• Asian Pacific Journal of Cancer Prevention
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• 제17권3호
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• pp.939-944
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• 2016

Background: Cell-free DNA circulating in blood is a candidate biomarker for malignant tumors. Unlike uniformly truncated DNA released from apoptotic non diseased cells, DNA released from necrotic cancer cells varies in size. Objectives: To measure the DNA integrity index in serum and the absolute DNA concentration to assess their clinical utility as potential serum biomarkers for colorectal carcinoma (CRC) compared to CEA and CA19-9. Materials and Methods: Fifty patients with CRC, 10 with benign colonic polyps and 20 healthy sex and age matched volunteers, were investigated by real time PCR of ALU repeats (ALU q-PCR) using two sets of primers (115 and 247 bp) amplifying different lengths of DNA fragments. The DNA integrity index was calculated as the ratio of q-PCR results of ALU 247/ALU 115bp. Results: Serum DNA integrity was statistically significantly higher in CRC patients compared to the benign and control groups (p<0.001). ROC curves for differentiating CRC patients from normal controls and benign groups had areas under curves of 0.90 and 0.85 respectively. Conclusions: The DNA integrity index is superior to the absolute DNA concentration as a potential serum biomarker for screening and diagnosis of CRC. It may also serve as an indicator for monitoring the progression of CRC patients. Combining CEA and CA19-9 with either of the genetic markers studied is better than either of them alone.

• Clinical and Experimental Reproductive Medicine
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• 제37권3호
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• pp.245-251
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• 2010

목 적: 인간 정자 핵 내의 DNA integrity는 배아의 발달 및 임신 유지에 중요한 역할을 하여 DNA integrity가 손상된 경우 불임과 유산의 원인이 된다고 하며, 정계정맥류는 DNA 손상을 일으키는 대표적인 원인 중 하나이다. 본 연구에서는 미세술기를 이용한 정계정맥류절제술로 정계정맥류를 교정을 하였을 때 정자 핵 내 DNA integrity가 어떠한 영향을 받는지에 대하여 알아보았다. 연구방법: 2006년 4월부터 2007년 4월까지 불임을 주소로 미세술기를 이용한 정계정맥류절제술을 받았던 18명의 환자에서 수술 전 후에 정액검사의 다른 지표들과 함께 정자 핵 내 DNA integrity가 어떻게 변화하였는지 조사하였다. 정자 핵 내 DNA integrity를 측정하는 방법으로 comet assay를 시행하였고, comet assay를 통한 DNA 손상 정도는 DNA fragmentation index (DFI)로 나타내었다. 결 과: 수술 후 4개월에 모든 환자에서 재발의 소견은 보이지 않았으며, DNA 손상 정도를 나타내는 평균 DFI는 수술 전에 19.3%, 수술 후에 13.7%로 유의한 변화를 보였다. 수술 전 DFI가 10 이상으로 비정상인 14명의 환자들 중 12명 (85%)에서 개선 소견을 보였으나, 수술 전 DFI가 10 미만인 정상 환자 4명에서는 1명 (25%)만이 개선 소견을 보였다. 수술 후 정자의 밀도, 운동성, 생존성에서 호전 양상을 보였으나 유의한 차이는 없었다. 결 론: 미세술기를 이용한 정계정맥류절제술을 통한 수술적 교정은 정액검사상의 다른 지표의 개선 뿐 아니라, 정자 핵 내 DNA 손상을 감소시킬 수 있다. 이상에서 정계정맥류의 수술적 교정으로 정자 핵 내 DNA integrity의 개선을 기대할 수 있으며, 이는 보다 양호한 정자를 많이 얻을 수 있어 자연임신이나 보조 생식술의 성공 가능성을 높일 수 있다는 점을 제시한다.

• Clinical and Experimental Reproductive Medicine
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• 제51권1호
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• pp.42-47
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• 2024

Objective: This study evaluated the effects of temperature and storage time on the quality and DNA integrity of freeze-dried sperm from individuals with normozoospermia. Methods: Normal sperm samples from 15 men aged 24 to 40 years were studied. Each sample was divided into six groups: fresh, freezing (frozen in liquid nitrogen), freeze-dried then preserved at room temperature for 1 month (FD-1m-RT), freeze-dried then preserved at room temperature for 2 months (FD-2m-RT), freeze-dried then preserved at 4 ℃ for 1 month (FD-1m-4 ℃), and freeze-dried then preserved at 4 ℃ for 2 months (FD-2m-4 ℃). The morphology, progressive motility, vitality, and DNA integrity of the sperm were evaluated in all groups. Results: In all freeze-dried groups, sperm cells were immotile after rehydration. The freeze-dried groups also showed significantly less sperm vitality than the fresh and frozen groups. Significantly more morphological sperm abnormalities were found in the freeze-dried groups, but freeze-drying did not lead to a significantly higher DNA fragmentation index (DFI). The DFI was significantly higher in the FD-2m-RT group than in the other freeze-dried groups. Conclusion: The freeze-drying method preserved the integrity of sperm DNA. The temperature and duration of storage were also identified as factors that influenced the DFI. Accordingly, more research is needed on ways to improve sperm quality in the freeze-drying process.

