• Journal of Plant Biotechnology
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• v.46 no.3
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• pp.228-235
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• 2019

Apple (Malus pumila) is one of the most economically important fruits in Korea. but virus infection has decreased the sustainable production of apples and caused serious problems such as yield loss and poor fruit quality. Virus or viroid infection including apple chlorotic leaf spot virus (ACLSV), apple stem pitting virus (ASPV), apple stem grooving virus (ASGV), apple mosaic virus (ApMV) and apple scar skin viroid (ASSVd) have been also reported in Korea. In many cases, as apple gets infected with virus and viroid with no specific symptoms, the damage and symptoms caused by the viruses are not detected. In our research, viruses in the rootstock were eliminated for a virus-free apple dwarfing rootstock of M.9 and M.26. The virus elimination methods were apical meristem culture, thermotherapy ($37^{\circ}C$, 6 weeks) and chemotherapy($Ribavirin^{(R)}$). The detection of apple viruses was accomplished by Enzyme-linked Immuno-Sorbent Assay (ELlSA) and reverse transcription-polymerase chain reaction (RT-PCR). RT- PCR method was 10 ~ 30% more sensitive than the ELISA method. The efficiency of virus elimination was enhanced in apical meristem culture method. The acquisition rate of virus-free apple dwarfing rootstocks was 30 ~ 40% higher in apical meristem culture. After the meristem culturing of M.9, the infection ratio of ACLSV, ASPV and ASGV was 45%, 60% and 50%, respectively. In the apple dwarfing rootstock of M.26, the infection ratio of ACLSV, ASPV and ASGV was 40%, 55% and 55%, respectively. Based on this study, the best method for the production of virus-free apple dwarfing rootstocks was the apical meristem culture.

• Journal of Life Science
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• v.29 no.8
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• pp.904-915
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• 2019

Prostate cancer (PCa) is one of the most metastatic tumor. Although hormone therapy or surgical castration is mostly conducted to treat PCa, it has a lot of side effects. Recently, many researchers have been exploring the tumor microenvironment to remedy these circumstances. Immune cells, especially macrophages, are an important composition of the tumor microenvironment. Under normal conditions, macrophages exhibit mild tumoricidal activity against tumors. However, once activated by interferon gamma or lipopolysaccharides, macrophages can kill cancer cells directly or indirectly by secreting cytokines and chemokines. In this study, murine macrophage RAW 264.7 cells were treated with Phellinus linteus extract. To analyze their pro-inflammatory phenotype, we were used several assays such as a real-time polymerase chain reaction, an enzyme-linked immunosorbent and nitric oxide assay. Prostate cancer cells were treated with the RAW 264.7-conditioned media, which was identified as a pro-inflammatory nature, for 48 h, and the expression of epithelial-mesenchymal transition (EMT)-related genes was determined. Not only N-cadherin, Snail, Twist, Slug, and Cadherin 11, which are mechenchymal-related proteins, were decrease, but epithelial marker of E-cadherin was increased. In addition, the mRNA level of vimentin, ccl2, and vegfa were decreased, as the EMT is closely related to the migration and invasion of cancer cells. In conclusion, the RAW 264.7-conditioned media stimulated with P. linteus extract inhibited migration and invasion and regulated the EMT pathway in human prostate cancer cells.

• Microbiology and Biotechnology Letters
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• v.47 no.1
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• pp.148-157
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• 2019

A 70% ethanol extract of Erigeron annuus (L.) Pers. was investigated for its whitening activity for application as a functional ingredient in cosmetic products. At the E. annuus extract concentration of $100{\mu}g/ml$, the electron-donating ability was found to be 67.83%, the tyrosinase inhibitory effect (related to skin-whitening) was 69%, the elastase inhibitory effect (related to skin-wrinkling) was 69%, and the astringent effect was 80%. The $ABTS^+$ radical-scavenging ability was 87% at the $500{\mu}g/ml$ concentration. In the cell viability test measured on melanoma cells, 96% of the cells treated with $100{\mu}g/ml$ of the extract were viable. According to the western blot results, the protein expression of the microphthalmia-associated transcription factor (MITF), tyrosinase, tyrosinase-related protein (TRP)-1, and TRP-2 was decreased by 60.22%, 47.83%, 54.79%, and 67.88%, respectively, at the extract concentration of $100{\mu}g/ml$. The protein expression of phosphorylated extracellular signal regulated kinase (p-ERK) and phosphorylated cAMP response element-binding protein (p-CREB) was decreased with increasing concentrations of the extract. Reverse transcription-polymerase chain reaction of the extract showed that the mRNA expression of MITF, tyrosinase, TRP-1, and TRP-2 was decreased by 86.51%, 85.22%, 74.26%, and 66.66%, respectively, at $100{\mu}g/ml$ extract concentration. The findings suggest that the 70% ethanol extract from E. annuus (L.) Pers. has potential as a cosmeceutical ingredient with whitening effect.

