• Journal of Life Science
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• v.29 no.8
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• pp.895-903
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• 2019

The aim of the present study was to improve the cell viability of human dental papilla derived single-induced pluripotent stem cells (iPSCs) using a Rho-associated protein kinase (ROCK) inhibitor, Y-27632. The iPSCs were produced using an episomal plasmid-based reprogramming method. After cell separation using trypsin, the iPSCs were treated with 0, 0.5, 1, 2.5, 5, 7.5, or $10{\mu}M$ Y-27632 for 5 d. Cell viability increased significantly following the $5{\mu}M$ Y-27632 treatment (p<0.05). When the iPSCs were exposed to medium containing $10{\mu}M$ Y-27632 for 0, 1, 2, 3, 4, and 5 d, the cell viability rate increased significantly in accordance with the cell viability rate (p<0.05). To evaluate the effect of the Y-27632 treatment on stemness characteristics, the expression of stem cell-specific transcripts and telomerase activity were investigated in the iPSCs treated with $10{\mu}M$ Y-27632 for 5 d. The expression levels of stem cell-specific transcripts, such as OCT-4, NONOG, and SOX-2, and telomerase activity were not significantly different in the iPSCs treated with $10{\mu}M$ Y-27632 as compared with those of untreated control iPSCs (p>0.05). Taken together, the results demonstrated that cell viability can be improved by treatment with the ROCK inhibitor Y-27632, without losing iPSC stemness characteristics.

• Journal of Life Science
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• v.31 no.3
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• pp.266-279
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• 2021

The present study examined the cytotoxic effects of a Smilax china L. extract (SCLE) in human cancer (A-549, MCF-7, MDA-MB-231, U87-MG, AGS, MKN-74, and SNU-601) and normal MRC-5 fibroblasts, as well as in mesenchymal stem cells derived from dental tissue (DSC). The 50% inhibitory concentration (IC50) values for SCLE were significantly (p<0.05) lower in the cancer cell lines (A-549, MCF-7, MDA-MB-231, U87-MG, AGS, MKN-74 and SNU-601) than in the MRC-5 and DSC cells. Cell growth was significantly (p<0.05) more inhibited in the cancer cell lines treated with 200 ㎍/ml SCLE than in the normal MRC-5 and DSC, and anoikis-like floating cell morphology was observed in the SCLE-treated cancer cells. The cells detached by SCLE treatment were retrieved daily and assayed for viability and telomerase activity. Cells retrieved at 4 days showed significantly decreased viability and telomerase activity (p<0.05), as well as apoptosis-like abnormal morphology, when compared to cells retrieved in the previous 3 days. The ratio of apoptosis and cells in the G1 phase was significantly (p<0.05) increased in the A-549, AGS, and MCF-7 cancer cells treated with SCLE for 4 days compared to untreated controls. However, after SCLE treatment, cell adhesion was not increased by application of an inhibitor of the associated protein kinase (ROCK) that mainly contributes to the increase in cell attachment. This suggests that the cellular detachment by SCLE is probably controlled by a Rho-independent mechanism(s). These observations indicate that SCLE readily induces anoikis in cancer cells and could serve as a potent agent for cancer chemotherapy.

• Asian Pacific Journal of Cancer Prevention
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• v.16 no.1
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• pp.77-81
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• 2015

Background: The aim of the study was to determine whether the expression of baseline phosphorus (P) and magnesium (Mg) levels were prognostic in terms of stage and overall survival (OS) in newly diagnosed non-small cell lung cancer (NSCLC) and small cell lung cancer (SCLC) patients. Materials and Methods: Retrospectively, 130 patients were selected at the time of diagnosis oflung cancer (100 with NSCLC and 30 with SCLC), before the initialization of any chemo-radiotherapy. The median age was 67 (range 29-92). IA, IB, IIA, IIB, IIIA, IIIB and IV stages were present in 3, 4, 19, 6, 25, 8, and 65 patients, respectively. After centrifugation, the levels of serum P and Mg were measured using the nephelometric method/ photometry and evaluated before any type of treatment. Results: Higher than normal levels of P were found in 127/130 patients, while only four patients had elevated Mg serum values. In terms of Spearman test, higher P serum values correlated with either stage (rho=- 0.334, p<0.001) or OS (rho=-0.212, p=0.016). Additionally, a significant negative correlation of Mg serum levels was found with stage of disease (rho=-0.135, P=0.042). On multivariate cox-regression survival analysis, only stage (p<0.01), performance status (p<0.01) and P serum (p=0.045) showed a significant prognostic value. Conclusions: Our study indicated that pre-treatment P serum levels in lung cancer patients are higher than the normal range. Moreover, P and Mg serum levels are predictive of stage of disease. Along with stage and performance status, the P serum levels had also a significant impact on survival. This information may be important for stratifying patients to specific treatment protocols or intensifying their therapies. However, larger series are now needed to confirm our results.

