• 제목/요약/키워드: microbial assay

검색결과 281건 처리시간 0.024초

Effect of Partial Replacement of Wheat Flour with High Quality Cassava Flour on the Chemical Composition, Antioxidant Activity, Sensory Quality, and Microbial Quality of Bread

  • Eleazu, Chinedum;Eleazu, Kate;Aniedu, Chinyere;Amajor, John;Ikpeama, Ahamefula;Ebenzer, Ike
    • Preventive Nutrition and Food Science
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    • 제19권2호
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    • pp.115-123
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    • 2014
  • In the current study, wheat flour was mixed with high quality cassava flour (HQCF) in several ratios: 90:10, 80:20, 70:30, and 60:40, and used to prepare 10%, 20%, 30%, and 40% National Root Crops Research Institute (NRCRI) cassava bread, respectively. 100% wheat bread was prepared as a control (100% wheat bread). Five bread samples were prepared per group. Antioxidant assays [i.e., 2,2-diphenyl-1-picrylhydrazyl radical (DPPH) scavenging assay, reducing power assay] revealed that the bread samples had considerable antioxidant capacities. Substitution of wheat flour with HQCF at various concentrations resulted in dose dependent decreases in the mineral and protein contents of the resulting bread samples. The crude fiber content of the bread samples was minimal, while the carbohydrate content of the bread samples ranged from 43.86% to 48.64%. A 20% substitution of wheat flour with HQCF yielded bread samples with a general acceptability that was comparable to that of 100% wheat bread. The mean bacteria counts of the bread samples ranged from $2.0{\times}10^3CFU/mL$ to $1.4{\times}10^4CFU/mL$, while the fungal counts ranged from 0 CFU/mL to $3{\times}10^3CFU/mL$. There was a positive correlation between the DPPH antioxidant activities and the reducing powers of the bread samples ($R^2=0.871$) and a positive correlation between the DPPH antioxidant activities and the flavonoid contents of the bread samples ($R^2=0.487$). The higher microbial load of the NRCRI cassava bread samples indicates that these bread samples may have a shorter shelf life than the 100% wheat bread. The significant positive correlation between total flavonoid content and reducing power ($R^2=0.750$) suggests that the flavonoids present in the lipophilic fractions of the bread samples could be responsible for the reductive capacities of the bread samples.

Development of Fecal Microbial Enzyme Mix for Mutagenicity Assay of Natural Products

  • Yeo, Hee-Kyung;Hyun, Yang-Jin;Jang, Se-Eun;Han, Myung-Joo;Lee, Yong-Sup;Kim, Dong-Hyun
    • Journal of Microbiology and Biotechnology
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    • 제22권6호
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    • pp.838-848
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    • 2012
  • Orally administered herbal glycosides are metabolized to their hydrophobic compounds by intestinal microflora in the intestine of animals and human, not liver enzymes, and absorbed from the intestine to the blood. Of these metabolites, some, such as quercetin and kaempherol, are mutagenic. The fecal bacterial enzyme fraction (fecalase) of human or animals has been used for measuring the mutagenicity of dietary glycosides. However, the fecalase activity between individuals is significantly different and its preparation is laborious and odious. Therefore, we developed a fecal microbial enzyme mix (FM) usable in the Ames test to remediate the fluctuated reaction system activating natural glycosides to mutagens. We selected, cultured, and mixed 4 bacteria highly producing glycosidase activities based on a cell-free extract of feces (fecalase) from 100 healthy Korean volunteers. When the mutagenicities of rutin and methanol extract of the flos of Sophora japonica L. (SFME), of which the major constituent is rutin, towards Salmonella typhimurium strains TA 98, 100, 102, 1,535, and 1,537 were tested using FM and/or S9 mix, these agents were potently mutagenic. These mutagenicities using FM were not significantly different compared with those using Korean fecalase. SFME and rutin were potently mutagenic in the test when these were treated with fecalase or FM in the presence of S9 mix, followed by those treated with S9 mix alone and those with fecalase or FM. Freeze-dried FM was more stable in storage than fecalase. Based on these findings, FM could be usable instead of human fecalase in the Ames test.

