• Asian Pacific Journal of Cancer Prevention
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• 제15권19호
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• pp.8319-8324
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• 2014

Although genetic markers identifying women at an increased risk of developing breast cancer exist, the majority of inherited risk factors remain elusive. Mutations in the BRCA1/BRCA2 gene confer a substantial increase in breast cancer risk, yet routine clinical genetic screening is limited to the coding regions and intronexon boundaries, precluding the identification of mutations in noncoding and untranslated regions. Because 3' untranslated region (3'UTR) polymorphisms disrupting microRNA (miRNA) binding can be functional and can act as genetic markers of cancer risk, we aimed to determine genetic variation in the 3'UTR of BRCA1/BRCA2 in familial and early-onset breast cancer patients with and without mutations in the coding regions of BRCA1/BRCA2 and to identify specific 3'UTR variants that may be risk factors for cancer development. The 3'UTRs of the BRCA1 and BRCA2 genes were screened by heteroduplex analysis and DNA sequencing in 100 patients from 46 BRCA1/2 families, 54 non-BRCA1/2 families, and 47 geographically matched controls. Two polymorphisms were identified. SNPs $c.^*1287C$ >T (rs12516) (BRCA1) and $c.^*105A$ >C (rs15869) (BRCA2) were identified in 27% and 24% of patients, respectively. These 2 variants were also identified in controls with no family history of cancer (23.4% and 23.4%, respectively). In comparison to variations in the 3'UTR region of the BRCA1/2 genes and the BRCA1/2 mutational status in patients, there was a statistically significant relationship between the BRCA1 gene polymorphism $c.^*1287C$ >T (rs12516) and BRCA1 mutations (p=0.035) by Fisher's Exact Test. SNP $c.^*1287C$ >T (rs12516) of the BRCA1 gene may have potential use as a genetic marker of an increased risk of developing breast cancer and likely represents a non-coding sequence variation in BRCA1 that impacts BRCA1 function and leads to increased early-onset and/or familial breast cancer risk in the Turkish population.

• Journal of Microbiology and Biotechnology
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• 제23권11호
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• pp.1519-1528
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• 2013

Bacillus pumilus SG2, a halotolerant strain, expresses two major chitinases designated ChiS and ChiL that were induced by chitin and secreted into the supernatant. The present work aimed to obtain a mutant with higher chitinolytic activity through mutagenesis of Bacillus pumilus SG2 using a combination of UV irradiation and nitrous acid treatment. Following mutagenesis and screening on chitin agar and subsequent formation of halos, the mutated strains were examined for degradation of chitin under different conditions. A mutant designated AV2-9 was selected owing to its higher chitinase activity. To search for possible mutations in the whole operon including ChiS and ChiL, the entire chitinase operon, including the intergenic region, promoter, and two areas corresponding to the ChiS and ChiL ORF, was suquenced. Nucleotide sequence analysis of the complete chitinase operon from the SG2 and AV2-9 strains showed the presence of a mutation in the catalytic domain (GH18) of chitinase (ChiL). The results demonstrated that a single base change had occurred in the ChiL sequence in AV2-9. The wild-type chitinase, ChiL, and the mutant (designated ChiLm) were cloned, expressed, and purified in E. coli. Both enzymes showed similar profiles of activity at different ranges of pH, NaCl concentration, and temperature, but the mutant enzyme showed approximately 30% higher catalytic activity under all the conditions tested. The results obtained in this study showed that the thermal stability of chitinase increased in the mutant strain. Bioinformatics analysis was performed to predict changes in the stability of proteins caused by mutation.

• Journal of Genetic Medicine
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• 제7권2호
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• pp.125-132
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• 2010

