• Title/Summary/Keyword: Dilution test

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Susceptibility Testing of Helicobacter pylori to Metronidazole and DNA Fingerprints of Resistant Strains in Singapore

  • HUA, JIESONG;KHAY-GUAN YEOH;PENGYUAN ZHENG;HAN CHONG NG;BOW HO
    • Journal of Microbiology and Biotechnology
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    • v.9 no.3
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    • pp.328-333
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    • 1999
  • Susceptibility of 61 strains of Helicobacter pylori to metronidazole was examined by both the disk diffusion method using a cut-off of 15㎜ for resistance and the E test with a cut-off of 8㎎/l. The MIC/sub 50/ and MIC/sub 90/ by the E test were 2 ㎎/l and 256㎎/l, respectively. Metronidazole resistance was found in 22 (36%) out of the 61 H. pylori strains by the E test and in three additional strains by the disk diffusion method. Amongst the latter three isolates, the MICs by the E test were 4 ㎎/l, 6㎎/l, and 6㎎/l, respectively. These figures are one log₂ or half log₂ dilution lower than the cut-off of 8㎎/l recommended as resistance for the E test. All 22 metronidazole resistant H. pylori isolates by the E test that were subjected to random amplified polymorphic DNA (RAPD) fingerprinting showed different DNA fingerprints. Interestingly, >90% of resistant isolates possess two common DNA bands of 0.4 and 0.9 kb. This study demonstrates that the results of the disk diffusion method for testing H. pylori susceptibility to metronidazole correlates well with that of the E test. The criteria for interpretation need to be internationally standardized so that the results from different centers can be compared.

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Emission Characteristics of Odorous Sulfur Gases from Food Types: A Case Study on Boiled Egg, Milk, Canned Meat, and Strawberry (음식물의 악취 황화합물 발생특성 조사: 계란, 우유, 고기통조림, 딸기에 대한 사례 연구)

  • Kim, Bo-Won;Ahn, Jeong-Hyeon;Kim, Ki-Hyun;Jo, Sang-Hee
    • Journal of Korean Society for Atmospheric Environment
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    • v.29 no.5
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    • pp.615-624
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    • 2013
  • In this study, the emission patterns of reduced sulfur compounds (RSC) were investigated using four different types of food samples (boiled egg, milk, canned meat and strawberry) between fresh and decaying stages. To this end, the concentrations of RSCs were measured at storage days of 0, 1, 3, 6, and 9 under room temperature. Four sulfur compounds ($H_2S$, $CH_3SH$, DMS and DMDS) were selected as target compounds along with two reference compounds ($CS_2$ and $SO_2$). Their concentrations were quantified using GC-PFPD equipped with thermal desorption (TD) system. The boiled egg showed the highest concentration of $H_2S$ (3,655 ppb) at D-1, while $CH_3SH$ reached its maximum value of 64.4~78.5 ppb after 3 days. In milk samples, concentration of $CH_3SH$, DMS, and DMDS went up to 487, 16.3, and 578 ppb, respectively with the progress of decay (D-9). In case of canned meat, concentration of $H_2S$ and $CH_3SH$ peaked in the beginning (D-0) such as 345 and 66.6 ppb. In case of strawberry, $CH_3SH$ and DMDS showed the maximum concentrations 135 and 50.5 ppb at D-1, respectively. The olfactometry dilution-to-threshold (D/T) ratio by air dilution sensory (ADS) test showed similar patterns when sum of odor intensity (SOI) was derived via conversion of odorant concentration data. The results of this study confirm that the time of strong RSC emissions is distinguished for each food type between fresh (e.g., strawberries) and decaying conditions (e.g., milk).

