• Title/Summary/Keyword: microbial assay

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Microbiological and Genotoxicological Safety of Gamma-Irradiated Chicken (감마선조사 닭고기의 미생물학적 및 유전독성학적 안전성 평가)

  • 곽희진;정차권;강일준
    • Korean journal of food and cookery science
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    • v.17 no.6
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    • pp.617-624
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    • 2001
  • Gamma irradiation (1-10 kGy) was applied to chicken for the evaluation of their microbiological safety and possible genotoxicity. In 3 kGy-irradiated sample, the growth of psychrophile was inhibited about 1.5 log cycles and no cells were recovered in total microbial counts. All kinds of contaminated microorganism were sterilized by 7 kGy-irradiation. Also, irradiation followed by freeze-storage at the same time was very effective in inhibiting bacterial growth. The genotoxicity of 10 kGy-irradiated chicken was evaluated by Salmonella Typhimurium reversion assay and in vivo micronucleus assay using mouse bone marrow cells. The results were negative in the bacterial reversion assay with S. Typhimurium TA98, TA100, TA1535, and TA1539. Clastogenic effects were not shown in vivo mouse micronucleus assay at 10 kGy-dose tested.

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Biofilm Formation of Food-borne Pathogens under Stresses of Food Preservation (식품 보존 스트레스에서의 식중독세균의 생체막 생성)

  • Lee, No-A;Noh, Bong-Soo;Park, Jong-Hyun
    • Korean Journal of Food Science and Technology
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    • v.38 no.1
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    • pp.135-139
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    • 2006
  • Most bacteria form biofilm as self-defence system, making efficient food sanitization, preservation, and instrument washing more difficult. Biofilm formation of Salmonella, E. coli, B. cereus, and S. aureus was observed during 24 hr food preservations by performing microtiter plate and glass wool assays. Most cells formed biofilm and attached onto glass wool. When biofilm formation and injury were analyzed on the microtiter plate, 10 and 20% acid-injured E. coli and S. aureus, respectively, 30-50% cold temperature $(4^{\circ}C)-injured$ B. cereus and E. coli, and 30-55% 6% sodium chloride solution-injured Salmonella showed significant biofilm formation. Results indicate biofilm formation level differed within species depending on type of stress.

A Study on the Anti-microbacterial Activity, Anti-inflammatory and Anti-allergic Effects of Samhwangseje gagambang(SHB) (삼황세제가감방(三黃洗劑加減方)의 항균, 항염 및 항알레르기 효과에 대한 실험적 연구)

  • Weon, Young-Ho;Shim, Eun-Ki;Ahn, Chan-Gn;Park, Min-Chul;Hwang, Chung-Yeon
    • The Journal of Korean Medicine Ophthalmology and Otolaryngology and Dermatology
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    • v.18 no.1
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    • pp.135-153
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    • 2005
  • Herbal mixture water extract of (Phellodendron amurense, Scuellaria baiklensis, Spphora flavescens, Lithospermum erythrorhizon, Mellaphis chinesis, Alumite, Zanthoxylum schinifolium, Glycyrrhiza uralensis), which exhibit several beneficial effects including acne and skin diseases, was tested for anti-microbial activity and anti-inflammation effects. The herbal mixture extract showed antimicrobial activity against Stapylococcus epidermis, Propionibacterium acne, and Malassezia furfur. The growth of Stapylococcus epidermis, Propionibacterium acne, and Malassezia furfur was inhibited allergy and LPS induced cyclooxygenase-2(COX-2)gene expression in RAW24.7macrophage. The results indicated th ear swelling and histamine release induced by compound 48/80 were dose-dependently reduced, ranging 11-38% and 11-56%, respectively. Furthermore the extract inhibited the expression of LPS-induced COX-2 proteins and mRNAs without an appreciable cytotoxic effects on RAW264.7 cells. The cytotoxicity of the extract using M7T assay showed the cytotoxicity of 7 and 18% against L929 cell line. Based on these results, it is concluded that the herbal mixture water extract can be applied to the acne and skin diseases therapy.

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A Study on the Anti-microbacterial Activity, Anti-inflammatory and Anti-allergic Effects of mixture of Rheum coreanum Scutellaria baikalensis Phellodenron amurense (대황(大黃), 황금(黃芩), 황백(黃柏) 복합 추출물의 항균, 항염 및 항알레르기 효과에 대한 실험적 연구)

