The C57BL/6J-fe/fe mouse is a coat color mutant. The coat color of the homozygote mouse becomes progressively lighter with advancing age. The faded gene (fe) of C57BL/6J-fe/fe was mapped in a 2.0 cM distal to D10mit191 by our group. To make a high-resolution map, we used the Korean wild mouse (KWHM) for a backcross panel, which was captured in 1995 and has been maintained as an inbred line by our laboratory. In the inter-specific backcross panel (N=400), the fe gene was mapped to 1.0 cM distal to D10mit156. The gene order was defined: centromere -D10mit3/85 (1.3+/-0.6 cM)-D10mit155 (1.3+/-0.6 cM)-D10mit191 (2.0+/-0.7 cM)-D10mit156 (1.0+/-0.5 cM)-fe-D10mit193 (1.3+/-0.6 cM)-D10mit54 (1.0+/-0.5 cM)-D10mit44 (8.5+/-1.4 cM)-D10mit42 (10.0+/-1.5 cM). The measured distance between D10mit191 and D10mit 44 differed in both inter-specific (DBA/2) and intra-specific (KWHM) backcross panels (14.2 vs 13.8 cM). Taken together, our high-resolution linkage map of the fe locus from an intra-specific backcross panel will provide a good entry point to isolate the fe gene.
Animals ; Centromere ; Chromosomes, Human, Pair 10 ; Gene Order ; Hair Color ; Homozygote ; Mice

Subcutaneous panniculitis-like T-cell lymphoma(SPTCL) is a rare subtype of cutaneous T-cell lymphoma and needs to be differentiated from benign causes of panniculitis and other cutaneous T-cell lymphomas involving subcutis, especially nasal type NK/T-cell lymphoma(NKTL) involving the panniculus. We report herein two cases of SPTCL which was distinguished from nasal type NKTL by the following findings; CD8+, CD56-, Ebstein Barr virus RNA- and clonal T cell receptor gene arrangement.
Gene Order ; Lymphoma, T-Cell* ; Lymphoma, T-Cell, Cutaneous ; Panniculitis ; Receptors, Antigen, T-Cell ; T-Lymphocytes*

The faded mouse is a coat color mutant that shows faded coat color and age-related loss of pigmentation. This mutation is transmitted by an autosomal recessive gene with 100% penetrance. In the present study, we carried out linkage analysis of the faded (fe) gene using intra-specific backcross panels. Affected faded mice were carefully confirmed by their faded coat color at about 4 weeks of age. In the intra-specific backcross between faded and CBA mice (n=198), the fe gene was mapped to a region 2.1 cM distal to D10mit191. Therefore, the gene order was defined as follows: centromere-D10mit51 (12.4+/-2.4 cM)-D10mit191 (2.1+/-1.0 cM)-fe-D10mit44 (13.3+/-2.4 cM)-D10mit42 (14.4+/-2.5 cM). This linkage map of the fe locus will provide a good entry point to isolate the fe gene. Since the faded mouse has pigmentary abnormalities, this mutant may be a useful model for studies of pigmentary abnormalities in humans.
Animals ; Chromosomes, Human, Pair 10 ; Gene Order ; Genes, Recessive ; Humans ; Mice ; Mice, Inbred CBA ; Penetrance ; Pigmentation

OBJECTIVETo establish a comprehensive and simple assay using denaturing high performance liquid chromatography (DHPLC) for the diagnosis of most common mutations and deletions of α-thalassemia gene in Southeast Asians and Southern Chinese.

METHODSThis assay has included a duplex polymerase chain reaction (PCR) followed by DHPLC analysis. An improved PCR was also performed followed by DHPLC analysis. With this assay, a blinded study of 160 samples was screened for three common mutations and three common deletions.

RESULTSThe duplex PCR-DHPLC combined with the improved PCR-DHPLC analysis has detected all mutations and the wild-type allele. The results were consistent with those by the original methods.

CONCLUSIONThis molecular assay may be used for the diagnosis of α-thalassemia patients from this geographical region. The method is accurate, rapid, semi-automatic and cost-effective, which makes it suitable for large-scale screening.

