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Isolation and Characterization of a Weizmannia coagulans Bacteriophage Youna2 and Its Endolysin PlyYouna2

  • Bokyung Son;Youna Kim;Booyoung Yu;Minsuk Kong
    • Journal of Microbiology and Biotechnology
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    • v.33 no.8
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    • pp.1050-1056
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    • 2023
  • Weizmannia coagulans (formerly Bacillus coagulans) is Gram-positive, and spore-forming bacteria causing food spoilage, especially in acidic canned food products. To control W. coagulans, we isolated a bacteriophage Youna2 from a sewage sludge sample. Morphological analysis revealed that phage Youna2 belongs to the Siphoviridae family with a non-contractile and flexible tail. Youna2 has 52,903 bp double-stranded DNA containing 61 open reading frames. There are no lysogeny-related genes, suggesting that Youna2 is a virulent phage. plyYouna2, a putative endolysin gene was identified in the genome of Youna2 and predicted to be composed of a N-acetylmuramoyl-L-alanine amidase domain (PF01520) at the N-terminus and unknown function DUF5776 domain (PF19087) at the C-terminus. While phage Youna2 has a narrow host range, infecting only certain strains of W. coagulans, PlyYouna2 exhibited a broad antimicrobial spectrum beyond the Bacillus genus. Interestingly, PlyYouna2 can lyse Gram-negative bacteria such as Escherichia coli, Yersinia enterocolitica, Pseudomonas putida and Cronobacter sakazakii without other additives to destabilize bacterial outer membrane. To the best of our knowledge, Youna2 is the first W. coagulans-infecting phage and we speculate its endolysin PlyYouna2 can provide the basis for the development of a novel biocontrol agent against various foodborne pathogens.

Antigenotoxic and Anticarcinogenic Effects of Styela plicata (오만둥이(Styela plicata)의 항유전독성 및 대장암 억제효과에 관한 연구)

  • Seo, Bo-Young;Kim, Jung-Mi;Lee, Seung-Cheol;Park, Eun-Ju
    • Journal of the Korean Society of Food Science and Nutrition
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    • v.38 no.7
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    • pp.839-845
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    • 2009
  • Colorectal cancer is the third most common malignant neoplasm in the world. Much attention has been focused on reducing colon cancer risk through medical properties of natural compound that could act as anticarcinogens. In this study, we evaluated the antioxidant and antigenotoxic effects of Styela plicata (S. plicata) from in vitro experiments. S. plicata extracts showed antioxidant activity measured by TRAP assay and antigenotoxic effect in $200{\mu}M$ $H_2O_2$ induced DNA damage in human leukocytes. Especially, freeze-dried S. plicata extracted with methanol showed the highest level of TRAP (0.225 mM) and inhibition of DNA damage (66.8%). Additionally we observed the effect of S. plicata on the formation of aberrant crypt foci (ACF) induced by dimethylhydrazine (DMH) and DMH induced DNA damage (by comet assay) in male SD rats. The animals were divided into three groups and fed high-fat and low fiber diet (100 g lard+20 g cellulose/kg diet) without (normal control and DMH control) or with a 3% (w/w) of lyophilized S. plicata powder (DMH+S. plicata). One week after beginning the diets, rats were treated with DMH (30 mg/kg, s.c.) for 6 weeks except for normal control group, which was treated saline instead; dietary treatments were continued for the entire experiment. Nine weeks after DMH injection, administration of S. plicata resulted in reduction of ACF numbers, to 82.7% of the carcinogen control value ($7.67{\pm}2.04$ vs. $1.33{\pm}0.53$: p<0.01). S. plicata supplementation induced antigenotoxic effect on DMH-induced DNA damage in the blood cell (% tail intensity: $6.79{\pm}0.26$ vs. $6.13{\pm}0.22$). These data indicate that S. plicata extract has antigenotoxic and anticarcinogenic effects from in vitro experiments and S. plicata exerts a protective effect on the process of colon carcinogenesis, possibly by suppressing the DMH-induced DNA damage in blood cell and the development of preneoplastic lesions in colon.

