• Title/Summary/Keyword: gram negative bacteria

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Evaluation of the Effects of Interfering Factors on the Bacterial Endotoxin Testing of Radiopharmaceuticals (방사성의약품의 박테리아 엔도톡신 시험에서 반응간섭인자들의 영향에 대한 평가)

  • Jun Young PARK
    • Korean Journal of Clinical Laboratory Science
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    • v.56 no.2
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    • pp.171-180
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    • 2024
  • The endotoxin test is based on the reaction between Limulus Amebocyte Lysate (LAL) and the lipopolysaccharides of Gram-negative bacteria. In this study, we sought to identify factors that interfere with the LAL testing of radiopharmaceuticals and evaluated acceptable ranges. A gel-clot LAL test and a chromogenic LAL test were used as endotoxin tests. We compared the performances of the Endosafe LAL and recombinant Endosafe Recombinant Cascade Reagent (rCR) cartridges for the chromogenic test. The factors that interfered with 68Ga-DOTATOC injection were pH, 4-(2-hydroxyethyl) piperazine-1-ethanesulfonic acid (HEPES) buffer, and organic solvents, especially ethanol. However, interference by these factors was overcome by diluting the 68Ga-DOTATOC injection tenfold. In addition, no interference was observed at pH values between 4 and 8, at a HEPES concentration of 2,000 ㎍/mL, or an ethanol concentration of <1%. Furthermore, results showed that interfering factors had similar effects on the performances of the Endosafe LAL and Endosafe rCR cartridges. The results of this study are expected to be useful for evaluating factors that interfere with the endotoxin testing of new radiopharmaceuticals.

MECHANISM IN ANTIBACTERIAL ACTIVITY OF POLYPHOSPHATES AGAINST PORPHYROMONAS ENDODONTALIS (Porphyromonas endodontalis에 대한 Polyphosphate의 항균기전에 관한 연구)

  • Choi, Sung-Baik;Park, Sang-Jin;Choi, Gi-Woon;Choi, Ho-Young
    • Restorative Dentistry and Endodontics
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    • v.25 no.4
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    • pp.561-574
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    • 2000
  • Poly-P has been used to prevent decomposition of foods and has been shown to have inhibitory effect on the growth of gram positive bacteria. The purpose of this study was to evaluate the effect of poly-P on the growth of Porphyromonas endodontalis, a gram negative obligate anaerobic rod, endodontopathic bacterium. P. endodontalis ATCC 35406 was in BHI broth containing hemin and vitamin K with or without poly-P. Inhibitory effect of each poly-P which was added at the beginning(lag phase) or during(exponential phase) the culture, MIC(minimum inhibitory concentration) was determined by measuring the optical density of the bacterial cell at 540nm. Viable cell counts were measured to determined whether poly-P has a bactericidal effect. Leakage of intracellular nucleotides from P. endodontalis was determined at 260nm and morphological change of P. endodontalis was observed under the TEM(transmission electron microscope). Binding of 32P-labeled poly-P to P. endodontalis was examined. SDS-polyacrylamide gel electrophoresis and zymography were performed to observe the changes in protein and enzyme profiles of P. endodontalis, respectively. The results from this study were as follows : 1. The minimal inhibitory concentration(MIC) of poly-P to P. endodontalis appeared to be 0.04~0.05%. 2. Poly-P added to the P. endodontalis culture during the exponential phase of P. endodontalis was as much effective as poly-P added at the begining of the culture, suggesting that the antibacterial effect of poly-P is not much dependent on the initial inoculum size of P. endodontalis. 3. Poly-P are bactericidal to P. endodontalis, demonstrating the decrease of the viable cell counts. 4. Intracellular nucleotide release from the P. endodontalis, was not increased in the presence of poly-P and was not reversed by the addition of divalent cations like $Ca^{2+}$ and $Mg^{2-}$. 5. Under the TEM, it was observed that fine electro-dense materials were prominent in the poly-P grown P. endodontalis, appearing locally in the cell, and the materials were more abundant and more dispersed in the cell as the incubation time with poly-P increased. In addition, highly electron dense granules accumulated in many poly-P grown cells, most of which were atypical in their shape. 6. Binding of 32P-labeled poly-P to P. endodontalis appeared to be 32.8 and 45.5 and 53.4% at 30 minutes, 1 hours and 2 hours, respectively. 7. In the presence of poly-P. the synthesis of proteins with apparent molecular masses of 25, 27, 35, 45 was lost or drastically decreased whereas expression of a protein with an apparent molecular mass of 75 was elevated. 8. Proteolytic activity of P. endodontalis was decreased by poly-P. The overall results suggest that use of poly-P may affect the growth of P. endodontalis, and the anti-bacterial activity of poly-P seems largely bactericidal. Changes in shape, protein expression, and proteolytic activity of P. endodontalis by poly-P may be directly and indirectly attributed to the antibacterial effect of poly-P. Further studies will be needed to confirm the effect of poly-P.

