포도 ‘My heart’의 기내증식과 기외이식 후 생장에 있어서 배양용기에 부착한 미세공극 Filter 처리가 건전한 유묘를 생 산하는데 효과적인 방법을 찾고자 실시하였다. 미세공극 Filter는 환기구 크기별로 White filter type (50.0 mm×3.5 mm)과 Green filter type (50.0 mm×7.0 mm) 으로 구분하여 밀폐 처리와 비교하였다. Shoot tip 배양에 있어서 Green filter type에서 shoot 분화율이 75%로 White filter type 77% 와 밀폐처리 80% 보다 낮았지만 투명화 shoot 발생율은 4% 로 White filter type 13.4%, 밀폐처리 24.7%에 비하여 9.4－ 20.7%가 적었다. 전체 배양묘의 54.9%가 투명화 발생이 되 었을 때 Green filter type으로 90일 동안 계대 배양하고 조사 한 결과 투명화율은 11.8%로 감소하였고 분화한 shoot 수는 89개에서 915개로 증가하였다. Filter type에 따라 IBA 2.0 mg·L-1를 첨가하여 30일 동안 배양을 하였을 때 Green filter type에서 기내 발근율 100%, 뿌리 수 7.3개, 엽수 10.0개로 White filter type과 밀폐 처리보다 좋았다. 기외 이식하고 15 일 후의 유묘 생존율도 Green filter type에서 100%로 다른 처 리에 비하여 1.5－29.5% 더 높았고 초장이 11.0cm, 생체중이 1.7g 으로 가장 양호하였다. 미세공극 Green filter 처리는 포 도 ‘My heart’의 기내배양에서 shoot 투명화를 감소시키고 shoot와 뿌리 생성을 촉진시키고 기외이식 후 생장은 통계적 으로 유의하게 확인되어 건전한 유묘 생산에 효과적이었다.

Background: mTeSR1 is a fully-defined, serum-free medium for the derivation and maintenance of Human embryonic stem cells (ESCs). This study investigates the impact of incorporating mTeSR1 supplement during in vitro culture (IVC) on blastocyst productivity, qualitative characteristics, and outgrowth potential of bovine blastocysts. Methods: In vitro fertilized (IVF) eggs were cultured in IVC medium (control) with the addition of mTeSR1 supplement at concentrations of 1%, 2%, and 5%, respectively. The development rates of fertilized eggs and gene expression patterns of blastocysts were assessed on day 9 of culture. For outgrowth culture, blastocysts were cultured on a mouse embryonic fibroblast feeder cells (MEFs) for 7 days. Results: In vitro development of bovine preimplantation embryos in the 2% mTeSR1 group was significantly higher than in the control (p < 0.05). The apoptotic index in the 2% mTeSR1 group was significantly lower compared to the control (p < 0.05). RTqPCR indicated that SRY-Box Transcription Factor 2 (Sox2) gene expression in the 5% mTeSR1 group was significantly higher than in the control (p < 0.05). The 5% mTeSR1 group also showed significantly higher BCL2 associated X (Bax) expression compared to the control and other mTeSR1 groups. On day 9 pi, blastocysts from the control and 2% mTeSR1 groups were cultured for 7 days. The 2% mTeSR1 group showed higher efficiency in forming dome-shaped colonies with stronger SOX2 expression compared to the control. Conclusions: The mTeSR1 supplement supports preimplantation embryo development and prevents apoptosis in blastocysts, leading to the efficient formation of domeshaped inner cell mass (ICM) colonies.

