PURPOSE: Endometriosis is a steroid dependent disease with a particular genetic background but the location of possible genomic aberrations are still poorly clarified. This study was designed to investigate the associations between the polymorphism of the progesterone receptor gene (PROGINS) and endometriosis. METHODS: 100 women with surgically diagnosed and histologically confirmed endometriosis were enrolled as a patient population and a total of 110 female control subjects undergoing health examination were enrolled as control population. DNA extraction and polymerase chain reaction (PCR) were used to genotype women for the presence of the PROGINS polymorphism in peripheral blood samples. The x2-test was used to compare genotype distributions between endometriosis and controls. RESULTS: T1/T2 heterozygote was found to be one patient in each group, and the rest of the subjects were all T1/T1 homozygotes. There was no difference in the genotype distribution between the endometriosis group and the control group. CONCLUSION: These results suggest that the progesterone receptor gene PROGINS is not associated with the risk for endometriosis.
DNA ; Endometriosis* ; Female ; Genotype ; Heterozygote ; Homozygote ; Humans ; Polymerase Chain Reaction ; Progesterone* ; Receptors, Progesterone*

No abstract available.
DNA*

Hereditary motor and sensory neuropathy (HMSN; Charcot-Marie-Tooth disease, CMT) was first described by Charcot and Marie in France and, independently, by Tooth in England in 1886. HMSN is the most common form of inherited motor and sensory neuropathy, and is a genetically heterogeneous disorder of the peripheral nervous system. Using positional cloning methods, the chromosomal localization (locus) of more than 40 inherited peripheral neuropathies was found in the last 15 years. However, these genetic analyses also show that many entities do not show linkage to the known loci. This issue deals with a clinical survey of inherited peripheral neuropathies regarding diagnostic approaches based on the molecular findings.
Charcot-Marie-Tooth Disease ; Clone Cells ; Cloning, Organism ; Diagnosis ; Diagnostic Tests, Routine* ; DNA* ; England ; France ; Hereditary Sensory and Motor Neuropathy* ; Peripheral Nervous System ; Peripheral Nervous System Diseases ; Tooth

Extensive research in this decade has led to detailed understanding of genetic changes underlying human cancers. Two major tumorigenic events are mutation and amplification of oncogenes and inactivation of tumor suppressor genes. These events then trigger a series of signal transduction cascades, activating expression of downstream genes that control various cellular activities including cell cycle progression, DNA synthesis, programmed cell death, DNA repair, and cell migration. Investigations of these molecular pathways has led to the identification of many targets for therapeutic intervention. Knowledge of the expression patterns and functions of all human genes wil l provide a frame work for future molecular, genetic medicine. During the past ten years, the human genome project has generated an enormous amount of sequencing information, and sequencing of the entire human genome may be completed by the year 2003 (1,2). One can envision that this will irreversibly transform the methodology of medical research and the practice of medicine. The search for new genes, which currently consumes the effort of many talented scientists, will become past history. Additionally, studying one gene at a time will be replaced by studying large number of genes simultaneously(3). Reductionistic approaches to human disease will be replaced by systemic approach. As a prelude to this revolution, tools used for parallel analysis of gene expression in the format of ordered gene arrays have been developed and are under continued expansion. In this technical tip, we will introduce the Atlas Human cDNA Expression Array system developed by Clontech Laboratories, Inc.(4). With this technology, a conventional laboratory can profile the expression of 588 human genes simultaneously in one simple experiment without the using of expensive equipment. We will demonstrate the profiling of 588 genes in a human glioblastoma cell line to exemplify the utility of this technique.
Aptitude ; Cell Cycle ; Cell Death ; Cell Line ; Cell Movement ; DNA ; DNA Repair ; DNA, Complementary* ; Gene Expression ; Genes, Tumor Suppressor ; Genome, Human ; Glioblastoma ; Glioma* ; Human Genome Project ; Humans* ; Oncogenes ; Signal Transduction