Chromatography, High Pressure Liquid ; methods ; DNA Mutational Analysis ; methods ; Gene Order ; Genotype ; Humans ; alpha-Globins ; genetics ; alpha-Thalassemia ; diagnosis ; genetics

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*

BACKGROUND: Mycosis fungoides(MF) is a representative of cutaneous T cell lymphoma and progresses through clinical stages, such as initial pre-mycotic(macule or patch), plaque, and tumor stage. MF is usually thought of as a disease of old adults, but it can develop at any age, including childhood. Despite the recognition that MF may occur in pediatric patients, there is scant literature regarding clinical feature, histopathologic finding and prognosis specially related to childhood onset. The purpose of this study was to examine the clinical, histopathologic and follow up observation of MF in children. METHODS:This study has been reviewed in the clinicopathologic findings of 14 MF patients who visited the Kosin University Gospel Hospital during about 10 years from January, 1991 to June, 2000. RESULTS: 1.Duration of cutaneous lesions was from 6 months to 10 years, with mean duration of 3.6 years. 2.The morphology of skin lesions showed various features(pityriasis lichenoides-like, hypopigmented, hyperpigmented, ichthyosis-like, inflammatory linear verrucous epidermal nevus-like). 3. Histopathologic features revealed epidermotropism, a perivascular infiltrate in all cases, haloed lymphocytes, lymphocytes aligned in the basal layer, and coarse collagen in the papillary dermis in most cases. Pautrier's microabscess was shown in 3 cases(23%). 4. In T cell receptor gamma gene arrangement using PCR(polymerase chain reaction), monoclonality was detected in 13 out of 14(93%). 5. Treatment including PUVA(psoralen and UVA), UVB, topical steroid agents and calcipotriol had good a response. 6. No patients had progressed to a more advanced disease. CONCLUSION: MF in children of this study showed a wide variety of skin lesions. Although no case of MF in this study progressed to a more aggressive status, MF in children should be carefully followed for development of more advanced cutaneous lesions and visceral involvement.
Adult ; Child* ; Collagen ; Dermis ; Follow-Up Studies ; Gene Order ; Humans ; Lymphocytes ; Lymphoma, T-Cell, Cutaneous ; Mycosis Fungoides* ; Prognosis ; Receptors, Antigen, T-Cell ; Skin

BACKGROUND: Mycosis fungoides(MF) is a representative of cutaneous T cell lymphoma and progresses through clinical stages, such as initial pre-mycotic(macule or patch), plaque, and tumor stage. MF is usually thought of as a disease of old adults, but it can develop at any age, including childhood. Despite the recognition that MF may occur in pediatric patients, there is scant literature regarding clinical feature, histopathologic finding and prognosis specially related to childhood onset. The purpose of this study was to examine the clinical, histopathologic and follow up observation of MF in children. METHODS:This study has been reviewed in the clinicopathologic findings of 14 MF patients who visited the Kosin University Gospel Hospital during about 10 years from January, 1991 to June, 2000. RESULTS: 1.Duration of cutaneous lesions was from 6 months to 10 years, with mean duration of 3.6 years. 2.The morphology of skin lesions showed various features(pityriasis lichenoides-like, hypopigmented, hyperpigmented, ichthyosis-like, inflammatory linear verrucous epidermal nevus-like). 3. Histopathologic features revealed epidermotropism, a perivascular infiltrate in all cases, haloed lymphocytes, lymphocytes aligned in the basal layer, and coarse collagen in the papillary dermis in most cases. Pautrier's microabscess was shown in 3 cases(23%). 4. In T cell receptor gamma gene arrangement using PCR(polymerase chain reaction), monoclonality was detected in 13 out of 14(93%). 5. Treatment including PUVA(psoralen and UVA), UVB, topical steroid agents and calcipotriol had good a response. 6. No patients had progressed to a more advanced disease. CONCLUSION: MF in children of this study showed a wide variety of skin lesions. Although no case of MF in this study progressed to a more aggressive status, MF in children should be carefully followed for development of more advanced cutaneous lesions and visceral involvement.
Adult ; Child* ; Collagen ; Dermis ; Follow-Up Studies ; Gene Order ; Humans ; Lymphocytes ; Lymphoma, T-Cell, Cutaneous ; Mycosis Fungoides* ; Prognosis ; Receptors, Antigen, T-Cell ; Skin