SPF 닭에서 재조합 H9N3 조류 인플루엔자 백신의 효능과 안전성 평가

  • Sin, Jeong-Hwa;Mo, In-Pil
    • Proceedings of the Korea Society of Poultry Science Conference
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    • 2006.11a
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    • pp.90-91
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    • 2006
  • To reduce the economic impact and control Low pathogenic avian influenza (LPAI), vaccination with inactivated vaccine has been considered in this country. We tried to develop inactivated vaccine with reassorted H9N3 AI virus which has different type of neuraminidase compare to those of field AI virus. Before reassorted vaccine was produced, we confirm the virus as master seed by limiting dilution, RT-PCR and sequencing method. Also, we evaluate the biological characteristics of the virus to find out the possibility of prevention against field infection of AI virus. Finally, we evaluate the safety and efficacy of the vaccine made of reassorted AI virus in the specific pathogen free (SPF) chickens. After limiting dilution, we choose RV7CE4 as a vaccine candidate and compare the gene sequence of this vaccine strain to those of AI05GA which is parents strain. Compared to amino acid sequences of specific gene of AI05GA and RV7CE4, exhibited a high degree of amino acid sequence homology. In the safety and efficacy test, there were no specific clinical signs or mortality. Reassorted H9N3 viruses were reisolated in cloaca swab on 5 days post inoculation. In the vaccine study, once or twice vaccination was performed and challenged with H9N2 field virus (01310). Vaccine has no adverse effect on birds and formed good immune capability which reduce viral shedding in the birds infected with 01310. Based on the above result, we developed reassorted H9N3 vaccine which will efficiently prevent the low pathogenic AIV (H9N2) infection in the poultry farms.

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Cytocompatibility and cell proliferation evaluation of calcium phosphate-based root canal sealers

  • Mestieri, Leticia Boldrin;Zaccara, Ivana Maria;Pinheiro, Lucas Siqueira;Barletta, Fernando Branco;Kopper, Patricia Maria Polli;Grecca, Fabiana Soares
    • Restorative Dentistry and Endodontics
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    • v.45 no.1
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    • pp.2.1-2.7
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    • 2020
  • Objectives: This study aimed to evaluate the cell viability and migration of Endosequence Bioceramic Root Canal Sealer (BC Sealer) compared to MTA Fillapex and AH Plus. Materials and Methods: BC Sealer, MTA Fillapex, and AH Plus were placed in contact with culture medium to obtain sealers extracts in dilution 1:1, 1:2 and 1:4. 3T3 cells were plated and exposed to the extracts. Cell viability and migration were assessed by 3-(4,5-dimethylthiazoyl)-2,5-diphenyl-tetrazolium bromide (MTT) and Scratch assay, respectively. Data were analyzed by Kruskal-Wallis and Dunn's test (p < 0.05). Results: The MTT assay revealed greater cytotoxicity for AH Plus and MTA Fillapex at 1:1 dilution when compared to control (p < 0.05). At 1:2 and 1:4 dilutions, all sealers were similar to control (p > 0.05) and MTA Fillapex was more cytotoxic than BC Sealer (p < 0.05). Scratch assay demonstrated the continuous closure of the wound according to time. At 30 hours, the control group presented closure of the wound (p < 0.05). At 36 hours, only BC Sealer presented the closure when compared to AH Plus and MTA Fillapex (p < 0.05). At 42 hours, AH Plus and MTA Fillapex showed a wound healing (p > 0.05). Conclusions: All tested sealers demonstrated cell viability highlighting BC Sealer, which showed increased cell migration capacity suggesting that this sealer may achieve better tissue repair when compared to other tested sealers.

Emission Characteristics of Odorous Gases with the Decay of Albumin and Yolk of Boiled Egg (삶은 달걀의 부패에 따른 부위별 냄새물질의 발생특성 연구)

  • Kim, Bo-Won;Kim, Ki-Hyun;Kim, Yong-Hyun;Ahn, Jeong-Hyeon
    • Journal of Korean Society for Atmospheric Environment
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    • v.30 no.2
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    • pp.95-109
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    • 2014
  • In this study, the concentration of odorants released from albumin (EA) and yolk (EY) portions of boiled egg samples were determined as a function of storage time. The concentrations were measured at storage days of 0, 1, 3, 6, and 9 under room temperature. As such, odorants produced during both fresh and decay conditions were measured through time. A total of 19 compounds were selected as the main target odorants along with 12 reference compounds. GC-MS (for VOC) and GC-PFPD system (for sulfur gases) equipped with thermal desorption (TD) system were employed for odorant analysis in this work. The initial concentrations measured from the chamber system were converted into flux terms ($ng{\cdot}g^{-1}{\cdot}min^{-1}$). The EA showed the highest concentration of $H_2S$ (234 $ng{\cdot}g^{-1}{\cdot}min^{-1}$) at EA-0, and the concentrations of AT (Acetone) was also seen clearly in the range of 11.7 (EA-0) to 58.6 $ng{\cdot}g^{-1}{\cdot}min^{-1}$ (EA-9). The EY showed similar patterns. EtAl (Ethyl alcohol) increased 9.47 (EA-1) to 96.7 $ng{\cdot}g^{-1}{\cdot}min^{-1}$ (EA-9) in EA samples. Ketone, alcohol, sulfur groups generally exhibited high concentrations compared to other odorants. These data were also compared in relation to olfactometry related dilution-to-threshold (D/T) ratio by air dilution sensory (ADS) test and sum of odor intensity (SOI).