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Comparative Study on the Antimicrobial Activity of Glycyrrhiza uralensis and Glycyrrhiza glabra Extracts with Various Countries of Origin as Natural Antiseptics (원산지별 감초 추출물의 항균 활성 비교 및 천연방부제로써의 효능 연구)

  • Kim, Hye Jin;Bae, Jeong Yun;Jang, Ha Na;Park, Soo Nam
    • Microbiology and Biotechnology Letters
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    • v.41 no.3
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    • pp.358-366
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    • 2013
  • The aim of this study was to evaluate the antimicrobial activities of Glycyrrhiza uralensis and Glycyrrhiza glabra extracts with various countries of origin. Three samples of licorice with various origins (Korea, China, and Uzbekistan) were evaluated for their antimicrobial activities against six skin microflora. The bioassay applied for determining the antimicrobial effects included the disc diffusion assay, minimum inhibitory concentration, and challenge test. The ethyl acetate fractions of G. uralensis and G. glabra extracts showed significant antimicrobial activities against two gram-positive (Bacillus subtilis, Propionibacterium acnes) and two gram-negative (Escherichia coli, Pseudomonas aeruginosa) bacteria. These samples had much more intensive antimicrobial activities than synthetic preservatives on B. subtilis, P. acnes, and P. aeruginosa, especially. Korean licorice showed the highest antimicrobial activity amongst the samples tested. In view of the observed inhibitory features of these G. uralensis and G. glabra extracts, it is suggested that they could be used as natural antiseptics against bacterial contamination in cosmetics and foods, instead of the common synthetic preservatives currently employed.

Antimicrobial Activity of Niaouli (Melaleuca quinquenervia) Leaf Extracts against Skin Flora (피부 상재균에 대한 니아울리 잎 추출물의 항균활성)

  • Jang, Ha Na;Park, Soo Nam
    • Journal of the Society of Cosmetic Scientists of Korea
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    • v.40 no.3
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    • pp.313-320
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    • 2014
  • In this study, the antimicrobial activity of niaouli leaf extracts was evaluated against skin flora. The skin flora used for experiments were three gram-positive bacteria such as Bacillus subtilis (B. subtilis), Staphylococcus aureus (S. aureus), Propionibacterium acnes (P. acnes), and two gram-negative, Escherichia coli (E. coli), Pseudomonas aeruginosa( P. aeruginosa), and the yeast, Plasmodium ovale (P. ovale). The bioassay applied for determining the antimicrobial effects of niouli leaf extracts or fraction included the disc diffusion assay and broth dilution assay. Minimum inhibitory concentration (MIC) values of 50% ethanol extract on B. subtilis, S. aureus, P. acnes, E. coli and P. aeruginosa were 0.25%, 0.50%, 1.00%, 0.13% and 0.25% respectively and the MIC values of water fraction were 0.25%, 0.25%, 4,00%, 0.25% and 0.25%. P. ovale did not show antimicrobial activities. The MIC values of methyl paraben used as positive control indicated 0.25%, 0.25%, 0.25%, 0.13% and 0.50%. Also, Minimum bactericidal concentration (MBC) values of 50% ethanol extract were 2.00%, 2.00%, 1.00%, 0.50% and 2.00% individually and the MBC values of water fraction were 0.50%, 0.25%, 4.00%, 0.50% and 1.00%. The MBC values of methyl paraben indicated 1.00%, 0.500%, 0.50%, 0.50% and 1.00%. These results showed that water fraction was as good as methyl paraben except for P. acnes. The 50% ethanol extract also showed activity similar with it. Thus, it is concluded that the 50% ethanol extract/fraction of niaouli could be applicable to cosmetics as a natural preservatives effective in antimicrobial activity against skin flora.

