• Korean Journal of Environmental Biology
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• v.38 no.2
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• pp.248-253
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• 2020

The level at which analyses of DNA content might contribute more significantly to the genetic mechanisms of evolution lies in the events of speciation. The object of this study was to investigate the DNA content of abalone (Haliotis discus hannai) and determine the optimal tissue samples for measuring the DNA content of abalone by flowcytometry without fixation. The DNA content (pg/nucleus) of gill tissue (2.5±0.08), which was contaminated with protozoa, was significantly lower than that of muscle tissue (3.2±0.02), mantle tissue (3.2±0.02) (p<0.05), and a standard reference standard, while the DNA contents of muscle tissue and mantle tissue were higher than that of the standard reference. Considering the results of this study, DNA content analysis with flowcytometry is an acute and rapid method by which muscle tissue and mantle tissue are the most appropriate sample for measuring the DNA content of abalone without fixation.

• Proceedings of the Korean Nuclear Society Conference
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• 1998.10a
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• pp.344-344
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• 1998

• Proceedings of the Korean Society of Life Science Conference
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• 2007.10a
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• pp.40.2-40.2
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• 2007

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• Reproductive and Developmental Biology
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• v.32 no.2
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• pp.123-126
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• 2008

The objective of this study was to determine the potential hazardous effects of sorting process by flowcytometry on the quality of boar spermatozoa by flowcytometer. Freshly collected boar semen was diluted and divided into two groups; control none sorted and sorted. Sperms in sorted group were processed with flowcytometer for cell sorting with $100\;{\mu}M$ nozzle under the 20 psi pressure. Measurements on each parameter were made at two time points, 0hr (right after sorting) and 24 hr post sorting. Although there was a tendency of lower viability in sorted group than none sorted control group, the percentage of live cells in control ($75.83{\pm}6.92\;&\;59.53{\pm}10.34$) was not significantly different from sorted ($59.70{\pm}7.37\;&\;43.97{\pm}3.76$) at both 0 and 24 hr post sorting. However, sorted sperm showed significantly lower mitochondrial function compared to the control at both 0 h ($79.37{\pm}3.22\;vs.\;63.50{\pm}10.05$) and 24 hr ($67.27{\pm}3.22$ vs. $46.97{\pm}5.37$) time points (p<0.007). Sperm DNA fragmentation rate was significantly lower in control ($22.0{\pm}7.04$) than that of sorted ($32.27{\pm}7.49$) at 24 hr time point (p<0.0002). Taken together, these data suggested thatsorting process by flowcytometer may have influenced sperm motility rather than viability. Also high speed sperm sorting by flowcytometer has significant effects on DNA fragmentation on elapsed time after sorting.

• Journal of Korean Biological Nursing Science
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• v.8 no.2
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• pp.5-12
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• 2006

목적 : 본 연구는 암환자 및 암의 유형에 따라 중요한 종양억제 조절인자로 알려져 있는 모노카인을 flowcytometry를 이용하여 분석, 비교하고자 하였다. 방법 : 연구대상은 고형종양(solid tumor)으로 진단받은 33세에서 76세 사이의 암환자 30명(유방암, 난소암, 폐암, 위암)을 대상으로 말초혈액 단구의 intracellular monokine 중 $TNF{\alpha}$, MIG, MIP를 분석한 유사실험설계 연구이다. 연구결과 : 암환자 군에서의 $TNF{\alpha}$, MIG, MIP 수치는 대조 군인 정상 군에 비해서 유의하게 증가되었으며 특히, 유방암과 난소암 환자 군에서의 $TNF{\alpha}$ 수치는 폐암과 위암의 대상자에 비해 의미 있는 차이를 보여주었다. 논의 : 본 연구에서 제시된 암환자 군에서의 모노카인 수치는 선행연구의 결과와 통일하게 종양 대상자의 면역에 중요한 역할을 하는 것으로 규명 되었으나, $TNF{\alpha}$는 고형종양 중에서도 여성생식기계 암환자 군에서 더 증가하였다. 이에 따라 종양 유형에 따른 모노카인의 역할과 호르몬과의 상호작용기전 규명에 대한 추후 연구가 필요하다.

• Korean Journal of Ichthyology
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• v.24 no.3
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• pp.151-159
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• 2012

Cross and reciprocal hybrids from intrageneric crosses between Rhodeus uyekii and R. notatus were analyzed morphometrically using truss and classical dimensions, erythrocyte size using blood smears, and cytogenetically using flowcytometry, then compared with the parental species. Although the morphometric assessment based on body proportions indicated that the hybrids were generally intermediate between the maternal and paternal species, the morphometric characters of cross and reciprocal hybrids included at least 14 paternal-like and 7 maternal-like, and 13 paternal-like and 5 maternal-like characters, respectively (P<0.05). The pigmentation of the hybrids was intermediate in some respects, and resembled that of the parental species. The DNA content of the cross and reciprocal hybrids, based on flow cytometry analysis, was significantly different from the parental species. The characterization of morphometric traits and the cytogenetic analysis of hybrids of R. uyekii and R. notatus used in this study may be useful for distinguishing genotypes in commercial Acheilognathinae aquaculture.