• Reproductive and Developmental Biology
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• v.35 no.2
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• pp.175-183
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• 2011

The present study compared the developmental potential, telomerase activity and transcript levels of X-linked genes (ANT3, HPRT, MeCP2, RPS4X, XIAP, XIST and ZFX) in the bovine somatic cell nuclear transfer (SCNT) embryos derived from different age and cell cycle of female donor nucleus. In experiment 1, the fusion rate, cleavage rate to 2-cell stage, developmental rate to blastocyst stage, and the mean number of total and ICM cells was slightly increased in embryos cloned with fetal fibroblasts compared to those with adult fibroblasts, but there was no significantly (p<0.05) differences. Telomerase activity was also similar in blastocysts cloned with fetal and adult fibroblasts. Up-regulated RPS4X and down-regulated MeCP2, XIAP, and XIST transcript level were observed in blastocysts cloned with adult fibroblasts, compared to those with fetal fibroblasts. In experiment 2, the fusion rate, cleavage rate to 2-cell stage, developmental rate to blastocyst stage, and the mean number of total and ICM cells was significantly (p<0.05) increased in embryos cloned with fetal fibroblasts at early G1 phase of the cell cycle, compared to those of fetal fibroblasts at late G1 phase. DNMT1 transcript was observed to significantly (p<0.05) increased in the fetal fibroblasts at 3 hrs after trypsin treatment of confluent culture. Further, level of telomerase activity and transcribed X-linked genes was also significantly (p<0.05) higher in the early G1 SCNT blastocysts than those of late G1. The results imply that fetal fibroblasts at early G1 phase induces the enhanced developmental potential and up-regulated telomerase activity and X-linked gene, but aberrant transcript pattern of X-linked genes may be displayed in the SCNT embryos.

• Korean Journal of Pharmacognosy
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• v.40 no.1
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• pp.6-12
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• 2009

Thirty five methanol extracts from 19 natural local plants, which have been used as traditional anti-cancer medicine, were prepared. They were analyzed the cytotoxic effects on primary fibroblast cells and carcinoma cells. The root extract of Solanum nigrum were highly toxic in both cell lines with $IC_{50}$ values of less than $0.01{\mu}g/{\mu}l$, and 26 of 35 extracts were toxic in all cells with $IC_{50}$ values of $0.1{\sim}2{\mu}g/{\mu}l$. Three extracts including the fruit extracts of Solanum nigrum and Morus alba had no cytotoxic activity in both cell lines. Five of 35 extracts were highly toxic in cancer cells than in primary cells. Because primary cells were more resistant on these extracts, the five extracts were selected for anti-cancer agent candidates. Apoptosis or programmed cell death has an essential role in chemotherapy-induced tumor cell killing. Recently, inducers of apoptosis have been used in cancer therapy. When two of 5 cancer cell-specific cytotoxic extracts (Ulmus parvifolia and Zelkova serrata) were treated in concentration of $0.02{\sim}0.1{\mu}g/{\mu}l$, apoptosis were increased at 3-5 times in cancer cell lines. Finally, the apoptotic effects of these extracts were confirmed by cleavages of both poly-(ADP-ribose)-polymerase and caspase-3 as apoptotic markers. In this report, we suggested that two of 35 medicinal herb extracts can be useful anti-cancer drug candidates inducing apoptosis in several carcinoma cell lines.

• Korean Journal of Clinical Laboratory Science
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• v.45 no.4
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• pp.149-153
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• 2013

Candidiasis is a fungal infection of the most common causes; generally, opportunistic infections occur often in patients with weakened immune systems. Because of high rates in fungal infection patients and increasing frequency of being isolated from clinical materials, quickly identifying of Candida albicans is critical. By identifying 404 yeast cell strains of referred samples via API 20C kits, NGL and PRO tests and Germ tube (GT) test were conducted and compared. In the 3.0 McFarland yeast cells, 0.1% ${\rho}-nitrophenyl-N-acetyl-{\beta}-D-galactosaminide$ (NGL) and 0.04% ${\small{L}}$-proline ${\beta}$-naphtylamide (PRO) were each put in test tubes and incubated at $35^{\circ}C$ for 15, 30, 60 and 90 minutes. Afterwards, 1 drop of 2% NaOH was applied, and if the color turned yellow; it was positive for NGL test. Afterwards, 1% ${\rho}$-dimethylaminocinnamaldehyde was applied, and if the upper layer turned pink or red, it was positive for PRO test. NGL and PRO tests were conducted for all C. albicans and identified accurately within 30 minutes. In NGL, PRO test, false-positive, negative were not seen, whereas, GT test showed false-positive in 1 strain and false-negative in 3 strains. Therefore, sensitivity and specificity of NGL, PRO tests were 100% and 99.5%, respectively, and positive and negative predictive rate were 99.5% and 100%, respectively. However, GT test sensitivity and specificity were 98.5% and 99.5%, respectively, and positive and negative predictive rates were 99.5% and 98.5%, respectively. In conclusion, NGL, PRO tests are better than GT tests for sensitivity and specificity, therefore, these reliable tests will be useful in clinical laboratories.