이 연구의 목적은 기금조성용 옥외광고 산업의 현재를 분석하고, 산업 의 발전을 저해하는 요인으로 평가받는 불법 도로변 야립광고에 대한 해 법을 제시하고자 하는 것이다. 이를 위하여 전문가를 대상으로 하는 심 층 인터뷰를 실시하여 그 내용을 분석하였다. 이를 통해 본 연구는 기금 조성용 옥외광고가 옥외광고 산업의 발전에 중요할 역할을 했음을 확인 하였으나, 시스템과 비즈니스 차원에서 해결해야 할 여러 문제점이 있음 을 발견하였다. 또 불법 도로변 야립광고의 난립을 방지하기 위해 법적, 기술적, 경제적, 사회적 해법에 대한 전문가들의 다양한 의견을 수집함으 로써 관련 산업 정책에 대한 의미있는 기초자료를 제공한다는데 의의가 있다.

In this review, the regulatory mechanisms of autophagy were described, and its interaction with apoptosis was identified. The role of autophagy in embryogenesis, tooth development, and cell differentiation were also investigated. Autophagy is regulated by various autophagy-related genes and those related to stress response. Highly active autophagy occurrences have been reported during cell differentiation before implantation after fertilization. Autophagy is involved in energy generation and supplies nutrients during early birth, essential to compensate for their deficient supply from the placenta. The contribution of autophagy during tooth development, such as the shape of the crown and root formation, ivory, and homeostasis in cells, was also observed. Genes control autophagy, and studying the role of autophagy in cell differentiation and development was useful for understanding human aging, illness, and health. In the future, the role of specific mechanisms in the development and differentiation of autophagy may increase the understanding of the pathological mechanisms of disease and development processes and is expected to reduce the treatment of various diseases by modulating the autophagic phenomenon.

새만금 내에서는 종종 식물플랑크톤이 증식하기에 알맞은 환경조건이 생성되며 일시에 식물플랑크톤 대증식이 발생하면서 조 류 관리기준을 초과하는 사례가 발생하고 있다. 이를 대비하기 위하여 과학적 예측기법을 토대로, 식물플랑크톤의 종별로 가장 효과적이 고 효율적인 녹조발생 억제 방안을 제안하기 위하여 식물플랑크톤 대증식 가능성을 예측하고, 제어할 수 있는 모델을 개발하였다. 즉, 하 천에서 유입하는 영양염(DIN, PO4-P)을 정책적으로 조절하고, 갑문운영을 통해 호 내 염분을 제어하는 것이다. 먼저 관측치로부터 인공신 경망 알고리즘을 이용해 식물플랑크톤 대증식 가능성을 예측 결과, 모델의 Kappa 수는 0.7889 ~ 1.0000의 범위로, good ~ excellent 수준이었 다. 다음으로 Garson 알고리즘을 이용하여 종별로 설명변수의 중요도를 평가하였고, 또한 DIN 및 염분 값의 변화에 따른 식물플랑크톤 대 량 증식 확률을 예측하였다. 그 결과, 각 종별로 식물플랑크톤의 대증식을 억제할 수 있는 DIN과 염분 농도를 정량적으로 예측할 수 있었 다. 따라서, 향후 새만금과 같은 거대한 인공 호수에서 식물플랑크톤의 대증식을 억제하기 위한 효율적이고 효과적인 대응방안을 마련할 수 있도록 녹조제어모델을 활용할 수 있을 것으로 판단된다.

Oral squamous cell carcinoma (OSCC) is the most common type of head and neck cancer and is associated with high recurrence, poor treatment, and low survival rates. Hypoxia-inducible factor-1α (HIF-1α) is a transcription factor that regulates the response to hypoxia, a major factor in the tumor microenvironment that affects tumor development and progression in various cancer types. However, microRNA (miRNA) sequence analysis revealed that only a few miRNAs targeting HIF-1α had been discovered. In the present study, we investigated HIF-1α expression in OSCC and the effect of HIF-1α-targeting miRNAs on the progression and metastatic potential of OSCC. We analyzed public databases to explore which miRNAs target HIF-1α expression. In addition, the expression of proteins involved in the cell cycle, proliferation, and apoptosis in HSC-2 cells was analyzed after miRNA-126 mimic treatment. Furthermore, to investigate the effect of miRNA-126 on the proliferation and invasion ability of OSCC cells, 5-ethynyl-2′-deoxyuridine and Transwell assays were performed. The activities of MMP-2 and MMP-9 were evaluated via gelatin zymography. Our results showed that miRNA-126, which targets HIF-1α, enhances OSCC cell proliferation by regulating the cell cycle and reinforces the cell mobility of OSCC via HIF-1α expression. These findings suggest that miRNA-126 may be a novel marker for OSCC treatment and the development of new tools for patients with OSCC.