• The Korean Journal of Malacology
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• v.29 no.1
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• pp.15-21
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• 2013

Nitric oxide (NO) is an important intra-intercellular signaling molecule that regulates many physiological processes and participates in the development some pathological conditions in animals. In this study, we compared different methods for determining NO concentration in the hemocytes of Manila clam Ruditapes philippinarum. For measuring the intracellular NO levels, we used the specific fluorescent probe 4,5-diaminofluorescein diacetate (DAF-2 DA), and the quantification methods that were compared were based on image analysis, spectrophotometry, and flow cytometry. NO concentration could be determined using all the 3 methods, and the concentration varied significantly depending upon the presence of NO regulators in the hemocytes; NO concentration increased in the presence of L-arginine, while it decreased in the presence of N-nitro-L-arginine methyl ester. In particular, it is found that estimation of NO using a flowcytometry is more economical, reliable and accurate compared to image analysis and spectrophotometry. Accordingly we believe that determining NO concentration by using flowcytometry will be useful in evaluating physiological and pathological conditions in marine bivalves.

• Journal of Acupuncture Research
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• v.23 no.5
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• pp.69-77
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• 2006

Background : Mesenchymal stem cells (MSCs) are present in most of the tissue matrix, taking part in their regeneration when injury or damage occurs. The aim of this study was to investigate the presence of cells with pluripotential characteristics in human subchondral bone and the capacity of these cells to differentiate to osteoblast. Methods : Human subchondral bone were digested with collagenase. Isolated cells were cultured with a-MEM, 15% FBS, 10-8M dexamethasone and 50 ng/mL ascoric acid. Cells from 0 day(isolated cells), 7 day (first subculture) and 14 days (third subculture) were used to carry out phenotypic characterization experiments flowcytometry analysis with 11 monoclonal antibodies) and osteogenic differentiation experiments. Osteogenic differentiation of cells was assessment by quantification of bone extracellular matrix components by following analysis: alkaline phosphatase(ALP) stains to detect ALP activity, RT-PCR and western blot to detect osteocalcin (OCN), osteopontin (OPN) and type I collagen(Col I), and Alizarin red stains to detect calcium deposition. Results : Flowcytometry analyses showed that in our population more than 98% of cells were positive for MSC markers: SH-2(CD105, 99%), CD29 (95%), CD73 (95%). Cells were negative for hematopoietic markers (CD11b, CD34, and CD45). Furthermore, cells showed positive stain to multipotent markers such as CDl17 (c-kit) (15.1%), and CD166 (74.9%), and cell adhesion molecules such as CD54 (78.1%) and CD106 (63.5%). The osteogenic specific marker analyses showed that the culture of these cells for 7 and 14 days stimulates ALP, OCN, OPN and Col I synthesis by RT-PCR and Western blot analysis. Also, after 14 days in the culture of MSCs induces mineralization by Arizarin red stain. Conclusion : In this work, we demonstrated a new and efficient method for osteoblastic differentiation of human subchondral bone stem cells. As MSCs takes part in reparative processes of adult tissues, these cells could play an important role in osteogenesis.

• IMMUNE NETWORK
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• v.1 no.1
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• pp.26-31
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• 2001

Background : Follicular dendritic cells (FDCs) play key roles during T cell-dependent humoral immune responses by allowing antigen-specific B cells to survive, proliferate, and differentiate within the FDC networks of secondary follicles, i.e., germinal centers (GC). Methods: A novel monoclonal antibody, 3C8, was generated by immunizing with an FDC line HK, in order to understand the molecular signals involved in the FDC-B cell interactions in the microenvironment of the GC. Results: The 3C8 antibody did not bind to mononuclear cells, including T cells, B cells, and monocytes. Murine L929 and human skin fibroblasts exhibited no or little reactivity to 3C8. However, 3C8 specifically recognized HK cells by flowcytometry. Furthermore, the antigen recognized by 3C8 was restricted to the GC of the human tonsil. Dendritic networks of the GC were intensely stained by 3C8, but cells outside the GC were not. Conclusion: Our results suggest that the antigen 3C8 may play some unique role on FDCs during the GC reactions.

• The Journal of Internal Korean Medicine
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• v.25 no.2
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• pp.214-223
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• 2004

The study examines the anti-cancer effects of the hot water extract of Mylabris phalerata(MP) using SNU-C5 cell lines. Microscopic analysis showed that 12 hours after MP treatment, the number of dead cells increased prominently. Significant cell death was observed 12, 24, and 48 hours after MP treatment through trypan blue exclusion testing. This suggests that MP is time-dependently cytotoxic. Mitotracker Red CMXRos staining and flowcytometry revealed that MP decreased mitochondrial membrane potentials. The absence of peaks on PI staining showed that DNA damage occurred in MP treated cells. Taken together, measurements suggest that MP has a strong anti-cancer effect on SNU-5 cell lines, and that this is likely to be due to the destruction of mitochondria and DNA damage.