• Title/Summary/Keyword: colony test

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Detection of Streptococcus mutans in Saliva using Monoclonal Antibodies (단일클론항체를 사용한 타액 내 Streptococcus mutans 검출)

  • Lee, Minjeong;Lee, Daewoo;Kim, Miah;Lee, Kyungyeol;Baik, Byeongju;Yang, Yeonmi;Kim, Jaegon
    • Journal of the korean academy of Pediatric Dentistry
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    • v.42 no.1
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    • pp.10-21
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    • 2015
  • The purpose of this study was to determine the usefulness of a detection method for Streptococcus mutans in saliva with monoclonal antibodies developed targeting Ag I/II and glucosyltransferases (gtf B, gtf C and gtf D) in Streptococcus mutans. In the three groups tested (adults, minors, and minors under orthodontic treatment), the results of the DMFT scores, the colony density (CFU/mL) in their saliva was measured using $Dentocult^{(R)}$-SM strip mutans, polymerase chain reaction was performed to test whether Streptococcus mutans and Streptococcus sobrinus were present, and Streptococcus mutans detecting tests performed in their saliva using four types of monoclonal antibody were collected. In conclusion, it was demonstrated that the Streptococcus mutants plays more important role in forming dental caries compared to Streptococcus sobrinus, and that the monoclonal antibodies against glucosyltransferases (gtf B, gtf C, gtf D) and Ag I/II of Streptococcus mutans are superior in detecting Streptococcus mutans to $Dentocult^{(R)}$-SM strip mutans or polymerase chain reaction.

Four-Week Repeated Intravenous Dose Toxicity and Toxicokinetic Study of TS-DP2, a Novel Human Granulocyte Colony Stimulating Factor in Rats

  • Lee, JooBuom;Lee, Kyungsun;Choe, Keunbum;Jung, Hyunseob;Cho, Hyunseok;Choi, Kiseok;Kim, Taegon;Kim, Seojin;Lee, Hyeong-Seok;Cha, Mi-Jin;Song, Si-Whan;Lee, Chul Kyu;Chun, Gie-Taek
    • Toxicological Research
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    • v.31 no.4
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    • pp.371-392
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    • 2015
  • TS-DP2 is a recombinant human granulocyte colony stimulating factor (rhG-CSF) manufactured by TS Corporation. We conducted a four-week study of TS-DP2 (test article) in repeated intravenous doses in male and female Sprague-Dawley (SD) rats. Lenograstim was used as a reference article and was administered intravenously at a dose of $1000{\mu}g/kg/day$. Rats received TS-DP2 intravenously at doses of 250, 500, and $1000{\mu}g/kg/day$ once daily for 4 weeks, and evaluated following a 2-week recovery period. Edema in the hind limbs and loss of mean body weight and body weight gain were observed in both the highest dose group of TS-DP2 and the lenograstim group in male rats. Fibro-osseous lesions were observed in the lenograstim group in both sexes, and at all groups of TS-DP2 in males, and at doses of TS-DP2 $500{\mu}g/kg/day$ and higher in females. The lesion was considered a toxicological change. Therefore, bone is the primary toxicological target of TS-DP2. The lowest observed adverse effect level (LOAEL) in males was $250{\mu}g/kg/day$, and no observed adverse effect level (NOAEL) in females was $250{\mu}g/kg/day$ in this study. In the toxicokinetic study, the serum concentrations of G-CSF were maintained until 8 hr after administration. The systemic exposures ($AUC_{0-24h}$ and $C_0$) were not markedly different between male and female rats, between the administration periods, or between TS-DP2 and lenograstim. In conclusion, TS-DP2 shows toxicological similarity to lenograstim over 4-weeks of repeated doses in rats.

Semaphoring mAb: a New Guide in RIT in Inhibiting the Proliferation of Human Skin Carcinoma