In vivo와 In vitro 평가모델을 利用한 韓藥抽出物의 毛髮成長 및 促進에 미치는 實驗的 硏究 (Extracts for the Hair Growth Stimulation using In vivo and In vitro Test Models)

  • 강학천;이수형;김남권;임홍진;황충연
    • 한방안이비인후피부과학회지
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    • 제15권2호
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    • pp.53-79
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    • 2002
  • To screen the effective materials for hair loss treatment, several natural extracts were tested using in vivo and in vitro test models. Firstly, all test materials were applicated onto the back skin of C57BL/6 mouse and then hair growth promoting effect was measured using hair growth index. As a result, Prunus mume, black bean, Brassica campestris subsp. black sesame and Rubi Fructus showed potent hair growth promoting effect, ranking as 1.5-2.0 of hair growth index. However, there were no plant extracts, which have remarkable potential of growth promotion of human hair dermal papilla cells cultured in vitro. In the experiments of 5${\alpha}$-reductase type Ⅱ inhibition assay, Prunus mume, Eriobotryae Folium showed effective potential to inhibit the activity of 5${\alpha}$-reductase type Ⅱ. To investigate the possible involvement of the effect of several plant extracts on the gene expression of growth factors in human hair dermal papilla cells, RT - PCR analyses were performed. However, there were no plant extracts, which have profound effect on the gene expression of several growth factors such as IGF-I, KGF, HGF and VEGF in the dermal papilla cells. Another tests for inhibition of microbial such as P. acne were also carried out to find whether these plant extracts have anti -microbial activities. Rubi Fructus showed anti -microbial effects on Propionibacterium acnes, which is believed as a pathogen of acne. Together, these results showed several plant extracts can be used for hair growth promotion.

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數種의 韓藥材가 毛髮成長에 미치는 影響 (Studies on the effects of medicinal plant extracts on the hair growth stimulation)

  • 최웅;최정화;김종한
    • 한방안이비인후피부과학회지
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    • 제15권2호
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    • pp.80-103
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    • 2002
  • To screen the effective materials for hair loss treatment, several natural extracts were tested using in vivo and in vitro test models. Firstly, all test materials were applicated onto the back skin of C57BL/6 mouse and then hair growth pormoting effect were measured using hair growth index As a result, Polygonum muitifiorum Thunb and Terrninalia chebula Retz. showed potent hair growth promoting effect, ranking as 1.5-2.0 of hair growth index. However, there were no plant extracts, which have remarkable potential of growth promotion of human hair dermal papilla cells cultured in vitro. In the experiments of 5${\alpha}$-reductase type Ⅱ inhibition assay, Morus alba L., Chaenomelis Fructus, Saussureae Radix, Angelicae Gigantis Radix, Polygonum multifiorum Thunb, and Angelica dahurica (Fischer) Bentham et Hooker f. showed effective potential to inhibit the activity of 5${\alpha}$-reductase type Ⅱ. To investigate the possible involvement of effects of several plant extracts on the gene expression of growth factors in human hair dermal papilla cells, RT-PCR analyses were performed. As a consequences, Mentha haplocalyx Briq., Cimicifuga foetida L., Eclipta prostrata (L.) L., Pinus densiflora S. et. Z, and Polygonum muitifiorum Thunb revealed the regulatory roles on the expression of growth factors such as IGF-I, KGF, HGF and VEGF in the dermal papilla cells. Another test for inhibition of microbial such as P. acne and P. ovale were also carried out to find whether these plant extracts have anti-microbial activities. Morus alba L. and Chaenomelis Fructus showed anti-microbial effects on Propionibacterium acnes, which is believed as a pathogen of acne. Together, these results showed several plant extracts can be used for hair growth promotion.

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Purification, Characterization and Immobilization of Lipase from Proteus vulgaris OR34 for Synthesis of Methyl Oleate

  • Misbah, Asmae;Koraichi, Saad Ibnsouda;Jouti, Mohamed Ali Tahri
    • 한국미생물·생명공학회지
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    • 제48권4호
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    • pp.491-505
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    • 2020
  • A newly isolated strain, Proteus vulgaris OR34, from olive mill waste was found to secrete an alkaline extracellular lipase at 11 U·ml-1 when cultivated on an optimized liquid medium. This lipase was purified 94.64-fold with a total yield of 9.11% and its maximal specific activity was shown to be 3232.58 and 1777.92 U·mg-1 when evaluated using the pH-stat technique at 55℃ and pH 9 and Tributyrin TC4 or olive oil as the substrate. The molecular mass of the pure OR34 lipase was estimated to be around 31 kDa, as revealed by SDS-PAGE and its substrate specificity was investigated using a variety of triglycerides. This assay revealed that OR34 lipase preferred short and medium chain fatty acids. In addition, this lipase was stable in the presence of high concentrations of bile salt (NaDC) and calcium ions appear not to be necessary for its activity. This lipase was inhibited by THL (Orlistat) which confirmed its identity as a serine enzyme. In addition, the immobilization of OR34 lipase by adsorption onto calcium carbonate increased its stability at higher temperatures and within a larger pH range. The immobilized lipase exhibited a high tolerance to organic solvents and retained 60% of its activity after 10 months of storage at 4℃. Finally, the OR34 lipase was applied in biodiesel synthesis via oleic acid mediated esterification of methanol when using hexane as solvent. The best conversion yield (67%) was obtained at 12 h and 40℃ using the immobilized enzyme and this enzyme could be reused for six cycles with the same efficiency.

