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Antibacterial Activity and Probiotic Potential of Lactobacillus plantarum HKN01: A New Insight into the Morphological Changes of Antibacterial Compound-Treated Escherichia coli by Electron Microscopy

  • Sharafi, Hakimeh;Maleki, Hadi;Ahmadian, Gholamreza;Zahiri, Hossein Shahbani;Sajedinejad, Neda;Houshmand, Behzad;Vali, Hojatollah;Noghabi, Kambiz Akbari
    • Journal of Microbiology and Biotechnology
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    • v.23 no.2
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    • pp.225-236
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    • 2013
  • Among several bacteria examined, an antibacterial-producing Lactobacillus strain with probiotic characteristics was selected and identified based on 16S rRNA gene sequencing. Subsequent purification and mode of action of the antibacterial compounds on target cells including E. coli were investigated. Maximum production of the antibacterial compound was recorded at 18 h incubation at $30^{\circ}C$. Interestingly, antibacterial activity remained unchanged after heating at $121^{\circ}C$ for 45 min, 24 h storage in temperature range of $70^{\circ}C$ to room temperature, and 15 min exposure to UV light, and it was stable in the pH of range 2-10. The active compounds were inactivated by proteolytic enzymes, indicating their proteinaceous nature, and, therefore, referred to as bacteriocin-like inhibitory substances. Isolation and partial purification of the effective agent was done by performing ammonium sulfate precipitation and gel filtration chromatography. The molecular mass of the GFC-purified active compound (~3 kDa) was determined by Tris-Tricine SDS-PAGE. To predict the mechanisms of action, transmission electron microscopy (TEM) analysis of ultrathin sections of E. coli before and after antibacterial treatment was carried out. TEM analysis of antibacterial compounds-treated E. coli demonstrated that the completely altered bacteria appear much darker compared with the less altered bacteria, suggesting a change in the cytoplasmic composition. There were also some membrane-bound convoluted structures visible within the completely altered bacteria, which could be attributed to the response of the E. coli to the treatment with the antibacterial compound. According to the in vivo experiments oral administration of L. plantarum HKN01 resulted in recovery of infected BALB/c mice with Salmonella enterica ser. Typhimurium.

Study of the Expression of FasL and of Apoptosis in Gastric Epithelial Dysplasia and Gastric Adenocarcinomas (위상피이형성과 위암종에서 FasL의 발현 및 Apoptosis에 관한 연구)

  • Park Gun Uk;Han Sang Young;Lee Jong Hun;Keum Dong Joo;Roh Myung Hwan;Choi Seok Ryeol;Kim Jong Seong;Roh Mee Sook
    • Journal of Gastric Cancer
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    • v.1 no.2
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    • pp.83-91
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    • 2001
  • Purpose: This study was to observe whether the apoptotic function of tumor-infiltrating lymphocytes (TIL) is induced in human gastric epithelial dysplasia and gastric adenocarcinoma according to the role of FasL expression. Materials and Methods: A total of 56 gastric epithelial dysplasia and gastric adenocarcinoma patients were enrolled in this study: 9 cases of gastric epithelial dysplasia, 18 cases of early gastric carcinomas (EGC) and 29 cases of advanced gastric carcinomas (AGC). Immunohistochemical staining was performed for FasL and CD45, and the terminal deoxynucleotidyl transferase mediated dUTP nick end labelling (TUNEL) method was used to detect cell death in tumor-infiltrating lymphocytes. Results: 1) Positive reactions of FasL to neoplastic cells were $88.9\%$ (8/9) in gastric epithelial dysplasia, $83.3\%$ (15/18) in EGC, and $75.9\%$ (22/29) in AGC. 2) Expression of TIL was decreased in the FasL positive region and was increased in the FasL negative region, and significant expression of TIL was observed in the AGC group (P=0.001). 3) Expression of apoptotic TIL was very similar to the FasL expression, and $100\%$ expression was observed in gastric epithelial dysplasia group. 4) Expression of apoptotic TIL was increased in the FasL positive region and decreased in the FasL negative region, and significant apoptotic expression was observed in the gastric epithelial dysplasia and EGC groups (P=0.0420, P=0.0263, respectively). Conclusion: These results suggest that FasL is a prevalent mediator of immune privilege in epithelial dysplasia and cancer of the stomach.

