• Title/Summary/Keyword: Human DNA

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Mitochondrial DNA polymorphism in the Cheju horses (제주마의 mitochondrial DNA 다형(多型)의 분석(分析))

  • Han, Bang-keun;Chang, Deuk-jee;Tsuchida, Shuichi;Ikemoto, Shigenori
    • Korean Journal of Veterinary Research
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    • v.34 no.2
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    • pp.243-247
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    • 1994
  • As a result of the detection of mitochondrial DNA(mtDNA) polymorphism to Thoroughbred and Percheron using 14 restriction enzymes, mtDNA polymorphism of Cheju horse observed in the Bam HI and Sac I. Only in both restriction enzymes two types were classified as of A type, which is high expression frequency and B type, which is low expression frequency. In the other 12 restriction enzymes mtDNA polymorphism was not detected. On the basis of this information mtDNA polymorphism of Cheju horse was examined but was not observed the polymorphism and only A type was expressed both Bam HI and Sac I restriction enzymes. Through this study Cheju horse was demonstrated that lower genetic variation was expressed from the detection of mtDNA polymorphism.

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Molecular Cloning, Identification and Characteristics of a Novel Isoform of Carbamyl Phosphate Synthetase I in Human Testis

  • Huo, Ran;Zhu, Hui;Lu, Li;Ying, Lanlan;Xu, Min;Xu, Zhiyang;Li, Jianmin;Zhou, Zuomin;Sha, Jiahao
    • BMB Reports
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    • v.38 no.1
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    • pp.28-33
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    • 2005
  • A gene coding a novel isoform of carbamyl phosphate synthetase I (CPS1) was cloned from a human testicular library. As shown by cDNA microarray hybridization, this gene was expressed at a higher level in human adult testes than in fetal testes. The full length of its cDNA was 3831 bp, with a 3149 bp open reading frame, encoding a 1050-amino-acid protein. The cDNA sequence was deposited in the GenBank (AY317138). Sequence analysis showed that it was homologous to the human CPS1 gene. The putative protein contained functional domains composing the intact large subunit of carbamoyl phosphate synthetase, thus indicated it has the capability of arginine biosynthesis. A multiple tissue expression profile showed high expression of this gene in human testis, suggesting the novel alternative splicing form of CPS1 may be correlated with human spermatogenesis.

Carcinogenic Potentials of HPV-16 and NNK in Human in Vitro Model (인체 세포 모델을 이용한 HPV-16과 NNK의 발암 잠재력에 관한 연구)

  • 양재호;이세영
    • Toxicological Research
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    • v.12 no.2
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    • pp.271-275
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    • 1996
  • Carcinogenic potential of HPV-16 DNA and NNK in a human keratinocyte cell line was assessed to study effects of viral-chemical interaction. Human cells were transfected with HPV-16 DNA and 6 clonal cell lines were subsequently obtained. Clonal line-3 and 6 at passage 7 showed characteristics of tumor cells such as increases of saturation density, soft-agar colony formation, cell aggregation and foci appearance. Among cells treated with 1$\mu M$, 10$\mu M$, 100$\mu M$ or 1 mM of NNK for 4 weeks, 100$\mu M$ treatment showed most tumorigenic characteristics at passage 7. These results indicate that either HPV-16 or NNK alone is tumorigenic in this in human in vitro model. When cells transfected with HPV-16 were subsequently exposed by 100 uM NNK for 4 weeks, all the clonal cells except clone-1 showed higher levels of tumor cell characteristics than HPV-16 DNA or NNK exposure alone. Clonal line-6, the most tumorigenic cells, showed higher transcriptional level of fibronectin and lower level of TGF-$\beta_1$, as compared to control cells, suggesting that alteration of growth factor or extracellular matrix may play a role in carcinogenesis process induced by HPV-16 and NNK. Taken together, the present study indicates that viral-chemical interactions between HPV-16 DNA and NNK enhance carcinogenic potentials of human cells and implies that smoking among people infected with human papillomavirus may pose an additional risk of causing cancer.