  • Son, Dae-Beom;Song, Seong-Pil;Hwang, Chi-Hwan;Hong, Seok-Hoon;Hwang, Chung-Yeon
    • The Journal of Korean Medicine Ophthalmology and Otolaryngology and Dermatology
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    • v.18 no.1
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    • pp.154-171
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    • 2005
  • Herbal mixture water extract of (Rheum coreanum Scutellaria baikalensis, Phellodendron amurese), which exhibit several beneficial effects including acne and skin diseases, was tested for anti-microbial activity and anti-inflammation effects. The herbal mixture extract showed antimicrobial activity against Stapylococcus epidermis and Propionbacterium acne. The growth of Stapylococcus epidermis and Propionibacterium acne was inhibited completely by addition of 1.0% of the extract. Also in the present study we examined the mixture extract on compound 48/80 induced allergy and LPS induced cyclooxygenase-2(COX-2) gene expression in RAW264.7 macrophage. The results indicated the ear swelling and histamine release induced by compound 48/80 were dose-dependently reduced, ranging 18-36% and 10-61%, respectively. Furthermore the extract inhibited the expression of LPS-induced COX-2 proteins and mRNAs without an appreciable cytotoxic effects on RAW264.1 cells. The cytotoxicity of the extract using MTT assay showed the cytotoxicity of 6% and 13% against L929 cell line. Based on these results, it is concluded that the herbal mixture water extract can be applied to this acne and skin diseases therapy.

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Induction of Lysozyme Gene Expression During Involution of Mouse Mammary Gland

  • Lee, M.J.;Han, O.;Back, K.;Choi, Y.J.;Baik, M.G.
    • Asian-Australasian Journal of Animal Sciences
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    • v.14 no.4
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    • pp.462-466
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    • 2001
  • To understand molecular mechanisms of mouse mammary gland involution, clones were isolated by differential screening of a cDNA library. Partial sequences of a clone showed 100% identity to cDNA sequences of mouse lysozyme P gene. Northern analysis was performed to examine expression levels of lysozyme mRNA in mammary gland at several physiological states. Expression of lysozyme gene was induced at involution day 5 compared with lactating stage. High levels of lysozyme mRNA were also detected at virgin tissues. Two types of separate genes, P and M lysozyme, have been known in mouse, and we found that both lysozyme P and M genes were expressed in mammary tissues by reverse transcriptase-polymerase chain reaction. The lysozyme enzyme activity determined by lysoplate assay was also higher in involuted mammary tissues compared with lactating tissues, showing a similar trend to its mRNA levels. Lysozyme is an antimicrobial protein and involved in host defense mechanism. The increase in lysozyme gene expression may help to prevent microbial infection during mammary gland involution at which stage the residual milk in the mammary gland provides good nutritional sources for microbial growth.

Microbial $9{\alpha}$-Hydroxylase:Epoxidation of 9(11)-dehydro-$17{\alpha}$-methyl-testosterone

  • Kang, Hee-Kyoung;Lee, Sang-Sup
    • Archives of Pharmacal Research
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    • v.20 no.6
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    • pp.525-528
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    • 1997
  • Steroid $9{\alpha}$.-hydroxylase is a key enzyme system in steroid nucleus degradation in company with ${\Delta}$-dehydrogenase. To examine $9{\alpha}$-hydroxylase activity during microbial transformation of steroids, 9(11)-dehydro-$17{\alpha}$-methyl-testosterone was adopted as a stable substrate for preventing the rupture of steroid nucleus. Using Nocardia restrictus ATCC 14887 capable of introducing a $9{\alpha}$-hydroxyl group into steroids, $9{\alpha}$,$11{\alpha}$-oxido-$17{\beta}$-hydroxy-$17{\alpha}$-methyl-4-androstene-3-one and $9{\alpha}$-hydroxyl group into steroids,$9{\alpha}$,$11{\alpha}$-oxido-$17{\beta}$-hydroxy-$17{\alpha}$-methyl-1,4-androstadiene-3- one were obtained. These microbiologically transformed products could be used as reference compounds in the enzyme assay.

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Drug adsorption and anti-microbial activity of functionalized multiwalled carbon nanotubes

  • Saxena, Megha;Mittal, Disha;Boudh, Richa;Kumar, Kapinder;Verma, Anita K.;Saxena, Reena
    • Advances in nano research
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    • v.11 no.6
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    • pp.667-678
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    • 2021
  • Multiwalled carbon nanotubes (MWCNTs) were first oxidized (O-CNTs) to introduce carboxylic group and then further functionalized (F-CNTs) with m-phenylenediamine, which was confirmed by FTIR and SEM. It was used as an effective adsorbent for the adsorptive removal of diclofenac drug from water. Under optimum conditions of pH 6, stirring speed 600 rpm, the maximum adsorption capacity obtained was 532 mg g-1 which is superior to the values reported in literature. The adsorption was quite rapid as 25 mg L-1 drug solution was adsorbed in only 3 minutes of contact time with 10 mg of adsorbent dose. The adsorption kinetics and isotherms were studied using various models to evaluate the adsorption process. The results showed that the data best fit in kinetics pseudo-second order and Langmuir isotherm model. Furthermore, the oxidized and functionalized MWCNTs were applied on gram-negative Escherichia coli and gram-positive Staphylococcus aureus using agar disc diffusion assay to validate their anti-microbial activity. Results were unique as both oxidized and functionalized MWCNTs were equally active against both E. coli and S. aureus. The newly synthesized F-CNTs have great potential in water treatment, with their dual action of removing drug and pathogens from water, makes it potential applicant to save environment.

