• Fisheries and Aquatic Sciences
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• 제5권1호
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• pp.36-42
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• 2002

Microarrayer is used to make DNA chip and microarray that contain hundreds to thousands of immobilized DNA probes on surface of a microscope slide. This paper shows the develop-ment results for a printing type of microarrayer. It realizes a typical, low-cost and efficient microarrayer for generating low density micro array. The microarrayer is developed by using a prependicular type robot with three axes. It is composed of a computer-controlled three-axes robot and a pen tip assembly. The key component of the arrayer is the print-head containing the tips to immobilize cDNA, genomic DNA or similar biological material on glass surface. The robot is designed to automatically collect probes from two 96-well plates with up to 12 pens at the same time. To prove the performance of the developed microarrayer, we use the general water types of inks such as black, blue and red. The inks are distributed at proper positions of 96 well plates and the three color inks are immobilized on the slide glass under the operation procedure. As the result of the test, we can see that it has sufficient performance for the production of low integrated DNA chip consisted of 96 spots within $1cm^2$ area.

• 대한두경부종양학회지
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• 제27권2호
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• pp.190-197
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• 2011

Background and Objective : Radiation resistance(RR) is one of main determinants of treatment outcome in oral squamous cell carcinoma(OSCC), but accurate prediction of RR is difficult. We aim to establish RR OSCC cell lines and identify genes related with RR by a measurement of altered gene expression after inducing RR. Material and Methods : OSCC cell lines, SCC15, SCC25 and QLL1, were treated with 2Gy radiation per session, and parts of them were alive in finally accumulated dosage of 60Gy through 30 times repetition of radiotherapy for inducing RR cell lines. We compared results of cDNA array and proteomics in non-radiated cell lines and RR cell lines to detect changes of gene expression. Western blot was used for the validation of results. Results : cDNA array revealed 265 commonly up-regulated genes and 268 commonly down-regulated genes in 3 RR cell lines comparing their non-radiated counterpart. Among them, 30 cancer related genes were obtained. Proteomics showed 51 commonly altered protein expressions in 3 RR cell lines and 18 cancer related proteins were obtained. Among the detected genes, we found NM23-H1 and PA2G4 were over-expressed in both cDNA array and proteomics. Western blot showed increased expression of NME1 in RR cell lines but not in PA2G4. Conclusion: We concluded that NM23-H1 may be a candidate of RR related gene and over-expression of NM23-H1 could be a biomarker to predict RR in OSCC.

• 대한두경부종양학회지
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• 제12권2호
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• pp.153-160
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• 1996

Regardless of the prognostic factors in papillary thyroid cancer, such as sex, age, size of tumor, extent of disease, and distant metastasis, the prognosis of papillary thyroid cancer is sometimes difficult to predict from clinical and microscopic analysis alone and additional prognostic indicators are needed. Recent studies of thyroid cancer have indicated that DNA aneuploidy may be correlated to the biological behavior of malignancy and inversely correlated to the prognosis, but it still remains contraversal. We performed this study to assess DNA ploidy patterns in relation with the previously known prognostic factors in AMES scoring system and lateral neck node metastasis in papillary thyroid cancer. A series of 132 patients with papillary thyroid cancer and 80 patients with benign thyroid tumor(27 follicular adenomas and 53 adenomatous goiters) as a control group from October 1993 to Feburary 1995 were analyzed and their nuclear DNA content was measured with flow cytometry using fresh tissue specimens. DNA aneuploidy was found in 8(6.1%) in papillary cancer and 8(10%) in benign tumor. S-phase traction(SFP) and proliferative index(PI) were higher in thyroid cancers, being 2.18$\pm$4.24%, 6.34$\pm$4.94% in the papillary thyroid cancers and 1.97$\pm$2.93%, 4.44$\pm$3.80% in the benign tumors, respectively. However there was no significant difference of values between two groups(p>0.05). Among variable prognostic factors studied(age, sex, size of tun or, extent of disease, distant metastasis in AMES scoring system and lateral neck node metastasis), DNA aneuploidy was found to be common in distant metastasis(p<0.001) and in lateral neck node metastasis(p>0.035), but there was no significant difference between the high risk and low risk group according to the AMES scoring system(p<0.08). In our study, DNA aneuploidy was not valuable in determining the presence of malignancy and did not correlate to the AMES scoring system. However, follow-up study of more cases will be needed for accurate information about the DNA ploidy as a independent prognostic factor.

