No abstract available.
Exons* ; Point Mutation* ; Steatocystoma Multiplex*

No abstract available.
Introns* ; Point Mutation* ; Protein-Tyrosine Kinases* ; Tyrosine*

Although uncommon, acquired hemophilia A (HA) is associated with a high rate of mortality due to severe bleeding. In spite of many hypotheses regarding the cause of acquired HA, there is as yet no established theory. In this study, we investigated the possibility that mutation(s) in the F8 gene may be correlated with the development of inhibitory autoantibodies. Direct sequencing analysis was performed on all 26 exons of the F8 gene of 2 patients exhibiting acquired HA. Both patients were found to share a common point mutation (c.8899G>A) in the 3'-untranslated region (3'-UTR) of exon 26. This is the first report on the genotyping of F8 in the context of acquired HA.
Autoantibodies ; Exons ; Hemophilia A ; Hemorrhage ; Humans ; Point Mutation

In order to understand the protein functions that are related to disease, it is important to detect the correlation between amino acid mutations and isease. Many mutation studies about disease-related proteins have been carried out through molecular biology techniques, such as vector design, protein engineering, and protein crystallization. However, experimental protein mutation studies are time-consuming, be it in vivo or in vitro. We therefore performed a bioinformatic analysis of known disease-related mutations and their protein structure changes in order to analyze the correlation between mutation and disease. For this study, we selected 111 diseases that were related to 175 proteins from the PDB database and 710 mutations that were found in the protein structures. The mutations were acquired from the Human Gene Mutation Database (HGMD). We selected point mutations, excluding only insertions or deletions, for detecting structural changes. To detect a structural change by mutation, we analyzed not only the structural properties (distance of pocket and mutation, pocket size, surface size, and stability), but also the physico-chemical properties (weight, instability, isoelectric point (IEP), and GRAVY score) for the 710 mutations. We detected that the distance between the pocket and disease-related mutation lay within 20 A (98.5%, 700 proteins). We found that there was no significant correlation between structural stability and disease-causing mutations or between hydrophobicity changes and critical mutations. For large-scale mutational analysis of disease-causing mutations, our bioinformatics approach, using 710 structural mutations, called "Structural Mutatomics," can help researchers to detect disease-specific mutations and to understand the biological functions of disease-related proteins.
Computational Biology ; Crystallization ; Humans ; Hydrophobic and Hydrophilic Interactions ; Isoelectric Point ; Molecular Biology ; Point Mutation ; Protein Engineering ; Proteins

Achondroplasia is abnormal intracartilagenous ossification that's caused by a genetic point mutation. Thoracic myelopathy in achondroplasia that is due to thoracolumbar kyphosis and spinal stenosis is a rare finding. There is no report available on this topic in Korea. We report here a case of achondroplasia with thoracic myelopathy due to thoracolumbar kyphosis and spinal stenosis, and we include a brief review of literature.
Achondroplasia* ; Korea ; Kyphosis* ; Point Mutation ; Spinal Cord Diseases* ; Spinal Stenosis*

AIMTo develop a simple, fast and inexpensive approach as well as an instrument for detection of gene mutation.

METHODSPyrosequencing based on bioluminometry assay was employed to detect gene mutation. Pyrosequencing is a method of sequencing by synthesis step-by-step using four enzymes, DNA-polymerase, ATP sulfurylase, luciferase and apyrase. The signal was produced by detecting pyrophosphate released during a dNTP incorporation. For mutation detection, a DNA fragment was amplified by PCR at first, followed by a single-stranded DNA preparation. In the second step, a short primer was annealed to the position just before the mutation point. Finally, specific dNTPs were added in terms of the template sequence. The mutation species can be readily determined by the sequence.

RESULTSA new instrument was developed for gene mutation detection by pyrosequencing. To iteratively inject small amount of each dNTP for the sequencing reaction, capillaries were used to connect dNTP reservoirs and the reaction chamber. Each dNTP was delivered by adding a gas pressure on the top of a corresponding dNTP reservoir, by which 0.2 microL of dNTP can be exactly added each time. It was theoretically proved that undesired liquid seep through the capillary did not affect the sequencing reactions in pyrosequencing. In addition, the three possible variants (wildtype, mutant and heterozygote) of a mutant point Cys275Ser in P53 gene exon 8 were determined by pyrosequencing using the instrument. A simple method was also described for rapidly distinguishing the type of a variant.

CONCLUSIONThe developed method is very simple, and the corresponding instrument is inexpensive and easy to operate, which can be used to detect many types of mutation.

Medullary thyroid carcinoma and papillary thyroid carcinoma are different subtypes of thyroid carcinoma. The concomitant occurrence of medullary thyroid carcinoma and papillary thyroid carcinoma as a collision tumor is rare. We describe five cases of medullary and papillary thyroid carcinoma as a collision tumor. Four women and one man underwent thyroidectomy for treatment of thyroid cancer. Collision tumor was then detected by histopathologic finding. Genetic testing, point mutation of the BRAF gene or mutation of the RET gene was performed in three cases. However, only one case had point mutation of the BRAF gene. Exact diagnosis of this uncommon event is important because the strategies for treatment of papillary thyroid carcinoma and medullary thyroid carcinoma are different.
Diagnosis ; Female ; Genetic Testing ; Humans ; Point Mutation ; Thyroid Neoplasms* ; Thyroidectomy

BACKGROUND: Mycobacterium abscessus is the most pathogenic and drug-resistant rapid-growing mycobacterium. Clarithromycin or azithromycin are the only regular oral antimycobacterial agents that have an effect on M. abscessus. We tried to detect the clarithromycin-resistant strains from the clinical isolates of M. abscessus. METHODS: We tried to isolate the clarithromycin-resistant strains from 220 clinical isolates of M. abscessus by performing using reverse hybridization assay (RHA) and the broth microdilution test (BMT). RESULTS: Seven resistant strains (3.2%) from all the tested clinical isolates were detected by BMT. Three of these resistant strains were also detected by RHA and it was confirmed that they had point mutants. CONCLUSION: These results showed that clarithromycin resistance in M. abscessus clinical isolates is related to a point mutation and other unknown mechanisms.
Anti-Bacterial Agents ; Azithromycin ; Chimera ; Clarithromycin ; Mycobacterium ; Point Mutation

No abstract available.
Animals ; Colon* ; Colonic Neoplasms* ; Oncogenes* ; Point Mutation* ; Rats*

No abstract available.
Colorectal Neoplasms* ; Humans* ; Oncogenes* ; Point Mutation* ; Polymerase Chain Reaction*