Actinomycetes, Gram positive soil bacteria, are valuable microorganisms which produce useful secondary metabolites including antibiotics, antiparasitic substances, anti-cancer drugs, and immunosuppressants. Although a major family of actinomycetes, known as streptomycetes, has been intensively investigated at the molecular level for several decades, a potentially valuable and only recently isolated non-streptomycetes rare actinomycetes (NSRA) family has been poorly characterized due to lack of proper genetic manipulation systems. Here we report that a PCR-based genome screening strategy was performed with approximately 180 independently isolated actinomycetes strains to isolate potentially valuable NSRA strains. Thanks to this simple PCR-based genome screening strategy we were able to identify only seven NSRA strains, followed by 16S rRNA sequencing for confirmation. Through further bioassays, one potentially valuable NSRA strain (tentatively named Nocardiopsis species MMBL010) was identified which possessed both antifungal and antibacterial activities, along with the presence of polyketide synthase and non-ribosomal peptide synthase genes. Moreover, Nocardiopsis species MMBL010, which was intrinsically recalcitrant to genetic manipulation, was successfully transformed via E. coli-driven conjugation. These results suggest that PCR-based genome screening, followed by the establishment of an E. coli-driven conjugation system, is an efficient strategy to maximize potentially valuable compounds and their biosynthetic genes from NSRA strains isolated from various environments.
Ju, Jung-Il;Paik, Seung-Woo;Yun, Tug-Sang;Park, Young-Chun;Lee, Bo-Hee;Son, Seung Wan
Korean Journal of Plant Resources
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v.33
no.5
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pp.436-445
/
2020
Goji berry (Lycium chinense Mill.) is one of medicinal plants. Its leaves has been used to manufacture the functional foods by replacing the dried-fruit because of low production costs. In order to produce the leaves of uniform quality, continuous selection and the establishment of cultivation techniques were required. Among the eleven recommended cultivars, 'Myeongan' showed high yielding and rapid regeneration after cutting. The dried-leaf yield was linearly increased from 106 kg/10a on May 16 to 287 kg/10a on June 20. An appropriate cutting date and cutting length (about 60-70 cm) was important factors for its efficient regeneration. The late cutting times were not suitable due to difficulties during plant harvesting because of stem rigidity and thorn generation. The betaine content of leaves ranged from 1.43 ~ 2.63% and significantly affected by the varieties and the cutting dates. The main physiological functionality of leaf was Angiotesin-1-converting enzyme (ACE) inhibitory activity, representing the anti-hypertensive. The other physiological functionalities, XOD inhibitory activity, antioxidant activity, tyrosinase inhibitory activity, SOD-like activity, α-glucosidase inhibitory activity and fibrinolytic activity, were not detected or less than 20%.
Kim, Kang Ju;Park, Bog Im;Min, Jae Hong;Chae, Min Suk;Lim, Jae You;Son, Hyeok Jin;Lee, Gi Hoon;An, So Youn;Jeon, Byung Hun;Choi, Na Young;You, Yong Ouk
Journal of Physiology & Pathology in Korean Medicine
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v.30
no.1
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pp.27-32
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2016
Streptococcus mutans plays a virtal role in trigering dental caries establishment due to its ability to synthesize two significant factors. The two factors are organic acids and glucans. The former demineralized dental enamel and the latter mediates the attachment of bacteria to tooth surface. It is believed that demineralization of dental enamel and attachment of bacteria are the crucial events that indicate and develop dental caries. For this reason, we studied the effect of the ethanol extracts of Radix Pulsatillae on the growth and acid production of S. mutans. Ethanol extracts of the Radix Pulsatillae showed concentration dependent inhibitory activity against the growth and acid production of S. mutans, and produced significant inhibition compared to the control groups (p<0.05). The extracts inhibited S. mutans adherence to hydroxyapatite treated with saliva, and cell adherence was repressed by Radix Pulsatillae. the ethanol extract of Radix Pulsatillae showed remarkable inhibition of glucosyltransferase, which synthesizes water insoluble glucan form sucrose. Phytochemical analysis showed Radix Pulsatillae contained major components such as phenolic compounds, glycosides, steroids, terpenoid, and saponin. These results suggest that Radix Pulsatillae may have anti-cariogenic properties, which may be related with major components such as phenolic compounds, glycosides, steroids, terpenoid, and saponin.
