Search PubMed⌕ Search

Biomedical subjects

D L Nelson

Publications and source records attributed to D L Nelson.

At least 181 records · Page 10Linked to original sources
PubMed1994

Reliability of diagnostic assessment of normal and premutation status in the fragile X syndrome using DNA testing.

Until recently, fragile X [fra(X)] syndrome was diagnosed by cytogenetic techniques and/or linkage analysis. Investigation of the mutation at the molecular level has shown that amplification of a polymorphic trinucleotide repeat (CGG) is diagnostic of this syndrome. Fu et al. [1991] observed that between 6-54 copies of the repeat were associated with alleles found in the general population, whereas 50-200 copies were associated with the premutation. In general, differences in copy number between the normal and premutated states are sufficiently large so that the probability of misclassification is, for all practical purposes, zero. However, there is a grey area in which members from both populations overlap. The purpose of our study was to determine the probability of misclassifying an individual from either the general or premutation population. DNA obtained from the general population and transmitting fra(X) females were analyzed from 3 centers in North America: Houston, Texas; Rochester, Minnesota; and Kingston, Ontario. The distribution of normal alleles from Houston was not significantly different from those obtained from Rochester. Therefore, these 2 samples were combined and the pooled distribution of normal alleles was compared with the pooled distribution of premutations. Results indicated that if 50 repeats were used as the cutoff criterion, sensitivity is 100%, specificity is 99.6%, and the probability that an individual has the fra(X) premutation given that the number of repeats is greater than 50 is 95%. Other cutoff criteria (45, 55, 60, 65) employed produced like findings, although 55 repeats appears to be a marginally superior criterion to 50. An independent sample from Kingston was used to verify the original assessments.(ABSTRACT TRUNCATED AT 250 WORDS)

Bayes Theorem↗
PubMed1994

Robust amplification and ethidium-visible detection of the fragile X syndrome CGG repeat using Pfu polymerase.

We report on a method for ethidium bromide detection of FMR1 normal and premutation-size CGG triplet repeats. A set of PCR conditions has been optimized for the polymerase of the hyperthermophilic bacterium, Pyrococcus furiosus. Using this protocol, normal-size alleles ranging from 5 to 50 repeats, as well as most of premutation-size alleles, varying from > 50 to approximately 200 repeats, could be detected by ethidium bromide staining of agarose gels. Since the protocol requires neither autoradiography nor polyacrylamide gel electrophoresis, it promises to provide a rapid means for the exclusion of fragile X syndrome in males with mental retardation.

Base Sequence↗
PubMed1994

Increased serum soluble interleukin-2 receptors in Caucasian and Korean schizophrenic patients.

Recent studies have identified immunologic abnormalities in some schizophrenic subjects. This experiment replicates previous findings that serum soluble interleukin-2 receptors (SIL-2Rs) are elevated in schizophrenic patients, and is the first study to describe this phenomenon in non-Caucasian patients. Despite differences between Korean and Caucasian schizophrenic patients in absolute serum SIL-2R levels, both groups were significantly elevated when compared with their respective ethnic control groups (477 +/- 171 U/ml versus 354 +/- 172 U/ml and 763 +/- 347 U/ml versus 567 +/- 231 U/ml, respectively). Neither age, gender, medication status, nor duration of illness correlated with SIL-2R levels. These findings are further evidence that immune activation is present, regardless of ethnic origin, in some schizophrenic patients.

Adult↗
PubMed1994

Purification and characterization of calmodulin (lysine 115) N-methyltransferase from Paramecium tetraurelia.