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The Effect of Carnosine Extracted from Eels Anguilla japonica on Oxidative DNA Damage Induced by Hydrogen Peroxide and the DNA Repair Capacity of Human Leukocytes (뱀장어(Anguilla japonica) 추출 Carnosine이 과산화수소로 유도된 인체 백혈구의 DNA 손상과 Repair에 미치는 효과)

  • Song, Ho-Su
    • Korean Journal of Fisheries and Aquatic Sciences
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    • v.50 no.5
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    • pp.520-526
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    • 2017
  • Carnosine was recently reported to protect against the DNA damage induced by oxidative stress. In this study, we investigated the protective effect of eel Anguilla japonica carnosine extracts prepared using different methods (heat treatment extracts, HTEs; ion exchange chromatography, IEC; ultrafiltration permeation, UFP) on leukocyte DNA damage using the comet assay. Human leukocytes were incubated with extracts of eel carnosine at concentrations (of 10, 50, $100{\mu}g/mL$), and then subjected to an oxidative stimulus [$200{\mu}M$ hydrogen peroxide ($H_2O_2$)]. Pretreatment of the cells for 30 min with carnosine significantly reduced the genotoxicity of $H_2O_2$ measured as DNA strand breaks. The protective effects of the three types of extract (HTE, IEC, and UFP) increased with concentration. At the highest concentration (100 g/mL). there were no statistical differences in oxidative damage between each extract treatment and PBS-treated negative controls. When leukocytes were incubated with carnosine for 30 min after exposure to $H_2O_2$. the protective ability of each extract changed. Therefore, eel carnosine inhibits the $H_2O_2$ induced damage to cellular DNA in human leukocytes, supporting the protective effect of this compound against oxidative damage.

Comparison of the Protective Effect of Antioxidant Vitamins and Fruits or Vegetable Juices on DNA Damage in Human Lymphocyte Cells Using the Comet Assay (Comet Assay를 이용한 항산화 비타민과 과일.야채즙의 인체 임파구 세포 DNA 손상 감소 효과 비교)

  • 전은재;박유경;김정신;강명희
    • Journal of Nutrition and Health
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    • v.37 no.6
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    • pp.440-447
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    • 2004
  • In this study the in vitro protective effects of several antioxidant vitamins (vitamin C, $\alpha$-tocopherol, $\beta$-carotene), fruits and vegetables (strawberry, tangerine, orange and 100% orange juice, carrot juice), on the levels of isolated human lymphocyte DNA damage was measured using Comet assay. Comet assay has been used widely to assess the level of the DNA damage in the individual cells. Lymphocytes were pre-treated for 30 minutes with antioxidant vitamins (10, 50, 100, 500 $\mu$M) or fruits$.$vegetables (10, 100, 500, 1000 $\mu$g/ml), an4 then oxidatively challenged with 100 $\mu$M $H_2O$$_2$ for 5 min at 4$^{\circ}C$. The protective effect of antioxidant vitamins against DNA damage at a concentration of 50 $\mu$M were 50% in vitamin C, 32% in $\alpha$-tocopherol, whereas, fJ-carotene showed a 55% protection at a dose as low as 10 $\mu$M. The inhibitory effects of DNA damage by strawberry, tangerine, orange, orange juices, carrot juices were 50 - 60% with wide ranges of doses. The results of the present study indicate that most the antioxidant vitamins and fruits$.$vegetables juices produced a significant reduction in oxidative DNA damage.

The Relationship between Papanicolaou Smear Test and Human Papilloma Virus DNA Chip Test in the Uterine Cervix

  • Lee, Young-Ju;Jung, Ji-Hun;Jung, Da-Young
    • Korean Journal of Clinical Laboratory Science
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    • v.43 no.1
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    • pp.26-31
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    • 2011
  • The genotypes of Human Papilloma Virus (HPV) are important in the carcinogenesis of uterine cervical cancer. Diagnosis of uterine cervical cancer screening has been executed using Papanicolau method (Pap) and HPV DNA Chip method. We researched the interrelation of HPV DNA genotypes in single and multiple infections and analyzed the results of Pap and HPV DNA Chip tests at Gunsan Medical Center (GMC). The correlation analysis was surveyed on collected results from 599 patients who have been tested with both Pap and HPV DNA chip tests from November 2004 to May 2010 at GMC. The inconsistency between Pap and HPV DNA Chip tests was 41.1%. The HPV DNA Chip genotype related with high risk cases were type 16 (13.5%), type 52 (10.5%), type 58 (10.1%), and type 18 (3.4%). Those related with low risk cases were type 70 (8.9%), type 6 (1.7%), type 40 (1.2%), type 11 (1.3%), and other types (14.3%). Among the 195 cases of HPV positive status, 161 cases were associated with single infection; 108 (67.1%) cases were related with high risk genotype; 19 (11.8%) cases were low risk genotype; 31 (21.1%) cases were related with other types. 29 cases were associated with double infections; 23 (79.3%) cases were high risks; 5 (17.2%) cases were mixed high and low risks; 1 (3.5%) case was low risk.