• 대한두경부종양학회지
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• 제16권1호
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• pp.14-19
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• 2000

Objectives: Epstein-Barr virus(EBV) is a B-lymphotrophic virus with a tumorigenic potential. EBV infection has been recognized as the main cause of nasopharyngeal carcinoma and Burkitt's lymphoma. Bcl-2 protein expression is known to be up-regulated by the EBV-latency associated antigen latent membrane protein(LMP). The aim of this study was to determine the incidence of EBV in squamous cell carcinomas of the larynx and the relationship between the presence of EBV and bcl-2 expression. Patients and Methods: From January 1994 to December 1977, 35 patients with primary squamous cell carcinoma of the larynx were studied. EBV genome DNA was surveyed by polymerase chain reaction(PCR) assay and then compared the results of in situ hybridization(ISH) for EBER1 using digoxigenin-tailed oligonucleotide probe. The expression of bcl-2 protein was studied by immunohistochemistry(IHC) using bcl-2 monoclonal antibody. Results: By PCR, EBV genome was detected in 22 of 35(62.9%) squamous cell carcinomas of the larynx. Nineteen of 35 cases(54.3%) showed a positive nuclear staining for EBER1 in tumor cells by ISH. Three cases showed positivity in inflammatory cells by ISH and one of them showed a positive staining of both tumor cells and inflammatory cells. Eighteen of 32 specimens(62.5%) were positive for bcl-2 protein. There was no significant correlations between the presence of EBV DNA and bcl-2 expression. Conclusions: It could be concluded that high incidence of EBV in the laryngeal cancer tissue may indicate a probable role of EBV in the development of laryngeal carcinoma.

• 한국어병학회지
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• 제34권2호
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• pp.161-168
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• 2021

Quantitative analysis of myxosporean parasites (Enteromyxum leei and Parvicapsula anisocaudata) were performed using real-time PCR on the internal organs (head kidney, body kidney, intestine, spleen, brain, liver, heart, muscle, blood, and eye) of emaciated Paralichthys olivaceus from farm-A. The highest DNA copy number of E. leei was shown in the intestine (1.3 × 10⁸ copies/mg tissue) of emaciatied P. olivaceus and DNA copy number in the other internal organs (1.3 × 10³~4.6 × 10⁵ copies/mg tissue) showed lower than in intestine. From the result of real-time PCR for P. anisocaudata, it was considered mildly infected, due to the low DNA copy numbers of the head kidney (1.3 × 10³ copies/mg tissue) and body kidney (9.1 × 10³ copies/mg tissue). In order to investigate whether myxosporean parasites can be detected in a non-invasive way, quantitative analysis of E. leei and P. anisocaudata from rearing water of three farms were performed by real-time PCR. The DNA copy number of E. leei from rearing water of farm-A and farm-B were 8 × 10⁴ and 5 × 10⁵ copies/L, respectively. However, it was not detected in farm-C. For P. anisocaudata from rearing water, farm-A, farm-B and farm-C showed 0, 2.0 × 10⁶ and 5.1 × 10⁶ copies/L, respectively.