Calmodulin (lysine 115) N-methyltransferase was purified from the cytosolic fraction of Paramecium tetraurelia by sequential dialysis, cellulose phosphate chromatography, Reactive Red 120 agarose chromatography, and calmodulin-Sepharose affinity chromatography. The enzyme was purified 6800-fold with a 15% yield. SDS-PAGE analysis of the purified enzyme invariably revealed a major protein of 37 kDa that was reproducibly obtained and minor proteins of 35 and 28 kDa that were sometimes obtained in variable yields. The enzyme formed a mixture of mono-, di-, and trimethyllysine residues at lysine 115 of calmodulin in vitro, had a Km for the methyl donor, S-adenosyl methionine (AdoMet), of about 1 microM and a pH optimum of about 7.5. The purified enzyme had an absolute requirement for the reductant DTT for activity, whereas the enzyme in crude fractions did not. The enzyme is a monomer with an estimated molecular mass of 33 kDa. Ca2+, Mg2+, Mn2+, and Ni2+ stimulated calmodulin N-methyltransferase activity but Zn2+ did not. Calmodulin N-methyltransferase was inhibited by its reaction product S-adenosyl homocysteine (SAH), but not by sinefungin and tubercidin. The calmodulin antagonists calmidazolium and mellitin were inhibitory but W7 was not. The enzyme was not stimulated by Triton X-100 nor by NaCl. Only calmodulins with an unmethylated lysine at residue 115, including cam2 calmodulin, were substrates. Histones and calcium-binding proteins from Paramecium other than calmodulin did not act as substrates for the purified calmodulin N-methyltransferase and no other substrates in the cytosolic fraction were observed.

Animals↗
PubMed1994

Advances in molecular analysis of fragile X syndrome.

Fragile X syndrome is a common cause of mental retardation that is inherited as an X-linked dominant disorder with reduced penetrance. Fragile X syndrome has been shown to be caused by an unstable CGG repeat within the fragile X mental retardation-1 (FMR1) gene. The repeat is normally polymorphic with six to 52 repeats, while affected males and females exhibit a massive expansion resulting in 230 to more than 1000 repeats. Such expansions, called "full mutations," are associated with abnormal methylation of the FMR1 gene leading to transcriptional suppression. The resulting absence of the encoded protein, FMRP, a cytosolic RNA-binding protein, is believed to result in the phenotype. Nonpenetrant male carriers and many female carriers exhibit premutation alleles of intermediate length (50 to 230 repeats), which are normally expressed. Male carriers transmit only unstable premutations while female premutation carriers can have carrier offspring with premutations or affected children with full mutations. The risk of having an affected child is directly related to the number of maternal repeats, with sequentially increasing probabilities of these alleles converting to full mutations as they are transmitted to subsequent generations. Advances have led to highly accurate laboratory diagnoses of both carrier and affected individuals as well as markedly improved prenatal diagnosis. In addition, a previously unrecognized class of mutation, later found responsible for several other important genetic diseases, has emerged.

DNA↗
PubMed1994

Dual x-ray absorptiometry forearm software: accuracy and intermachine relationship.

An ash study was performed to analyze the accuracy of forearm measurements at the one-third site of three dual x-ray absorptiometry (DXA) systems using 20 cadavers. A Hologic QDR-2000, a Hologic QDR-1000/W, and a Lunar DPX-L system were used. The correlations between ash weight and DXA BMC were excellent for the three instruments (r > 0.97, p < 0.001), with accuracy errors < 5.2%. To perform a forearm cross-calibration of bone mineral content (BMC) and density (BMD) between the scanners, 10 healthy volunteers were additionally scanned at the distal one-third radius. The correlations among the DXA machines were excellent (r > 0.95); the absolute BMC and BMD values were significantly different between the two Hologics and the Lunar machine. The slope and intercept of both the BMC and BMD between the two Hologic systems were close to unity. In conclusion, the DXA forearm software packages provide accurate methods for assessing bone mineral content and density. The conversion of data among different manufacturers should be performed by careful cross-calibration measurements.

Absorptiometry, Photon↗
PubMed1994

Racial variation in serum-soluble interleukin-2 receptor levels: a population-based study of healthy smokers and nonsmokers.

To investigate the influence of race and cigarette smoking on serum levels of soluble interleukin-2 receptors (sIL-2R), we studied a population-based cohort of 282 white and 173 black adults, ages 20-69 years. Serum sIL-2R concentrations in this healthy population ranged from 146 to 2623 U/ml. Whites had significantly higher sIL-2R levels than blacks (455 versus 365 U/ml; P < 0.001), and cigarette smokers had significantly higher levels than nonsmokers (508 versus 420 U/ml; P = 0.01). White smokers had the highest levels (550 U/ml); white nonsmokers and black smokers had intermediate levels (455 and 450 U/ml, respectively); and black nonsmokers had the lowest levels (365 U/ml). Smoking cessation appeared to normalize sIL-2R levels; exsmokers who had not smoked for at least 2 years had sIL-2R levels similar to those of never smokers. These data demonstrate the wide range of serum sIL-2R concentrations found in normal healthy adults and the significant influence of race and cigarette smoking. Further investigation of this natural heterogeneity may provide insights into the mechanisms underlying genetic and environmental influences on this important immunologic parameter.