Tumor necrosis factor (TNF)-related apoptosis-inducing ligand (TRAIL/Apo2L) is a recently identified member of the TNF ligand family that can initiate apoptosis through the activation of their death receptors. TRAIL has been paid attention as a potential anti-cancer drug, because it selectively induces apoptosis in tumor cells in vitro and in vivo but not in most normal cells. However, recent studies have shown that some cancer cells including malignant renal cell carcinoma and hepatocellular carcinoma, are resistant to the apoptotic effects of TRAIL. Therefore, single treatment with TRAIL may not be sufficient for the treatment of various malignant tumor cells. Understanding the molecular mechanisms of TRAIL resistance and identification of sensitizers capable of overcoming TRAIL resistance in cancer cells is needed for the establishment of more effective TRAIL-based cancer therapies. Chemotherapeutic drugs induce apoptosis and the upregulation of death receptors or activation of intracellular signaling pathways of TRAIL. Numerous chemotherapeutic drugs have been shown to sensitize tumor cells to TRAIL-mediated apoptosis. In this study, we summarize biological agents and drugs that sensitize tumors to TRAIL-mediated apoptosis and discuss the potential molecular basis for their sensitization.
As Korean ginseng is hybrid, an individual variation is very severe, and it takes long times in new breeding because it is required 4 years to pick the seed. But, transformation technique makes the high-functional breeding in short time. The focus of these ginseng studies is to find and secure the useful gene. And it is urgent to accumulate the fundamental data for the molecular breeding and secure the useful genes. Therefore, transformation and soil acclimatization technique are necessary to molecular breeding in use of the introduction of functional genes. In this study, it add to secure of new regulation gene and useful gene as to accumulate the fundamental data for the place where it will contribute to raise the national competitive power. To analyze the useful genes in large scale, we constructed CDNA libraries with various tissues, species, and treated tissue. EST analysis of ginseng perform in large scale and build the EST database of ginseng. We perform the full length sequencing about the selected lots of clones that include the entire open reading frame of the amino acid residues and construct cDNA chip with the parental EST clones. Establishment of the transformation and a soil acclimatization system throuth the re-introduction of the selected ginseng gene that related with the secondary metabolism and anti-stress into the ginseng.
Park Sun-Hee;Won Sung Yong;Park Soo-Young;Yoon Sung Wook;Han Jin Hyun;Jeong Yong Seok
Proceedings of the Microbiological Society of Korea Conference
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2000.05a
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pp.23-36
/
2000
Japanese encephalitis virus (JEV) is the causative agent of a mosquito-borne encephalitis and is transmitted to human via persistently infected mosquito vectors. Although the virus is known to cause only acute infection, there were reports that showed neurological sequelae, latent infection in peripheral mononuclear cells, and recurrence of the disease after acute encephalitis. Innate resistance of certain cell lines, abnormal SN1 expression of the virus, and anti-apoptotic effect of cullular bcl-2 have been suggested as probable causes of JEV persistence even in the absence of defective interfering (DI) particles. Although possible involvement of DI particles in JEV persistence was suggested, neither has a direct evidence for DI presence nor its molecular characterization been made. Two questions asked in this study are whether the DI virus plays any role in JEV persistent infection if it is associated with and what type of change(s) can be made in persistently infected cells to avoid apoptosis even with the continuous virus replication, DI-free standard stock of JEV was infected in BHK-21, Vero, and SW13 cells and serial high multiplicity passages were performed in order to generate DI particles. There different-sized DI RNA species which were defective in both structural and nonstructural protein coding genes. Rescued ORFs of the DI genome maintained in-frame and the presence of replicative intermediate or replicative form RNA of the DI particles confirmed their replication competence. On the other hand, several clones with JEV persistent infection were established from the cells survived acute infections during the passages. Timing of the DI virus generation during the passages seemed coincide to the appearance of persistently infected cells. The DI RNAs were identified in most of persistently infected cells and were observed throughout the cell maintenance. One of the cloned cell line maintained the viral persistence without DI RNA coreplication. The cells with viral persistence released the reduced but continuous infectious JEV particle for up to 9 months and were refractory to homologous virus superinfection but not to heterologous challenges. Unlike the cells with acute infection these cells were devoid of characteristic DNA fragmentation and JEV-induced apoptosis with or without homologous superinfection. Therefore, the DI RNA generated during JEV undiluted serial passage on mammalian cells was shown to be biologically active and it seemed to be responsible, at least in part, for the establishment and maintenance of the JEV persistence in mammalian cells. Viral persistence without DI RNA coreplication, as in one of the cell clones, supports that JEV persistent infection could be maintained with or without the presence of DI particles. In addition, the fact that the cells with JEV persistence were resistant against homologous virus superinfection, but not against heterologous one, suggests that different viruses have their own and independent pathway for cytopathogenesis even if viral cytopathic effect could be converged to an apoptosis after all.
