• Title/Summary/Keyword: Microbial agents

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Permanent Mycoplasma Removal Removel from Tissue Culture Cells: A Genetic Approach

  • Motr, Gabriele;Preininger, Alexandra;Himmelspach, Michele;Plaimauer, Barbara;Arbesser, Christine;York, Heinz;Dorner, Friedrich;Schlokat, Use
    • Biotechnology and Bioprocess Engineering:BBE
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    • v.5 no.2
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    • pp.84-91
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    • 2000
  • Mycopasma contamination of tissue culture cells easily evades detection and, thus, represents a continous therat to cell biologists. In case where infected cell can not simply be replaced, attempts have to be made to eradicate mycoplacma from the tissue culture cells. A variety of anti-microbial agents have been shown to be toxic to mycoplasma strains ; however, cell associated mycoplasma are often protected from antibiotics at concentrations shown to be effective in vitro. Antibiotic concentrations high enough to be lethal to cell as sociated mycoplasmas frequently are also detrimentrations to the host cells, while moderately increased antibiotic levels tolerated by the host cells often lead to only temporary growth suppression and/or to the emergence of mycoplasma strains resistanct even to high concentrations of the antibiotis applied. Hare, a genetic approach for the elimination of mycoplasma from tissue culture cells that overcomes thens limitations is described. By expression of a selection marker conferring resistance to an otherwise toxic agent, Acholeplasma laidlawii infected BHK-21 cells used as the model system were enabled to temporarily tolerate antibiotic concentrations high enough to be lethal to cell associated mycopalsma while leaving the host cells unharmed. Upon successful mycoplasma eradicated, cultvation of the cured host cells in the absence of the selective agent yielded revertant cell clones that had regained susceptibillity to the toxic agent. Cressation of the selection marker expression was shown to result from the loss of the selection marker DNA, which is a consequence of the fact that the stable and permanent integration of foreign DNA in eucaryotic cell chrosomes is highly inefficient. Thus, the cells were cured from mycoplasma yet remained biochemically unaltered.

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Characteristics of Permeation and Fouling of UF/MF Hollow Fiber Membranes for Drinking Water Treatment (정수처리 적용을 위한 UF/MF 중공사막의 투과성능과 오염현상)

  • 이주형;김정학;이용택
    • Membrane Journal
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    • v.10 no.2
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    • pp.75-82
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    • 2000
  • This study, which was tarried out to investigate the characteristics of permeation rates and fouling phenomena in drinking-water treating processes with MF membrane, showed that pressure drop was dependent on the length of membrane module and operating pressure; the pressure drop increased with the length of membrane module and operating pressure, operation at a relatively low pressure(0.5kg/$cm^2$) is better than that at a relatively (2.0kg/$cm^2$), since high operating pressure accelerates the clogging. In case of out-in permeation type, almost same flux was obtained after a certain operating time regardless of membrane length and operating pressure. In order to understand, the microbial fouling, chemical cleaning was carried out to the forced contaminated hollow-fiber membrane with chemicals($H_2O_2$, NaOCl, and NaOH). Chemical cleanings with $H_2O_2$and NaOCl, which are oxidizing agents, are better for sterilizing and desorbing the microbes than those with NaOH.

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Amino Acid-Based Material for the Complementary Therapy of Decubitus Ulcers

  • Nogueira, Frederico;Gouveia, Isabel C.
    • Journal of Microbiology and Biotechnology
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    • v.27 no.4
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    • pp.747-758
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    • 2017
  • Chronic wounds, pressure sores, lesions, and infections of microbial origin in bedridden, paralyzed, or malnutrition patients remain the object of study of many researchers. A variety of factors behind the development of these disorders are related to the patient's immune system, making it unable to respond effectively to the treatment of the wound. These factors can be properly controlled, giving particular importance to the ethiology and stage of the wound, as well as the time periods corresponding to the replacement of the dressings. The present research reports a novel foam/soft material, ${{\small}L}$-Cys-g-PCL, with an application for decubitus/pressure ulcers, especially for wounds with a difficult healing process due to infections and constant oxidation of the soft tissues. During this work, the interactions between S. aureus and ${{\small}L}$-Cys-g-PCL foam were studied under conditions that simulate decubitus ulcers; namely, pH and exudate. The effects of duration of grafting (1 or 8 h) and pH (7.0 and 8.9) on wettability, surface energy, swelling, and porosity were also evaluated. Results showed an effective microbicidal activity exhibiting an inhibition ratio of 99.73% against S. aureus. This new ${{\small}L}$-Cys-g-PCL soft material showed saftey to contact skin, ability to be shaped to fill in sunken holes (craters) - pressure ulcers stage III - and to act as a smart material responsive to pH, which can be tailored to develop better swelling properties at alkaline pH where exudates are normally higher, so as to address exudate self-cleaning and prevention of desiccation.