목 적: 착상전 유전진단(preimplantation genetic diagnosis, PGD)은 유전질환을 가진 부부들을 대상으로 체외수정을 통해 획득한 배아에서 유전진단을 하는 방법이다. 다양한 유전질환을 가진 부부에게 그 질환에 맞는 PGD의 설계가 진행되어야 하기 때문에 PGD 시행 전유전상담을 시행하는 것은 PGD 설계에 있어서 매우 중요하다. 이에, PGD 시행 시 필요한 유전상담의 내용에 대해 환자 및 가족과 전문가의 구체적인 의견을 수렴하고 분석하고자 하였다. 대상 및 방법: 본 연구는 PGD에 있어서 유전상담의 필요성과 중요성에 대한 의견을 알아보고자, 2010년 2월 3일부터 4월 30일까지 PGD를 실시 또는 실시 예정인 부부들과 PGD 관련 전문가들을 대상으로 이메일과 직접 설문지를 배포하여 설문조사를 실시하였다. 결 과: 환자 60명과 전문가 31명을 포함하여 총 91명이 설문조사에 응답하였으며, 환자들은 염색체 이상 질환 49명(81.7%)과 단일유전자 이상 질환 11명(18.3%) 이었다. 설문에 응답한 환자와 전문가 모두 유전상담이 PGD의 의료서비스 일환으로 반드시 필요하다고 답하였다. 환자의 충분한 이해를 위하여 필요한 유전상담의 시간에 대해 환자와 가족 그리고 전문가 의견을 수렴한 결과, 각각 45명(75.0%)과 23명(74.2%)이 적정한 유전상담시간을 30분 이상이라고 응답하였다. 하지만, 현 의료시스템에서는 짧은 진료시간 내 진료와 유전상담을 동시에 진행함으로써 환자에게 완벽한 정보제공이 이루어지지 않는 것으로 나타났다. 한편, 전문가 그룹에서는 진료시간의 부족과 유전질환의 정보 부족이 유전상담의 어려운 점이라고 답하였으며, 이에 비 의사(non-MD) 전문유전상담사가 필요하다는 의견이 30명(96.7%)으로 높게 나타났다. 환자와 가족들은 PGD 시술 시 예기치 못한 결과의 가능성, 환자가 가진 유전질환의 위험을 예방할 수 있는 선택사항, 환자가 가지고 있는 유전질환의 위험도 평가, 유전자 검사 시 검사의 목적 설명 및 검사기술의 한계점과 오진률의 설명, PGD 시술 전반에 관한 기술적인 정보 등에 대하여 관심을 가지고 있으며 더 자세한 설명을 필요로 하는 것으로 나타났다. 이에 대한 전문가 의견 역시 환자 및 가족이 관심 있고 자세한 설명을 원하는 정보와 대부분 일치하였다. 이에 따라 환자의 요구와 의견으로 나타난 위의 결과들을 향후 PGD를 위한 유전상담의 지침(guide-line) 구축 시 반영하여야 할 것으로 사료된다. 결 론: 본 연구에서 유전진단과 생식의학 기술의 발전과 더불어 PGD의 적용과 효율성 등에 대한인식이 높아짐에 따라, PGD를 시행함에 있어서 구체적이고 체계적인 유전상담이 필요하다는 것을 확인할 수 있었다. 본 연구의 설문조사 결과가 향후 PGD를 위한 유전상담 지침서(guideline)에 반영되어 적절한 PGD의 설계, 실시, 사후관리에 큰 도움이 되기를 기대한다.

• Asian-Australasian Journal of Animal Sciences
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• 제19권11호
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• pp.1519-1523
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• 2006

Through a screening and selection approach method, decamer random markers were used in a technique called random amplified polymorphic DNA (RAPD) assay with 252 genomic DNAs isolated from four major Haimen chicken populations: Rugao (62), Jiangchun (62), Wan-Nan (63) and Cshiqishi (65). A total of 3-score decamer random primers (S241-S260, S1081-S1100 and S1341-S1360) were employed in the preliminary RAPD-polymerase chain reaction (RAPD-PCR) assay with 50 random template DNA samples from all the populations. Four (6.67%) of the primers that produced obvious polymorphic patterns, interpretable and reproducible bands were selected and used with both the individual DNAs from each population and with pooled DNA samples of the four populations in subsequent analyses. The selected primers produced a total of 131 fragments with molecular size ranging from 835 to 4,972 base pairs (bp) when used with the individual DNAs; 105 (80.15%) of these fragments were polymorphic. With the pooled DNAs, 47 stable and characteristic bands with molecular size ranging from 840 to 4,983 bp, of which 23 (48.94%) polymorphic, were also generated. The band-sharing coefficient (BSC) calculated for the individuals in the population and among populations of bulked samples was between 0.8247 (Rugao) and 0.9500 (Cshiqishi); for pairwise populations, it was between 0.7273 (Rugao vs. Wan-Nan) and 0.9367 (Jiangchun vs. Cshiqishi) chicken populations. Using the BSC for individual and pairwise populations, the Nei's standard genetic distances between the chicken populations were determined and ranged from 0.0043 (Jiangchun vs. Cshiqishi) to 0.1375 (Rugao vs. Cshiqishi). The reconstructed dendrogram linked the Jiangchun and Cshiqishi chickens as closely related populations, followed by Wan-Nan, while the Rugao was the most genetically distant among the populations.

