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L J Elsas

Publications and source records attributed to L J Elsas.

At least 73 records · Page 4Linked to original sources
PubMed1991

A simple method for evaluation of Rb+ transport and Na(+)-K+ pump stoichiometry in adherent cells.

This report describes a method based on flame photometry for the evaluation of transmembrane Rb+ transport and Na(+)-K+ pump stoichiometry in adherent cells. In monolayers of cultured fibroblasts, the rates of 86Rb+, an isotope widely used as a K+ congener in transport studies, and nonradioactive Rb+ influx were equivalent when measured in the absence and presence of the transport inhibitors ouabain and bumetanide. Ouabain- and bumetanide-sensitive Rb+ fluxes were also equal with the two methods. Flame photometry allowed the simultaneous determination of intracellular [Na+] in the same sample in which Rb was measured. The incubation of human fibroblasts with ouabain for 5 min promoted a significant increase in intracellular [Na+]. Under appropriate experimental conditions, the ratio between the rate of ouabain-promoted increase in intracellular [Na+] and ouabain-sensitive Rb+ influx was 1.4, close to the theoretical value of 1.5 corresponding to a Na(+)-K+ pump stoichiometry of 3 Na+ extruded from the cell in exchange for 2 K+.

Biological Transport, Active↗
PubMed1991

Assignment of Emery-Dreifuss muscular dystrophy to the distal region of Xq28: the results of a collaborative study.

Emery-Dreifuss muscular dystrophy (EDMD) is an X-linked humeroperoneal dystrophy associated with cardiomyopathy that is distinct from the Duchenne and Becker forms of X-linked muscular dystrophy. Linkage analysis has assigned EDMD to the terminal region of the human X chromosome long arm. We report here further linkage analysis in two multigenerational EDMD families using seven Xq28 marker loci. Cumulative lod scores suggest that EDMD is approximately 2 cM from DXS52 (lod = 15.67) and very close to the factor VIII (F8C) and the red/green color pigment (R/GCP) loci, with respective lod scores of 9.62 and 10.77, without a single recombinant. Several recombinations between EDMD and three proximal Xq28 markers suggest that the EDMD gene is located in distal Xq28. Multipoint linkage analysis indicates that the odds are 2,000:1 that EDMD lies distal to DXS305. These data substantially refine the ability to perform accurate carrier detection, prenatal diagnosis, and the presymptomatic diagnosis of at-risk males for EDMD by linkage analysis. The positioning of the EDMD locus close to the loci for F8C and R/GCP will assist in future efforts to identify and isolate the disease gene.

Chromosome Mapping↗
PubMed1990

Identification of a common mutation in patients with medium-chain acyl-CoA dehydrogenase deficiency.

Medium-chain acyl-CoA dehydrogenase (MCAD) deficiency is one of the most common recessively inherited metabolic diseases in man. We have studied fibroblast cultures obtained from three patients with MCAD deficiency by sequencing the entire coding region of MCAD mRNA. A single A to G nucleotide replacement which resulted in lysine329-to-glutamic acid329 substitution of the MCAD protein was identified in all cultures. Furthermore, this point mutation was present in 91% (31 of 34) of mutant MCAD alleles, indicating that the majority of cases with MCAD deficiency are caused by this type of mutation.

Acyl-CoA Dehydrogenase↗
PubMed1990

Insulin-receptor autophosphorylation and kinase activity are constitutively increased in fibroblasts cultured from a patient with heritable insulin-resistance.

Mutations in the insulin receptor gene have been described in families with the inherited insulin-resistant syndrome leprechaunism. At a cellular level, these mutations result in decreased insulin binding and impaired insulin stimulation of receptor autophosphorylation and sugar transport. By contrast, we previously found that fibroblasts cultured from leprechaun patient Atl had constitutively increased sugar transport, even though insulin binding was markedly reduced. Here we report that these fibroblasts have basal insulin-receptor autophosphorylation and kinase activity constitutively increased above insulin-stimulated control cells.

Cells, Cultured↗
PubMed1990

Selective urinary screening for mucopolysaccharidoses.

Two methods for analysis of urinary glycosaminoglycans (GAGs) have been modified and improved for efficient screening of mucopolysaccharidoses. Urinary excretion of GAGs was estimated by spectrophometric measurement of alcian blue complex formation and was used in conjunction with qualitative analysis by thin layer chromatography. After normal variation in GAG excretion was established using 120 urine samples, these screening methods were applied to a total of 2057 urine samples over 4 years. Six patients with abnormal urinary GAG excretion had a mean of 50.5 +/- 20.6 mg GAG/mmol creatinine compared to 3.4 +/- 2.9 for age-matched controls. Qualitative analysis by thin layer chromatography using alternating solvent systems identified the GAGs excreted in excess and facilitated selection of specific enzyme assays for final confirmation. Six cases were diagnosed prospectively and demonstrate these quantitative and qualitative methods to be economical, efficient, and suitable for clinical use.

