacting with T antigen are not the TATA or CAT box binding proteins, but the proteins interacting with upstream regulatory sequences.

Adenovirus Early Proteins↗
PubMed1989

Haplotypes in tribal Indians bearing the sickle gene: evidence for the unicentric origin of the beta S mutation and the unicentric origin of the tribal populations of India.

To determine the origin of sickle cell anemia (SS) in India, we analyzed haplotypes of the beta gene cluster in beta S-carrying individuals belonging to tribal populations living in the Nilgiris region of southern India and complemented the available data on tribes of east-central India. We found that in the Nilgiris tribes chromosomes bearing the beta S gene are linked in 91% of the cases to the "Asian" (Arab-Indian) haplotype (although 25% of the haplotypes had the epsilon polymorphic site negative, making the 5' portion of the haplotype identical with the African Senegal haplotype). These XmnI (+) chromosomes were associated with high G gamma expression (67.2 +/- 5.9%) and a high percentage of Hb F (15.5 +/- 7.9%; range, 6-25.3%). We have similar findings for tribal groups from west-central India (Gujarat). In east-central India we have confirmed the data of others, finding the same haplotype linked to beta S in tribes living in the east (Orissa, Andhra Pradesh). We conclude that the beta S gene in presently isolated and disperse tribal populations in India is associated with one predominant typical haplotype, suggesting a unicentric origin of the mutation in India. In addition, this finding implies a unicentric origin of the tribal populations themselves: The gene must have arisen and spread before tribal dispersion. Furthermore, we find extremely high frequencies of the (-alpha) haplotype in the Nilgiris (0.89) and in Gujarat (0.95). The beta S gene linkage to a high Hb F-expressing haplotype and the high incidence of alpha-thalassemia predict a mild phenotypical expression of sickle cell anemia in India.

Anemia, Sickle Cell↗
PubMed1988

A simple strategy to amplify specifically the HLA-DQ beta gene region with genomic DNA as template.

The nature of codon 57 in the HLA-DQ beta gene was recently reported as a potential marker of genetic susceptibility to insulin-dependent diabetes mellitus. When exploring the relevance of this marker by using genomic DNA amplification, we encountered difficulties resulting from the coamplification of the homologous DX beta region. A simple strategy is proposed to amplify the DQ beta region exclusively. It involves the preliminary digestion of genomic DNA with a restriction enzyme which cleaves DX beta specifically, leaving intact the DQ beta sequence. The amplified material is suitable for dot blot analysis and restriction enzyme digestion. This strategy is of general interest when homologous sequences impair the specificity of enzymatic DNA amplification.

DNA↗
PubMed1988

Analysis of ligand-binding data without knowledge of bound or free ligand molar concentration.

A method is proposed to set the parameters in a nonlinear regression procedure to determine the equilibrium dissociation constant (Kd) and the high affinity receptor concentration (Bmax) of systems consisting of one ligand, one high affinity receptor, and n low affinity binding sites. This method is suitable when neither bound or free ligand formal concentrations nor the maximum of the binding signal can be deduced from the experimental data. The method makes use of (i) the abscissa of the first inflection point of the plot of any signal proportional to the binding of ligand to receptors versus the logarithm of the total ligand concentration, and (ii) the initial slope of the saturation curve plotted in direct coordinates. We first demonstrate that when such an inflection point exists, its abscissa lies between Bmax/2 + Kd(1 + d) and Bmax + 2Kd(1 + d), where d is a parameter representative of the binding to the low affinity sites. Second, we demonstrate that the initial slopes of two saturation curves in direct coordinates, where Bmax varies by a known factor, allows an estimation of the Bmax/Kd ratio, within certain limits. From these two sets of data it is subsequently possible to define a precise window for the values of both Bmax and Kd. The performance of the method has been evaluated in representative cases using Monte Carlo studies. The results establish conditions for the existence of an inflection point as well as the influence of low affinity binding, whether or not proportional to Bmax.

Binding Sites↗
PubMed1988

Functional characterization of thrombin Salakta: an abnormal thrombin derived from a human prothrombin variant.