• Journal of Genetic Medicine
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• 제16권2호
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• pp.49-54
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• 2019

Purpose: Potocki-Lupski syndrome (PTLS), is a recently identified, rare genomic disorder. The patients are affected by infantile hypotonia, poor growth and developmental delay. Facial dysmorphism may not be obvious in some patients. PTLS is associated with microduplication at chromosome 17p11.2. In the current study, three Korean patients are reported with their clinical and genetic features. Materials and Methods: The clinical findings of each patient were reviewed. Karyotyping and multiplex ligation-dependent probe amplification (MLPA) analyses were done for genetic diagnoses. Results: All the patients did not have the characteristic dysmorphic features, such as broad forehead, triangular face, asymmetric smile and palpebral fissures. On the other hand, all three patients were affected by variable degree of developmental delay, poor oral intake, failure to thrive, and language development disorders. Chromosome 17p11.2 duplication was identified by conventional karyotyping analysis only in one patient, whereas the other confirmed by MLPA analyses. Conclusion: Delayed development was mostly commonly observed in our patients without distinct dysmorphic facial features. In this respect, genomic screening in patients with developmental delay would identify more cases with PTLS to understand their long-term clinical courses with the development of adequate psychological and rehabilitation education program.

• Genomics & Informatics
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• 제17권4호
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• pp.42.1-42.6
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• 2019

Robust identification of genetic alterations is important for the diagnosis and subsequent treatment of tumors. Screening for genetic alterations using tumor tissue samples may lead to biased interpretations because of the heterogeneous nature of the tumor mass. Liquid biopsy has been suggested as an attractive tool for the non-invasive follow-up of cancer treatment outcomes. In this study, we aimed to verify whether the mutations identified in primary tumor tissue samples could be consistently detected in plasma cell-free DNA (cfDNA) by digital polymerase chain reaction (dPCR). We first examined the genetic alteration profiles of three colorectal cancer (CRC) tissue samples by targeted next-generation sequencing (NGS) and identified 11 non-silent amino acid changes across six cancer-related genes (APC, KRAS, TP53, TERT, ARIDIA, and BRCA1). All three samples had KRAS mutations (G12V, G12C, and G13D), which were well-known driver events. Therefore, we examined the KRAS mutations by dPCR. When we examined the three KRAS mutations by dPCR using tumor tissue samples, all of them were consistently detected and the variant allele frequencies (VAFs) of the mutations were almost identical between targeted NGS and dPCR. When we examined the KRAS mutations using the plasma cfDNA of the three CRC patients by dPCR, all three mutations were consistently identified. However, the VAFs were lower (range, 0.166% to 2.638%) than those obtained using the CRC tissue samples. In conclusion, we confirmed that the KRAS mutations identified from CRC tumor tissue samples were consistently detected in the plasma cfDNA of the three CRC patients by dPCR.

• 한국버섯학회지
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• 제1권1호
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• pp.44-47
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• 2003

버섯으로부터 새로운 암전이 억제물질을 개발하기 위하여 먼저 7종 52균주의 자실체 (또는 균사체)에 대한 다양한 추출물들을 제조한 후 이들의 암전이 억제에 관련된 혈관신생 억제활성을 수정란을 이용하여 Choriollantoic membrane (CAM) assay로 조사하였다. 7종 52균주의 버섯에 대한 물, 에탄올, 메탄올 추출물 중에서 장수버섯(Fomitella fraxinea) ASI 17003과 17009의 물 추출물과 비늘버섯(Pholiota sp.) ASI 24008과 잎새버섯(Grifola frondosa) ASI 9017의 에탄올 추출물 그리고 차가버섯(Inonotus obliquus) ASI 74012의 메탄올 추출물에서 $10{\mu}g/egg$농도일 때 62.5%~68.8%의 높은 혈관신생 억제활성을 보였다. 그러나 추출 수율과 안전성 등을 고려하여 비늘버섯 ASI 24008을 혈관신생 억제효과 우수버섯으로 최종 선정하여 현재 이들의 암전이 억제 기작 규명을 위한 정제 실험을 실시하고 있다.