Inhibitory effects of Prunus persica flower extracts on UV-induced skin damage

  • Lee, Kang-Tae;Yoo, Young-Kyoung;Kim, Sung-Woo;Jeong, Ji-Hean;Jo, Byoung-Kee;Kim, Young-Ha;Yang, Hye-Eum;Heo, Moon-Young;Kim, Hyun-Pyo
    • Journal of the Society of Cosmetic Scientists of Korea
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    • v.27 no.1
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    • pp.73-81
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    • 2001
  • For an attempt to develop safe materials protecting UV-induced skin damage, plant extracts were evaluated for their antioxidative and free radical scavenging activities. From the results of these screening procedures, the ethanol extract of the flowers of Prunus persica was selected for further study. It was found that Prunus persica (50-200 $\mu\textrm{g}$/㎖) inhibited UVB-induced DNA damage measured by tail moment in the Single Cell Gel Electrophoresis(COMET assay) and inhibited UV-induced lipid peroxidation, expecially against UVB-induced peroxidation at higher than 10 $\mu\textrm{g}$/㎖. Also P.persica(100∼1,000 $\mu\textrm{g}$/㎖) inhibited the amount of $\^$14/C-arachidonic acid metabolites release from UVB-irradiated keratinocytes and it possessed the protective activity against UV-induced cytotoxicity of keratinocytes. All these results indicate that the flowers of P. persica extract may be beneficial for protection UV-induced skin damage when topically applied.

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Cloning and Sequence Analysis of the Full-length cDNA of Coxsackievirus B3 Isolated in Korea (한국에서 분리된 콕사키 바이러스 B3 cDNA의 클로닝 및 전체 염기서열 분석)

  • Chung, Yoon-Seok;Kim, Ki-Soon;Park, Jeong-Koo;Lee, Yoon-Sung;Shin, Soo-Youn;Cheon, Doo-Seong;Jee, Young-Mee;Kim, Moon-Bo;Na, Byoung-Kuk;Yoon, Jae-Deuk;Lee, Kwang-Ho;Song, Chul-Yong
    • The Journal of Korean Society of Virology
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    • v.30 no.1
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    • pp.71-81
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    • 2000
  • We have determined and analyzed the full-length cDNA sequence of a coxsackievirus B3 (CVB3) Korean isolate (CVB3-Korea/97) which has been known as a general human pathogen. The whole genome contains 7,400 nucleotides and has a single large open reading frame with 6,555 nucleotides that encodes a potential polyprotein precursor of 2,185 amino acids. The genome also contains a 5' non-coding region (NCR) of 741 bases and a 3' NCR of 104 bases followed by poly(A) tail. Sequence homologies of nucleotides and deduced amino acids between the CVB3-Korea/97 strain and the prototype (Nancy strain) were 81.7% and 91.5%, respectively. The genes encoding the functional proteins including viral protease and RNA dependent RNA polymerase showed higher homology than those encoding the structural proteins. We have further analyzed the sequences of 5' NCR, VP1 and VP2 of CVB3-Korea/97, which are known as cardiovirulent determining factors at the nucleotide and amino acid levels. Although the CVB 3-Korea/97 strain was isolated from an aseptic meningitis patient without cardiomyopathy, its 234th nucleotide and 165th amino acid were uracil and Asn as same as those of other cardiovirulent strains one. However, the 155th amino acid of VP1, which closely associated with cardiovirulence, was replaced with $Arg^{155}$ by single nucleotide substitution from $A^{2916}$ to $T^{2916}$. Moreover, additional amino acid substitutions were observed in the flanking region of $Asp^{155}$. Taken together, amino acid(s) substitution in VP1 may playa critical role in determining cardiovirulence of the CVB3-Korea/97 strain rather than individual nucleotide replacements in the 5' NCR and/or an amino acid substitution in VP2.

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Complete Mitochondrial Genome Sequences of Chinese Indigenous Sheep with Different Tail Types and an Analysis of Phylogenetic Evolution in Domestic Sheep