Disruption of Established Bacterial and Fungal Biofilms by a Blend of Enzymes and Botanical Extracts

  • Gitte S. Jensen;Dina Cruickshank;Debby E. Hamilton
    • Journal of Microbiology and Biotechnology
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    • v.33 no.6
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    • pp.715-723
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    • 2023
  • Microbial biofilms are resilient, immune-evasive, often antibiotic-resistant health challenges, and increasingly the target for research into novel therapeutic strategies. We evaluated the effects of a nutraceutical enzyme and botanical blend (NEBB) on established biofilm. Five microbial strains with known implications in chronic human illnesses were tested: Candida albicans, Staphylococcus aureus, Staphylococcus simulans (coagulase-negative, penicillin-resistant), Borrelia burgdorferi, and Pseudomonas aeruginosa. The strains were allowed to form biofilm in vitro. Biofilm cultures were treated with NEBB containing enzymes targeted at lipids, proteins, and sugars, also containing the mucolytic compound N-acetyl cysteine, along with antimicrobial extracts from cranberry, berberine, rosemary, and peppermint. The post-treatment biofilm mass was evaluated by crystal-violet staining, and metabolic activity was measured using the MTT assay. Average biofilm mass and metabolic activity for NEBB-treated biofilms were compared to the average of untreated control cultures. Treatment of established biofilm with NEBB resulted in biofilm-disruption, involving significant reductions in biofilm mass and metabolic activity for Candida and both Staphylococcus species. For B. burgdorferi, we observed reduced biofilm mass, but the remaining residual biofilm showed a mild increase in metabolic activity, suggesting a shift from metabolically quiescent, treatment-resistant persister forms of B. burgdorferi to a more active form, potentially more recognizable by the host immune system. For P. aeruginosa, low doses of NEBB significantly reduced biofilm mass and metabolic activity while higher doses of NEBB increased biofilm mass and metabolic activity. The results suggest that targeted nutraceutical support may help disrupt biofilm communities, offering new facets for integrative combinational treatment strategies.

Validation of Synovial Fluid Clinical Samples for Molecular Detection of Pathogens Causing Prosthetic Joint Infection Using GAPDH Housekeeping Gene as Internal Control

  • Jiyoung Lee;Eunyoung Baek;Hyesun Ahn;Youngnam Park;Geehyuk Kim;Sua Lim;Suchan Lee;Sunghyun Kim
    • Biomedical Science Letters
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    • v.29 no.4
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    • pp.220-230
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    • 2023
  • Identification of the pathogens causing infection is important in terms of patient's health management and infection control. Synovial fluids could be used as clinical samples to detect causative pathogens of prosthetic joint infections (PJIs) using molecular diagnostic assays, therefore, normalization and validation of clinical samples are necessary. Microbial culture is considered the gold standard for all infections, including PJIs. Recently, molecular diagnostic methods have been developed to overcome the limitation of microbial culture. Therefore, guideline for validating clinical samples to provide reliable results of molecular diagnostic assays for infectious diseases is required in clinical field. The present study aimed to develop an accurate validating method of synovial fluid clinical samples using GAPDH gene as an internal control to perform the quantitative PCR TaqMan probe assay to detect pathogens causing PJIs.

Fibrinolytic, Immunostimulating, and Cytotoxic Activities of Microbial Strains Isolated from Kochujang (고추장 분리 균주의 혈전용해능, 면역활성능 및 세포독성 효과 조사)

  • Seo, Mi-Young;Kim, Seung-Ho;Lee, Cheol-Ho;Cha, Seong-Kwan
    • Korean Journal of Food Science and Technology
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    • v.39 no.3
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    • pp.315-322
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    • 2007
  • This study was carried out to investigate the functional activities of microorganisms isolated from kochujang, such as fibrinolytic, immunostimulating, and cytotoxical activities, and to apply these microorganisms to kochujang products. Ninety-one microbial strains with proteolytic activity were selected from 294 strains isolated from traditional and commercial kochujang. Three strains (TPP 0014, TPP 6013, and TPP 6015) with high fibrinolytic activity were tested for their immunostimulating and cytotoxical activities. For the assessment of macrophage activation, cytokines such as tumor necrosis factor, $interleukin-1{\alpha}$ and nitrogen oxide were measured with the murine macrophage cell line RAW 264.7. In addition, the cytotoxical activities of the three strains were examined by MTT assay on the colon cancer cell line SNU-C4 and normal cell line CHO-K1. Using an API identifying kit, two of the microbial strains (TPP 0014 and TPP 6015) were identified as Bacillus stearothermophilus and the other strain (TPP 6013) was identified as B. amyloliquefacience.