• Restorative Dentistry and Endodontics
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• 제21권1호
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• pp.1-18
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• 1996

There are many advantages when using IIF and DNA probe methods over anaerobic culture method in that they are time-and effort-saving, more precise and more sensitive. Furthermore, in IIF and DNA probe methods, the detection is possible only with small amount of bacteria, the quantitative analysis is possible, and the cell viability is not necessary. The purpose of this study is to observe the incidence of P.endodontalis by carrying out anaerobic culture, IIF and colony lift using DNA probe method respectively, and to compare these 3 methods in terms of effectiveness and sensitivity in order to identify the most effective detection method. 30 teeth with at least one clinical symptoms, with single canal, and with pulp necrosis were sampled. For sampling bacteria, access cavity was prepared after disinfecting tooth and its surroundings. Then the paper point was inserted up to the periapical area, leave there for a while, and finally it was placed into PRAS Ringer's sol. and PBS sol. In anaerobic culture method, P.endodontalis was identified by biochemical tests after subculturing black and brown colonies which were produced after 7 days of incubation on BAP and Brucella BAP in anaerobic chamber. To identify P.endodontalis in IIF method, species-specific polyclonal rabbit-antisera of P.endodontalis(ATCC 35406) was reacted with sampled PBS sol. dispensed onto glass slide, and then P.endodontalis was examined by phase contrast microscopy after incubating with Goat anti-rabbit lgG conjugated to Fluorescein isothiocyanate. For colony lift using DNA probe method, membranes were laid over colonies on the surface of BAP and were hybridized with cloned DNA probe of P.endodontalis. The existence of P.endodontalis was then identified by the methods of chemiluminescent detection and color metric detection. Black colony was found in 11 teeth out of 30 teeth and P.endodontalis was detected in 6 teeth (20 %) by anaerobic culture method, 16 teeth (53 %) by IIF method, and 7 teeth (23 %) by DNA probe method. IIF method is significantly better in detecting P.endodontalis than DNA probe method and anaerobic culture method. There was no significant differences between DNA probe method and anaerobic culture method. There was significant correlation between the formation of black colony and the existence of P.endodontalis. The probability of detecting P.endodontalis when black colony being present is 2.89 times higher than when not being present. There was significant relationship between the foul odor of clinical symptoms and P.endodontalis. The sensitivity of existing P.endodontalis when foul odor being present was 93.75 %, while the specificity of not existing P.endodontalis when foul odor not being present was 28.57 %. These results suggested that the probes of P.endodontalis will be used to decide the method and prognosis in endodontic treatments.

• 한국연초학회지
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• 제15권2호
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• pp.130-136
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• 1993

Protoplasts of palisade cells were aseptically isolated from leaves of Nicotiono apicona Merxm. by the one step enzymatic method. Efficiency of colony formation were depended on cell density and light condition during incubation, but an intensity of 47 ft-c during a period of 2 weeks after isolation of the protoplasts in the Nagata and Takebe's medium promoted the planting efficiency. Protoplast - derived calli of N. africana can be differentiated into shoot when cultured on Murashige and skoog's medium containing IAA(0.Smg/$\ell$) and zeatin(5.0mg/$\ell$).

• Mycobiology
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• 제34권4호
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• pp.196-199
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• 2006

Effects of various preservation periods and subcultures on fruiting body formation of Cordyceps militaris were investigated using EFCC C-10995 single ascospore strains. Fruiting body formation by original strains was profuse when preserved at $4^{\circ}C$ for $5{\sim}6$ months. Fruiting from subcultures was stable till second to sixth subcultures, after which it decreased sharply. The more the colony color of subcultures changed, the less the fruiting bodies formed. Liquid inoculum preparation of single ascospore strains in the same or separate broths did not affect fruiting body formation. Similarly, two strains C-10995-3 and C-10995-6 in different numbers during liquid inoculum preparation produced similar fruiting bodies.

• BMB Reports
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• 제43권8호
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• pp.524-529
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• 2010

Glucocorticoids (GCs) are useful drugs for the treatment of various diseases, but their use for prolonged periods can cause severe side effects such as osteoporosis. GCs have a direct effect on bone cells, where they can arrest bone formation, in part through the inhibition of osteoblast. On the other hand, GCs potently suppress osteoclast resorptive activity by disrupting its cytoskeleton based on the inhibition of RhoA, Rac and Vav3 in response to macrophage colony-stimulating factor. GCs also interfere with microtubule distribution and stability, which are critical for cytoskeletal organization in osteoclasts. Thus, GCs inhibit microtubule-dependent cytoskeletal organization in osteoclasts, which, in the context of bone remodeling, further dampens bone formation.

• 약학회지
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• 제35권2호
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• pp.135-141
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• 1991

The general pharmacological tests with rhGM-CSF indicated that it had no influences on rotarod and locomotor activity tests, but shortened hexobarbital-sleeping time at the large dose of 3 mg/kg s.c. in mice. It elicited no hypothermic, analgesic and antiepileptic action. No influences on blood pressure and respiration in rabbits were observed at the dose of 1 mg/kg, i.v. and it did neither affect the receptors of adrenaline, acetylcholine, serotonin, histamine, kinin and oxytocin, nor antagonize the actions of histamine, serotonin and oxytocin at its concentrations of 1$\times$$10^{-6}$g/ml. However, this substance was demonstrated to stimulate the formation of leucocytes in rats.