The genetic variant prothrombin Salakta has been described in a patient presenting with a normal level of prothrombin antigen but reduced prothrombin activity. Initial studies indicated that factor Xa-catalyzed cleavages proceed normally but lead to the production of a thrombin molecule with an altered enzymatic activity. To characterize the functional abnormality of thrombin Salakta more precisely, it was purified by chromatography on heparin-Sepharose and diethylaminoethyl-Sephadex. The purified variant does not differ from normal thrombin by size, as judged by sodium dodecyl sulfate-polyacrylamide gel electrophoresis, and is 93.1% +/- 7.6% active by titration with p-nitrophenyl-p'-guanidinobenzoate. Its activity, however, is altered to various extents toward the following substrates: H-D-phenylalanyl-L-pipecolyl-L-arginine paranitroanilide (S 2238), fibrinogen, factor V, protein C, and antithrombin III. The Michaelis constant (Km) of thrombin Salakta for S 2238 is higher (12.2 +/- 3.3 mumol/L) than normal (2.8 +/- 0.7 mumol/L), whereas the turnover number (Kcat) is normal (84.4 +/- 6.6 s-1 v 85.9 +/- 14.0 s-1 for normal thrombin). The interaction of thrombin Salakta with benzamidine is also altered as evidenced by an increased inhibition constant (Ki = 3.5 mmol/L v 0.28 mmol/L for normal thrombin). The inability of fibrinogen to act as a competitor in the inactivation of thrombin Salakta by diisopropylfluorophosphate clearly indicates that fibrinogen binding to the fibrinopeptide groove is drastically impaired. In contrast, interactions involving sites remote from the active site such as those with fibrin and thrombomodulin are only slightly impaired. These results indicate that thrombin Salakta exhibits a specific pattern of functional alterations different from those reported for other variants. The structural defect seems to affect essentially the primary substrate binding site and to a lesser extent recognition site(s) remote from the catalytic site such as those for fibrin and thrombomodulin.

Catalysis↗
PubMed1987

Measurement of glycated albumin in diabetic patients by biospecific affinity chromatography.

The percentage of glycated plasma albumin was measured by a procedure involving ammonium sulphate precipitation and Affi-Gel-Blue and phenylboronate chromatographies. The value correlates well with the amount of ketoamine-bound sugars determined by colorimetric assay (r = 0.98, n = 39). The normal mean value is 3.9 +/- 0.3% (mean +/- S.D., coefficient of variation = 7.7%, n = 32) and varies from 3.9 to 21% in diabetics (n = 54). A good correlation is found with the mean blood glucose value of the preceding twenty days (r = 0.92, n = 57). Because of its relative ease of determination, glycated albumin constitutes a good short-term glycemic index and an alternative to glycated haemoglobin in some specific cases.

Blood Glucose↗
PubMed1987

Coronary arteriography in the intact rabbit: demonstration of coronary vasomotor and electrocardiographic effects of ergonovine and indomethacin in rabbits after abrupt cessation of prolonged nitroglycerin treatment.

Abrupt withdrawal of chronic nitroglycerin treatment may predispose the coronary circulation to spasm. To test this hypothesis directly, we developed a technique for performing selective coronary arteriography in the intact rabbit with images obtained by video-based methods or cineangiographic film. Experiments were then conducted in rabbits treated three times daily with topical nitroglycerin ointment for 6 weeks and in age- and sex-matched control animals. Forty hours after cessation of treatment, animals were anesthetized, and ECG and coronary vasoactive effects (determined by coronary arteriography) of ergonovine, 0.2 mg/kg (intravenous) and indomethacin, 25 mg/kg (intravenous) were assessed. Of six nitroglycerin-treated rabbits, one died of ventricular fibrillation prior to arteriographic study. The remaining five developed ECG abnormalities (single ventricular premature beats, nonsustained ventricular tachycardia, and ST segment deviation) upon challenge with ergonovine or indomethacin. Neither agent evoked ECG changes in control rabbits. In contrast, the degree of luminal diameter reduction in epicardial coronary arteries provoked by ergonovine or indomethacin did not differ between control and nitroglycerin-treated animals. Focal coronary artery spasm was not observed in any rabbit. Our results demonstrate that selective coronary arteriography in rabbits is feasible and that changes in vessel caliber may be assessed from images thus obtained. Data from this study indicate that ergonovine- and indomethacin-induced ECG abnormalities observed in nitroglycerin-treated rabbits cannot be ascribed to epicardial coronary artery spasm.

Animals↗
PubMed1987

Contrast echo washout curves from the left ventricle: application of basic principles of indicator-dilution theory and calculation of ejection fraction.

Time-intensity curves can be obtained from contrast echocardiography of the left ventricle. The purposes of this study were: 1) to verify whether these curves conform to the basic principles of indicator-dilution theory; and 2) to derive indexes of left ventricular ejection fraction from curve analysis. In seven closed chest dogs, 31 doses of the polysaccharide agent SHU-454 were injected into the left ventricular cavity during apical four chamber two-dimensional echocardiography. Data were obtained at different levels of ejection fraction, which were induced by changes in preload, afterload and contractility, and measured by single plane Simpson's rule analysis of digital subtraction left ventriculograms. In a subset of two dogs, eight incremental doses (from 1 to 8 ml) of SHU were injected in the basal state. Contrast echocardiograms were digitized off-line, the mean gray level/pixel of a region of interest inside the left ventricular cavity was measured, and the average value for three systolic frames of each beat was used to obtain time-intensity curves. A good correlation was observed between the peak of the time-intensity curve and the quantity of contrast injected (correlation coefficient r = 0.91 by a logarithmic fit). The echo intensities observed in each animal were subsequently transformed in quantity of contrast according to these functions and their natural logarithm was calculated both with and without background subtraction. All curves relating time and the natural logarithm of the corrected intensity exhibited a descending rectilinear portion (washout) in which the correlation was very good (r = 0.97 +/- 0.02 = mean +/- SD) and which was not significantly affected by background subtraction. The validity of this fit was also unaffected by heart rate (55 to 158 beats/min) and angiographic ejection fraction (22 to 74%), and only minimally influenced by duration of contrast washout (3.3 to 14.6 seconds). Ejection fraction was calculated by an algorithm derived from indicator-dilution theory: ejection fraction = [1 - e(-bd)] X 100, where b = slope of the curve and d = cardiac cycle duration. Linear regression analysis between values of ejection fraction derived by angiography and contrast echo yielded r = 0.73. A second index, based on b and d, was derived by multiple regression analysis. Linear regression analysis of this index and angiographic ejection fraction yielded a correlation of r = 0.87.(ABSTRACT TRUNCATED AT 400 WORDS)

