Amplification Failure of the Amelogenin X Gene Caused by a Rare Mutation in the Primer-Binding Region.

The study of gender markers is essential in forensic genetic analysis. Mutations in the X or Y homologs of the amelogenin gene can be misleading, resulting in serious mistakes in forensic genetic analysis. We recently discovered two male cases of the X homolog of the amelogenin (AMELX) allelic dropout while analyzing short tandem repeat genotypes obtained from crime scene evidence. Subsequently, we evaluated the molecular characteristics of AMELX allelic dropout in this study. We used two previously reported amelogenin primers to verify a half level of amelogenin gene amplification intensity in the two male cases, which we confirmed was caused by AMELX allelic dropout. We then characterized the point mutation using Sanger sequencing and designed mutation-specific primers that could overcome AMELX allelic dropout. Short tandem repeat genotyping analysis confirmed that the AMELX allelic dropout was recovered by the mutation-specific primer designed specifically for this case. The sequencing of the AMELX allele revealed a single-point variant from A→G at base position 7 downstream from the 3' end in the amelogenin forward primer-binding region. This point mutation was identically found in two different male cases, resulting in AMELX allelic dropout. To our knowledge, these mutations and the X homolog amplification failure of amelogenin have not been reported in the Korean population. Our study provides a reliable approach to AMELX allelic dropout due to rare case mutations and could enable the better interpretation of gender markers for forensic samples.

Development of diagnostic SNP markers and a novel SNP genotyping assay for distinguishing opium poppies.

The opium poppy acts as an important natural pain reliever but is also responsible for increased rates of severe drug abuse and addiction owing to its characteristic psychoactive effect. Non-medical illicit use of the poppy plant is markedly increasing worldwide, thereby highlighting the need for a robust species identification strategy. In this study, we identified SNPs within the region of two universal DNA barcodes, matK (maturase K) and the trnL-trnF (tRNA-Leu [3'exon]-tRNA-Phe [exon] intergenic spacer, that are forensically applicable for distinguishing opium poppy species based on a genetic analysis of 164 samples of family Papaveraceae obtained from locations spanning Jeolla-do and Jeju Island, Republic of Korea. A comparative analysis of the DNA barcode sequences for two narcotic types of the Papaver species (Papaver somniferum, Papaver somniferum subs. setigerum) to eight non-narcotic species revealed three unique nucleotide substitution events. Newly identified SNPs were located at position 255 of matK and at positions 305 and 306 of trnL-trnF; the narcotic species contained C, A, and T, whereas non-narcotic species contained T, G, and C at these positions. Phylogenetic analysis demonstrated that newly identified SNPs, which we named PsMAT255 and PsLF305/306, could be used to clearly differentiate between the narcotic and non-narcotic types of Papaver species based on the patterns of nucleotide variation. These results indicate that the nucleotide differences between the narcotic and non-narcotic species may influence genetic markers. We, therefore, developed a novel SNP-based allelic genotyping assay using the RT-PCR system that can reliably differentiate the narcotic type of the Papaver species. In summary, our findings suggest that the newly identified species-specific SNPs of both matK and trnL-trnF can be used as identification markers of narcotic Papaver species. Furthermore, a newly developed TaqMan allelic discrimination assay may be used as a practically applicable diagnostic method to survey several illicit narcotic specimens carrying the type-specific SNP.

Identification of psychrophile Shewanella sp. KMG427 as an eicosapentaenoic acid producer.

An isolate from holothurians was identified as an eicosapentaenoic acid (EPA)-producing bacterium KMG427, which is characterized by EPA synthesis efficiency, by thin layer and gas chromatographic analyses. The EPA production was maximized to more than 10% of the total fatty acids by incubation at 4o degrees C after cell proliferation at 20 degrees C. The isolated bacterium was categorized as Gramnegative, rod-shaped, aerobic, and motile with a single polar flagellum. According to phylogenetic analysis based on morphological and physiological specificities as an EPA-producing bacterium, the isolate KMG427 was found to belong to the genus Shewanella. The 16S rDNA of KMG427 was revealed to have 100% of sequence identity to that of S. hanedai CIP 103207T. Therefore, the isolate might be classified and identified as Shewanella sp. KMG427.

Intensive psychotherapy training in korean psychiatric residency programs.

OBJECTIVE: The authors investigated the current practice of intensive psychotherapy by residents in the department of psychiatry. METHODS: We mailed a questionnaire to 126 fourth-year psychiatry residents in order to obtain data on their clients' sociodemographic characteristics, the settings in which psychotherapy is being conducted, the effects of psychotherapy, the difficulties associated with psychotherapy, the state of supervision and the level of clients' satisfaction. RESULTS: Approximately 51.5% of the residents completed the questionnaires. The average number of clients was 4.9+/-3.8, the average number of psychotherapy sessions was 26.2+/-20.1, and 69.4% of the residents had performed insight-oriented psychotherapy. Approximately 69.8% of the fourth-year residents had received some form of supervision, and 58.7% agreed to increase the frequency of supervision. Approximately 74.2% of the cases were supervised. The average number of supervisions per case was 9.2+/-10.5. CONCLUSION: The setting in which psychotherapy is conducted, number of clients, and type of supervision varied greatly among the training institutes surveyed. Based on these findings, we expect to create better psychotherapy training programs for psychiatric residents.

SHN-1, a Shank homologue in C. elegans, affects defecation rhythm via the inositol-1,4,5-trisphosphate receptor.

Protein localization in the postsynaptic density (PSD) of neurons is mediated by scaffolding proteins such as PSD-95 and Shank, which ensure proper function of receptors at the membrane. The Shank family of scaffolding proteins contain PDZ (PSD-95, Dlg, and ZO-1) domains and have been implicated in the localizations of many receptor proteins including glutamate receptors in mammals. We have identified and characterized shn-1, the only homologue of Shank in Caenorhabditis elegans. The shn-1 gene shows approximately 40% identity over 1000 amino acids to rat Shanks. SHN-1 protein is localized in various tissues including neurons, pharynx and intestine. RNAi suppression of SHN-1 did not cause lethality or developmental abnormality. However, suppression of SHN-1 in the itr-1 (sa73) mutant, which has a defective inositol-1,4,5-trisphosphate (IP(3)) receptor, resulted in animals with altered defecation rhythm. Our data suggest a possible role of SHN-1 in affecting function of IP(3) receptors in C. elegans.