• Asian-Australasian Journal of Animal Sciences
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• v.32 no.7
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• pp.922-929
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• 2019

Objective: Mutations in low-density lipoprotein receptor (LDLR), which encodes a critical protein for cholesterol homeostasis and lipid metabolism in mammals, are involved in cardiometabolic diseases, such as familial hypercholesterolemia in pigs. Whereas microRNAs (miRNAs) can control LDLR regulation, their involvement in circulating cholesterol and lipid levels with respect to cardiometabolic diseases in pigs is unclear. We aimed to identify and analyze LDLR as a potential target gene of SSC-miR-20a. Methods: Bioinformatic analysis predicted that porcine LDLR is a target of SSC-miR-20a. Wild-type and mutant LDLR 3'-untranslated region (UTR) fragments were generated by polymerase chain reaction (PCR) and cloned into the pGL3-Control vector to construct pGL3 Control LDLR wild-3'-UTR and pGL3 Control LDLR mutant-3'-UTR recombinant plasmids, respectively. An miR-20a expression plasmid was constructed by inserting the porcine premiR-20a-coding sequence between the HindIII and BamHI sites in pMR-mCherry, and constructs were confirmed by sequencing. HEK293T cells were co-transfected with the miR-20a expression or pMR-mCherry control plasmids and constructs harboring the corresponding 3'-UTR, and relative luciferase activity was determined. The relative expression levels of miR-20a and LDLR mRNA and their correlation in terms of expression levels in porcine liver tissue were analyzed using reverse-transcription quantitative PCR. Results: Gel electrophoresis and sequencing showed that target gene fragments were successfully cloned, and the three recombinant vectors were successfully constructed. Compared to pMR-mCherry, the miR-20a expression vector significantly inhibited wild-type LDLR3'-UTR-driven (p<0.01), but not mutant LDLR-3'-UTR-driven (p>0.05), luciferase reporter activity. Further, miR-20a and LDLR were expressed at relatively high levels in porcine liver tissues. Pearson correlation analysis revealed that porcine liver miR-20a and LDLR levels were significantly negatively correlated (r = -0.656, p<0.05). Conclusion: LDLR is a potential target of miR-20a, which might directly bind the LDLR 3'-UTR to post-transcriptionally inhibit expression. These results have implications in understanding the pathogenesis and progression of porcine cardiovascular diseases.

• Journal of Life Science
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• v.31 no.8
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• pp.710-718
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• 2021

Placenta-derived mesenchymal stem cells (PD-MSCs) are promising candidates for cell-based therapy in regenerative medicine. The migration and homing potential of PD-MSCs to injured sites is a critical property of MSC engraftment. MicroRNAs (miRNAs) have recently been shown to regulate the critical functions of MSCs, such as proliferation, survival, and migration. The objective of the present study was to identify the miRNA and target genes involved in PD-MSCs homing in a bile duct ligation (BDL) rat model. We selected candidate miRNAs targeting genes for PD-MSCs homing based on microarray analysis. PD-MSC engraftment in BDL-injured rat liver was identified by immunofluorescence assay and human-specific Alu gene expression by quantitative real-time polymerase chain reaction (qRT-PCR) one week after transplantation. Compared with migrated naïve PD-MSCs under hypoxic and normoxic conditions (Hyp/Nor), the transplanted group with PD-MSCs (Tx) showed distinct differences in miRNA expressions in BDL-injured rat liver. We also validated the miRNAs and their target genes for PD-MSCs homing. The expressions of integrin α4 (ITGA4) and integrin α5 (ITGA5) target genes for miR-199a-5p and miR-148a-3p were significantly upregulated in the Tx group (p<0.05). In addition, integrin β1 (ITGB1) and integrin β8 (ITGB8) were upregulated by suppressing miR-183-5p and miR-145-5p, respectively. These results demonstrated that PD-MSCs regulate miRNA expression related to the integrin family for their homing effects on the BDL-injured rat liver. The findings further suggest that miRNA-mediated regulation of the integrin family contributes to the therapeutic efficacy of PD-MSCs in the rat hepatic fibrosis model by BDL.