Laser therapy in peri-implantitis treatment: literature review (임플란트주위염 처치에서 레이저의 이용: 문헌고찰)

  • Lee, Kyung-Joong;Lee, Jong-Ho;Kum, Kee-Yeon;Lim, Young-Jun
    • Journal of Dental Rehabilitation and Applied Science
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    • v.31 no.4
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    • pp.340-348
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    • 2015
  • Peri-implantitis is the most common reason for a late failure and can occur even after years of successful osseointegration. The role of microbial plaque accumulation in the development of peri-implantitis has been well documented. On the other hand, the ideal method of implant surface decontamination to re-establish the health of peri-implant tissue remains to be determined. Removal of bacterial deposits is essential in the treatment of peri-implant infections, and various therapeutic approaches have been described in the literature, including mechanical debridement, disinfection with chemotherapeutic agents, and laser therapy. Recently, there has been a plenitude of scientific data regarding the use of laser irradiation to achieve titanium surface decontamination. Thus, research is focusing on lasers' potential use in the treatment of peri-implantitis. The aim of this literature review is to analyze and evaluate the efficacy of laser therapy for the treatment of peri-implantitis.

Enhancement of β-cyclodextrin Production and Fabrication of Edible Antimicrobial Films Incorporated with Clove Essential Oil/β-cyclodextrin Inclusion Complex

  • Farahat, Mohamed G.
    • Microbiology and Biotechnology Letters
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    • v.48 no.1
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    • pp.12-23
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    • 2020
  • Edible films containing antimicrobial agents can be used as safe alternatives to preserve food products. Essential oils are well-recognized antimicrobials. However, their low water solubility, volatility and high sensitivity to oxygen and light limit their application in food preservation. These limitations could be overcome by embedding these essential oils in complexed product matrices exploiting the encapsulation efficiency of β-cyclodextrin. This study focused on the maximization of β-cyclodextrin production using cyclodextrin glucanotransferase (CGTase) and the evaluation of its encapsulation efficacy to fabricate edible antimicrobial films. Response surface methodology (RSM) was used to optimize CGTase production by Brevibacillus brevis AMI-2 isolated from mangrove sediments. This enzyme was partially purified using a starch adsorption method and entrapped in calcium alginate. Cyclodextrin produced by the immobilized enzyme was then confirmed using high performance thin layer chromatography, and its encapsulation efficiency was investigated. The clove oil/β-cyclodextrin inclusion complexes were prepared using the coprecipitation method, and incorporated into chitosan films, and subjected to antimicrobial testing. Results revealed that β-cyclodextrin was produced as a major product of the enzymatic reaction. In addition, the incorporation of clove oil/β-cyclodextrin inclusion complexes significantly increased the antimicrobial activity of chitosan films against Staphylococcus aureus, Staphylococcus epidermidis, Salmonella Typhimurium, Escherichia coli, and Candida albicans. In conclusion, B. brevis AMI-2 is a promising source for CGTase to synthesize β-cyclodextrin with considerable encapsulation efficiency. Further, the obtained results suggest that chitosan films containing clove oils encapsulated in β-cyclodextrin could serve as edible antimicrobial food-packaging materials to combat microbial contamination.