Alpha-linolenic acid is an important polyunsaturated fatty acid that exhibits anticancer, anti-inflammatory, and antioxidative effects. In this study, we investigated the protective effects of alpha-linolenic acid on the cell proliferation and differentiation of C2C12 cells under essential amino acid-deficient conditions. Different concentrations of alpha-linolenic acid and essential amino acids were added to the growth and differentiation media. The concentrations of 10 μM of alphalinolenic acid and 2% essential amino acid were chosen for subsequent experiments. Supplementation with alpha-linolenic acid and essential amino acids improved the proliferation and differentiation of C2C12 cells and significantly increased the mRNA levels of catalase, superoxide dismutase, B-cell lymphoma-2, and beclin-1 as well as the protein levels of PPARγ coactivator-1α compared to those in the controls. Moreover, supplementation with alpha-linolenic acid and essential amino acids reduced the levels of phosphorylated H2A.X variant histone, Bcl-2-associated X, p53, and light chain 3 during C2C12 cell proliferation, and increased the expression levels of myogenic factors 4 (myogenin) and 5 during C2C12 cell differentiation. Overall, we determined that alpha-linolenic acid and essential amino acids maintained the cell proliferation and differentiation of C2C12 cells via their anti-oxidative, anti-apoptotic, and anti-autophagic effects.

Salivary gland adenocarcinoma(AdCa NOS) is one of the major causes of mortality among malignant salivary gland tumors. New therapeutic measure are needed to improve the outcome for patients with AdCa NOS because current therapy does not significantly improve survival rates. Transglutaminase 2(TGase 2) was implicated in forming cross-linked protein polymer, apoptosis and matrix interaction. And also TGase 2 expression is up-regulated in proliferation, migration, invasion, and metastasis of cancer cells. shRNA which has emerged as an effective method to target specific genes for silencing has provided new opportunities for cancer therapy. But there has been rarely reported using shRNA-TGase 2 transfection in AdCa NOS. The purpose of this study were to examine the specific inhibition of TGase 2 mRNA and protein expression by siRNA transfection of TGase 2 through RT-PCR and immunoslot blotting, and to study proliferation, migration and invasion assay of SGT cell line from AdCa NOS. Cell cycle analysis showed that the downregulation of shRNA-TGase 2 caused the accumulation of cells in the sub-G0/G1 phase. In migration assay, suppressing shRNA-TGase 2 inhibited the capacity of the cells to migrate compared to parental cells. In invasion assay, cells transfected with shRNA-TGase 2 decreased in invasion when compared to SGT and vector transfected cells. shRNA-TGase 2 expressing plasmids efficiently downregulated TGase 2 mRNA and TGase 2 protein expression. It suggested that the shRNA-TGase 2 targeting system against TGase 2 could have a therapeutic potentiality for malignant salivary gland tumors, especially in inhibiting and/or preventing cancer cell proliferation, migration and invasion.

Vinpocetine induces anti-inflammatory effects in various inflammatory diseases via the inhibition of phosphodiesterase type-1-independent nuclear factor-κB signaling pathway and the release of inflammatory cytokines. In this study, we investigated the effect of vinpocetine on the proliferation of colon cancer cells and its underlying molecular mechanisms. Our data showed that vinpocetine inhibits the viability and proliferation of colon cancer cells. Vinpocetine treatment induced cell death in HCT116 cells, which the percentages of sub-G1 phase were significantly increased, and the apoptosis-related genes were regulated after HCT116 cells were treated with vinpocetine. In sum, our findings indicated that vinpocetine could be a therapeutically useful candidate in the treatment of colon cancer.