Adult↗
PubMed1994

Factors influencing serum neopterin and beta 2-microglobulin levels in a healthy diverse population.

Sera and questionnaire data from a population-based random sample of healthy adults was used to evaluate factors influencing neopterin and beta 2-microglobulin (beta 2m) values. Both neopterin and beta 2m levels increased with age and were higher among white than blacks (mean values for whites and blacks: neopterin, 5.06 vs 4.49 nmol/L; beta 2m, 1.36 vs 1.28 mg/L). Gender differences were noted for beta 2m but not neopterin values (beta 2m males vs females: 1.37 vs 1.29 mg/L). Neopterin values were lower among current smokers than among nonsmokers (4.32 vs 5.16 nmol/L) and were higher among users of antihistamines (5.46 among users vs 4.65 nmol/L among nonusers). Neopterin and beta 2m were correlated in this healthy adult population (adjusted r = 0.53, P = 0.001), yet no other interrelationships with numerous biologic markers except between beta 2m and serum-soluble interleukin-2 receptor levels (adjusted r = .41, P = 0.05) were observed. These findings provide important baseline information to consider before planning or evaluating studies utilizing neopterin or beta 2m levels.

Adult↗
PubMed1994

Molecular genetic analysis of the primary immunodeficiency disorders.

Exciting progress in the search for the genetic basis of the primary immune deficiency disorders continues to shed insight into functioning of the immune system in health and disease. The molecular genetic causes of 7 of the 17 WHO-recognized primary immune deficiency diseases have been defined. Additional studies will shed more insight into congenital and acquired immune deficiency disorders.

Chromosome Mapping↗
PubMed1994

An assessment of serum and urine soluble interleukin-2 receptor concentrations during renal transplant rejection.

During rejection, renal transplant recipients have increased concentrations of soluble interleukin-2 receptors (sIL-2R) in their serum and urine. However, the clinical application of this measurement in the diagnosis of rejection or the assessment of treatment efficacy is limited by the variance of the measurement in sample populations. We examined the serum and urine sIL-2R concentrations in 20 renal transplant recipients, 12 of whom experienced 13 episodes of allograft rejection. There was no statistical difference in the mean serum sIL-2R concentration at the time of rejection compared with the baseline value (2,817 +/- 801 v 1,943 +/- 255 U/mL). By contrast, the urinary excretion rate, expressed as units of sIL-2R per milligram creatinine, was 26.2 +/- 6.4 compared with 14.2 +/- 2.5 (P < 0.05). Furthermore, when urinary sIL-2R was expressed as a fractional excretion (FE), both the absolute measurement (4.4% +/- 1.7%) and the percent increase (+245%) at the time of rejection provided the greatest degree of discrimination of rejection from those values during allograft stability (1.2% +/- .2% and +2.5%, respectively; P < 0.005). We conclude that (1) serum and urine sIL-2R concentrations are affected by a number of factors during rejection; (2) FE calculations of sIL-2R improve discrimination of rejection from graft stability; and (3) serial measurement of sIL-2R excretion may be a useful adjunct to the diagnosis of rejection and, possibly, the subsequent assessment of response to immunotherapy.

Adult↗
PubMed1994

Length of uninterrupted CGG repeats determines instability in the FMR1 gene.

Analysis of 84 human X chromosomes for the presence of interrupting AGG trinucleotides within the CGG repeat tract of the FMR1 gene revealed that most alleles possess two interspersed AGGs and that the longest tract of uninterrupted CGG repeats is usually found at the 3' end. Variation in the length of the repeat appears polar. Alleles containing between 34 and 55 repeats, with documented unstable transmissions, were shown to have lost one or both AGG interruptions. These comparisons define an instability threshold of 34-38 uninterrupted CGG repeats. Analysis of premutation alleles in Fragile X syndrome carriers reveals that 70% of these alleles contain a single AGG interruption. These data suggest that the loss of an AGG is an important mutational event in the generation of unstable alleles predisposed to the Fragile X syndrome.

Base Sequence↗
PubMed1994

Isolation of a GCC repeat showing expansion in FRAXF, a fragile site distal to FRAXA and FRAXE.