Aroma Characteristics of Neungee(Sarcodon aspratus) (능이버섯의 향기특성)

  • Jeong, Ok-Jin;Yoon, Hyang-Sik;Min, Young-Kyoo
    • Korean Journal of Food Science and Technology
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    • v.33 no.3
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    • pp.307-312
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    • 2001
  • Flavor compounds in Neungee (sarcodon aspratus) were extracted by simutaneous distillation and extraction (SDE), supercritical fluid extraction (SFE) and headspace method. Flavor compounds obtained by various extraction methods were analyzed with GC and GC-MS. The funtionality of flavor compounds were determined by aroma extract dilution analysis (AEDA) of GC-ofactometry methods. Fifty one flavor compounds were totally identified in Neungee mushroom. However, the numbers of flavor extracted SDE, SFE and headspace were 33, 26 and 17 respectively. The major flavor compounds obtained by SDE, SFE and headspace were 1-octen-3-ol, 1-octen-3-one, 3-octanone, 2-octen-1-ol, 3-octanol, 1-octanol and benzenealdehyde. As the results of sniffing test, the major flavor compounds were found to be fresh mushroom flavor, wood flavor, refreshing sweet flavor, mold flavor, bitter-mushroom and metalic-flavor.

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Fungicide selections for control of chili pepper stem rot caused by Sclerotium rolfsii using an agar dilution method

  • Lee, Soo Min;Min, Jiyoung;Kim, Heung Tae
    • Korean Journal of Agricultural Science
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    • v.44 no.3
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    • pp.339-347
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    • 2017
  • Sclerotium rolfsii causing southern blight on numerous vegetable and fruit crops was isolated from stems of chili peppers showing wilting symptoms. The pathogen was identified by morphological observation and DNA sequencing analysis of ITS region. To select an effective fungicide for control of southern blight, we investigated the inhibition efficacy of thirty fungicides included in nine groups of fungicides with different mechanisms of action. A fungal growth inhibition assay was conducted through an agar dilution method by using mycelial discs and sclerotia of the pathogen as inoculum, respectively. When mycelial discs were used as an inoculum, several fungicides showed good inhibitory activity against the mycelial growth of S. rolfsii 12-6. All DMI fungicides tested had a good inhibition except for prochloraz which had low inhibitory effect. All strobilurin fungicides tested except for kresoxim-methyl and all SDHI fungicides tested except for boscalid and fluopyram, had a good inhibition. Also, fludioxonil, a protective fungicide and fluazinam had a good inhibitory effect. Interestingly, when sclerotia were used as an inoculum, inhibition efficacy was increased for fluopyram, a SDHI fungicide, and for some protective fungicides such as propineb, chlorothalonil, dithianon, and folpet. All the fungicides selected in this study should be tested in the field for their control activities against stem rot for practical use in chili pepper cultivation.

The Antimicrobial Effects of Natural Aromas for Substitution of Parabens (합성 항균제를 대체하기 위한 천연물질의 항균 효과)

  • 조춘구;김봉남;홍세흠;한창규
    • Journal of the Society of Cosmetic Scientists of Korea
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    • v.28 no.1
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    • pp.166-185
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    • 2002
  • Aroma oils extracted from the natural material have antibacterial, antivirus, antiinflammatory, and preservative effect. The preserve efficacy testing between aroma oils and parabens as an artificial preservative had been performed and then it had been suggested that aroma oil was possibile to apply to the cosmetics. Aroma oils were pine, rosemary, lemon and eucalyptus, and parabens were methylparaben, blitylparaben. Antiseptic concentrations of aroma oils and parabens having 0.0, 0.1, 0.2, 0.4, 0.8, 1.0wt% were tested respectively. Escherichia coil(ATCC No.8739), Pseudomonas aeruginosa(ATCC No. 9027) which are gram-negative and Staphylococcus aureus (ATCC No. 6538), Bacillus subtilis(ATCC No. 6633) which are gram-positive were used as the test organisms. Disk paper and broth dilution methods were used as the methods of preservative efficacy testing. The antibacterial activity of aroma oils and parabens for gram-positive were better than that for gram-negative. For the antibacterial activity aroma oils were better than parabens. Among the aroma oils, rosemary and pine having superior antibacterial activity were selected and blended to illuminate if there is any synergy, There was synergical effect and optimum ratio of aroma blend is 3 : 1(rosemary pine) in this study.