Chromatography, Thin Layer↗
PubMed1990

Human fibroblasts express the insulin-responsive glucose transporter (GLUT4).

The transport of glucose across the plasma membrane of nonepithelial cells is mediated by a family of facilitative glucose transporters. One glucose transporter is insulin-responsive (GLUT4) and is found in muscle, heart, and fat cells, differentiated cells which are difficult to maintain and study in culture. Cultured dermal fibroblasts, on the other hand, also are insulin-responsive, can be easily maintained in culture, and retain the genetic complement of the donor. In this paper, we evaluate RNA isolated from cultured human fibroblasts for the expression of four different glucose transporters. Northern blot analysis indicated that human fibroblasts expressed the erythrocyte (GLUT1), the fetal skeletal muscle (GLUT3), and the insulin-responsive (GLUT4) glucose transporters, but not the liver glucose transporter (GLUT2). To confirm the presence of GLUT4 mRNA, cDNA was synthesized from human fibroblast RNA, amplified using primers specific for GLUT4 by the polymerase chain reaction, and sequenced. The sequence was identical to that of GLUT4 cDNA. These data indicate that cultured human fibroblasts express at least three genetically distinct facilitative glucose transporters.

Base Sequence↗
PubMed1990

Sequence of a cDNA encoding human galactose-1-phosphate uridyl transferase.

We report a revised sequence of a cDNA that encodes a human galactose-1-phosphate uridyl transferase. The cDNA is 1295 bases in length and encodes a 43,000 Mr protein. The sequence was derived from a cDNA clone isolated from a transformed human lymphoblast cell line and amplified in a polymerase chain reaction. The revised sequence reveals a higher degree of amino acid conservation between the human enzyme and the homologous enzymes from Escherichia coli and yeast than was previously thought to exist.

Amino Acid Sequence↗
PubMed1989

Prenatal ascertainment of triploidy by maternal serum alpha-fetoprotein screening.

We report our experience in ascertaining fetal triploidy during routine maternal serum alpha-fetoprotein (MSAFP) screening. Three cases were identified after elevated MSAFP tests. Two of the three had normal amniotic fluid alpha-fetoprotein (AFAFP). The third had amniocentesis too late for AFAFP interpretation. Three additional cases were detected by amniocentesis without prior MSAFP screening and none had an elevated AFAFP. A literature review revealed eight triploid fetuses detected as a result of an elevated MSAFP. Of the five with AFAFP quantitation, only one had an abnormal value and the elevation was minimal. In these 14 cases from our own and other reports, ultrasound findings of placental and fetal abnormalities were often noted, but a pattern diagnostic of triploidy was not present. We conclude that, for optimal prenatal detection of triploidy, fetal karyotyping should be included when an amniocentesis is performed for elevated MSAFP.

Adult↗
PubMed1989

EEG mean frequencies are sensitive indices of phenylalanine effects on normal brain.

We previously reported that changes in plasma phenylalanine (PHE) concentrations of 1000 microM or more adversely affected cognitive function and reduced mean frequency of the EEG power spectrum. In the present study, we characterized EEG effects of changes in plasma PHE from physiological to supraphysiological concentrations. Subjects were mentally normal children and adult volunteers with 3 different genotypes for phenylalanine hydroxylase (PHY): homozygous deficient, heterozygous, and homozygous normal. Double-blinded crossover studies were performed at equilibrium during PHE restriction and supplementation. The mean frequency of the power spectrum and the mean across a set of alpha-theta factors showed highly significant, reversible, generalized EEG slowing during PHE supplementation in subjects homozygous for PHY deficiency. Smaller but significant changes in EEG mean frequencies occurred in the heterozygous and normal subjects. Spectral profiles of EEG change were similar in both groups; the mean alpha-theta was more sensitive in the second group. Demonstration of EEG changes with PHE supplementation in normal individuals has important dietary implications.

Adult↗
PubMed1989

Increased glucose transport by human fibroblasts with a heritable defect in insulin binding.

Insulin and IGF-I binding and their regulation of hexose transport were evaluated in skin fibroblasts cultured from a family (Atl) whose proband had leprechaunism, hypoglycemia, and severe insulin resistance. High affinity insulin binding to proband Atl cells was absent, and partially, but equally, impaired in fibroblasts from his related parents. IGF-I binding to his cultured fibroblasts was within the normal range. Cells from proband Atl had insulin receptor mRNAs similar to control fibroblasts. 3-O-Methyl-D-glucose (OMG) transport by proband Atl was threefold higher than in control fibroblasts (37.7 v 7.6-11 nmol/mL/s) and was insulin-insensitive. Proband Atl fibroblasts had a threefold increase in the Vmax for OMG entry and a concomitant increase in the number of D-glucose-inhibitable cytochalasin B binding sites on their plasma membrane. Similar levels of glucose transporter mRNA were observed in control and proband Atl fibroblasts. These results suggest that fibroblasts from patient Atl have a genetically transmitted mutation in the alpha subunit of their insulin receptor. In the homozygous affected proband, this mutation impairs insulin binding and causes elevated, insulin-insensitive glucose transport. The dysfunction resulting from this mutation is similar to that introduced in Chinese hamster ovary cells by transfection with a truncated alpha subunit.