  • Fan, Hongying;Zhao, Fuping;Zhu, Caiye;Li, Fadi;Liu, Jidong;Zhang, Li;Wei, Caihong;Du, Lixin
    • Asian-Australasian Journal of Animal Sciences
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    • v.29 no.5
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    • pp.631-639
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    • 2016
  • China has a long history of sheep (Ovis aries [O. aries]) breeding and an abundance of sheep genetic resources. Knowledge of the complete O. aries mitogenome should facilitate the study of the evolutionary history of the species. Therefore, the complete mitogenome of O. aries was sequenced and annotated. In order to characterize the mitogenomes of 3 Chinese sheep breeds (Altay sheep [AL], Shandong large-tailed sheep [SD], and small-tailed Hulun Buir sheep [sHL]), 19 sets of primers were employed to amplify contiguous, overlapping segments of the complete mitochondrial DNA (mtDNA) sequence of each breed. The sizes of the complete mitochondrial genomes of the sHL, AL, and SD breeds were 16,617 bp, 16,613 bp, and 16,613 bp, respectively. The mitochondrial genomes were deposited in the GenBank database with accession numbers KP702285 (AL sheep), KP981378 (SD sheep), and KP981380 (sHL sheep) respectively. The organization of the 3 analyzed sheep mitochondrial genomes was similar, with each consisting of 22 tRNA genes, 2 rRNA genes (12S rRNA and 16S rRNA), 13 protein-coding genes, and 1 control region (D-loop). The NADH dehydrogenase subunit 6 (ND6) and 8 tRNA genes were encoded on the light strand, whereas the rest of the mitochondrial genes were encoded on the heavy strand. The nucleotide skewness of the coding strands of the 3 analyzed mitogenomes was biased toward A and T. We constructed a phylogenetic tree using the complete mitogenomes of each type of sheep to allow us to understand the genetic relationships between Chinese breeds of O. aries and those developed and utilized in other countries. Our findings provide important information regarding the O. aries mitogenome and the evolutionary history of O. aries inside and outside China. In addition, our results provide a foundation for further exploration of the taxonomic status of O. aries.

Physical Characteristics and Microsatellite Polymorphisms in Miryang Native Dogs (밀양지방 토종개의 형태학적 특징 및 유전적 다양성 연구)

  • Cho, Byung-Wook;Cho, Gil-Jae
    • Journal of Life Science
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    • v.16 no.4
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    • pp.626-631
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    • 2006
  • This study was carried out to investigate the physical characteristics (height, body lenght, chest depth, head type, ear type, body color, eye type and tail type) and genetic diversity using 15 microsatellite DNA markers (PEZ 1, 5, 8, 10, 11, 12, 13, 15, 16, 17, 20, 21, FHC 2010, FHC 2054 and FHC 2079) in 44 random Miryang native dogs(6 months${\sim}$12 years old). The height, body lenght, and chest depth of Miryang native dogs were 43-55 cm(mean 49.5 cm), 45-60 cm(mean 54.3 cm), and 50-64 cm(mean 57.9 cm), respectively. Miryang native dog was medium sized. The head and eye type were reverse-triangle(100%), triangle (90.9%) and newborn moon(9.1%), respectively. Most of body color had white coat color(93.2%), light pink tongue color(100%), light black anal color(90,9%) and pink claw color(100%). The ear type showed erect ear(100%), and half-curled(56.8%), upward(34.1%), curled(9.1%) in tail type, respectively. Number of alleles observed at a single locus ranged from 2 (PEZ 21 and FHC 2010) to 14 (PEZ 13), with average number of alleles per locus of 6.13. The expected heterozygosities of 15 microsatellite loci were estimated based on gene frequencies. The highest expected heterozygosity, 0.863 was estimated in PEZ 13 locus and the lowest, 0.455 in PEZ 21 and FHC 2010 locus. And the mean expected heterozygosity of 15 microsatellite markers was calculated as 0.635. Polymorphic information content (PIC) values were ranged from 0.348 (PEZ 21 and FHC 2010) to 0.837 (PEZ 13), and the mean PIC value was calculated as 0.570. Of the 15 markers, PEZ 10, PEZ 13, PEZ 17 and FHC 2054 loci have relatively high PIC value (> 0.7) in Miryang native dog. In order to determine the efficieney of parentage control, exclusion probabilities (PE) were calculated for each allele. The highest PE 1 and PE 2 in PEZ 13 locus was caculated to 0.548 and 0.710, respectively. And the total exclusion power in PE 1 and PE 2 was calculated to 0.9895 and 0.9996, respectively. These results can give basic information for perservation and research in Miryang native dog, and phylogenetic relationships of the Korean native dog and Asian dog breeds.