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Fermentation characteristics of persimmon wine by the mixed culture of Pichia anomala JK04 and Saccharomyces cerevisiae Fermivin cells (Pichia anomala JK04와 Saccharomyces cerevisiae Fermivin의 혼합발효에 의한 감와인의 발효 특성)

  • Kwon, Soo-Jin;Choi, Jun-Su;Yeo, Soo-Hwan;Park, Heui-Dong
    • Food Science and Preservation
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    • v.22 no.5
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    • pp.768-777
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    • 2015
  • Persimmon contains high levels of vitamins and phenolic compounds, as well as soluble solids, necessary for the fermentation of persimmon wine. Co-fermentation of persimmon wine was carried out using a mixed culture of Pichia anomala JK04, a Korean indigenous yeast that improves wine quality and flavor, and Saccharomyces cerevisiae Fermivin, an industrial wine yeast, in the following ratios: 9:1 (v/v), 5:5 (v/v), 1:9 (v/v) and 0:10 (v/v). During fermentation, the alcohol contents increased more slowly in samples of mixed culture than in samples of the single culture of S. cerevisiae Fermivin. The alcohol contents of all samples reached 12~13% (v/v) after 15 days. All samples of the mixed culture showed greater variety in flavor and taste than S. cerevisiae Fermivin only. In the sensory evaluation, mixed culture samples had higher scores in terms of flavor and overall preference than the single culture samples. Therefore, P. anomala JK04 is thought to improve the wine flavor of Korean domestic persimmon wine.

Preparation and Quality Characteristics of Yogurt Added with Cultured Ginseng (배양인삼 요구르트의 제조 및 품질 특성)

  • Lee, In-Seon;Paek, Kee-Yoeup
    • Korean Journal of Food Science and Technology
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    • v.35 no.2
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    • pp.235-241
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    • 2003
  • Yogurt base was prepared from whole and skin milk added with $0.5{\sim}2.0%\;(w/v)$ of cultured ginseng ethanol extract and fermented with lactic acid bacteria (Streptococcos thermophilus : Lactobacillus bulgaricus=1:1) at $37^{\circ}C$ for $24{\sim}30\;h$. Quality characteristics of the yogurt were evaluated in terms of acid production, number of viable cells, viscosity, and sensory property during lactic acid fermentation. Total contents of amino acids and some organic acids were analyzed. Addition of cultured ginseng extract stimulated the growth of lactic acid bacteria, and enhanced acid production and viscosity of yogurt. Total contents of amino acids of 0.5% cultured ginseng-added yogurt were higher than control group before fermentation, whereas glutamic acid, cysteine, valine, and phenyalaine contents increased after 30 h incubation. Contents of lactic, citric, and formic acids of 0.5% cultured ginseng-added yogurt increased during fermentation for 24 h. whereas decreased thereafter. Sensory scores of yogurts added with 0.5 and 1 % cultured ginseng extract were significantly higher than other groups in taste and overall acceptability. When cultured ginseng yogurt was kept at $5^{\circ}C$ for 15 days, its quality-keeping property was relatively good.

Genetic Stability of Magnaporthe oryzae during Successive Passages through Rice Plants and on Artificial Medium