• Fisheries and Aquatic Sciences
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• v.9 no.1
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• pp.30-37
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• 2006

We performed experiments on the uptake and excretion of dissolved organic phosphorus (DOP) using two toxic dinoflagellates, Alexandrium tamarense Lebour (Balech) and Gymnodinium catenatum Graham, isolated from Hiroshima Bay, Japan. ATP (adenosine triphosphate), UMP (uridine-5-monophosphate), G-6-P (glucose-6-phosphate) and Glycero-P (glycerophosphate) were used as DOP sources in preliminary uptake experiments. ATP was selected as the DOP species for the short-tenn uptake experiment because preliminary experiments showed it to be the DOP source used by both species. Although the $K_s$ values of A. tamarense and G. catenatum (5.63 and $7.61{\mu}M$, respectively) obtained from the short-term experiments for ATP were only slightly higher than those reported for dissolved inorganic phosphorus (DIP), the ${\rho}_{max}$ values (5.04 pmol/cell/h and 13.4 pmol/cell/h, respectively) were much higher. The DOP excretion rate in batch-culture experiments was estimated at 0.084 pmol/cell/h for A. tamarense and 0.012 pmol/cell/h for G. catenatum, accounting for about 30% and 25%, respectively, of the assimilated phosphorus. Our results suggest that the DIP-depleted conditions of Hiroshima Bay favor these two species by supporting their ability to use DOP.

• Proceedings of the Korean Society of Embryo Transfer Conference
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• 2003.10a
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• pp.140-140
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• 2003

• Journal of the Korean Crystal Growth and Crystal Technology
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• v.28 no.1
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• pp.1-8
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• 2018

The 23 wt% polyvinylidene fluoride (PVDF)/15 wt% polyurethane (PU) fibers were electrospun using the conjugated nozzle at a flow rate of 1.0 mL/h and an electric field of 1 kV/cm. The formation of ${\beta}$ crystal phase in the PVDF and the PVDF/PU fibers was confirmed by Fourier transform infrared spectroscopy. After electrospinning, the asspun fibers were immersed in a boiling water and then dried at $100^{\circ}C$ in a convection oven to make a crimp phenomenon. The crimps with a diameter of $2.0{\pm}0.08{\mu}m$ were observed for the PVDF/PU fibers after hydrothermal treatment without sacrificing the extent of ${\beta}$ crystal phase. All the PU, PVDF and PVDF/PU fibers exhibited average cell viability of more than 98 %. The cell proliferation results suggested that L-929 cells adhered well to the PU, PVDF and PVDF/PU fibers and proliferated continuously with increasing time, indicating that the PVDF/PU fibers are highly applicable to the biomedical applications.

• Reproductive and Developmental Biology
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• v.35 no.1
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• pp.105-113
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• 2011

The present study investigated the nuclear remodeling, development potential with telomerase activity and transcription level of X-linked genes (ANT3, HPRT, MeCP2, RPS4X, XIAP, XIST and ZFX) in the bovine somatic cell nuclear transfer (SCNT) embryos using two different fusion and activation methods. Female adult fibroblasts were injected into perivitelline space of in vitro matured oocytes. The oocyte-nucleus complexes were fused and followed by immediately either activated (Group 1), or activated at 1 h post-fusion (hpf) (Group 2), respectively. The incidence of normal premature chromosome condensation (PCC) at 1 hpf was slightly increased in the Group 2, compared to those of Group 1, but there was no significant (p<0.05) difference. The incidence of normal pronucleus (PN) and chromosome spread at 5 and 18 hpf were significantly (p<0.05) higher in the Group 2 than those of Group 1. The cleavage rate to 2-cell stage, developmental rate to blastocyst stage, and the mean number of total and ICM cell numbers were significantly (p<0.05) higher in the Group 2, compared to those of Group 1. Level of telomerase activity was significantly (p<0.05) higher in the SCNT blastocysts of Group 2, compared to those of Group 1. Transcript levels of HPRT, MeCP2 and XIST were not significantly (p<0.05) different between blastocysts of Group 1 and 2. However, transcript level of ANT3, RPS4X, XIAP and ZFX were significantly (p<0.05) up-regulated in the SCNT blastocysts of Group 2, compared to those of Group 1. Taken together, it is concluded that oocyte activation at 1 hpf induces the enhanced developmental potential by efficient nuclear remodeling and subsequent facilitation of the nuclear reprogramming of bovine SCNT embryos.