Neuroprotective Effects of Cheongnoemyeongsin-hwan against Hydrogen Peroxide-induced DNA Damage and Apoptosis in Human Neuronal-Derived SH-SY5Y Cells (인체 신경세포에서 청뇌명신환(淸腦明神丸)의 산화적 스트레스에 대한 세포보호 효과)

  • Pi, Guk Hyun;Hwang, Won Deuk
    • Herbal Formula Science
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    • v.25 no.1
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    • pp.51-68
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    • 2017
  • Objectives : Oxidative stress due to excessive accumulation of reactive oxygen species (ROS) is one of the risk factors for the development of several chronic diseases, including neurodegenerative diseases. Methods : In the present study, we investigated the protective effects of cheongnoemyeongsin-hwan (CNMSH) against oxidative stress‑induced cellular damage and elucidated the underlying mechanisms in neuronal-derived SH-SY5Y cells. Results : Our results revealed that treatment with CNMSH prior to hydrogen peroxide (H2O2) exposure significantly increased the SH-SY5Y cell viability, indicating that the exposure of the SH-SY5Y cells to CNMSH conferred a protective effect against oxidative stress. CNMSH also effectively attenuated H2O2‑induced comet tail formation, and decreased the phosphorylation levels of the histone ${\gamma}H2AX$, as well as the number of apoptotic bodies and Annexin V‑positive cells. In addition, CNMSH exhibited scavenging activity against intracellular ROS generation and restored the mitochondria membrane potential (MMP) loss that were induced by H2O2, suggesting that CNMSH prevents H2O2‑induced DNA damage and cell apoptosis. Moreover, H2O2 enhanced the cleavage of caspase-3 and degradation of poly (ADP-ribose)-polymerase, a typical substrate protein of activated caspase-3, as well as DNA fragmentation; however, these events were almost totally reversed by pretreatment with CNMSH. Furthermore, CNMSH increased the levels of heme oxygenase-1 (HO-1), which is a potent antioxidant enzyme, associated with the induction of nuclear factor-erythroid 2-related factor 2 (Nrf2). According to our data, CNMSH is able to protect SH-SY5Y cells from H2O2-induced apoptosis throughout blocking cellular damage related to oxidative stress through a mechanism that would affect ROS elimination and activating Nrf2/HO-1 signaling pathway. Conclusions : Therefore, we believed that CNMSH may potentially serve as an agent for the treatment and prevention of neurodegenerative diseases caused by oxidative stress.

Antidiabetic Effect of Korean Red Ginseng by Puffing Process in Streptozotocin-Induced Diabetic Rats (Streptozotocin 유발 당뇨쥐에서 팽화가공 처리한 홍삼의 항당뇨 효과)

  • Kim, Shin-Hee;Kang, Ju-Seop;Lee, Sang-Jun;Chung, Young-Jin
    • Journal of the Korean Society of Food Science and Nutrition
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    • v.37 no.6
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    • pp.701-707
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    • 2008
  • Antidiabetic effect of Korean red ginseng (RG) processed by puffing in streptozotocin (STZ)-induced diabetic (DM) rats was investigated. Five week-old SD rats were divided into four groups; normal control (NC) group, DM group, red ginseng (RG) group and puffed red ginseng (PG) group. The RG and PG groups were orally provided with RG or PG dissolved in water (500 mg/kg) respectively for seven weeks after single injection of STZ (50 mg/kg, i.v.) followed by identification of DM. NC group received saline vehicle instead of STZ. At the end of feeding of RG or PG, the changes of fasting blood glucose, serum insulin and amylase level and serum lipid profiles were evaluated. Also, oral glucose tolerance test (OGTT), comet assay and histopathological examination were performed. At 7th week, the fasting blood glucose levels of the RG and PG groups were reduced compared to the DM group by 11.54% and 20.22%, respectively. The result of OGTT did not show significant differences among DM and two red ginseng groups. While serum insulin and TG levels were predominantly improved in PG group (p<0.05), serum amylase level was increased in RG group. Alkaline comet assay for checking the oxidative damage of DNA showed that TL (tail length, ${\mu}m$) and TM (tail moment) in the blood lymphocyte of PG group significantly decreased in contrast with DM group. Histopathological results of pancreas showed that destruction of exocrine as well as endocrine might be cured by the administration of RG and PG. These results suggest that PG could exert more protection against STZ-induced toxicity than RG group.