In order to examine whether microdeletions on the Y chromosome exist or not and observe the aspects of expression of DAZ which is suggested to be essential in spermatogenesis in testicular tissue, tissues of 21 patients with azospermia were analyzed by using PCR methods and reverse transcription-PCR. The primers used for the analysis of the microdeletions on the Y chromosome were gene-specific. According to the results of the PCR with genomic DNA of the peripheral blood extracted from each patient, of the 21 men with azospermia 2 displayed microdeletions of the DAZ gene in the Y chromosome but none of HSP70A and HSP70B. And the reverse transcription-PCR of the RNA extracted from the testicular tissue of the patients gave results which found no amplified products of the mRNA of DAZ in the patients with microdeletions of that gene as expected, and confirmed patterns of expression of the mRNA of HSP70B in 5 whereas none could be seen with regard to HSP70A. Furthermore, the sperm of these 5 men were discovered t In accordance with the results previously described, one can see that the microdeletions of DAZ are associated with spermatogenesis and contemplate that HSP70B plays an important part in the maturation process of sperm. But it is considered that there is no correlation between the genes DAZ and HSP and since factors associated with the process of spermatogenesis are being continously discovered more studies on this should be advanced.
Azoospermia ; DNA ; Humans ; Male ; Polymerase Chain Reaction ; RNA ; RNA, Messenger ; Spermatogenesis ; Spermatozoa ; Y Chromosome

Painting probe for HSRs in IMR-32 Neuroblastoma, a pediatric malignant neoplasm of neural crest origin, has a wide range of clinical virulence. The mechanisms contributing to the development of neuroblastomas are largely unclear, but non-random chromosomal changes identified over the past years suggest the involvement of genetic alterations. Amplification of the human N-myc proto-oncogene is frequently seen either in extrachromosomal double minutes or in homogeneously staining regions(HSRs) of aggressively growing neuroblastomas. N-myc maps to chromosome 2 band 24, but HSR have never been observed at this band, suggesting transposition of N-myc during amplification. We have constructed and analyzed the region-specific painting probe for HSR in neuroblastoma IMR-32 to determine the derivative chromosomes. Microdissection was performed on HSR using an inverted microscope with the help of microglass needles and an micromanipulator. We pretreated the microdissected fragments with Topoisomerase I which catalyzes the relaxation of supercoiled DNA, and performed two initial rounds of DNA synthesis with T7 DNA polymerase followed by conventional PCR to enable the reliable preparation of Fluorescent in situ hybridization probe from a single microdissected chromosome. With this method, it was possible to construct the region-specific painting probe for HSR. The probe hybridized specifically to the HSRs of IMR-32, and to 2p24, 2p13 of normal chromosome. Our results suggest there was coamplification of N-myc together with DNA of the chromosome 2p24 and 2p13. Moreover, the fluorescent signals for the amplified chromosomal regions in IMR-32 cells were also easily recognized at a Thus this painting probe can be applied to detect the similar amplification of N-myc in neuroblastoma tissue, and the probe pool for HSR may be used to identify the cancer-relevant genes.
Cell Line* ; Chromosomes, Human, Pair 2 ; DNA ; DNA Topoisomerases, Type I ; DNA, Superhelical ; Genes, myc ; Humans* ; In Situ Hybridization, Fluorescence ; Microdissection ; Needles ; Neural Crest ; Neuroblastoma* ; Paint* ; Paintings* ; Polymerase Chain Reaction ; Relaxation ; Virulence

The mitochondrial myopathy, encephalopathy, lactic acidosis, and stroke-like episodes (MELAS) is inherited maternally, in which the MTTL1*MELAS 3243 mutation has been most commonly found as a heteropla smy of A to G point mutation in the tRNALeu(UUR) gene. The MTTL1*MELAS 3271 mutation is known to be the second common mutation, though clinical features of both mutations are not remarkably different. Recently, a variety of minor mutations have been reported in patients with MELAS. In this study, major efforts have been made to investigate the allele frequency of major three mutations including MTTL1*MELAS 3243, 3252, 3271 in 10 Korean families with MELAS probands. The PCR and subsequent direct sequencing of the PCR product in the regions spanning these three mutation sites were employed to identify the mutation in each proband. All family members have been screened for the presence of these three mutations by PCR-RFLP assay using Apa I, Acc I and Bfr I restriction enzymes. The MTTL1*MELAS 3243 mutation was most commonly found (7 out of 10 families tested) followed by the MTTL1*MELAS 3271 which was identified in 1 out of 10 families. In the remaining 2 families none of three mutations were found, indicating the presence of either nuclear mutation or yet unidentified mitochondrial DNA mutation in these families.
Acidosis, Lactic* ; DNA, Mitochondrial ; Gene Frequency ; Humans ; MELAS Syndrome* ; Point Mutation ; Polymerase Chain Reaction ; Stroke*