  • Liu, Yuan;Ma, Jing-Yue;Luo, Su-Ju;Sun, Chen-Wei;Shao, Li-Li;Liu, Quan-Zhong
    • Asian Pacific Journal of Cancer Prevention
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    • v.16 no.3
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    • pp.941-945
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    • 2015
  • Semaphoring is a transmembrane receptor which participates in many cytokine-mediated signal pathways that are closely related to the angiogenesis, occurrence and development of carcinoma. The present study was designed to access the effect of mono-antibody (mAb) guided radioimmunotherapy (RIT) on skin carcinoma and investigate the potential mechanisms. Semaphoring mAb was acquired from mice (Balb/c), purified with rProtein A column; purity, concentration and activity were tested with SDS-PAGE and indirect ELISA; specificity and expression on the cutanuem carcinoma line and tissue were tested by Western blotting; morphology change was assessed by microscopy. MTT assay and colony inhibition tests were carried out to test the influence on the proliferation of tumor cells; Western blotting was also carried out for expression of apoptosis-associated (caspase-3, Bax, Bcl-2) and proliferation-related (PI3K, p-Akt, Akt, p-ERK1/2, ERK1/2) proteins and analyse the change in signal pathways (PI3K/Akt and MEK/ERK). The purity of purified semaphorin mAb was 96.5% and the titer is about $1{\times}10^6$. Western blotting showed semaphoring mAb to have specifically binding stripes with semaphoring b1b2 protein, B16F10, and A431 cells at 39KDa, 100KDa and 130KDa, respectively. Positive expression was detected both in cutanuem carcinoma line and tissue and it mostly located in cell membranes. MMT assay revealed dose-relate and time-relate inhibitory effect of semaphorin mAb on A431 and B16F10. Colony inhibition tests also showed dose-relate inhibitory effects. Western blotting demonstrated the expression of apoptosis and proliferation-related protein and changes in signal pathway. In conclusion, we demonstrated that semaphorin is highly expressed on the tumor cell-surfaces and RIT with semaphorin mAb has effect in i nhibiting proliferation and accelerating apoptosis of tumor cells.

Pharmacological Effects of Proteoglycans Extracted from Fruiting Bodies of Fomitella fraxinea (장수버섯 자실체의 열탕추출액으로부터 분리한 단백다당체의 약리적 효과)

  • Yoon, Sang-Hong;Lim, Jae-Hyeon;Kim, Yang-Seop;Kim, Chang-Han;Jo, Joon-Hyeong;Hwang, Young-Soo
    • The Korean Journal of Mycology
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    • v.26 no.4 s.87
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    • pp.511-518
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    • 1998
  • Anti-complementary assay for immuno-stimulating polysaccharide screening, human tumor colony-forming assay for discovering anti-tumor drugs, and toxic assay against mouse were performed to examine pharmacological activities of polysaccharides extracted from fruiting bodies of Jang-soo mushroom (Fomitella fraxinea). Hot water $(100^{\circ}C,\;FF-I)$, 1% ammonium oxalate solution $(80^{\circ}C,\;FF-II)$, and 5% sodium hydroxide solution $(80^{\circ}C,\;FF-III)$ were used for extraction of three polysaccharides from fruiting bodies of it. Anti-complementary activity of FF-I was more effective than the others. FF-I was further fractionated into three groups of polysaccharide by DEAE-Sephadex A25 column chromatography (FF-NP, FF-AP1, and FF-AP2). FF-AP1 showed not only the highest anti-complementary activity but also the growth-inhibitory activity against Snu-I (human stomach cancer cell) among 9 kinds of human tumor cell lines. But FF-AP2 exhibited its activity against Hep-2(larynx cancer) and KB(mouth epidermal cancer) cell lines at $500\;{\mu}g/ml$ although its anti-complementary activity was lower than it of FF-AP1. When FF-I was orally administrated to mice with dosage of 5000 mg/kg, no remarkable changes were observed in viewpoint of tissue-pathology.

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Selection of Cadmium Resistant Cell Line from Calli of Nicotiana tabacum cv. BY4 (담배(Nicotiana tabacum L. cv. BY4)캘러스로부터 카드뮴 저항성 세포주의 선발)

  • 오승철;소웅영;조덕이;양덕춘
    • Korean Journal of Plant Tissue Culture
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    • v.24 no.6
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    • pp.361-367
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    • 1997
  • This study was carried out to select cadmium resistant cell lines from leaf-derived calli of diploid and haploid of Nicotiana tabacum cv. BY4, for understanding adaptation mechanism of plants in cadmium contaminated environment. suspended cell clumps were plated onto selection medium containing 0 to 2,000 $\mu$M cadmium. Cadmium resistant colonies were formed on the selection medium after 3 or 4 weeks of culture. The minimum inhibition concentration (MIC) of cadmium on colony formation were 300 $\mu$M in diploid and 200 $\mu$M in haploid plants, respectively. In order to test the resistance to cadmium, selected cell line on MIC were transferred to medium containing high concentration of cadmium. The selected cell lines, especially haploid cell line, were resistant an the high concentration of cadmium. And dry weight, ash weight, and cadmium contents of cell were increased. These results indicated that the selected cell lines showed higher resistance of cadmium than control cells, and haploid plant is more resistant than diploid plant on medium with cadmium.