  • Park, Sook-Young;Chi, Myoung-Hwan;Milgroom, Michael G.;Kim, Hyo-Jung;Han, Seong-Sook;Kang, Seog-Chan;Lee, Yong-Hwan
    • The Plant Pathology Journal
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    • v.26 no.4
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    • pp.313-320
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    • 2010
  • Genetic instability of the rice blast fungus Magnaporthe oryzae has been suggested as a major factor underlying the rapid breakdown of host resistance in the field. However, little information is available on the mechanism of genetic instability. In this study, we assessed the stability of repetitive DNA elements and several key phenotypic traits important for pathogenesis after serially transferring two isolates though rice plants and an artificial medium. Using isolate 70-15, we obtained a total of 176 single-spore isolates from 10 successive rounds of culturing on artificial medium. Another 20 isolates were obtained from germ tubes formed at the basal and apical cells of 10 three-celled conidia. Additionally, 60 isolates were obtained from isolate KJ201 after serial transfers through rice plants and an artificial medium. No apparent differences in phenotypes, including mycelial growth, conidial morphologies, conidiation, conidial germination, appressorium formation, and virulence, or in DNA fingerprints using MGR586, MAGGY, Pot2, LINE, MG-SINE and PWL2 as probes were observed among isolates from the same parent isolate. Southern hybridization and sequence analysis of two avirulence genes, AVR-Pita1 and AVR-Pikm, showed that both genes were also maintained stably during 10 successive generations on medium and plants. However, one reversible loss of restriction fragments was found in the telomere-linked helicase gene (TLH1) family, suggesting some telomere regions may be more unstable than the rest of the genome. Taken together, our results suggest that phenotype and genotype of M. oryzae isolates do not noticeably change, at least up to 10 successive generations on a cultural medium and in host plants.

Experimental Assessment of Hemostatic Agents: Comparison with New Developed Chitosan-Based Material (신개발 키토산 제재의 지혈 효과에 대한 비교)

  • Cho, Young-Kyoo;Lee, Sang-Yun;Kim, Tae-Jung;Lim, Hyun-Ju;Oh, Eun-Jung;Lee, Soo-Bok;Choi, Kang-Young;Yang, Jung-Dug;Cho, Byung-Chae;Chung, Ho-Yun
    • Archives of Plastic Surgery
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    • v.38 no.4
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    • pp.369-375
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    • 2011
  • Purpose: Many hemostatic agents and dressings have been tested with variable degree of success. Chitosan has a positive charge, it attracts red blood cells, which have a negative charge. Our goal is to test the efficacy of new developed chitosan-based hemostatic materials in providing durable hemostasis in a high-flow arterial wound model. Methods: We compared each group with SD rats motality tests and in vitro blood compatibility test by blood clotting index (BCI). We devided the SD rats into 6 groups (N =15) by type of hemostatic agents. A: 100% nonwoven chitosan (degree of the deacetylation: 90%). B: 50% N-acetylation on nonwoven of chitosan gel (degree of the deacetylation: 50%). C: 60% N-acetylation on nonwoven of chitosan ge (degree of the deacetylation: 40%)l. D: Cutanplast$^{(R)}$. E: HemCon$^{(R)}$ F: Gauze. In vivo test, a proximal arterial injury was created in unilateral femoral arteries of 90 anesthetized SD rats. Each materials was made same size and thickness then applied to the injury site for 3 minutes. In vitro test, we compared each group with BCI in human blood. Results: In vivo test, group A showed lower motality rate of 46% than any other groups, Group B and C showed lower motality rate of 60% than group D and E's motality rate of 66%. In vitro test, BCI of group A ($30.6{\pm}1.2$) and B ($29.3{\pm}1.0$) were showed nearly about group D ($29.1{\pm}1.8$) and E ($27.4{\pm}1.6$). Group C ($37.1{\pm}2.0$) showed higher BCI than group A and B, it means group C decreased blood clotting. Conclusion: In conclusion, this study suggests a newly developed chitosan-based hemostatic materials induced durable hemostasis and increased blood clotting, and are considered as effective biologic hemostatic agents.