Evaluation for the Antimicrobial, Antioxidant and Anti-thrombosis Activity of Korean Traditional Liquors (시판 전통주의 항균, 항산화 및 항혈전 활성 평가)

  • Ryu, Hee-Young;Kum, Eun-Joo;Bae, Kyung-Hwa;Kim, Young-Kwan;Kwun, In-Sook;Sohn, Ho-Yong
    • Microbiology and Biotechnology Letters
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    • v.35 no.3
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    • pp.238-244
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    • 2007
  • To identify the useful biological activities and the superiority in quality of Korean traditional liquors, we prepared the alcohol-free concentrates from different types of commercial traditional liquors, Takju ($T1{\sim}T3$), Yakju/Cheongju ($Y-1{\sim}Y-5$), Fruit wine (F-1) and Soju (S-1, S-2), respectively. We investigate their compositions as well as antimicrobial, antioxidant, and anti-thrombosis activity. Among the 11 traditional liquors, Y-3, Y-4, Y-5 and F-1 showed higher total-polyphenol, total-flavonoids and reducing sugars than the others. The strong antibacterial and anti-thrombosis activities were identified in Y-3, Y-4, Y-5 and F-1, and a minor antioxidant activity was found in F-l. The antibacterial activity of the Y-3, Y-4, Y-5 and F-1 alcohol-free concentrates showed a broad-spectrum, and growth inhibition was found in gram-positive, gram-negative, and ampicillin-resistant bacteria. The sequential solvent fractionation of Y-3, Y-4, Y-5 and F-1, and following analysis showed that ethyl acetate fractions of Y-3, Y-4, Y-5 and F-1 possess strong antibacterial and anti-thrombosis activity. Especially, the ethyl acetate fractions of Y-3, Y-4 and F-1 showed superior anti-thrombosis activity compared than that of aspirin. Our results suggest that the useful substances are produced from substrates and edible plant added during the fermentation, and the Korean traditional liquors could be developed as strong antibacterial and anti-thrombosis agents.

Antioxidative, Antimicrobial, and Anti-proliferative Activities of the Floret and Stalk of Broccoli (Brassica oleracea L.) (브로콜리 꽃송이 및 줄기의 항산화, 항균 및 대장암 세포 생육억제효과)

  • Kim, Mi-Sun;Lee, Ye-Seul;Kwon, Ha-Young;Kim, Jong Sik;Sohn, Ho-Yong
    • Microbiology and Biotechnology Letters
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    • v.42 no.1
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    • pp.58-66
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    • 2014
  • In the course of study for a use for non-edible parts of broccoli (Brassica oleracea L), and the development of processed food utilizing these parts, edible floret and non-edible stalk were extracted with ethanol and different organic solvent fractions were prepared. With 10 different extracts and fractions, their useful components and various biological activities, such as antioxidant, antimicrobial and anti-proliferation activity, were investigated. The stalk has more abundant water soluble carbohydrate when compared with the floret, and floret has higher hexane-soluble pigments. Analysis of total flavonoid and total polyphenol contents showed that the floret has 1.5~1.99 times higher concentrations than the stalk. Among the fractions, ethylacetate (EA) fractions have the highest amount of total flavonoid and total polyphenol. The stalk and floret possessed 9.45 and 42.01 mg-total flavonoid/g, respectively. In the antioxidation activity assay, the EA fraction of floret showed strong radical scavenging activity and reducing power, while the n-hexane fraction of the stalk exhibited nitrite scavenging activity. In the antimicrobial activity assay, the EA fraction of floret showed a strong and broad-range of antibacterial activity, irrespective of gram positive or gram negative bacteria. In a while, the hexane and EA fractions revealed anti-proliferative effects against the human colorectal cancer cell HCT-116. Strong anti-proliferative activities were found in the hexane fraction of stalk (18.4% of cell viability), and the n-butanol fraction of floret (6.9% of cell viability). Our results suggest that the further study of the characterization of active fractions and the identification of active components from different parts of broccoli are needed to develop functional foods or novel plant-derived medicines.