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Physiological Activities of Phellinus ribis Extracts (찔레 영지버섯(Phellinus ribis) 추출물의 생리활성)

  • Song, Jae-Hwan;Lee, Hyun-Sook;Hwang, Jin-Kook;Chung, Tae-Young;Hong, Sung-Ryul;Park, Ki-Moon
    • Korean Journal of Food Science and Technology
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    • v.35 no.4
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    • pp.690-695
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    • 2003
  • Physiological activities of 40% ethanol extracts of Phellinus ribis were studied by employing several biological and biochemical assays. The extracts of Phellinus ribis displayed nitrate-scavenging activities (NSA) at pH 1.2 as with 64% NSA with 1.0 mg/mL of the extracts. They also had 91% 1,1-diphenyl-2-picrylhydrazyl (DPPH) radical scavenging activities at the concentration of 1.0 mg/mL. Antioxidant activities of the extracts (at 0.5 mg/mL) on the autoxidation of linoleic acid (p<0.001) were also observed.. The inhibitory effect of the extracts on angiotensin converting enzyme was 11%. Cytotoxic effects of Phellinus ribis extracts against human cancer cell tines were also examined using MTT assay. The extracts (at 50 mg/mL) had severe growth inhibitory effects on A549, Hela, AGS, and SK-Hep-1, which were 8, 44, 76 and 42%, respectively. Ames test indicated that the extracts had no mutating effects on Salmonella typhimurium TA98 and TA100.

Application of a PCR Method for the Detection of Mycoplasma in Veterinary Live Viral Vaccines (동물용 생 바이러스 백신에서 Mycoplasma 검출을 위한 PCR 기법 적용)

  • Jeon Woo-Jin;Kim Byoung-Han;Jung Byeong-Yeal;An Dong-Jun;Yi Chul-Hyun;Jang Hwan;Chung Gab-Soo
    • Korean Journal of Microbiology
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    • v.41 no.4
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    • pp.269-274
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    • 2005
  • We evaluated the PCR assay and two commercialized PCR kits for the detection of mycoplasma in veterinary via live vaccines. The PCR assay could specifically detect all the tested Mycoplasma spp. and Acholeplasma spp., whereas two commercialized PCR kits did not. Also, the specificity of the PCR assay showed that 4 reference strains and 7 field isolates belonging to avian mycoplasma species could be all detected. The sensitivity of the PCR assay was determined using pure cultured Mycoplasma spp. and Acholeplasma spp. with a range of 1 to 100 colony forming units/ml in 9 CFR Mycoplasma broth. To test the availability of the PCR assay for veterinary live viral vaccines, A. laidlawii was artificially inoculated into the swine transmissible gastroenteritis-rota virus combined vaccine and canine parvovirus vaccine, respectively and the sensitivity of the PCR assay was similar with the result of cultured samples. In this study, the PCR assays could be used as rapid and sensitive methods for the detection of mycoplasma in veterinary live viral vaccines.

Transcription Factor E2F7 Hampers the Killing Effect of NK Cells against Colorectal Cancer Cells via Activating RAD18 Transcription

  • Bingdong Jiang;Binghua Yan;Hengjin Yang;He Geng;Peng Li
    • Journal of Microbiology and Biotechnology
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    • v.34 no.4
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    • pp.920-929
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    • 2024
  • As a pivotal defensive line against multitudinous malignant tumors, natural killer (NK) cells exist in the tumor microenvironment (TME). RAD18 E3 Ubiquitin Protein Ligase (RAD18) has been reported to foster the malignant progression of multiple cancers, but its effect on NK function has not been mined. Here, the study was designed to mine the mechanism by which RAD18 regulates the killing effect of NK cells on colorectal cancer (CRC) cells. Expression of E2F Transcription Factor 7 (E2F7) and RAD18 in CRC tissues, their correlation, binding sites, and RAD18 enrichment pathway were analyzed by bioinformatics. Expression of E2F7 and RAD18 in cells was assayed by qRT-PCR and western blot. Dual-luciferase assay and chromatin immunoprecipitation (ChIP) assay verified the regulatory relationship between E2F7 and RAD18. CCK-8 assay was utilized to assay cell viability, colony formation assay to detect cell proliferation, lactate dehydrogenase (LDH) test to assay NK cell cytotoxicity, ELISA to assay levels of granulocyte-macrophage colony-stimulating factor (GM-CSF), tumor necrosis factor-α (TNF-α) and interferon-γ (IFN-γ), and immunofluorescence to detect expression of toxic molecules perforin and granzyme B. High expression of RAD18 and E2F7 was found in CRC tissues and cells. Silencing RAD18 could hamper the proliferation of CRC cells, foster viability and cytotoxicity of NK cells, and increase the secretion of GM-CSF, TNF-α, IFN-γ as well as the expression of perforin and granzyme B. Additionally, ChIP and dual-luciferase reporter assay ascertained the binding relationship between RAD18 promoter region and E2F7. E2F7 could activate the transcription of RAD18, and silencing RAD18 reversed the inhibitory effect of E2F7 overexpression on NK cell killing. This work clarified the inhibitory effect of the E2F7/RAD18 axis on NK cell killing in CRC, and proffered a new direction for immunotherapy of CRC in targeted immune microenvironment.