Animals↗
PubMed1987

ADP-induced platelet aggregation depends on the conformation or availability of the terminal gamma chain sequence of fibrinogen. Study of the reactivity of fibrinogen Paris 1.

Fibrinogen Paris I contains a mutant gamma chain that is longer than the normal chain, resulting in altered fibrin polymerization and cross-linking. Because these functions involve the carboxy-terminal region of the gamma chain, we decided to determine whether fibrinogen Paris I or the isolated Paris I gamma chain supports normal ADP-induced platelet aggregation, a function that requires the ultimate 12 residues of the normal gamma chain (400 through 411). Aggregation of ADP-stimulated normal platelets was defective with fibrinogen Paris I and markedly depressed with the gamma Paris I chain. These findings prompted us to characterize the carboxy-terminal structure of the region of the gamma Paris I chain responsible for this activity. The carboxy-terminal cyanogen bromide (CNBr) peptide of the normal gamma chain (385 through 411) or that from gamma Paris I was isolated by differential adsorption to triethylene-tetramine resin or by reverse-phase high-performance liquid chromatography (HPLC). The CNBr peptide from the Paris I gamma chain was identical to that of the normal gamma chain in its retention time on HPLC, its amino acid composition, and its sequence. Thus, the primary structure of the gamma Paris I chain from residue 384 through 411 is normal, indicating that a peptide insertion has occurred upstream from residue 384, resulting in an impairment of those physiologic functions attributable to the carboxy-terminal end of the gamma chain from position 384 (ie, cross-linking, ADP-induced platelet aggregation, and at least a portion of the gamma chain polymerization site). These observations demonstrate that the gamma chain platelet recognition site in the fibrinogen molecule is necessary but not alone sufficient to support normal ADP-induced platelet aggregation. There appears to be an additional requirement for normal conformation of the gamma chain or availability of its terminal sequence during the interaction of fibrinogen with platelets.

Adenosine Diphosphate↗
PubMed1986

Differences in DNase I sensitivity and methylation within the human beta-globin gene domain and correlation with expression.

We have analysed the chromatin features of DNA regions encompassing human epsilon, G gamma, A gamma, delta and beta globin structural genes in fetal and adult erythroid cells on the one hand and adult lymphocytes on the other. Highly purified nuclei from these cells were submitted to DNase I digestion and the kinetic data were obtained from the percentage of residual hybridization of defined regions in Southern blots. Our results, as others have shown by different approaches, indicate that the structural genes of the beta-globin cluster are generally more sensitive to DNase I in the erythroid cells than in non-erythroid cells. Thus a domain of DNase I sensitivity related to the committed state is defined. In addition we show that within this DNase-I-sensitive beta cluster domain, individual genes of the cluster are arranged in subdomains of differential DNase I sensitivity, which correlate with their expression status. Furthermore the differential expression of the two fetal genes in the fetal stage is shown to be directly proportional to the degree of hypomethylation of these genes.

Adult↗
PubMed1985

Purification, immunological and biochemical characterization of rat 28 kDa cholecalcin (cholecalciferol-induced calcium-binding proteins). Identity between renal and cerebellar cholecalcins.

The cholecalcins are intracellular vitamin D-dependent calcium-binding proteins. High concentrations of 28 kDa cholecalcins have been found in the kidneys and cerebella of birds and mammals. However, whereas the synthesis of the renal protein is vitamin D-dependent, that of the cholecalcin in the cerebella of young growing chicks and rats is apparently not. In the present study a range of immunological, physicochemical and structural characteristics of renal and cerebellar cholecalcins isolated from a single species, the rat, has been examined to ascertain what, if any, differences there are between them, other than the vitamin D-dependence. Both proteins behaved in exactly the same way during purification and showed complete immunocross-reactivity in Ouchterlony double immunodiffusion and radioimmunoassay. No difference could be detected between them on electrophoresis under denaturing conditions or on two-dimensional isoelectric focusing/electrophoresis. Their amino acid compositions were very similar, as were their u.v.-absorption spectra and their chymotryptic and tryptic peptide maps. The N-terminal sequence was found to be Gly-Gly-Val-Ser... for both cholecalcins. We have thus been unable to show any significant difference between the two proteins and suggest that the 28 kDa cholecalcins of the rat cerebellum and kidney are extremely similar, if not identical.

Amino Acids↗