Spinal muscular atrophy(SMA) is the second most common fatal disease of childhood with autosomal dominant mode of inheritance, and in its less severe form the third most common neuromuscular disease of childhood after Duchenne muscular dystrophy. The genetic defect was found to be on the long arm of chromosome 5(5q11.2-q13.3) where many genes and microsatellite markers were missing. One of the most important genes is the Survival Motor Neuron(SMN) gene which is homozygously missing in 90% of SMA patients. Another important gene, the Neuronal Apoptosis Inhibitory Protein(NAIP) gene was found to be defective in 67% of SMA type I patients. Studies so far suggest SMA occurs when the genes on the long arm of chromosome 5 are mutated or deleted. Recently our hospital encountered 2 SMA patients of type I and II respectively. These patients both had homozygously defective SMN genes but intact NAIP genes. We are reporting these cases with bibliographic review and discussion. Korean SMA patients presumably have defects in SMN genes similar to those found in European patients, although the siginificance of NAIP genes remains to be established. SMN gene defects can be easily diagnosed using PCR and restriction enzymes, and this method could be applied towards convenient prenatal diagnosis and towards screening for family members at risk.
Apoptosis ; Arm ; Chromosomes, Human, Pair 5 ; Diagnosis* ; Humans ; Mass Screening ; Microsatellite Repeats ; Muscular Atrophy, Spinal* ; Muscular Dystrophy, Duchenne ; Neuromuscular Diseases ; Neurons ; Polymerase Chain Reaction ; Prenatal Diagnosis ; Wills

Steroid 21 hydroxylase deficiency is a major cause of congenital adrenal hyperplasia(CAH) and is caused by genetic impairment (CYP21B) of this enzyme. In the human genome, CYP21B is located within MHC class III region on the short arm of chromosome 6. Most of the genes in this region are highly polymorphic and crowded. Also the CYP21B gene is accompanied by its pseudogene (CYP21A) and tandemly arranged with two genes of fourth component of complement. This highly complex gene arrangement in this area may predispose genetic unstability of CYP21 genes,i.e. mutations. In the current study, we tried to investigate the frequency of duplication and deletion of CYP21 genes and pattern of the genetic alteration of these genes by RFLPs. We also compared the genetic alteration of CYP21 in normal subjects with those of the CAH patients. According to our study, 15% of the normal Korean population have duplication or deletion of CYP21. There was one normal subject with heterozygous deletion of CYP21B gene. Of the 5 CAH patients examined, we found abnormal patterns in 2 patients. One was a large scale gene conversion and the other was a deletion of CYP21B and C4 locus II genes with gene conversion. These results suggest that high frequency of duplication and deletion of CYP21 and C4 in the general population may provide the genetic pool of instable CYP21 genes and these duplicated or deleted genes may result in gene conversions between CYP21A(pseudogene) and CYP21B(true gene) by preventing the normal recombination event.
Adrenogenital Syndrome* ; Arm ; Chromosomes, Human, Pair 6 ; Complement System Proteins ; Gene Conversion ; Gene Order ; Genome, Human ; Humans ; Polymorphism, Restriction Fragment Length ; Pseudogenes ; Recombination, Genetic ; Steroid 21-Hydroxylase*

Myotonic dystrophy(DM) is caused by the expansion of CTG trinucleotide repeat near the 3' end of the gene encoding for a member of protein kinase gene family (DMPK). The normal range of the CTG repeat was determined in 178 nomal individuals (141 unrelated individuals and 37 of 9 families) by polymerase chain reaction (PCR), polyacrylamide gel electrophoresis and silver staining method. And the expansion of the CTG repeats in a DM family was analyzed with Southern analysis. In normal population, the range of CTG repeat is between 5 and 34 and 19 different alleles were obserbed in that range, and (CTG)11-14 alleles were predominant. 4 members of an affected family showed the 0.5 - 2.0 kb size expansion of CTG repeats. In this study we could predict the incidence of DM in Korea as 1 in 20,000 and we could establish the diagnostic procedure for myotonic dystrophy.
Alleles ; Electrophoresis, Polyacrylamide Gel ; Humans ; Incidence ; Korea ; Myotonic Dystrophy* ; Polymerase Chain Reaction ; Protein Kinases ; Reference Values ; Silver Staining ; Trinucleotide Repeats