Antimicrobial Activities of Natural Medicinal Herbs on the Food Spoilage or Foodborne Disease Microorganisms (식품부패 및 병원성미생물에 대한 천연약용식물 추출물의 항균효과)

  • Oh, Deog-Hwan;Ham, Seung-Shi;Park, Boo-Kil;Ahn, Cheol;Yu, Jin-Young
    • Korean Journal of Food Science and Technology
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    • v.30 no.4
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    • pp.957-963
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    • 1998
  • Several medicinal herbs, which are nontoxic and have been used widely in traditional folk medicine, were extracted and antimicrobial activity of the extracts was investigated against various foodborne pathogens or food poisioning microorganisms. The ethanol extract of Sansa, Hwangryun, Cheukbaek, and Seokchangpo showed strong antimicrobial activities against Gram postive and Gram negative bacteria, whereas those of Sakunja, Sukjihwang and Baekji had little antimicrobial activities on microorganisms tested. Among medicinal herb extracts, ethanol extract of Coptis chinensis Franch (hwangryun) showed the strongest antimicrobial activity. Antimicrobial activity of ethanol extract of Coptis chinensis Franch was not destroyed by heating at $100^{\circ}C$ for 60 min and at $121^{\circ}C$ for 30 min, which is very stable over heat. The pH effect on minimal inhibitory concentrations (MIC) of the extract of Coptis chinensis Franch indicated that MIC was reduced with increasing the pH value of the medium. The inhibitory effect of partially purified substance from the ethanol extract of Coptis chinensis Franch on the growth of Listeria monocytogenes and Staphylococcus aureus was investigated. Growth of those strains occurred at the concentration of $100{\;}{\mu}g/mL$ and were inhibited at $500{\;}{\mu}g/mL$, whereas those strains was completely inactivated in the presence of $1000{\;}{\mu}g/mL$.

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Optimal Lactic Acid Fermentation Conditions and Quality Properties for Rubus coreanus Miquel (Bokbunja) and Chlorella Mixtures (복분자와 클로렐라 혼합물의 젖산발효 최적조건 및 품질 특성)

  • Kim, Jae-Young;Lee, Sang-Uk;Kim, Na-Hyung;Moon, Kwang-Hyun;Baek, Seung-Hwa
    • Journal of the Korean Society of Food Science and Nutrition
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    • v.45 no.3
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    • pp.386-395
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    • 2016
  • To develop a functional fermentation food from Rubus coreanus Miquel (Bokbunja) and chlorella mixtures, optimal lactic acid fermentation conditions were established, and quality properties based on physicochemical evaluation such as chemical compositions, free sugars, organic acids, and antibacterial activities were investigated. Regarding optimal fermentation strain selection, formation of lactic acid was best in Lactobacillus plantarum among the experimental strains (10 kinds), and the optimal fermentation temperature was $37^{\circ}C$. In addition, overall acceptability in the sensory evaluation was highest in the 5% chlorella mixture sample. Therefore, quality properties of the prepared sample under the established optimal fermentation conditions were investigated. Moisture, total sugar (dry basis), crude fiber (dry basis), and pH of fermented Rubus coreanus Miquel juice (RCM) with 5% chlorella mixture (RCM-C5) were reduced by 4.90%, 14.15%, and 0.32%, respectively, as compared with non-fermented RCM. Meanwhile, crude protein, crude fat, and crude ash (dry basis) of RCM-C5 were elevated by 13.75%, 0.18%, and 0.73%, respectively, as compared with RCM. The yellowness (b value) of color values was greater in RCM-C5 compared to RCM. The free sugar and organic acid contents of RCM-C5 were elevated by 0.97% and 616.30 mg%, respectively, as compared with RCM. In addition, the gram positive bacterium Staphylococcus aureus was elevated by 5.83% while gram negative bacteria Escherichia coli and Salmonella Typhimurium were elevated by 2.94% and 4.67%, respectively, as compared with RCM. In conclusion, the quality properties of RCM and chlorella lactic acid fermentation mixtures were improved compared with the general RCM product. Consequently, it is possible to apply fermented RCM as a functional fermentation food.