Structural Basis for Recognition of L-lysine, L-ornithine, and L-2,4-diamino Butyric Acid by Lysine Cyclodeaminase

  • Min, Kyungjin;Yoon, Hye-Jin;Matsuura, Atsushi;Kim, Yong Hwan;Lee, Hyung Ho
    • Molecules and Cells
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    • v.41 no.4
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    • pp.331-341
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    • 2018
  • L-pipecolic acid is a non-protein amino acid commonly found in plants, animals, and microorganisms. It is a well-known precursor to numerous microbial secondary metabolites and pharmaceuticals, including anticancer agents, immunosuppressants, and several antibiotics. Lysine cyclodeaminase (LCD) catalyzes ${\beta}$-deamination of L-lysine into L-pipecolic acid using ${\beta}$-nicotinamide adenine dinucleotide as a cofactor. Expression of a human homolog of LCD, ${\mu}$-crystallin, is elevated in prostate cancer patients. To understand the structural features and catalytic mechanisms of LCD, we determined the crystal structures of Streptomyces pristinaespiralis LCD (SpLCD) in (i) a binary complex with $NAD^+$, (ii) a ternary complex with $NAD^+$ and L-pipecolic acid, (iii) a ternary complex with $NAD^+$ and L-proline, and (iv) a ternary complex with $NAD^+$ and L-2,4-diamino butyric acid. The overall structure of SpLCD was similar to that of ornithine cyclodeaminase from Pseudomonas putida. In addition, SpLCD recognized L-lysine, L-ornithine, and L-2,4-diamino butyric acid despite differences in the active site, including differences in hydrogen bonding by Asp236, which corresponds with Asp228 from Pseudomonas putida ornithine cyclodeaminase. The substrate binding pocket of SpLCD allowed substrates smaller than lysine to bind, thus enabling binding to ornithine and L-2,4-diamino butyric acid. Our structural and biochemical data facilitate a detailed understanding of substrate and product recognition, thus providing evidence for a reaction mechanism for SpLCD. The proposed mechanism is unusual in that $NAD^+$ is initially converted into NADH and then reverted back into $NAD^+$ at a late stage of the reaction.

Formulation of a New Bacillus thuringiensis Strain NT0423. (새로운 Bacillus thuringiensis NT0423 균주의 제제화)

  • 김호산;노종열;이대원;장진희;제연호;우수동;김주경;유용만;강석권
    • Microbiology and Biotechnology Letters
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    • v.26 no.4
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    • pp.358-364
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    • 1998
  • New microbial-control agents were prepared with B. thuringiensis strain NT0423 having unique properties which are different with other B. thuringiensis strains belonging to serotype 7[Kor. J. Appl. Entomol. 32: 426-432.]. Three B. thuringiensis formulations designated as BioBact 10%, 20% and 40%, were made with various combinations of adjuvants. These formulations showed good physical properties in wettability, suspensibility, particle size and adherence. In addition the result of SDS-PAGE analysis indicated that $\delta$-endotoxins remain stably in all formulations. Among the tested formulations, two wettable powder formulations, BioBact 20% and 40%, comprising 20% and 40% of B. thuringiensis technical powder showed the effective control against diamondback moth larvae (Plutella xylostella) in laboratory and field tests. Especially, when compared with commercial B. thuringiensis formulations (A and B commercial formulations) in field evaluation, BioBact 20% and 40% formulations showed equal activity up to 80% lethality and a good persistence effect which remain on leaves at least 7 days.

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Effect of Catechins, Green tea Extract and Methylxanthines in Combination with Gentamicin Against Staphylococcus aureus and Pseudomonas aeruginosa - Combination therapy against resistant bacteria -