• 생명과학회지
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• 제27권11호
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• pp.1331-1339
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• 2017

미토콘드리아 게놈에 존재하는 시토크롬C 산화효소 서브유닛 I (cytochrome C oxidase subunit I, COI) 유전자의 DNA 염기서열을 기반으로 하는 종 판별은 수산물 자원의 지속적인 개발과 어류 다양성 보존을 위해 폭넓게 적용되고 있다. 본 연구에서는 한국에서 소비되는 옥돔과 가짜 옥돔으로 둔갑하는 옥두어의 종 판별을 위한 분석법을 개발하였다. 옥돔과 옥두어, 두 종의 종 판별과 검증을 위해 미토콘드리아 게놈의 DNA 염기서열 차이를 이용하여 real-time PCR법에 의해 분석하였다. 미토콘드리아 DNA 서열의 생물정복학적 분석에서 옥돔과 형태학적 옥돔 유사종인 옥두어, 두 종 사이에 COI 유전자 내에서 상당히 유사한 DNA 서열 부분과 일부 서열 변화 부분이 확인되었다. 명확하게 종 판별을 하기 위해 COI 유전자 내에서 일부 변화된 서열에서 종 특이적 프라이머를 디자인하였다. 10 개체의 옥돔과 옥두어에서 게놈 DNA을 추출하여 옥돔과 옥두어의 종 특이적 프라이머를 이용하여 real-time PCR 시스템에 의해 분석되었다. 이러한 real-time PCR 시스템을 이용한 genomic DNA 기반의 분자 기술은 동물 조직의 분류학적 분류를 위한 신뢰할 수 있는 방법을 제공한다. 옥돔판별을 위해, 옥돔 DNA에서 옥돔 종 특이적 프라이머를 이용한 Ct 평균값($21.85{\pm}3.599$)과 옥두어 DNA에서 옥돔 종 특이 프라아머를 이용한 Ct 평균값($33.49{\pm}1.183$) 차이를 나타내었다. 그리고 옥두어판별을 위해, 옥두어 DNA에서 옥두어 종 특이적 프라이머를 이용한 Ct 평균값($22.49{\pm}0.908$)과 옥돔 DNA에서 옥두어 종 특이 프라아머를 이용한 Ct 평균값($33.93{\pm}0.479$)을 통해 옥돔과 옥두어의 각 종 특이 프라이머의 효율성, 특이성 및 교차 반응성 측정은 통계적으로 유의한 차이를 보여 주었다. 제안된 방법은 10개의 상용 샘플로 검증이 되었다. 따라서, threshold cycle (Ct) value와 같은 real-time PCR 결과 분석에 의해 종 판별이 가능하였다.

• 대한두경부종양학회지
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• 제20권2호
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• pp.147-155
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• 2004

Objectives: Head and neck squamous cell carcinoma (HNSCC) is the most common head and neck malignant tumor. The molecular genetic changes involving both oncogenes and tumor suppressor genes are known to be involved in head and neck squamous cell carcinogenesis, but the roles of the known tumor suppressor genes in carcinogenesis are not fully elucidated. The objectives of this study are to demonstrate the genetic alterations including the loss of heterozygosity (LOH) , amplification, and microsatellite instability of known tumor suppressor genes in HNSCC and to evaluate the relationship between genetic alterations of tumor suppressor genes and clinicopathologic features. Materials and Methods: Genetic alterations of 10 micro satellite markers of the 6 known tumor suppressor genes (APC, EXT1, DPC4, p16, FHIT, and PTEN) were analysed by DNA-PCR in paraffin-embedded histologically confirmed HNSCC specimens. Results: The genetic alterations of tumor suppressor genes were found frequently. Among the genetic alterations, LOH was most frequently found one. LOH was found frequently in APC (45.4%), EXT1 (36.4%), DPC4 (54.5%), and p16 (50%), but not found in FHIT. Also, the author found that abnormalities of APC gene was related to cervical lymph node metastasis and recurrence and that abnormalities of EXT1 gene were coexisted with those of APC gene or DPC4 gene. But these coexistences had no correlation with clinical features. Conclusion: These results suggested that APC, EXT1, p16, and DPC4 genes might play important roles and multiple tumor suppressor genes may participate dependently or independently in the carcinogenesis of HNSCC. These results also suggested that APC gene might relate to prognosis.