Riboflavin Inhibits Growth of Helicobacter pylori by Down-regulation of polA and dnaB Genes

  • Kwon, Hye Jin;Lee, Min Ho;Kim, Hyun Woo;Yang, Ji Yeong;Woo, Hyun Jun;Park, Min;Moon, Cheol;Kim, Sa-Hyun;Kim, Jong-Bae
    • Biomedical Science Letters
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    • v.26 no.4
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    • pp.288-295
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    • 2020
  • Infection of Helicobacter pylori on gastric mucosa is associated with various gastric diseases. According to the WHO, H. pylori causes gastric cancer and has been classified as a class I carcinogen. Riboflavin is an essential vitamin which presents in a wide variety of foods. Previous studies have shown that riboflavin/UVA was effective against the growth inhibition of Staphylococcus aureus, S. epidermidis and multidrug-resistant Pseudomonas aeruginosa and had the potential for antimicrobial properties. Thus, we hypothesized that riboflavin has a potential role in the growth inhibition of H. pylori. To demonstrate inhibitory concentration of riboflavin against H. pylori, we performed agar and broth dilution methods. As a result, we found that riboflavin inhibited the growth of H. pylori. The MIC was 1 mM in agar and broth dilution test. Furthermore, to explain the inhibitory mechanism, we investigated whether riboflavin has an influence on the replication-associated molecules of the bacteria using RT-PCR to detect mRNA expression level in H. pylori. Riboflavin treatment of H. pylori led to down-regulation of polA and dnaB mRNA expression levels in a dose dependent manner. After then, we also confirmed whether riboflavin has cytotoxicity to human cells. We used AGS, a gastric cancer cell line, and treated with riboflavin did not show statistically significant decrease of cell viability. Thus, these results indicate that riboflavin can suppress the replication machinery of H. pylori. Taken together, these findings demonstrate that riboflavin inhibits growth of H. pylori by inhibiting replication of the bacteria.

Biocompatibility and Bioactivity of Four Different Root Canal Sealers in Osteoblastic Cell Line MC3T3-El

  • Jun, Nu-Ri;Lee, Sun-Kyung;Lee, Sang-Im
    • Journal of dental hygiene science
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    • v.21 no.4
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    • pp.243-250
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    • 2021
  • Background: Endodontic sealers or their toxic components may become inflamed and lead to delayed wound healing when in direct contact with periapical tissues over an extended period. Moreover, an overfilled sealer can directly interact with adjacent tissues and may cause immediate necrosis or further resorption. Therefore, the treatment outcome conceivably depends on the endodontic sealer's biocompatibility and osteogenic potential. This study aimed to evaluate the cell viability and osteogenic effects of four different sealers in osteoblastic cells. Methods: AH Plus (resin-based sealer), Pulp Canal Sealer EWT (zinc oxide-eugenol sealer), BioRoot RCS (calcium silicate-based sealer), and Well-Root ST (MTA-based calcium silicate sealer) were mixed strictly according to the manufacturer's instructions, and dilutions of sealer extracts (1/2, 1/5 and 1/10) were determined. Cell viability was measured using the water-soluble tetrazolium-8 (WST-8) assay. Differentiation was assessed by alkaline phosphatase (ALP) activity and mineralized nodule formation by Alizarin Red S staining. Results: The cell viability of the extracts derived from the sealers excluding Well-Root ST was concentration dependent, with sealer extracts having the least viability at a 1/2 dilution. At sealer extract dilution of 1/10, the test groups showed the same survival rate as that control group, with the exception of BioRoot RCS. Among all experimental groups, BioRoot RCS showed the highest cell viability after 48 hours. The ALP activity was significantly higher in a concentration-dependent manner. Furthemore, all four materials promoted ALP activity and mineralized nodule formation compared to the control at 1/10 dilutions. Conclusion: This is the first study to highlight the differences in biological activity of these four materials. These results suggest that the composition of root canal sealers appears to alter the form of biocompatibility and osteoblastic differentiation.