This study aimed to examine the effect of a mild elevation in serum cholesterol level in a porcine coronary overstretch restenosis model using a balloon angioplasty catheter or drug-eluting coronary stent. Pigs were divided into two groups and were fed a commercial normal diet (CND, n = 4) or a high-fat diet (HFD, n = 4) for 5 weeks. Coronary overstretch injury by balloon angioplasty or stent implantation was induced in the left anterior descending and left circumflex artery after 1 week of feeding. Histopathological analysis was performed at 4 weeks after coronary injury. During the experiment, the total cholesterol level in the HFD group increased by approximately 44.9% (from 65.9 ± 3.21 mg/dL at baseline to 95.5 ± 9.94 mg/dL at 5 weeks). The lumen area in the CND group was reduced in comparison with that in the HFD group after balloon angioplasty. After stent implantation, the injury score showed no significant difference. There were significant differences in the neointimal area (2.7 ± 0.33 mm2 in the CND group vs. 3.3 ± 0.34 mm2 in the HFD group, p<0.05), lumen area (2.6 ± 0.54 mm2 in the CND group vs. 2.0 ± 0.33 mm2 in the HFD group, p<0.05), and percent area stenosis (52.0 ± 7.96% in the CND group vs. 62.4 ± 5.15% in the HFD group, p<0.05). Body weight change was not different between the two groups. Increased serum cholesterol level activated vascular smooth muscle cell proliferation in the porcine coronary overstretch model.

The purpose of this study was to investigate the inhibitory effect of enzyme activity and anti-proliferation of human cancer cell lines (HCT 116, NCI-H460 and MCF-7) of peanut skin depending on cultivars (Arachis hypogaea L. cv. K-Ol, cv. Sinpalkwang, cv. Daan, cv. Heuksaeng) and extraction solvent. Peanut skin was extracted with 80% ethanol, 80% methanol, 80% acetone, and distilled water, followed by analysis of the enzyme inhibitory activity and anticancer activity. Methanol extract of Daan cultivar most effectively inhibited α-gluosidase (65.08%, 0.025 mg/mL), tyrosinase (82.49%, 2 mg/mL) and ACE (73.61%, 10 mg/mL). The inhibitory effect of peanut skin extracts on colon cancer cell (HCT-116), lung cancer cell (NCI-H460) and breast cancer cell (MCF-7) growth were investigate using MTT assay. The highest anti-proliferation of cancer cell line of peanut skin extracts was observed in the methanol extract of Daan cultivar. The cell viability on HCT 116, NCI-H460 and MCF-7 cell lines of methanol extracts from peanut skin of Daan cultivar was 48.13%, 41.03%, and 36.02% at 200 μg/mL, respectively. These results suggest that peanut skin extracts may mediate physiological activity, and provide valuable information for the use of peanut byproduct as a functional food material.