Effects of Dry Roasting on the Vitamin E Content and Microstructure of Peanut (Arachis hypogaea)

  • Eitenmiller, Ronald R;Choi, Sung-Gil;Chun, Jiyeon
    • Journal of agriculture & life science
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    • v.45 no.4
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    • pp.121-133
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    • 2011
  • Effects of roasting on vitamin E content, color, microstructure and moisture of peanuts, and vitamin E content in peanut oils prepared from the roasted peanuts were investigated. Runner-type peanuts were roasted at 140, 150, and $160^{\circ}C$ for 10-20 min. As roasting temperature and time increased, the CIELAB $L^*$ value of peanuts decreased while $a^*$ and $b^*$ values increased, resulting in formation of the golden brown color of roasted peanuts. Moisture ratio (M/Mo) and color $b^*$ value of peanuts roasted at 140 to $160^{\circ}C$ showed a correlation of $b^*=21.61\;(M/Mo)^2-40.62\;(M/Mo)+34.12$ ($R^2=0.9123$). Overall changes in the tocopherol contents of peanuts and peanut oils were significantly affected by roasting temperature and time (p<0.05). Roasting at $140^{\circ}C$ caused a slight increase in the levels of tocopherols of peanuts over roasting time up to 20 min (p<0.05). There was no significant change in the tocopherol levels of peanuts during roasting at $150^{\circ}C$ for 20 min (p>0.05). At $160^{\circ}C$, the levels of tocopherols significantly decreased during the initial 10 min of roasting (p<0.05) while there was no extended loss after 10 min, resulting in about 5, 12, 20, and 10% losses of ${\alpha}$-, ${\beta}$-, ${\gamma}$- and ${\delta}$-T, respectively. After 20 min, total tocopherols decreased by 18%. However, tocopherol contents of pressed peanut oils significantly decreased at all roasting temperatures (p<0.05). After roasting peanuts at $160^{\circ}C$ for 20 min, about 84% of initial ${\alpha}$-T in peanut oils was retained. ${\alpha}$-T was the most stable to roasting while ${\gamma}$-T was the least. Swollen epidermal cells on the inner surface and broken cell walls of parenchyma tissue of peanut cotyledon were observed in peanuts after roasting at $160^{\circ}C$ for 15 min. Severe changes in microstructure of peanut by roasting would contribute to vitamin E stability because of exposure of oil droplets in peanuts to oxygen.

Effect of Astragali Radix Pharmacocupuncture for Wound Healing of Full-Thickness Skin Wound in Mouse (황기약침이 전층피부손상 마우스의 치유에 미치는 영향)

  • Hong, Chang-Ho;Kang, Kyung-Hwa;Yoon, Hwa-Jung;Yoon, Hyoun-Min;Song, Choon-Ho;Kim, Cheol-Hong
    • The Journal of Korean Medicine Ophthalmology and Otolaryngology and Dermatology
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    • v.31 no.1
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    • pp.22-31
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    • 2018
  • Objectives : In this study, we investigated whether Astragali Radix Pharmacocupuncture (ARP) has an effective on the full thickness defect wound healing process of mouse. Methods : A total of 50 mice (ICR mouse, 7 week-old male) were divided into control group and ARP group. A single full thickness skin defect was made on the dorsal side of the each mouse using an 8mm diameter biopsy punch. Control group were treated with 0.2㎖ saline and ARP group were treat with 0.2ml ARP at 8 points around the wound every three days total 4 times during the experimental period. The change in wound size, contraction rate, healing rate, and epithelization rate was measured by digital images taken on days 3, 6, 9, and 13, and evaluated using a digital image analysis program. Tissues were collected for histological analysis, RT-PCR, and Western blot on days 3, 6, 9, and 15. Results : The results are as follows. ARP group accelerated the rate of wound contraction, wound healing and epithelization compared to the control group. ARP group showed the decrease of inflammatory cells in early inflammatory phase compared to the control group. ARP group upregulated PECAM-1 mRNA and protein expression compared to the control group. ARP group inhibited the scar width and area compared to the control group. Conclusions: ARP showed positive effects on wound healing through the inhibition of inflammatory reaction and increase of PECAM-1 expression related to the wound healing process.