• Journal of Life Science
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• v.29 no.11
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• pp.1227-1234
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• 2019

Prunus mume, also known as maesil, is a popular fruit consumed in East Asia (Korea, Japan, and China). It contains high amounts of organic acids, minerals, and polyphenols and has been used as a medication for fever, vomiting, and detoxification. In this study, the anti-proliferative and apoptotic effects of solvent fractions from maesil were evaluated using sulforhodamine B (SRB) assays, morphological evaluations, Hoechst 33258 staining, and western blotting. Addition of the maesil methanol fraction (MMF) and the maesil butanol fraction (MBF) significantly and dose-dependently decreased the cell viability of HT-29 human colon cancer cells. Colony-forming assays confirmed that the MMF and MBF treatments decreased colony numbers when compared with untreated control cells. Treatment of HT-29 cells with MMF and MBF caused a distortion of the cell morphology to a shrunken cell mass. Treatment with MMF and MBF also dose-dependently increased nuclear condensation and the formation of apoptotic bodies in HT-29 cells. Treatment with MMF and MBF significantly and dosedependently increased the expression of Bax (a pro-apoptotic protein), caspase-3, and poly ADP-ribose polymerase (PARP) and decreased the expression of Bcl-2 (an anti-apoptotic protein). MMF significantly and dose-dependently inhibited cell proliferation induced by bisphenol A, an environmental hormone. Therefore, MMF may have potential use as a functional food and as a possible therapeutic agent for the prevention of colon cancer.

• Journal of Life Science
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• v.31 no.1
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• pp.10-16
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• 2021

The consumption of fresh-cut agricultural materials is increasing due to increased public interest in health and the increase of single-person households. Most fresh-cut agricultural materials can be eaten without heating, thus easily exposing the consumer to food-borne pathogens. As a result, food-borne diseases are increasing worldwide. In the analysis of food-borne pathogens, it is important to detect the strains, but this is time consuming and laborious. Alternative detection methods that have been introduced, include polymerase chain reaction-denaturing gradient gel electrophoresis (PCR-DGGE), which is performed without prior culturing. Samples of fresh-cut agricultural materials, such as vegetables, were analyzed by the culture-based method. In 129 samples, non-pathogenic Escherichia coli (3.9%), Bacillus cereus (31.8%), Clostridium perfringens (5.4%), Yersinia enterocolitica (0.8%), and enterohemorrhagic E. coli (0.8%) were detected. Eight samples contaminated with bacteria were randomly selected, further analyzed by PCR-DGGE, and compared with the culture-based method. Two cases detected non-pathogenic E. coli by PCR-DGGE only, despite a lack of detection by the culture method. It was supposed there was possibility of sample loss during its 10-fold dilution for appropriate cultivation. In the detection of high-risk food-borne pathogens, it was found that the detection limit was lower in PCR-DGGE than in the culture-based method (10 CFU/g). This suggests that PCR-DGGE can be alternatively used to detect strains. On the other hand, low-risk food-borne pathogens seem to have higher detection limits in PCR-DGGE. Consequentially, this study contributes to the improvement of food-borne pathogen detection and the prevention of its related-diseases in fresh-cut agricultural materials.