화학소독제 처리 후 가타파차 콘의 멸균 효과 및 표면 성상의 변화 평가 (Assessment of decontamination of gutta-percha cone and the change of surface texture after rapid chemical disinfection)

  • 방난심;정일영;유윤정;금기연
    • Restorative Dentistry and Endodontics
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    • 제31권2호
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    • pp.133-139
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    • 2006
  • 본 연구의 목적은 첫째 임상에서 진료실에 노출된 가타파차 콘 표면의 오염 균종을 중합효소연쇄반응법 (polymerase chain reaction, PCR)을 이용해 동정하고, 둘째 이들 세균으로 오염시킨 가타파차 콘에 대해 2종의 소독제의 rapid sterilization 효과를 비교하였다. 또한 이들 소독제에 5분간 처리된 가타파차 콘 표면 성상의 변화를 주사전자현미경으로 관찰하였다. 진료실에 수 개월간 노출된 가타파차 콘 100개를 수거하여 배양지에 넣어 배양 후 universial primer를 사용한 PCR assay 를 통해 오염 균종을 동정하였다. 실험실 상에서 이 균종을 다시 배양하여 소독된 가타파차 콘에 접종하고 1주일간 배양한 후 2종의 소독제(5% NaOCl, 2% Chlorhexidine)에 1, 3, 5, 10 분간 담근 후 각 소독제의 종류와 적용시간에 따른 멸균 효과를 turbidity test 와 subculture 를 이용하여 평가하였다. 또한 각 소독제에 5분간 처리된 가타파차 콘 표면 성상의 변화를 주사전자현미경으로 관찰하였다. 중합효소연쇄반응법의 분석결과 17개의 가타 파차 콘이 오염된 것으로 나타났고 대부분이 Staphylococcus 계통이었으며, 2종의 소독제 모두 이들 균종에 대해 1분내에 멸균 효과를 나타냈다. 주사전자현미경상 NaOCl로 소독된 가타파차 콘 표면에는 cuboidal crystal 의 침전물이 전반적으로 관찰되었다. 본 연구 결과 2종의 소독제 모두 근관충전 전 가타파차 콘의 rapid sterilization 을 위해 유용하였으나 클로헥시딘으로 처리된 가타파차 콘이 크리스탈 침전물이 없는 좀더 깨끗한 표면을 갖는 것으로 나타났다.

생물의약품 제조공정에서 마이코플라스마 정량 검출을 위한 TaqMan Probe Real-Time PCR (TaqMan Probe Real-Time PCR for Quantitative Detection of Mycoplasma during Manufacture of Biologics)

  • 이재일;김인섭
    • KSBB Journal
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    • 제29권5호
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    • pp.361-371
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    • 2014
  • Mycoplasma is well recognized as one of the most prevalent and serious microbial contaminants of biologic manufacturing processes. Conventional methods for mycoplasma testing, direct culture method and indirect indicator cell culture method, are lengthy, costly and less sensitive to noncultivable species. In this report, we describe a new TaqMan probe-based real-time PCR method for rapid and quantitative detection of mycoplasma contamination during manufacture of biologics. Universal mycoplasma primers were used for mycoplasma PCR and mycoplasma DNA was quantified by use of a specific TaqMan probe. Specificity, sensitivity, and robustness of the real-time PCR method was validated according to the European Pharmacopoeia. The validation results met required criteria to justify its use as a replacement for the culture method. The established real-time PCR assay was successfully applied to the detection of mycoplasma from human keratinocyte and mesenchymal stem cell as well as Vero cell lines artificially infected with mycoplasma. The overall results indicated that this rapid, specific, sensitive, and robust assay can be reliably used for quantitative detection of mycoplasma contamination during manufacture of biologics.

