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H S Yoon

Publications and source records attributed to H S Yoon.

At least 55 records · Page 3Linked to original sources
PubMed1992

Screening for mutations by RNA single-strand conformation polymorphism (rSSCP): comparison with DNA-SSCP.

Single-strand conformation polymorphism (SSCP) is a simple method for detecting the presence of mutations in a segment of DNA, but the fraction of all mutations detected is unclear. We have evaluated SSCP for the detection of single-base mutations in the factor IX gene. Multiple conditions were examined including electrophoresis temperature, electrophoresis buffer concentration, acrylamide to bis-acrylamide ratio, and water-cooled versus fan-cooled gel apparatuses. Depending on conditions, 10-11 of 12 known mutations were detected in a 183 bp segment whereas only 11-14 of 22 known mutations were detected in a 307 bp segment. We hypothesized that single stranded RNA should have a larger repertoire of secondary structure because shorter hairpins form stable duplexes and the 2' hydroxyl group is available for sugar-base and sugar-sugar hydrogen bonds. By incorporating phage promoter sequences into PCR primers, RNA-SSCP (rSSCP) could be compared directly with standard DNA SSCP. rSSCP was generally superior to SSCP, especially for the 307 bp segment. In addition, the abundance of transcript produced as a result of rSSCP allows the rapid, nonradioactive detection of mutations by staining the gel with ethidium bromide. To gauge the utility of the method in a prospective manner, a blinded study was performed in which SSCP, rSSCP, and direct genomic sequencing were compared in 28 patients with hemophilia B. A total of 2.6 kb of factor IX genomic sequence was examined in nine regions ranging from 180 to 497 nucleotides of factor IX sequence. Sequence changes at 20 different sites were detected by direct genomic sequencing; 70% of these were detected by rSSCP while only 35% were detected by SSCP.

Base Sequence↗
PubMed1992

Dideoxy fingerprinting (ddE): a rapid and efficient screen for the presence of mutations.

We describe dideoxy fingerprinting (ddF), a hybrid between dideoxy sequencing and SSCP that can detect the presence of single base and other sequence changes in PCR-amplified segments. As implemented herein, ddF involves a Sanger sequencing reaction with one dideoxynucleotide followed by nondenaturing gel electrophoresis. When ddF was used to examine segments of the human factor IX gene, 84 of 84 different mutations were detected with a very low rate of false positive signals. The approximate locations of the sequence changes could be determined from the ddF pattern and samples with different sequence changes had different fingerprints. In addition, large segments could be amplified and rapidly screened by ddF in multiple smaller subsegments. The patterns observed with ddF are instructive in that they suggest an inherent limitation in the detection of certain mutations by SSCP.

DNA Fingerprinting↗
PubMed1991

Stimulation of transcript elongation requires both the zinc finger and RNA polymerase II binding domains of human TFIIS.

The eukaryotic transcriptional factor TFIIS enhances transcript elongation by RNA polymerase II. Here we describe two functional domains in the 280 amino acid human TFIIS protein: residues within positions 100-230 are required for binding to polymerase, and residues 230-280, which form a zinc finger, are required in conjunction with the polymerase binding region for transcriptional stimulation. Interestingly, a mutant TFIIS with only the polymerase binding domain actually inhibits transcription, whereas a mutant in which the polymerase binding and zinc finger domains are separated by an octapeptide is only weakly active. The zinc finger itself has no effect on transcription, but in contrast to the wild-type protein, it binds to oligonucleotides. These findings suggest that TFIIS may interact with RNA polymerase II such that the normally masked zinc finger can specifically contact nucleotides in the transcription elongation zone at a position juxtaposed to the polymerization site.

Amino Acid Sequence↗
PubMed1991

Cloning, expression and characterization of the human transcription elongation factor, TFIIS.

The cDNA for the human elongation factor, TFIIS, has been cloned and expressed in E. coli with the T7 expression system. This 280-amino acid TFIIS protein is shorter by 21 residues than that of the mouse. The missing 21 residues are located in the amino-terminal region, which is not thought to be required for transcriptional stimulation. Apart from this gap, human and mouse proteins reveal 96% overall identity and 98.5% sequence similarity if conservative substitutions are taken into account. The bacterially expressed human protein and the purified calf thymus proteins are indistinguishable in their ability to stimulate transcript elongation by purified RNA polymerase II. Estimation of the native molecular size of the human protein in solution indicates that it exists as a dimer.