• 대한의생명과학회지
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• 제28권3호
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• pp.157-169
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• 2022

DNA typing is the typical technology in the forensic science and plays a significant role in the personal identification of victims and suspects. Short tandem repeat (STR) is the short tandemly repeated DNA sequence consisting of 2~7 bp DNA units in specific loci. It is disseminated across the human genome and represents polymorphism among individuals. Because polymorphism is a key feature of the application of DNA typing STR analysis, STR analysis becomes the standard technology in forensics. Therefore, the DNA database (DNA-DB) was first introduced with 4 essential STR markers for the application of forensic science; however, the number of STR markers was expanded from 4 to 13 and 13 to 20 later to counteract the continuously increased DNA profile and other needed situations. After applying expanded STR markers to the South Korean DNA-DB system, it positively affected to low copy number analysis that had a high possibility of partial DNA profiles, and especially contributed to the theft cases due to the high portion of touch DNA evidence in the theft case. Furthermore, STR marker expansion not only contributed to the resolution of cold cases but also increased kinship index indicating the potential for improved kinship test accuracy using extended STR markers. Collectively, the expansion of the STR locus was considered to be necessary to keep pace with the continuously increasing DNA profile, and to improve the data integrity of the DNA-DB.

• Clinical and Experimental Reproductive Medicine
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• 제49권1호
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• pp.40-48
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• 2022

Objective: As the associations of sperm DNA fragmentation with morphology have not been examined in detail, this study aimed to investigate the relationship between abnormalities of morphological details and DNA integrity in human sperm. Methods: In this cross-sectional study, men from infertile couples were enrolled at Hue Center for Reproductive Endocrinology and Infertility, Vietnam. Conventional semen parameters, including morphological details, were analyzed following the World Health Organization 2010 criteria. Sperm DNA fragmentation was evaluated using a sperm chromatin dispersion assay. The relationships and correlations between semen parameters, sperm morphology, and the type of halosperm and the DNA fragmentation index (DFI) were analyzed. Results: Among 130 men in infertile couples, statistically significant differences were not found in the sperm halo type between the normal and abnormal sperm morphology groups. The percentage of round-head spermatozoa was higher in the DFI >15% group (16.98%±12.50%) than in the DFI ≤15% group (13.13% ±8.82%), higher values for amorphous heads were found in the DFI >15% group, and lower values for tapered heads were observed in the DFI ≤15% group; however, these differences were not statistically significant. Small-halo sperm and the DFI were positively correlated with round-head sperm (r=0.243, p=0.005 and r=0.197, p=0.025, respectively). Conclusion: The rate of general sperm morphological abnormalities in semen analysis was not related to sperm DNA integrity. However, round sperm heads were closely associated with sperm DNA fragmentation.

• Asian Pacific Journal of Cancer Prevention
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• 제17권6호
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• pp.2989-2993
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• 2016

The commonest cancer in Egyptian females occurs in the breast cfDNA is a non-invasive marker for tumor detetion and prognostic assessment in many types of cancer including breast cancer. This study aimed to assess the role of cfDNA and its fragmentation pattern in breast cancer prognosis and treatment response. Forty female patients with malignant breast tumors and a comparable group of healthy blood donors were enrolled prospectively. cfDNA levels and fragmentation patterns were investigated after cfDNA extraction, gel electrophoresis and gel analysis. The percentage of breast cancer patients positive for cfDNA (92.5%) was significantly higher than that of controls (55%). Also, mean concentration of cfDNA was significantly higher than in the control group (P<0.05). Most Her-2 positive patients had long cfDNA fragments, this being significant as compared to Her-2 negative patients (P<0.05). Metastasis was also positively linked to significantly higher cfDNA (P<0.05) and the mean cfDNA integrity index was significantly higher in non-responders compared to treatment responders (P<0.05). In conclusion, both qualitative and quantitative aspects of cfDNA and its different fragments in breast cancer patients could be related to prognosis, metastasis and treatment response. Long cfDNA fragments could be particularly useful for prediction purposes.