Beauvericin, a cyclic peptide, inhibits inflammatory responses in macrophages by inhibiting the NF-κB pathway

  • Yoo, Sulgi;Kim, Mi-Yeon;Cho, Jae Youl
    • The Korean Journal of Physiology and Pharmacology
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    • 제21권4호
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    • pp.449-456
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    • 2017
  • Beauvericin (BEA), a cyclic hexadepsipeptide produced by the fungus Beauveria bassiana, is known to have anti-cancer, anti-inflammatory, and anti-microbial actions. However, how BEA suppresses macrophage-induced inflammatory responses has not been fully elucidated. In this study, we explored the anti-inflammatory properties of BEA and the underlying molecular mechanisms using lipopolysaccharide (LPS)-treated macrophage-like RAW264.7 cells. Levels of nitric oxide (NO), mRNA levels of transcription factors and the inflammatory genes inducible NO synthase (iNOS) and interleukin (IL)-1, and protein levels of activated intracellular signaling molecules were determined by Griess assay, semi-quantitative reverse transcriptase-polymerase chain reaction (RT-PCR), luciferase reporter gene assay, and immunoblotting analysis. BEA dose-dependently blocked the production of NO in LPS-treated RAW264.7 cells without inducing cell cytotoxicity. BEA also prevented LPS-triggered morphological changes. This compound significantly inhibited nuclear translocation of the $NF-{\kappa}B$ subunits p65 and p50. Luciferase reporter gene assays demonstrated that BEA suppresses MyD88-dependent NF-${\kappa}B$ activation. By analyzing upstream signaling events for $NF-{\kappa}B$ activation and overexpressing Src and Syk, these two enzymes were revealed to be targets of BEA. Together, these results suggest that BEA suppresses $NF-{\kappa}B$-dependent inflammatory responses by suppressing both Src and Syk.

계면활성제가 돌연변이원성 물질의 활성에 미치는 영향 (Effect of Synthetic Surfactants on the Activity of Mutagens)

  • 최윤호;정용
    • Environmental Analysis Health and Toxicology
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    • 제7권3_4호
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    • pp.69-79
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    • 1992
  • Recently, concerns of water pollution and health risks caused by synthetic detergents have emerged, as the use of various detergents has increased It has been suggested that some surfactants are cocarcinogens. The surfactants tested were linear alkylbenzene sulfonate, sodium lauryl sulfate, polyoxyethylene sorbitan monooleat (tween 80), and the mutagens were 1-nitropyrene, N -methyl- N'-nitro-N -nitrosoguanidine, benzo (a) pyrene, and aflatoxin B$_1$. This study was undertaken to investigate the effects of surfactants on the activity of mytagens using the Ames mutagenic assay with Salmonella typhimurium TA98, TA100. The results were summarized as follows: 1. The surfactants have no mutagenic activity of themselves. 2. Higher doses of surfactants than 100 $\mu\textrm{g}$/plate reduced the number of revertants. It is assumed that the reduction would inhibited cell growth. 3. When the comutagenic ratio is defined as the ratio between mutagenic activity itself and the activity with mutagen and surfactant (drinking water quality standard), LAS showed the comutagenic ratio 0.86-1.17 and SLS 0.74-1.10 as well. According to the comparisons, it could not be recognised for the comutagenicity of drinking water quality standard of surfactant. 4. As increasing the amount of mutagens, the designated amount of surfactant did not affected the mutagen's activity statistically. From the above result, syunthetic surfactants do not present mutagenicity and comutagenicity in the microbial assay.

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Anti-microbial Activity of Saussurea lappa C.B. Clarke Roots

  • Chang, Kyung-Mi;Choi, Soo-Im;Chung, Sophia J.;Kim, Gun-Hee
    • Preventive Nutrition and Food Science
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    • 제16권4호
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    • pp.376-380
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    • 2011
  • We investigated the total phenolic and flavonoid contents and the antimicrobial activity of ethanol extracts obtained from Saussurea lappa C.B. Clarke. The ethanol extracts of S. lappa C.B. Clarke were fractionated with various solvents (n-hexane, chloroform, and n-butanol). The antimicrobial activity of S. lappa C.B. Clarke was examined by disc-diffusion and micro-dilution susceptibility assays with six food-borne pathogens, and compared to that of the synthetic antibiotics. It is found that the S. lappa C.B. Clarke ethanol extract and n-hexane fraction have strong activity against B. cereus and V. parahaemolyticus strains compared to ampicillin. The inhibitory concentration ($IC_{50}$) values of hexane fraction against L. monocytogenes, B. cereus, and B. subtilis were 62.5, 250 and 500 ppm, respectively. Therefore, these data suggest that S. lappa C.B. Clarke may be useful as antimicrobial agents against food-borne pathogens.