Amino Acid Sequence↗
PubMed1991

Missense mutations and evolutionary conservation of amino acids: evidence that many of the amino acids in factor IX function as "spacer" elements.

We report 31 point mutations in the factor IX gene and explore the relationship between the level of evolutionary conservation of an amino acid and the probability of a mutation causing hemophilia B. From our total sample of 125 hemophiliacs and from those reported by others, we identify 95 independent missense mutations, 94 of which occur at amino acids that are evolutionarily conserved in the available mammalian factor IX sequences. The likelihood of a missense mutation causing hemophilia B depends on whether the residue is also conserved in the factor IX-related proteases: factor VII, factor X, and protein C. Most of the possible missense mutations in generically conserved residues (i.e., those conserved in factor IX and in all the related proteases) should cause disease. In contrast, missense mutations in factor IX-specific residues (i.e., those conserved in human, cow, dog, and mouse factor IX but not in the related proteases) are sixfold less likely to cause disease. Missense mutations at nonconserved residues are 33-fold less likely to cause disease. At least three models are compatible with these observations. A comparison of sequence alignments from four and nine species of factor IX and an examination of the missense mutations occurring at CpG residues suggest a model in which most residues fall on opposite ends of a spectrum. In about 40% of residues, virtually any missense mutation in a minority of the residues will cause disease, while virtually no missense mutations will cause disease in most of the remaining residues. Thus, many of the residues in factor IX are spacers; that is, the main chains are presumably necessary to keep other amino acid interactions in register, but the nature of the side chain is unimportant.

Amino Acid Sequence↗
PubMed1991

Why does the human factor IX gene have a G + C content of 40%?

The factor IX gene has a G + C content of approximately 40% in all mammalian species examined. In human factor IX, C----T and G----A transitions at the dinucleotide CpG are elevated at least 24-fold relative to other transitions. Can the G + C content be explained solely by this hot spot of mutation? Using our mathematical model, we show that the elevation of mutation at CpG cannot alone lower the G + C content below 45%. To search for other hot spots of mutation that might contribute to the reduction of G + C content, we assessed the relative rates of base substitution in our sample of 160 families with hemophilia B. Seventeen independent single-base substitutions are reported herein for a total of 96 independent point mutations in our sample. The following conclusions emerge from the analysis of our data and, where appropriate, the data of others: (1) Transversions at CpG are elevated an estimated 7.7-fold relative to other transversions. (2) The mutation rates at non-CpG dinucleotides are remarkably uniform; none of the observed rates are either more than twofold above the median for transitions or more than threefold above the median for transversions. (3) The pattern of recent mutation is compatible with the pattern during mammalian evolution that has maintained the G + C content of the factor IX gene at approximately 40%.

Base Composition↗
PubMed1990

The pattern of factor IX germ-line mutation in Asians is similar to that of Caucasians.

To begin documenting the pattern of germ-line mutations in different human races, we have delineated the mutation in nine Korean families with hemophilia B by direct genomic sequencing of the regions of likely functional significance in the factor IX gene. An evaluation of these mutations in combination with previously described point mutations in the factor IX gene of Asians indicates that transitions predominate followed by transversions and microdeletions/insertions. Transitions at the dinucleotide CpG are a dramatic hot spot of mutation. This pattern of mutation is very similar to that observed in Caucasians with hemophilia B, despite the many differences between Asians (mostly Koreans) and Caucasians in diet, environment and cultural life-styles. The similarity may reflect the predominance of endogenous processes or ubiquitous mutagens rather than specific mutagens in the environment. The following additional conclusions emerge: (1) The missense mutations in Asians occur at evolutionarily conserved amino acids. When combined with the previous data this makes it likely that more than two-thirds of the missense mutations which could possibly occur at nonconserved amino acids do not cause hemophilia B. (2) Surprisingly, a change in the sixth base of the intron 2 donor splice-junction sequence is associated with severe disease in HB 74/77. (3) Direct carrier testing of nine Korean families demonstrates that the stability of DNA at ambient temperature in blood with the anticoagulant ACD solution B makes it feasible for a diagnostic laboratory to perform such testing at a distance of 7,000 miles. Carrier testing revealed that the mutation in HB78 arose in his mother's germ-line.(ABSTRACT TRUNCATED AT 250 WORDS)

Alleles↗
PubMed1989

[A survey on elements of an effective hospital infection control program for Korea].