• Clinical and Experimental Reproductive Medicine
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• 제43권4호
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• pp.199-206
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• 2016

Objective: This study was carried out to investigate the correlations of the sperm DNA fragmentation index (DFI) with semen parameters and apoptosis, and to investigate the effects of density-gradient centrifugation (DGC) and magnetic-activated cell sorting (MACS) on reducing the proportion of sperm with DNA fragmentation and protamine deficiency. Methods: Semen analysis and a sperm DNA fragmentation assay were performed to assess the correlations between semen parameters and the DFI in 458 semen samples. Sperm with progressive motility or non-apoptosis were isolated by DGC or MACS, respectively, in 29 normozoospermic semen samples. The effects of DGC or MACS alone and of DGC and MACS combined on reducing the amount of sperm in the sample with DNA fragmentation and protamine deficiency were investigated. Results: The sperm DFI showed a significant correlation (r=-0.347, p< 0.001) with sperm motility and morphology (r=-0.114, p< 0.05) but not with other semen parameters. The DFI ($11.5%{\pm}2.0%$) of semen samples was significantly reduced by DGC ($8.1%{\pm}4.1%$) or MACS alone ($7.4%{\pm}3.9%$) (p< 0.05). The DFI was significantly further reduced by a combination of DGC and MACS ($4.1%{\pm}1.3%$, p< 0.05). Moreover, the combination of DGC and MACS ($1.6%{\pm}1.1%$, p< 0.05) significantly reduced the protamine deficiency rate of semen samples compared to DGC ($4.4%{\pm}3.2%$) or MACS alone ($3.4%{\pm}2.2%$). Conclusion: The combination of DGC and MACS may be an effective method to isolate high-quality sperm with progressive motility, non-apoptosis, high DNA integrity, and low protamine deficiency in clinical use.

• Journal of Plant Biotechnology
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• 제5권4호
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• pp.201-207
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• 2003

Tissue culture techniques arguably are an important approach for ex situ conservation of rare and endangered plant species. However, there is utmost importance on maintaining the genetic integrity of the introduced plants especially in tree species. To examine the genetic integrity of the micropropagated plants, we randomly screened few hardened plants of Syzygium travancoricum, a critically endangered tree taxon, using Randomly Amplified Polymorphic DNA (RAPD) markers. Twenty-three random. primers were tried and twenty-five polymorphic loci were identified. The dendrogram based on the Unweighted Pair-Group Method Arithmetic Average and Nei's similarity index depicted about 97% homology between the mother plants and micropropagated plants. Further, an attempt was made to reintroduce the micropropagated plants in the wild. Over three hundred small trees could be successfully established.

• Clinical and Experimental Reproductive Medicine
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• 제38권1호
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• pp.10-17
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• 2011

Objective: To explore potential relationships between sperm DNA integrity and both semen parameters and clinical outcomes. Methods: Semen analysis of 498 samples was performed according to the 2010 criteria of the World Health Organization. The sperm DNA fragmentation Index (DFI) of the semen samples was assessed using a neutral comet assay. Results: Sperm DFI showed a significant correlation with semen parameters, including the patient's age, sperm viability, motility, morphology, and number of leukocytes (p<0.05). The sperm DFI values for asthenozoospermic (15.2%), oligoteratozoospermic (18.3%), asthenoteratozoospermic (17.5%), and oligoasthenoteratozoospermic semen samples (21.3%) were significantly higher than that observed in normozoospermic semen samples (10.5%, p<0.05). A sperm DFI value of 14% was used as a threshold of sperm DFI in assessing whether DNA was highly damaged. In 114 IVF-ET cycles, the fertilization rate of the sperm DFI <14% group (70 cycles, 61.7%) was significantly higher than that observed for the ${\geq}14%$ group (44 cycles, 55.3%), but there was no difference in the other clinical outcomes between the two groups. In the ${\geq}14%$ group, the pregnancy rates of the ICSI cycles (40.0%) and half-ICSI (44.0%) were higher than conventional IVF cycles (30.7%), but the difference was not statistically significant. Conclusion: Along with the conventional semen analysis, the sperm DFI assessed using the comet assay was shown to improve the quality of the semen evaluation. To evaluate the precise effect of ICSI on pregnancy rates in the patients who demonstrate high sperm DFI values, further study is necessary.

• 생태와환경
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• 제43권1호
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• pp.24-34
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• 2010

This study was carried out to analyze some influences on ecological health conditions, threaten by various stressors such as physical, chemical and biological parameters. We collected samples in 2008 from three zones of upstream, midstream and downstream, Gap Stream. We applied multi-metric fish assessment index (MFAI), based on biotic integrity model to the three zones along with habitat evaluations based on Qualitative Habitat Evaluation Index (QHEI). We also examined fish fauna and compositions, and analyzed relations with MFAI values, QHEI values, and various guild types. Chemical parameters such as oragnic matter (BOD, COD), nutrients (TP, $NH_3$-N), coli-form number (as MPN), and suspended solids (SS) were analyzed to identify the relationship among multiple stressor effects. Using the sentinel species of Zacco platypus, the population structures and condition factors were analyzed along with DNA damages related with genotoxicant effects by comet assay. This study using all these parameters showed that stream condition was degraded along the longitudinal gradient from upstream to downstream, and the downstream, especially, was impacted by nutrient enrichment and toxicant exposure from the point source, wastewater treatment plant. Overall results indicated that our approaches applying various parameters may be used as a cause-effect technique in the stream health assessments and also used as a pre-warning tool for diagnosis of ecological degradation.