Mitochondrial genomes have been extensively studied for phylogenetic purposes and to investigate intra- and interspecific genetic variations. In recent years, numerous groups have undertaken sequencing of platyhelminth mitochondrial genomes. Haplorchis taichui (family Heterophyidae) is a trematode that infects humans and animals mainly in Asia, including the Mekong River basin. We sequenced and determined the organization of the complete mitochondrial genome of H. taichui. The mitochondrial genome is 15,130 bp long, containing 12 protein-coding genes, 2 ribosomal RNAs (rRNAs, a small and a large subunit), and 22 transfer RNAs (tRNAs). Like other trematodes, it does not encode the atp8 gene. All genes are transcribed from the same strand. The ATG initiation codon is used for 9 protein-coding genes, and GTG for the remaining 3 (nad1, nad4, and nad5). The mitochondrial genome of H. taichui has a single long non-coding region between trnE and trnG. H. taichui has evolved as being more closely related to Opisthorchiidae than other trematode groups with maximal support in the phylogenetic analysis. Our results could provide a resource for the comparative mitochondrial genome analysis of trematodes, and may yield genetic markers for molecular epidemiological investigations into intestinal flukes.
Animals ; Asia ; Codon, Initiator ; DNA, Mitochondrial/chemistry/genetics ; Gene Order ; Genes, Helminth ; *Genome, Mitochondrial ; Heterophyidae/*genetics/isolation & purification ; Humans ; Molecular Sequence Data ; Sequence Analysis, DNA

OBJECTIVETo develop a method for elucidating genetic basis of 21-hydroxylase deficiency.

METHODSSanger sequencing of entire 21-hydroxylase coding gene CYP21A2 was carried out to detect point mutations, and multiplex ligation-dependent probe amplification (MLPA) and locus-specific PCR/enzyme restriction method were used to detect large deletions and conversion mutations.

RESULTSNine children were analyzed. Point mutations of the CYP21A2 gene have been identified as: IVS2 13A/C>G (9 alleles), p.Arg356Trp (1 allele), Cluster E6 (1 allele), p.Gln318X (1 allele), and Prom conv (1 allele). While the former 4 mutations are pathogenic, the role of Prom conv mutation in the pathogenesis was uncertain. Three cases had entire CYP21A2 gene deletions (3 alleles), three had CYP21A1P/CYP21A2 chimeric mutations (3 alleles). The genotypes of all patients were determined. And all of the mutations were inherited from parents.

CONCLUSIONA rational method for detecting point mutations and large deletions/conversions of CYP21A2 gene has been established.

Adrenal Hyperplasia, Congenital ; diagnosis ; genetics ; Alleles ; Base Sequence ; Child ; Child, Preschool ; Female ; Gene Order ; Genotype ; Humans ; Infant ; Male ; Multiplex Polymerase Chain Reaction ; Steroid 21-Hydroxylase ; genetics

Lysosomal dysfunction is a common pathological feature of neurodegenerative diseases. GTP-binding protein type A1 (GBA1) encodes beta-glucocerebrosidase 1 (GCase 1), a lysosomal hydrolase. Homozygous mutations in GBA1 cause Gaucher disease, the most common lysosomal storage disease, while heterozygous mutations are strong risk factors for Parkinson's disease. However, whether loss of GCase 1 activity is sufficient for lysosomal dysfunction has not been clearly determined. Here, we generated human neuroblastoma cell lines with nonsense mutations in the GBA1 gene using zinc-finger nucleases. Depending on the site of mutation, GCase 1 activity was lost or maintained. The cell line with GCase 1 deficiency showed indications of lysosomal dysfunction, such as accumulation of lysosomal substrates, reduced dextran degradation and accumulation of enlarged vacuolar structures. In contrast, the cell line with C-terminal truncation of GCase 1 but with intact GCase 1 activity showed normal lysosomal function. When alpha-synuclein was overexpressed, accumulation and secretion of insoluble aggregates increased in cells with GCase 1 deficiency but did not change in mutant cells with normal GCase 1 activity. These results demonstrate that loss of GCase 1 activity is sufficient to cause lysosomal dysfunction and accumulation of alpha-synuclein aggregates.
Cell Line ; Enzyme Activation/genetics ; Gene Knockout Techniques ; Gene Order ; Genetic Loci ; Glucosylceramidase/genetics/*metabolism ; Humans ; Lysosomes/*metabolism ; Mutation ; *Protein Aggregation, Pathological/genetics ; Protein Binding ; Zinc Fingers ; alpha-Synuclein/chemistry/*metabolism