• Journal of Life Science
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• v.31 no.2
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• pp.199-208
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• 2021

Euonymus porphyreus, a species of plant in the Celastraceae family, is widely distributed in East Asia, especially in Southern China. The botanical characteristics of E. porphyreus have been reported, but its antioxidative and anticancer activities remain unclear. In this study, we evaluated the antioxidative and anticancer effects of ethanol extracts of E. porphyreus (EEEP) and the molecular mechanism of its anticancer activity in human lung adenocarcinoma A549 cells. The total polyphenol and flavonoid compound contents from EEEP were 115.42 mg/g and 23.07 mg/g, respectively. EEEP showed significant antioxidative effects with a concentration at 50% of the inhibition (IC50) value of 11.09 ㎍/ml, as measured by 2,2-diphenyl-1-picrylhydrazyl (DPPH) radical scavenging assay. EEEP showed cytotoxic activity by increasing the SubG1 cell population of A549 cells in a dose-dependent manner. Apoptosis in A549 cells treated with EEEP was evident due to increased apoptotic cells and apoptotic bodies, as detected by Annexin V and 4,6-diamidino-2-phenylindole (DAPI) staining, respectively. EEEP-induced apoptosis resulted in increased expression of the First apoptosis signal (Fas), p53, and Bax, with decreased expression of Bcl-2 and subsequent activation of caspase-8, -9, and caspase-3, leading to cleavage of poly (ADP-ribose) polymerase (PARP). Collectively, these results suggest that EEEP may exert an anticancer effect by inducing apoptosis in A549 cells through both intrinsic and extrinsic pathways.

• Journal of Life Science
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• v.31 no.3
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• pp.305-313
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• 2021

This study verified the antioxidant and whitening activities of a Pinus koraiensis extract (PK) and a Hibiscus cannabinus L. extract (HC), and further evaluated the interaction of the extract ingredients when mixed at a 1:1 ratio (PKHC). The electron-donating and ABTS+ radical scavenging activities of the PKHC extract at 1,000 ㎍/ml concentration were 93.7% and 94%, respectively, indicating a higher efficacy than achieved with either extract alone. Measurements of the tyrosinase the activities in response to PK, HC, and PKHC extracts at 1,000 ㎍/ml concentrations showed inhibitions of 40%, 27.5%, and 43%, respectively, confirming a higher efficacy of the mixture due to the synergistic action of the ingredients. The cell toxicity values in melanoma cells treated with PK, HC, and PKHC at 1,000 ㎍/ml concentration were 87.4%, 80.2%, and 98%, confirming a higher viability in cells treated with the mixture due to antagonism. The expression of microphthalmia-associated transcription factor (MITF), tyrosinase-related protein-1 (TRP-1), tyrosinase-related protein-2 (TRP-2), and tyrosinase protein expression determined by Western blotting decreased by 53.9%, 64.8%, 67.3%, and 56.1%, respectively, when PKHC was administered at a concentration of 100 ㎍/ml. Reverse transcription-polymerase chain reaction (RT- PCR) results also showed that PKHC at a concentration of 100 ㎍/ml inhibited the mRNA expression of MITF, TRP-1, TRP-2, and tyrosinase mRNA by 54.4%, 64.9%, 66.6%, and 63.1%, respectively. Taken together, the data confirmed the antioxidant and whitening effect of the PKHC extract and verified the possibility that this extract mixture has great potential as a cosmetic ingredient.

• Research in Plant Disease
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• v.27 no.1
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• pp.1-7
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• 2021

In August 2020, necrotic ringspots on leaves were observed on 20 from 143 Plantago asiatica plants in open fields in Eumseong, Chungcheongbuk-do. Eight symptomatic Plantago asiatica plants were subjected to investigation on viral infection by reverse transcription and polymerase chain reaction. Impatiens necrotic spot virus, tomato spotted wilt virus and cucumber mosaic virus were detected from the symptomatic plants. Two impatiens necrotic spot virus (INSV) isolates ('INSV-plantain kr1' and '-plantain kr5') were sequenced and analyzed by comparing L genomic segment, nucleoprotein (N) gene and non-structural protein S (NSs) gene sequences. The nucleotide sequence of 'INSV-plantain kr1' isolate (MW114834) was most closely related to that of a 'Phalaenopsis' isolate (GQ336991) from China in the L genomic segment. 'INSV-plantain kr1' and '-plantain kr5' isolates shared the highest identities with those from 'Pepe' isolate (LC384872) and 'J' isolate (AB109100) in the NSs gene, respectively, and with that from 'YSMi-SH' isolate (FN400773) in the N gene. Phylogenetic analysis based on L genomic segment grouped the INSV-plantain kr1 isolate together with isolates from Korea (LC384870), China (GU112505, GQ336991), and Italy (DQ425094). This is the first report on INSV in P. asiatica from Korea.