L-carnitine은 라이신과 메티오닌으로 생합성되며 골격근 과 심근을 포함한 다양한 동물조직에서 발견된다. L-carnitine이 포함된 식품으로는 양고기, 소고기, 돼지고기 등이 있고 근육발달에 도움을 주며 뼈를 강화하거나 대사작용을 도와주는 기능을 하여 영양 보조제로 많이 섭취하는 것으로 알려져 있다. 최근 L-carnitine은 제 2형 당뇨병, 골다 공증, 대사성 신경증후군 등의 다양한 질병의 약물로도 연구 되고 있으며 암에서는 치료 보조제로 개발되어있다. 하지만 대장암에서의 L-carnitine에 대한 효과 및 기전에 대해서는 명확하지 않고 연구된 바가 없기 때문에 본 연구에서 저자들은 L-carnitine의 효능을 인간대장암세포주 HCT116에서 규명하고자 하였다. L-carnitine은 세포 내 활성산소종 (ROS)를 높은 수준으로 증가시켜 세포 증식을 억제하였다. 또한, 세포 증식과 죽음에 관련한 단백질 ERK1/2와 p38을 유의적으로 활성화 시킨다는 것을 입증 하였다. 이때, ERK1/2 억제제(PD98059)를 처치하여 ERK1/ 2의 활성화가 활성산소종 발생 및 세포사멸에 중요하다는 것을 밝혔다. 따라서, 본 연구 결과는 L-carnitine이 대장 암세포주의 증식을 억제 할 수 있고 이는 대장암의 치료에 있어 잠재적인 치료 물질이 될 수 있음을 시사하며 이 과정에 관여하는 신호전달기전을 조사하여 항암의 치료기 전에서 활성산소종이나 ERK1/2, p38 단백질의 활성화의 중요성을 제시하였다.

It is well known that lymph node metastasis is a major prognostic factor in patients with oral squamous cell carcinoma (OSCC). 30-40% of patients with OSCC have already undergone regional metastasis at diagnosis. The survival rate of patients with metastasis is reduced by more than 50%. Therefore, prevention and early detection of metastasis are very important to increase the survival rate of patients. Many investigators have studied the molecular mechanism of metastasis and tried to develop the molecules to inhibit any step of metastatic cascade. Among those molecules, an interest in the metastasis suppressor gene has been increasing. Expression of metastasis suppressor KiSS-1 has shown to be significantly related to poor clinical outcome and worse survival rate of patient in various malignancies of different organs. In addition, our previous study in OSCC also revealed that downregulation of KiSS-1 expression correlated with the presence of cervical lymph node metastasis, one part of tumor progression. Therefore, further investigation was needed to identify the molecular function of KiSS-1 using OSCC cell line and to evaluate the possibility of KiSS-1 as a new therapeutic target.

Autophagy is recently receiving the spotlight as the development strategy for promising anticancer drugs. In particular, the majority of anticancer drugs originating from natural products are known to induce autophagy. Saururus chinensis has been used for treating various inflammatory diseases. Recent research has revealed that the extract of Saururus chinensis possess cytotoxicity for various types of human cancer cells. However, the exact action mechanism of Saururus chinensis extract for oral squamous cell carcinoma (OSCC) has not been studied yet. Therefore, the authors of this research aim to study the effect of methanol extract of S. chinensis (MESC) on OSCC cells. To observe the cell proliferation inhibitory effect of MESC on HSC3 cells, the authors conducted the trypan blue exclusion assay. Also, the action mechanism of MESC was studied by conducting the cell cycle analysis, acidic vesicular organelle (AVO) staining and flow cytometry analysis, monodansylcadaverine (MDC) staining, propidium iodide staining, and Western blotting on MESC-treated HSC3 cells. When HSC3 cells were treated in MESC, the cell proliferation was suppressed in time-dependent and dose-dependent manners. Also, the number of sub-G1 arrested cells increased in a dose-dependent manner. MDC punctate and AVO puncta significantly increased respectively. Western blot analysis demonstrated the expression of autophagy-related proteins increased, but apoptotic proteins were not observed. Also, the pAkt protein was reduced, while the p-p38 protein and pERK protein increased. According to our results, MESC induced autophagy and accompanied changes in the cell cycle in HSC3 cells. Also, the alteration in Akt, ERK, and p38 pathways were confirmed. This result suggested the possibility of MESC as the new promising adjuvant for treating OSCC patients.