Purpose: Delayed intentional replantation was introduced as a new alternative to treat the teeth with severe periodontal involvement. The purpose of this study was to elucidate the possibility of delayed intentional replantation and establish theoretical backgrounds. Materials and Methods: Studies were performed into the following two subjects; (1)Clinical evaluation of patients who underwent delayed intentional replantation using clinical and radiographic data. Severe periodontitis involved teeth were carefully extracted and proper time for delayed replantation was evaluated by analyzing inflammation markers (IL-6, TNF-${\alpha}$). (2) Theoretical studies for efficacy of delayed intentional replantation using (-)-Epigallocatechin-3-gallate (EGCG) for preservation of periodontal ligament cells on root surface by minimizing inflammation and treatment of inflammatory extraction sockets. Results: Meaningful success ratio and survival rate were found in delayed intentional replantation showing reduced bone loss and maintained bone level. Additionally, viability of EGCG applied periodontal ligament cells was much higher than control group. Also, EGCG promoted healing of inflammatory extraction sockets by inhibiting inflammatory cell proliferation. Conclusion: Within the limitations of this study, 1-2 weeks after extraction is an appropriate time to do delayed intentional replantation. Also, EGCG provides helpful effects on viability of periodontal ligament cells and periodontium.
Lee, Young Ju;Kim, Ae Kyung;Kim, Ouk Hee;Lee, Chun Young;Lee, Hyun Kyung;Jung, Sun Ok;Lee, Sae Ram;Kim, Hee Sun;Kim, Il Young;Yu, In Sil;Jung, Kweon
Korean Journal of Medicinal Crop Science
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v.24
no.2
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pp.121-128
/
2016
Background: Turmeric (Curcuma longa L.) that is used as a food material has antioxidant, anticancer and anti-inflammatory properties. Recently the demand for functional foods and drugs has increased. The present study was carried out to determined of contents of residual sulfur dioxide, heavy metals, ash, acid insoluble ash and curcuminoids in turmeric from the Seoul Yak-ryeong market. Methods and Results: A total of 31 samples were obtained. Residual sulfur dioxide was not detected in any samples. Heavy metals (arsenic, cadmium, lead and mercury) were analyzed by ICP-MS (Inductively Coupled Plasma Mass Spectrometer) and a mercury analyzer and were detected in the ranges of 0.00 - 0.28, 0.00 - 0.07, 0.00 - 0.29 and 0.002 - 0.027mg/g respectively. No significant difference were observed between the average heavy metal contents of domestic and imported tumeric. However, average content of ash in domestic samples (7.8%) were significantly higher than that in imported samples (6.1%), whereas that of curcuminoids was significantly higher in imported samples (47.6mg/g) than in domestic samples (11.2mg/g). The average content of acid insoluble ash was not significantly different between two sample types (0.9% in each). Conclusions: There are no specific standards for turmeric used as food materials. Therefore, this study can be provided as basic data for the establishment of quality standards for turmeric.
This study was conducted to establish the extraction conditions for the optimization of the biological activities of black garlic using the response surface methodology (RSM). The extraction conditions were based on the central composite design, with 15 kinds of variations in the extraction temperature (50-$90^{\circ}C$ ), extraction time (3-15 hrs), and ethanol concentration (0-100%). The total phenol, flavonoids, thiosulfinate contents, and anti-oxidant activity of black garlic extract were significantly higher at the J condition ($90^{\circ}C$ for 9 hrs with a 50% ethanol concentration). In this condition, the biological activities such as DPPH radical scavenging (66.10%), ABTS radical scavenging activity (75.02%), and reducing power by of FRAP (375.4 ${\mu}M/mL$) were excellent. Overall, the extraction conditions for the optimized biological activities of black garlic via RSM were expected to be at $89.68^{\circ}C$ for 9.79 hrs with a 55.72% ethanol concentration. The actual values were 96.4-114.8% of the predicted values.
The study was conducted to identify the relationship between the e-cigarette smoking rate and demographic factors, health status and health behavior factors within the last one month of adults over 19 years of age, and to provide basic data for cessation education and health policy establishment to reduce the e-cigarette smoking rate. This is a secondary data analysis study using survey data of 22,908 people over the age of 19 who participated in the 2018 Community Health Survey in Seoul and the SAS program was used for analysis. Smoking rate of e-cigarettes within the last one month was 4.5%, and as a result of multivariate logistic regression analysis, factors related to e-cigarette smoking were sex, age, education level, occupation and general cigarette smoking status. In order to prevent e-cigarette smoking and reduce the smoking rate, education should first be given to professionals with educational backgrounds of college graduates or higher to inform the dangers of e-cigarettes. In addition, public relations and campaigns to inform the general public that e-cigarette smoking is no different from general cigarette smoking are required, and deployment of the same anti-smoking policy that treats e-cigarettes as general cigarettes is required.
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