Anticancer and Immune Effects of Chungpae-tang on the Metastasis of Lung Cancer Cell (청폐탕(淸肺湯)의 실험적 폐전이암에 대한 항암 및 면역효과에 관한 연구)

  • Lee Dong-Ju;Kim Myung-Dong;Kim Young-Sam;Yoo Yeong-Min;Lee Seon-Goo
    • Journal of Physiology & Pathology in Korean Medicine
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    • v.19 no.5
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    • pp.1281-1291
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    • 2005
  • Chungpae-tang is suggested to have the antitumor activity on lung cancer. This study was peformed to investigate apoptotic effect in vitro and antitumor effect and immune response after injection of B16-F10 melanoma cells and Chungpae-tang into a tail vein of C57BL/6 mice and administratition of Chungpae-tang in A549 human lung cancer cell line in vivo, respectively. Experimental studies were obtained by measuring the median survival time and cytokine expression through RT-PCR, and ELISA assay. The results were summarized as follows: 5 mg/ml of Chungpae-tang causing DNA fragmentation, caspase-3 enzyme activation, PARP fragmentation, and cytochrome c release, suggested that Chungpae-tang has in vitro apoptotic effect in A549 human lung cancer cell line in the apoptosis-induced experiment. The median survival time of the Chungpae-tang treated group was 21 days and that of control group was 22 days, suggesting that the median survival time between the Chungpae-tang treated group and the control group was not significant. Cytokine expression between the Chungpae-fang treated group and the control group was noticeable, but was not significant in the RT-PCR. In the ELISA assay, IL-2 productivity in the Chungpae-tang treated group was to increase more than that in the normal group (p<0.05) and was no significant between the Chungpae-tang treated group and the control group. $INF-\gamma$ productivity of the control group decreased more than that of the normal group (p<0.05) and that of the Chungpae-tang-treated group increased more than that of the control group (p<0.05). IL-12 productivity of the control group increased more than that of the normal group (p<0.05) and that of the Chungpae-tang-treated group decreased more than that of the control group (p<0.05) and the normal group. IL-4 productivity of the Chungpae-tang-treated group increased more than that of the normal group and the control group (p<0.05). IL-10 productivity of the Chungpae-tang-treated group increased more than that of the normal group and the control group (p<0.05). Accordingly the results show Chungpae-tang could induce apoptosis in A549 human lung cancer cell line and bring to antitumor effect and immune response against injection of B16-F10 melanoma cells into a tail vein of C57BL/6 mice but it needs more research on the precise mechanism of such effects.

Effects of Bacillus Calmette-Guerin (BCG) on the DNA Synthesis of Mouse Duodenal Mucosal Epithelial Cells Inoculated with Ehrlich Carcinoma Cells (Bacillus Calmette-Guerin (BCG) 투여가 Ehrlich 종양세포를 이식한 생쥐 샘창자 상피세포의 DNA합성에 미치는 영향)

  • Ko, Jeong-Sik;Kim, Heung-No;Park, Kyung-Ho;Park, Dae-Kyoon;Kim, Duk-Soo
    • Applied Microscopy
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    • v.40 no.4
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    • pp.193-200
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    • 2010
  • This experiment was performed to evaluate the morphological responses of the duodenal epithelial cells of the mouse, inoculated with Ehrlich carcinoma cells in the inguinal area, following administration of Bacillus Calmette-Guerin (BCG). In the experimental groups, each mouse was inoculated with $1{\times}10^7$ Ehrlich carcinoma cells subcutaneously in the inguinal area. From next day after inoculations, 0.2 mL of saline or BCG (0.5 mL/25 g B.W.: $0.03{\times}10^8\sim0.32{\times}10^8$ CFU) were injected subcutaneously to the animals every other day, respectively. The day following the 7th injection of saline or BCG, each mouse was injected with a single dose of $0.7{\mu}Ci$/g of methyl-$^3H$-thymidine (25 Ci/mmol, Amersham Lab, England) through tail vein. Seventy minutes after the thymidine injection, animals were sacrificed, and duodenal tissues were taken and fixed in 10% neutral formalin. Deparaffinized sections were coated with autoradiographic emulsion EM-1 (Amersham Lab, England) in a dark room and dried and were placed in a light-tight box. The number of labeled epithelial cells in the duodenal mucosae (mean number of labeled epithelial cells per 3.5 mm length of mucosa) were observed and calculated. On the light microscopic study, duodenal mucosae had no severe morphological changes following the injection of BCG. In the tumor control and BCG treated groups, a number of small lymphocytes and eosinophile leucocytes are slightly increased as compared with those of the normal control ones. On the autoradiographic study, number of the labeled cells of normal control, tumor control and BCG-treated mice were 632.3 (${\pm}14.47$), 761.7 (${\pm}27.65$) and 505.0 (${\pm}17.09$) respectively. From the above results, BCG may suppress the DNA synthesis of the duodenal epithelial cells, but does not results severe structural defect on the duodenal mucosae. And it is suggested that BCG may greatly improve the anticancer therapy on certain kind of cancer.