3-O-Methylglucose↗
PubMed1989

Restriction fragment length polymorphisms of the insulin receptor gene in families with insulin resistance and leprechaunism.

Restriction fragment length polymorphisms (RFLPs) for the insulin receptor gene were compared among controls and families Atl and Ark-1, whose probands have extreme insulin resistance and leprechaunism. Previous studies indicated that fibroblasts cultured from patients Atl and Ark-1 had less than 10% of normal insulin binding. Cells cultured from the related parents of Atl had partial reduction in insulin binding, suggesting that patient Atl inherited the same mutant allele for the insulin receptor from both his parents. By contrast, the parents of Ark-1 were unrelated and their cells had different degrees of impaired insulin binding, suggesting that patient Ark-1 inherited two different, non-complementing alleles for the insulin receptor. To test these hypotheses, RFLPs were defined for the insulin receptor gene and their transmission was studied in families Atl and Ark-1. Patients affected with leprechaunism had no private polymorphisms in the insulin receptor gene, indicating that gross rearrangements were not present. A polymorphism generated with EcoRI at the 5' end of the insulin receptor gene gave fragments of 16 and 20 kb and was the only one informative in family Atl. Both parents were heterozygous, presenting a 16 and a 20 kb fragment, and transmitted the 20 kb allele to their affected offspring. Ark-1's parents were heterozygous for three different RFLPs, including this EcoRI polymorphism, and patient Ark-1 inherited a different allele for the insulin receptor from each parent. These results support the genetic hypotheses suggested by insulin-binding studies and indicate that RFLPs can be used to identify transmission of the insulin receptor gene in families with insulin resistance.

DNA Probes↗
PubMed1988

Mitochondrial DNA mutation associated with Leber's hereditary optic neuropathy.

Leber's hereditary optic neuropathy is a maternally inherited disease resulting in optic nerve degeneration and cardiac dysrhythmia. A mitochondrial DNA replacement mutation was identified that correlated with this disease in multiple families. This mutation converted a highly conserved arginine to a histidine at codon 340 in the NADH dehydrogenase subunit 4 gene and eliminated an Sfa NI site, thus providing a simple diagnostic test. This finding demonstrated that a nucleotide change in a mitochondrial DNA energy production gene can result in a neurological disease.

Animals↗
PubMed1988

Verification of the fetal valproate syndrome phenotype.

We have evaluated 19 children who were exposed to valproic acid (VPA) in utero to look for manifestations of a fetal valproate syndrome (FVS), as proposed by Di Liberti et al. [1984]. We found no consistent alterations of pre- or postnatal growth with exposure to VPA monotherapy. Postnatal growth deficiency and microcephaly were present however, in two thirds of children exposed to VPA in combination with other anticonvulsants. Developmental delay or neurologic abnormality was found in 71% of those exposed to VPA monotherapy, and in 90% of those exposed to VPA and other anticonvulsants. Craniofacial anomalies, which can be seen with other anticonvulsant exposures, including midface hypoplasia, short nose with a broad and/or flat bridge, epicanthal folds, minor abnormalities of the ear, philtrum or lip, and micrognathia were also found in infants whose mothers used VPA. Prominent metopic ridge and outer orbital ridge deficiency or bifrontal narrowing and certain major anomalies such as tracheomalacia, talipes equinovarus (with intact spine) and lumbosacral meningomyelocele seem to be peculiar to infants with VPA exposure. Other defects such as urogenital anomalies, inguinal or umbilical hernias, and minor digital anomalies that are common to other prenatal anticonvulsant exposures are also occasionally found in those exposed to VPA. Heart defects have been found in infants exposed to nearly every class of anticonvulsant although the types of defects associated with maternal VPA use may be clarified when classified by pathogenetic mechanism. Our findings overall are in agreement with the report of Di Liberti et al. [1984].

Abnormalities, Drug-Induced↗
PubMed1988

Fine mapping of glycerol kinase deficiency and congenital adrenal hypoplasia within Xp21 on the short arm of the human X chromosome.

We have studied patients with Duchenne muscular dystrophy (DMD), DMD together with glycerol kinase (GK) deficiency, or DMD together with both GK deficiency and congenital adrenal hypoplasia (AHC). Analysis of deletions in these patients allows the mapping of these mutations in Xp21. The following order is proposed: Xpter - L1 - AHC - GK - DMD - Xcen. One of the boys with DMD, GK, and AHC is shown by pulsed-field-gel electrophoresis to have a deletion which has a proximal endpoint at least 500 kb distal from the pERT87 (DXS164) locus.

Acid Phosphatase↗