  • Bazzaz, Bibi Sedigheh Fazly;Sarabandi, Sahar;Khameneh, Bahman;Hosseinzadeh, Hossein
    • Journal of Pharmacopuncture
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    • v.19 no.4
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    • pp.312-318
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    • 2016
  • Objectives: Bacterial resistant infections have become a global health challenge and threaten the society's health. Thus, an urgent need exists to find ways to combat resistant pathogens. One promising approach to overcoming bacterial resistance is the use of herbal products. Green tea catechins, the major green tea polyphenols, show antimicrobial activity against resistant pathogens. The present study aimed to investigate the effect of catechins, green tea extract, and methylxanthines in combination with gentamicin against standard and clinical isolates of Staphylococcus aureus (S. aureus) and the standard strain of Pseudomonas aeruginosa (P. aeruginosa). Methods: The minimum inhibitory concentration (MIC) and the minimum bactericidal concentration (MBC) values of different agents against bacterial strains were determined. The interactions of green tea extract, epigallate catechin, epigallocatechin gallate, two types of methylxanthine, caffeine, and theophylline with gentamicin were studied in vitro by using a checkerboard method and calculating the fraction inhibitory concentration index (FICI). Results: The MICs of gentamicin against bacterial strains were in the range of $0.312-320{\mu}g/mL$. The MIC values of both types of catechins were $62.5-250{\mu}g/mL$. Green tea extract showed insufficient antibacterial activity when used alone. Methylxanthines had no intrinsic inhibitory activity against any of the bacterial strains tested. When green tea extract and catechins were combined with gentamicin, the MIC values of gentamicin against the standard strains and a clinical isolate were reduced, and synergistic activities were observed (FICI < 1). A combination of caffeine with gentamicin did not alter the MIC values of gentamicin. Conclusion: The results of the present study revealed that green tea extract and catechins potentiated the antimicrobial action of gentamicin against some clinical isolates of S. aureus and standard P. aeruginosa strains. Therefore, combinations of gentamicin with these natural compounds might be a promising approach to combat microbial resistance.

Antifungal Activity and Exoenzyme Production of Several Bacteria Antagonistic to Trichoderma spp. Causing Green Mold Disease (버섯 푸른곰팡이균에 대한 길항세균의 항균활성과 세포외 분비효소 생성능)

  • Hyun, Soung-Hee;Min, Bong-Hee
    • The Korean Journal of Mycology
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    • v.30 no.2
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    • pp.147-151
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    • 2002
  • Trichoderma spp. are the aggressive causal agents for green mold disease on oyster mushroom (Pleurotus spp.) cultivation. Antifungal bacteria (KATB 99121, KATB 99122 and KATB 99123 strains) were isolated from the compost for Pleurotus ostreatus. Among these bacterial strains, KATB 99121 strain showed an excellent inhibitory activity to the pathogens for green molds such as T. harzianum, T. viride and T. hamatum and an animal pathogen, Candida albicans, but did not affect on the culture of Pleurotus ostreatus (2209, Chunchu 2 and Wonhyung strains). KATB 99121 strain secreted amylolytic, proteolytic and cellulolytic exoenzymes. KATB 99122 and KATB 99123 strains excreted amylolytic, proteolytic, cellulolytic, lipolytic exoenzymes and showed ${\beta}$-glucosidase activity. Further studies will be conducted on the development of microbial fungicides using the antagonistic bacteria for the control of green mold disease on Pleurotus spp.

Immunogenomics approaches to study host innate immunity against intestinal parasites

  • Lillehoj, Hyun S.
    • Proceedings of the Korea Society of Poultry Science Conference
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    • 2006.11a
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    • pp.7-16
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    • 2006
  • Poultry products including meat and eggs constitute a major protein source in the American diet and disease - causing pathogens represent major challenges to the poultry industry. More than 95 % of pathogens enter the host through the mucosal surfaces of the respiratory, digestive and reproductive tracts and over the past few decades, the two main mechanisms used to control diseases have been the use of vaccines and antibiotics. However, in the poultry industry, there are mounting concerns over the ability of current vaccines to adequately protect against emerging hyper - virulent strains of pathogens and a lack of suitable, cost effective adjuvants. Thorough investigation of the immunogenetic responses involved in host-pathogen interactions will lead to the development of new and effective strategies for improving poultry health, food safety and the economic viability of the US poultry industry. In this paper, I describe the development of immunogenomic and proteomic tools to fundamentally determine and characterize the immunological mechanisms of the avian host to economically significant mucosal pathogens such as Eimeria. Recent completion of poultry genome sequencing and the development of several tissue-specific cDNA libraries in chickens are facilitating the rapid application of functional immunogenomics in the poultry disease research. Furthermore, research involving functional genomics, immunology and bioinformatics is providing novel insights into the processes of disease and immunity to microbial pathogens at mucosal surfaces. In this presentation, a new strategy of global gene expression using avian macrophage (AMM) to characterize the multiple pathways related to the variable immune responses of the host to Eimeria is described. This functional immunogenomics approach will increase current understanding of how mucosal immunity to infectious agents operates, and how it may be enhanced to enable the rational development of new and effective strategies against coccidiosis and other mucosal pathogens.

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