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Effect of Microsurgical Varicocelectomy on Human Sperm Nucleus DNA Integrity (미세술기를 이용한 정계정맥류절제술이 정자 핵 내 DNA Integrity에 미치는 효과)

  • Kim, Gi-Young;Lee, Jae-Seok;Chi, Hee-Jun;Kim, Jong-Hyun
    • Clinical and Experimental Reproductive Medicine
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    • v.37 no.3
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    • pp.245-251
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    • 2010
  • Objective: Human sperm nucleus DNA damage may negatively affect pregnancy outcome, and the spermatozoa of infertile men have more DNA damage than that of fertile men. The aim of this study was to evaluate the effect of microsurgical varicocelectomy on human sperm nucleus DNA integrity. Methods: We reviewed the medical records of 18 subfertile male patients who underwent microsurgical varicocelectomy at our hospital from April 2006 to April 2007. Varicocele was diagnosed by physical examination and Doppler ultrasound. Standard semen analysis was performed in 18 patients before and 4 months after microsurgical varicoceletcomy using a computer assisted semen analyzer. Sperm nucleus DNA integrity was assessed by a single-cell gel electrophoresis (comet assay). Results: No recurrence of varicocele was observed after 4 months later. The DNA fragmentation index improved after varicocelectomy compared with pre-operatively (19.3 versus 13.7%, respectively, p<0.05). Semen analysis parameters (total count, concentration, motile sperm, viability, strict morphology) increased after varicocelectomy, but the difference did not reach statistical significance. Conclusion: Our data suggest that microsurgical varicocelectomy can improve semen analysis parameters and human sperm nucleus DNA integrity in infertile men with varicocele.

Phylogenetic Analysis of Bacterial Diversity of Enhanced Biological Phosphorus Removal Activated Sludge by Isolation and Cloning of 16S rDNA

  • Nakamura, Kazunori;Hanada, Satoshi;Kamagata, Yoichi;Kawaharasaki, Mamoru
    • Proceedings of the Korean Society for Applied Microbiology Conference
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    • 2000.10a
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    • pp.109-117
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    • 2000
  • Bacterial community structure composing enhanced biological phosphorus removal (EBPR) activated sludge was analyzed phylogenetically by cloning 165 rDNA after direct DNA extraction. Then, this result was compared with 165 rDNA sequences of randomly isolated bacterial species. The results clearly showed that there are no coincidence between the sequences retrieved directly from activated sludge and those of isolated strains, suggesting that many important bacteria are hidden in activated sludge because of the difficulty in isolation and culture of them.

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Gender Determination of X and Y-Specific Alphoid Repeat Sequences by PCR (PCR에 의한 X,Y-Specific Alphoid Repeat Sequences의 분석)

  • Choi, Dong-Ho;Kang, Pil-Won;Lee, Yang-Han;Han, Myun-Soo
    • Analytical Science and Technology
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    • v.12 no.1
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    • pp.80-83
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    • 1999
  • Recently, it has been possible to the alphoid gene, which has X and Y specificity, and determine the sex from human physical evidence using PCR methods. Samples from single sources, PCR method applied to the alphoid gene results in highly sensitive and accurate results even when only 60 pg of the genomic DNA was available for sex determination. Even for samples containing DNA from more than one gender source where the female DNA was present in the amount 10 times than that of the male, sex determination was possible. Therefore, this result suggests that alphoid gene, which has X and Y specificity, could be used effectively for sex determination in case of mixed DNA samples from biological evidence.

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Applied Research of Ultra Sonication for Ancient DNA Preparation of Excavated Human Skeletal Remains (초음파를 이용한 출토 인골 DNA 추출법 연구)

  • Kim, Yun-Ji;Jee, Sang-Hyun;Hong, Jong-Ouk
    • 보존과학연구
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    • s.29
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    • pp.137-148
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    • 2008
  • Analyses of ancient DNA (aDNA) from archaeological and historical skeletal material are characterized by low quality. Many soil contaminants such as humic acid, fulvic acid, and bone collagen are often co-extracted with aDNA and inhibit amplification by polymerase chain reaction (PCR). In this study, we compared with two methods of DNA extraction by phenolchloroform extraction and silica-bead extraction. In addition, we applied new protocol, ultra sonication based silica-bead extraction method to extract aDNA from some ancient human skeletal remains. This method was more effective by both mitochondrial DNA (mtDNA) and amelogenin gene amplification.

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