THE RELATIONSHIP BETWEEN ORAL STATUS AND DENTAL CARIES ACTIVITY ON MENTAL RETARDATION STUDENTS (정신지체 학생의 구강실태 및 우식활성도에 관한 연구)

  • Han, Soo-Keong;Kim, Nam-Soon;Cho, Hong-Kyu;Song, Ho-Jun;Choi, Choong-Ho;Choi, Nam-Ki
    • Journal of the korean academy of Pediatric Dentistry
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    • v.34 no.1
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    • pp.19-26
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    • 2007
  • The purpose of this study is to evaluate oral health status, caries-inducing bacterial activity of the mental retardation students, and developing motivation method for improving oral hygiene management by clarifying the relationship between caries-inducing bacterial activity and oral health status of mental retardation students. Caries experience indices, caries susceptibility test, gingival health evaluation, and oral hygiene management and oral health survey were performed. Results were as follows : 1. The DMFT index of disabled students(12 years) was 2.07. 2. The gingival inflammation was occurred more frequently in older ages. 3. There also was a high positive correlation between caries incidences and the results of caries-inducing bacterial activity test especially in deciduous teeth. This result suggested that the Dentocult SM mutans test as a caries activity test is a reliable method for measuring the status of dental caries in mental retardation students. Because it would motivate the mental retardation students to care more actively for their oral hygiene if they learn how much dental caries-inducing bacteria are living in their mouth by observing the bacterial colony on the cultured test strip, it can become a possibly efficient educational tool for the mental retardation students.

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Isolation and Identification of Microorganisms Producing the Soy Protein-Hydrolyzing Enzyme from Traditional Mejus (전통메주로부터 대두단백질 가수분해효소 생산성 미생물의 분리 및 동정)

  • Kang, Min-Jung;Kim, Seong-Ho;Joo, Hyun-Kyu;Lee, Gap-Sang;Yim, Moo-Hyun
    • Applied Biological Chemistry
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    • v.43 no.2
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    • pp.86-94
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    • 2000
  • In order to develop the enzymatic hydrolysis system concerned with taste and flavor, strains having the high hydrolyzing activity on the soy protein were selected from some traditional Mejus. Two molds and one bacterium producing enzymes which were different in character of hydrolysis were isolated and identified. Leucine and azodye enzyme activities of both M4 and M5 were relatively high among in the isolated molds. And, leucine enzyme activity of B16 was the lowest in the isolated bacteria. These strains were isolated as microorganisms having a dissimilar hydrolysis pattern on the soy protein by enzymatic reactions. Mold M4 on the culture solid media was mycelium colors of white and its sclerotia colors were changed from white to black. According to the result of slide culture, radial conidial head, subclavate vesicle, conidia of subglobose, stipes of uncolored with smooth walls and metula and phialides were existed. Because M4 was taxonomically similar to the characteristics of Aspergillus oryzae (ahlburg) species, M4 was identified and named as Aspergillus oryzae M4.Mold M5 showed white and black mycelium on the MEA medium. Mold M5 colony exhibited grayish-green color and have long(7 mm) sporangiophores at slide culture. Sporangia became brownish-gray and the wall of larger sporangia was broken to form small collars, and smaller sporangia were fomed continually from large basal membrane. Columella is globose and hyaline, and sporangiospores are ellipsoidal of small diameter$(80\;{\mu}m)$. Because M5 was taxonomically similar to the Mucor circinelloides of zygomycetes, M5 was was identified and named as Mucor circinelloides M5. Bacteria B16 colony was opaque white, circular and lobate, and had rod shaped endospore. B16 was found positive in stain, catalase, ${\beta}-glucosidse$ and V-P tests. B16 was found to utilize D-fructose, ${\alpha}-D-glucose$, maltose, D-mannose, D-raffinose, stachyose and sucrose. By the morphological and physiological results, the characteristics of B16 was thought to correspond to that of Bacillus megaterium. However, fatty acid composition was similar to Paenibacillus marcerans, requiring further study for the definite identification. Accordingly, Bacteria B16 was provisionally classified and named as Bacillus megaterium B16.

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