• 한국수산과학회지
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• 제56권6호
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• pp.916-922
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• 2023

An abnormal shape of Bathyraja brachyurops was first reported from the catch of a bottom trawl in the southwest Atlantic Ocean in June 2022. Both pectoral fins of the specimen did not fuse with the head, resulting in a horn-like structure separated from the sides of the eyes. Analysis of mitochondrial DNA cytochrome c oxidase subunit I sequences showed that our specimen was perfectly matched to Bathyraja brachyurops registered with the National Center for Biotechnology Information. Our specimen possessed the following morphological features: a pair of flexible but elongated and pointed horns on the head; rough dorsal disc, densely covered with numerous small denticles on the head, anterior margin of pectoral fins and median line of the disc; a thorn between the first and second dorsal fins; and a pair of large ocelli at the base of pectoral fins. Unlike the normal B. brachyurops, our specimen had a slender clasper and no nuchal thorns, which may be related to the morphological abnormality. The horn-like structure on the head may be owing to the lack of fusion between the pectoral fins and head during early embryonic development.

• Journal of Korean Neurosurgical Society
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• 제29권2호
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• pp.188-195
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• 2000

Objective : Essential tremor(ET) is the most common movement disorder, however, there has been little agreement in the neurologic literature regarding diagnostic criteria for ET. Familial ET is an autosomal dominant disorder presenting as an isolated postural tremor. The main feature of ET is postural tremor of the arms with later involvement of the head, voice, or legs. In previous studies, it was reported that ET susceptibility was inherited in an autosomal dominant inheritance. As previous results, it would suggest that ET might be associated with defect of mitochondrial or nuclear DNA. Recent studies are focusing on molecular genetic detection of movement disorders, such as essential tremor and restless legs syndrome. Moreover, authors have analysed mitochondrial DNA(mtDNA) from the blood cell of positive control(PC) and ET patients via long and accurate polymerase chain reaction(LA PCR). Materials & Methods : Blood samples were collected from PC and 9 ET patients. Total DNA was extracted twice with phenol followed by chloroform : isoamylalcohol. For the analysis of mtDNA, LA PCR was performed by mitochondrial specific primers. Results : With this technique, deletions of large quantities were detected within several regions of mtDNA in ET patients except for D-loop and CO I regions. Conclusion : The authors believe that ET is a genentic disorder with deficiency of mitochondrial DNA multicomplexes and mitochondiral dysfunction could be one of major causative factors of ET. Mitochondrial dysfunction may play an important role in the pathogenesis and possibility of disease progression among familial group with ET patients.

• 대한두경부종양학회지
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• 제22권1호
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• pp.8-14
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• 2006

Objectives: The p53 tumor suppressor gene has a key role in cellular control mechanisms involving apoptosis and DNA repair, leading to the G1 arrest following DNA damage. Its mutation is one of the most frequent alterations in human cancers. Ki-67 is identified in replicating cells of both benign and malignant lesions, so it can be the predictor of proliferative activity. The aim of this study is to evaluate the expression of p53 and Ki-67 in salivary gland tumors. Materials and Methods: Immunohistochemical analysis was used to detect expression of p53 and Ki-67 in paraffin-embedded samples from 31 benign and 27 malignant salivary gland tumors. Results were analyzed between benign and malignant tumors and compared with the clinical parameters such as stage and recurrence in malignant tumors. Results: p53 overexpression was detected in 19.6% of benign tumors and 40.7% of malignant tumors, but there was no statistical significance. p53 was significantly expressed in Warthin's tumor(45.5%) compared with pleomorphic adenoma(5.9%). Only 5.9% of pleomorphic adenoma were positive for p53, while 60% of carcinoma ex pleomorphic adenoma were positive for p53. Ki-67 was expressed in 3.2% of benign tumors and 51.9% of malignant tumors, which showed significant higher expression in malignant tumors. In malignant tumors, p53 and Ki-67 expressions bore no correlation to stage and recurrence. Conclusion: p53 overexpression is not associated with the progression of malignant tumors, and Ki-67 overexpression can be used as biologic indicator of malignant salivary gland tumors.