Dabigatran Toxicity Secondary to Acute Kidney Injury (급성신손상으로 인해 발생한 dabigatran 독성)

  • Moon, Hyoung Ho;Lee, Seung Eun;Oh, Dong Jun;Jo, Hee Bum;Kwon, Ki Hwan;Kim, Yoon Jin;Kim, Kyung Soo;Shin, Sung Joon
    • Journal of The Korean Society of Clinical Toxicology
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    • v.12 no.2
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    • pp.92-96
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    • 2014
  • Dabigatran is the first oral direct thrombin inhibitor approved by the US Food and Drug Administration (FDA) for prevention of stroke and systemic embolism in patients with nonvalvular atrial fibrillation. Because dabigatran is excreted mainly by the kidneys, serum levels of dabigatran can be elevated to a supratherapeutic range in patients with renal failure, predisposing to emergent bleeding. We describe the case of a 66-year-old man taking dabigatran 150 mg twice daily for atrial fibrillation and cerebral infarction who presented with hematochezia and disseminated intravascular coagulation. Laboratory evaluation showed a hemoglobin level of 6.3 g/dL, platelets of $138,000/mm^3$, activated partial thromboplastin time (aPTT) of 10 s, and an international normalized ratio (INR) of 8.17. Colonoscopy showed a bleeding anal fissure. Hemostasis was provided by hemoclips and packed red blood cells and fresh frozen plasma were transfused. Since then, there was no further hematochezia, however, bleeding including oral mucosal bleeding, hematuria, and intravenous site bleeding persisted. At presentation, his serum creatinine was 4.96 mg/dL (baseline creatinine, 0.9 mg/dL). Dabigatran toxicity secondary to acute kidney injury was presumed. Because acute kidney injury of unknown cause was progressing after admission, he was treated with hemodialysis. Fresh frozen plasma transfusion was provided with hemodialysis. At 15 days from admission, there was no further bleeding, and laboratory values, including hemoglobin, partial thromboplastin time, and prothrombin time were normalized. He was discharged without bleeding. After 2 months, he undergoes dialysis three times per week and no recurrence of bleeding has been observed.

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Peri-implantitis, systemic inflammation, and dyslipidemia: a cross-sectional biochemical study

  • Blanco, Carlota;Linares, Antonio;Dopico, Jose;Pico, Alex;Sobrino, Tomas;Leira, Yago;Blanco, Juan
    • Journal of Periodontal and Implant Science
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    • v.51 no.5
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    • pp.342-351
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    • 2021
  • Purpose: The aim of this study was to compare the inflammatory and lipid profile of patients with and without peri-implantitis. Methods: A cross-sectional biochemical study was carried out in which blood samples were collected from 16 patients with peri-implantitis and from 31 subjects with healthy implants. Clinical peri-implant parameters were obtained from all subjects. Levels of tumor necrosis factor-alpha and interleukin-10 (IL-10) were measured in serum. Lipid fractions, glucose and creatinine levels, and complete blood count were also assessed. Results: After controlling for a history of periodontitis, statistically significant differences between peri-implantitis patients and controls were found for total cholesterol (estimated adjusted mean difference, 76.4 mg/dL; 95% confidence interval [CI], 39.6, 113.2 mg/dL; P<0.001), low-density lipoprotein (LDL) cholesterol (estimated adjusted mean difference, 57.7 mg/dL; 95% CI, 23.8, 91.6 mg/dL; P<0.001), white blood cells (WBC) (estimated adjusted mean difference, 2.8×103/µL; 95% CI, 1.6, 4.0×103/µL; P<0.001) and IL-10 (estimated adjusted mean difference, -10.4 pg/mL; 95% CI, -15.8, -5.0 pg/mL; P<0.001). The peri-implant probing pocket depth (PPD) was modestly positively correlated with total cholesterol (r=0.512; P<0.001), LDL cholesterol (r=0.463; P=0.001), and WBC (r=0.519; P<0.001). A moderate negative correlation was observed between IL-10 and PPD (r=0.609; P<0.001). Conclusions: Otherwise healthy individuals with peri-implantitis showed increased low-grade systemic inflammation and dyslipidemia.