본 연구에서는 발아와 고압처리에 따른 검정콩 조사포닌 추출물의 in-vitro 항암 활성을 확인하기 위하여 검정콩을 발아시킨 후 고압처리하고, 조사포닌 추출물을 제조한 후 인체 유래 유방암(MCF-7), 대장암(HCT-116), 전립선암(PC-3) 및 위암(AGS) 세포주에 대한 증식억제효과를 검토하였다. 대조구의 조사포닌 추출물은 400 μg/mL 농도에서 4종의 암세포주에 대하여 67.02~91.70% 범위의 생존율을 보여 항암 효과가 낮았지만, 발아 4일차 콩을 150 MPa의 압력에서 고압처리한 검정콩의 조사포닌 추출물은 23.94~57.37% 범위의 생존율을 보여 발아와 고압처리에 의해 암세포 성정억제효과가 증가하였다. 특히, 위암세포(AGS)의 경우 다른 세포주에 비해 저농도에서도 세포증식효과를 보여 가장 높은 암세포 증식억제 효과를 나타내었다. 이상의 결과로부터 항암 활성을 나타내는 Soyasaponin, B group의 soyasaponin 및 soyasapogenol의 함량은 발아와 고압처리에 의해 증가하였고, 고압처리 발아콩으로부터 항암 활성을 나타내는 사포닌 추출물의 개발이 가능할 것으로 생각된다. 또한, 추후 항암 활성 물질의 분리동정과 메커니즘 규명에 대한 연구가 수행되어야 할 것으로 판단된다.

Previous studies have shown that kisspeptin (Kp-10) is expressed in mammalian ovaries; however, the expression and role of Kp-10 in bovine ovarian granulosa cells are still unclear. In this study, we assessed the expression of Kp-10 and its effects on the proliferation and apoptosis of bovine granulosa cells. Immunohistochemical analysis showed that Kp-10 was expressed in the cytoplasm of bovine ovarian granulosa cells. Moreover, MTT (3-[4,5-dimethyl-2-thiazolyl]-2,5-diphenyl-2- H-tetrazolium bromide) assays showed that 100 nM Kp-10 significantly inhibited the viability of granulosa cells (P<0.01). Flow cytometry analysis showed that Kp-10 could significantly increase accumulation of cells in the G1 phase, decrease accumulation of cells in the S phase, and promote apoptosis in bovine granulosa cells (P<0.05). Additionally, Kp-10 decreased the mRNA levels of Bcl-2, an anti-apoptotic gene; increased the mRNA levels of caspase-3, a pro-apoptotic gene; and increased the mRNA levels of Fas and Fasl, two membrane surface molecule genes (P<0.05). Thus, our findings demonstrated for the first time that Kp-10 inhibited proliferation and promoted apoptosis in bovine ovarian granulosa cells. These findings provide insights into our understanding of the role of Kp-10 in mediating the proliferation of bovine granulosa cells.

Induced pluripotent stem cell (iPSC)-derived mesenchymal stem cells (iMSCs) serve as a unique source for cell therapy. We investigated whether exosomes from iMSCs promote the proliferation of human keratinocytes (HaCaT) and human dermal fibroblasts (HDFs). iPSCs were established from human Wharton’s jelly MSCs and were allowed to differentiate into iMSCs. Exosomes were collected from the culture supernatant of MSCs (MSC-exo) and iMSCs (iMSC-exo), and their characteristics were investigated. Both exosome types possessed basic characteristics of exosomes and were taken up by skin cells in vitro and in vivo. A significant increase in HaCaT proliferation was observed with iMSC-exo, although both exosomes increased the viability and cell cycle progression in HaCaT and HDFs. No significant difference was observed in the closure of wound scratch and the expression of reparative genes between cells treated with the two exosome types. Both exosomes enhanced the secretion of collagen in HaCaT and HDFs; however, an increase in fibronectin level was observed only in HaCaT, and this effect was better with iMSC-exo treatment. Only iMSC-exo increased the phosphorylation of extracellular signal-regulated kinase (ERK)-1/2. Our results indicate that iMSC-exo promote the proliferation of skin cells by stimulating ERK1/2 and highlight the application of iMSCs for producing exosomes.