Biosynthesis of Silver Nanoparticles Using Microorganism (미생물을 이용한 은 나노입자 생합성)

  • Yoo, Ji-Yeon;Jang, Eun-Young;Hong, Chang-Oh;Kim, Keun-Ki;Park, Hyean-Cheal;Lee, Sang-Mong;Kim, Young-Gyun;Son, Hong-Joo
    • Journal of Life Science
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    • v.28 no.11
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    • pp.1354-1360
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    • 2018
  • The aim of this study was to develop a simple, environmentally friendly synthesis of silver nanoparticles (SNPs) without the use of chemical reducing agents by exploiting the extracellular synthesis of SNPs in a culture supernatant of Bacillus thuringiensis CH3. Addition of 5 mM $AgNO_3$ to the culture supernatant at a ratio of 1:1 caused a change in the maximum absorbance at 418 nm corresponding to the surface plasmon resonance of the SNPs. Synthesis of SNPs occurred within 8 hr and reached a maximum at 40-48 hr. The structural characteristics of the synthesized SNPs were investigated by various instrumental analysis. FESEM observations showed the formation of well-dispersed spherical SNPs, and the presence of silver was confirmed by EDS analysis. The X-ray diffraction spectrum indicated that the SNPs had a face-centered cubic crystal lattice. The average SNP size, calculated using DLS, was about 51.3 nm and ranged from 19 to 110 nm. The synthesized SNPs exhibited a broad spectrum of antimicrobial activity against a variety of pathogenic Gram-positive and Gram-negative bacteria and yeasts. The highest antimicrobial activity was observed against C. albicans, a human pathogenic yeast. The FESEM observations determined that the antimicrobial activity of the SNPs was due to destruction of the cell surface, cytoplasmic leakage, and finally cell lysis. This study suggests that B. thuringiensis CH3 is a potential candidate for efficient synthesis of SNPs, and that these SNPs have potential uses in a variety of pharmaceutical applications.

Genetic Identification and Biochemical Characteristics of Edwardsiella Strains Isolated from Freshwater Fishes Cultured in Korea (내수면 양식 어류에서 분리된 Edwardsiella 속 균주들의 유전학적 동정 및 생화학적 특성)

  • Jang, Mun Hee;Kim, Keun-Yong;Lee, Yu Hee;Oh, Yun Kyung;Lee, Jeong-Ho;Song, Jun-Young
    • Journal of fish pathology
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    • v.33 no.2
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    • pp.111-118
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    • 2020
  • The genus Edwardsiella belonging to the family Enterobacteriaceae is a member of Gram-negative rod-shaped bacteria that cause disease in diverse aquatic organisms such as fish, amphibians and reptiles as well as avians and mammals including human throughout the world. This genus had been composed of three species, E. hoshinae, E. ictaluri and E. tarda, but recent researches erected two novel species, E. anguillarum and E. piscicida that were conventionally identified as E. tarda. In this study, we isolated seven strains belonging to the genus Edwardsiella from freshwater fishes that had been reared at inland fish farms in South Korea and investigated their biochemical characteristics and molecular phylogenetic relationships. The seven strains showed typical characteristics of four Edwardsiella species, E. anguillarum, E. ictaluri, E. piscicida and E. tarda, by biochemical analyses of Gram staining, indole and hydrogen sulfide (H2S) production, and API (Analytic Profile Index) 20E test. Molecular phylogenetic analyses inferred from DNA sequence data of both 16S ribosomal RNA (rRNA) and DNA gyrase subunit B (gyrB) genes were congruent with the biochemical characteristics. As a result, both biochemical and molecular phylogenetic analyses identified four strains isolated from three Anguilla species as E. anguillarum, E. piscicida and E. tarda, two strains from Pelteobagrus fulvidraco and Silurus asotus as E. ictaluri, and one strain from Moroco oxycephalus as E. piscicida. In this study, we isolated and successfully identified recently newly erected species, E. anguillarum and E. piscicida in addition to historically notorious pathogenic species, E. ictaluri and E. tarda. In the future study, systematic and comprehensive monitoring of the four Edwardsiella species are required for studying differences in pathogenicity among freshwater fishes.