The main purpose of this survey was to identify for elements of an effective hospital infection control program for Korea. Nurses and doctors who had participated in an education program for infection control or were responsible for hospital infection control were selected as data informers. The data were collected from 51 subjects by employing a Delphi technique in a series of three rounds from September 1, 1987, to March 31, 1988. In each round the responses to questionnaires were analyzed and the results were communicated back to the individuals. Finally 32 elements of effective hospital infection control program were reduced to 10 elements. 10 elements are as follows; 1) Hospital administrator's knowledge of importance and necessity for HIC (hospital infection control) 2) Supporting HIC administratively 3) Constituting infection control committee and role of ICC 4) Developing an educational program and providing a work manual for the hospital infection control staff 5) Educating and informing medical staff about hospital infection 6) Surveillance for hospital infection 7) Developing patient care technique 8) Controlling the hospital environment 9) Executing regular health examinations of all medical staff 10) Recruiting the medical staff sufficiently Three rankings of response rate about 32 elements are as follows: 1) Hiring a full time staff member for the HIC (66%) 2) Establishing a hospital policy and standards for the HIC (66%) 3) Activating the infection control committee and taking administrative action to support the ICC (63%) In addition the rankings of importance score by Likert 5 scale are as follows: 1) Washing hands scrupulously (4.88) 2) Nurses participation as key members of the ICC (4.75) 3) Reviewing and evaluating all ongoing aseptic techniques (4.69) In conclusion, first of all, administrative support must be given to hiring a full time staff member and to organization of infection control committee for the HIC in Korea.

Cross Infection↗
PubMed1986

Ultrasonic wave propagation and attenuation in wet bone.

The propagation of ultrasonic longitudinal waves in bovine plexiform and human Haversian bone has been studied over the range 0.5-16 MHz. In wet bone little velocity dispersion was observed, in contrast to the results of earlier studies on dry bone. Large values of attenuation were observed.

Animals↗
PubMed1984

The effects of remodeling on the elastic properties of bone.

Cortical bone can be modeled as a complex hierarchical composite interrelating both structure and material properties on four levels of structural organization: molecular, ultrastructural, microscopic, and macroscopic. In young animals, the microstructural systems are long parallel lamellar units, plexiform bone, which in older or more mature animals converts by internal remodeling into multiple concentric lamellar units, secondary osteons, forming haversian bone. Ultrasonic wave propagation measurements performed on both plexiform and haversian bone clearly show a definitive relationship with microstructure; haversian bone can be described as a transversely symmetric material whereas plexiform bone appears to be orthotropic in nature. The anisotropy of the elastic constants are found to reflect the tissue symmetry; moreover, plexiform bone is stiffer and more rigid in all directions than is haversian bone. Similar experiments were performed on osteoporotic and osteopetrotic bone. While the results for osteoporotic bone are understandable in terms of the increased porosity, the results for the osteopetrotic bone are anomalous with respect to its density. Since Wolff, the remodeling of bone has been interpreted as a way of altering the mechanical properties to suit some need. For haversian remodeling from plexiform bone, the argument that adaptation occurs to optimize properties requires additional clarification since haversian bone appears to have inferior mechanical properties to plexiform bone.

Animals↗
PubMed1983

Slow compressional wave propagation in wet human and bovine cortical bone.

Ultrasonic wave propagation in bovine plexiform and human Haversian bone was studied in the range 0.5 to 15 megahertz. A new longitudinal wave was observed which traveled more slowly than the ordinary longitudinal wave. The slow wave was associated with the dynamics of fluid motion in the pores of bone.

Animals↗