• 대한바이러스학회지
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• 제30권3호
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• pp.161-170
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• 2000

Continuous efforts are being made to find effective therapeutic agents against HIV-1, the causative agents of AIDS. In this study, we developed a cell-based assay system employing a trans-complementation for production of recombinant viruses which are capable of undergoing one round of replication in CD4+ T cells. This assay system was tested for ability to screen the agents that act at late stage of HIV-1 life cycle. The effect of a protease inhibitor on the trans-complementation assay was assessed. Recombinant HIV-1 viruses were prepared from a trans-complementation in the presence of various concentrations of protease inhibitor. Inhibition of single round infection of these recombinant viruses by protease inhibitor was observed to be a dose-dependent manner. Inhibitory effects of a protease inhibitor on HIV-1 Gag polyprotein processing by HIV-1 protease was detected at concentrations of the protease inhibitor compatible with inhibition of virus infection, confirming that the corresponding step was involved in the inhibitory mechanism of this compound. Together, these results provide evidence that a cell-based assay system established in this study can be used to screen the agents that target the late stage of HIV-1 life cycle.

• 한국균학회지
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• 제39권3호
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• pp.194-197
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• 2011

항고혈압 효능이 우수하며 부작용이 없는 새로운 고혈압 예방 소재를 개발하고자 한국 식품연구원에서 분리, 동정한 48종의 효모들의 PD broth 배양 농축물들을 대상으로 angiotensin 전환효소(ACE) 저해활성을 조사하여 활성이 우수한 효모들을 선정한 다음 ACE 저해물질의 생산 최적 조건을 검토하였다. ACE 저해활성은 Saccharomyces cerevisiae Y183-3 균주의 PD broth 배양 농축물이 71.8%의 가장 높은 ACE 저해활성을 보여 최종 우수 균주로 선발하였다. 그리고 이 Y183-3 효모를 $30^{\circ}C$에서 36시간 PD broth에 배양했을 때 ACE 저해물질이 가장 많이 생산되었다.

• 한국컴퓨터정보학회논문지
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• 제17권12호
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• pp.251-258
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• 2012

과거 종이로 기록되던 환자의 의료정보는 점점 현대과학의 눈부신 진화와 발전으로 현재는 종이를 대신하는 전자적 시스템인 전자의무기록시스템 형태로 발전되었다. 또한 병원에서 활용되는 모든 의료정보들이 전자적시스템을 이용하여 활용되고 있는 것이 현실이다. 그러나 과학기술의 발전과 더불어 의료정보의 피해 사례는 계속 증가할 것으로 보인다. 보험사기 예방을 위해 의료정보를 공유해야한다는 보험단체들의 주장, 개인정보를 얻기 위해 금전적 불법거래가 성행되고 있다는 뉴스 등을 접할때면 의료정보 보호에 관한 대책 마련이 절실함을 의미한다. 따라서 의료정보와 관련된 유출상의 문제점을 정확히 파악하는 것은 의료정보 유출의 부작용을 예방하고 의료정보 보호와 미비점을 보완하는 선결과제인 것이다. 따라서 현재의 의료법 및 개인정보보호법제만으로는 의료정보의 표준화와 의료정보보호 등을 규율할 수 없고, 의료정보에서 발생되는 여러 문제들을 보호하기에는 미흡하다고 판단된다. 따라서 본 논문은 미흡한 의료정보관련 법제의 연구를 위한 선결 과제로 의료정보의 유출로 발생될 수 있는 문제점을 파악하는 것이 무엇보다 중요하다고 판단되며, 이에 의료정보의 유출로 인한 문제점을 파악하고 의료정보를 보호할 수 있는 입법방안을 중심으로 연구하고자 한다.