Three folate-sensitive fragile sites, termed FRAXA, FRAXE and FRAXF, have been identified on the distal end of chromosome Xq. The first two contain expanded, hypermethylated and unstable CGG (or GCC) repeats within CpG islands. We now report the isolation of similar sequences responsible for the third fragile site, FRAXF. A 5-kilobase EcoRI fragment derived from a cosmid coincident with the cytogenetic anomaly detects expanded, methylated and unstable sequences in five individuals who exhibit fragile sites in distal Xq; these individuals have normal repeat lengths at both FRAXA and FRAXE. By sequence analysis, the expanded region contains a GCC repeat. PCR and sequence analysis of chromosomes from the general population indicates that the repeat is polymorphic (6 to 29 triplets), and is stable upon transmission.

Alleles↗
PubMed1994

Loss of mutation at the FMR1 locus through multiple exchanges between maternal X chromosomes.

The mutation observed in the fragile X syndrome, an X-linked inherited disorder causing mental retardation, is almost exclusively an expanded CGG repeat in the first exon of the FMR1 gene. Here we describe a daughter of a female carrier, who inherited the fragile X premutation chromosome based on haplotype analysis using flanking markers. However, the CGG repeat sequence and the intragenic polymorphic marker FMRb showed the normal maternal alleles, while two other intragenic markers, FMRa and FRAXAC2 and other, more distant markers, showed the risk haplotype. Since FMRa and FRAXAC2 are located in between the markers CGG and FMRb, this results in patches of normal and fragile X sequences in the FMR1 gene of the daughter. This observation is very likely due to gene conversion. As this daughter received a normal CGG repeat region, we expect that her risk to have affected offspring is the same as the population risk. The observed phenomenon would therefore represent a back mutation at the FMR1 locus.

Base Sequence↗
PubMed1994

Frequency and stability of the fragile X premutation.

Although considered the most common heritable cause of neurodevelopmental disability, precise prevalence figures for the FMR1 mutation in the general population are lacking. Since no fragile X premutation alleles have yet been observed to originate from FMR1 alleles within the normal size range, there is also little information available about the origin of the fragile X premutation and mechanisms leading to instability of the FMR1 trinucleotide repeat region. In this study, 977 genetically unrelated individuals from families unselected for mental retardation or fragile X were analyzed with Southern blot analysis for the presence of FMR1 mutations. A subgroup of subjects with evidence of a large CGG repeat number, and any available relatives, were further studied with PCR to investigate the stability of the trinucleotide repeat segment of FMR1. One subject had a 75 repeat length which was unstable (increased in size) when passed to subsequent generations. This includes one male descendent who had a premutation/full mutation mosaic pattern. Two other alleles with > or = 46 repeats from different subjects were also found to be unstable and increased in size in subsequent generations. Considering all three unstable alleles to be indicative of an evolving or actual premutation, the estimated frequency of the fragile X premutation is one in 510 X chromosomes. However, since 11 other alleles with > or = 46 repeats were found to be stable through at least one meiotic transmission, repeat length appears to be an important but not sufficient condition leading to instability of the FMR1 gene.

Adolescent↗
PubMed1994

Radiographic absorptiometry for bone mineral measurement of the phalanges: precision and accuracy study.

PURPOSE: To evaluate the accuracy and precision of a radiographic absorptiometry (RA) method for assessment of bone mineral of the middle phalanges. MATERIALS AND METHODS: Nineteen cadaveric hands were radiographed with an aluminum wedge, once at 50 kVp and 400 mA and once at 60 kVp and 300 mA. Bone mineral content (BMC) and bone mineral density (BMD) of the second to fourth middle phalanges, expressed in arbitrary units (BMC-AU and BMD-AU), were analyzed and averaged in each hand. RESULTS: The precision error of this method was 1.0% for BMC-AU and 0.6% for BMD-AU. A 2.0%-2.4% reduction in BMD-AU seen on radiographs obtained through ethanol thicknesses of 5 and 6 mm compared with that seen on controls was statistically significant (P < .01). The correlation between BMC-AU and forearm BMC determined with dual x-ray absorptiometry was good (r = .887), and that between BMC-AU and ash weight in the phalanges was excellent (r = .983). CONCLUSION: The RA method is precise and accurate for bone mineral assessment of the peripheral appendicular skeleton.

Absorptiometry, Photon↗