• Archives of Plastic Surgery
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• 제34권5호
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• pp.551-556
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• 2007

Purpose: Pseudomonas aeruginosa is an etiologic agent in serious wound infection. Pseudomonas aeruginosa infection is problematic because this organism is resistant to many antimicrobial drugs. The purpose of this study was to compare the bactericidal effect of commonly used topical agents and their effect on wound healing. Methods: Pseudomonas aeruginosa-infected full-thickness skin defect was developed on the mouse to compare 3 commonly used topical agents-Betadine, 2% Gentamicin solution and 0.3% Acetic acid with the control group. Wound size change, bacterial colony counts and histologic findings of each groups were analyzed. Results: The wound size decreased in all treated groups as compared with the control group. However, there was no statistical difference. Gentamicin solution group was showed the lowest bacterial colony count and statistically significant difference compared with the control group(p=0.032). Other treated groups were also effective against Pseudomonas aeruginosa, but not different statistically. Histologic findings revealed that epithelialization, granulation tissue formation and microvessel proliferation were increased and necrosis and inflammation were decreased in all treated groups compared to the control group, but not different statistically. Betadine group significantly increased granulation tissue formation compared to the control group (p= 0.041). Conclusion: There is no universal topical agent that enhances most aspects of wound healing while simultaneously decreasing the bacterial concentration. However, Gentamicin solution may be an optimal topical agent for Pseudomonas aeruginosa infected wound. Further study should experiment on human with Gentamicin solution to confirm a effect on Pseudomonas aeruginosa infected wound for clinical applications.

• Asian Pacific Journal of Cancer Prevention
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• 제13권9호
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• pp.4363-4368
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• 2012

The epidermal differentiation complex (EDC) contains a large number of gene products which are crucial for the maturation of the human epidermis and can contribute to skin diseases, even carcinogenesis. It is generally accepted that activation of oncogenes and/or inactivation of tumor suppressor genes play pivotal roles in the process of carcinogenesis. Here, NICE-3, a novel EDC gene, was found to be up-regulated in human hepatocellular carcinoma (HCC) by quantitative real-time RT-PCR. Furthermore, overexpression of exogenous NICE-3 by recombinant plasmids could significantly promote cell proliferation, colony formation and soft agar colony formation in Focus and WRL-68 HCC cell lines. Reversely, NICE-3 silencing by RNA interference could markedly inhibit these malignant phenotypes in YY-8103 and MHCC-97H cells. Moreover, cell cycle analysis of MHCC-97H transfected with siRNA by flow cytometry showed that NICE-3 knockdown may inhibit cell growth via arrest in G0/G1 phase and hindering entry of cells into S phase. All data of our findings indicate that NICE-3 may contribute to human hepatocellular carcinoma by promoting cell proliferation.

• 한국수정란이식학회지
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• 제31권3호
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• pp.179-183
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• 2016

Even though klotho deficiency in mice exhibits multiple aging-like phenotypes, studies using large animal models such as pigs, which have many similarities to humans, have been limited due to the absence of cell lines or animal models. The objective of this study was to generate homozygous klotho knockout porcine cell lines and cloned embryos. A CRISPR sgRNA specific for the klotho gene was designed and sgRNA (targeting exon 3 of klotho) and Cas9 RNPs were transfected into porcine fibroblasts. The transfected fibroblasts were then used for single cell colony formation and 9 single cell-derived colonies were established. In a T7 endonuclease I mutation assay, 5 colonies (#3, #4, #5, #7 and #9) were confirmed as mutated. These 5 colonies were subsequently analyzed by deep sequencing for determination of homozygous mutated colonies and 4 (#3, #4, #5 and #9) from 5 colonies contained homozygous modifications. Somatic cell nuclear transfer was performed to generate homozygous klotho knockout cloned embryos by using one homozygous mutation colony (#9); the cleavage and blastocyst formation rates were 72.0% and 8.3%, respectively. Two cloned embryos derived from a homozygous klotho knockout cell line (#9) were subjected to deep sequencing and they showed the same mutation pattern as the donor cell line. In conclusion, we produced homozygous klotho knockout porcine embryos cloned from genome-edited porcine fibroblasts.

• 생약학회지
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• 제18권2호
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• pp.127-135
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• 1987

In vitro sensitivity testing was performed for 21 kinds putative anticancer drugs selected from references and information. Cellular damage of P815 mastocytoma cells following exposure to water extracts of drugs was evaluated by colony formation assay. Highly effective drugs with more than 50% inhibition of colony formation were seven (Houttuyniae Herba, Sanguisorbae Radix, Nepetae Herba, Manitis Squama, Lonicerae Flos, Amomi Semen, Polyporus), though not more effective than BCNU. According to the results of $^3H-thymidine$ incorporation assay for determination of selective cytotoxicity, 3 of these drugs (Houttuyniae Herba, Polyporus, Manitis Squama) were found to be low cytotoxic to normal mouse lymphoid cells. These findings suggest that the above 3 drugs may be used for effective anticancer drugs in vivo.

• 한국식품과학회지
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• 제49권2호
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• pp.146-150
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• 2017

본 연구에서는 지금까지 거의 보고된 바 없는 corosolic acid의 항균활성에 대해 대표적 식중독균 중 하나인 S. aureus를 대상으로 연구하였다. 대수기의 S. aureus에 corosolic acid를 처리하여 생장 저해 여부를 확인하였고, broth micro-dilution 방법과 agar disc diffusion 방법을 통해 corosolic acid의 S. aureus에 대한 항균활성을 측정해본 결과, corosolic acid의 농도에 비례하여 S. aureus 생장이 저해되는 것으로 확인되었다. 또한 corosolic acid의 존재 조건에서 S. aureus의 바이오필름 형성능이 MIC 보다 높은 농도뿐 아니라 낮은 농도에서 역시 저해되는 것으로 관찰되었다. 따라서 천연물질인 corosolic acid는 S. aureus에 의한 식품 오염을 억제하고 식중독 예방 및 잠재적 치료제로서 개발 될 가능성이 있는 것으로 판단되며, 나아가 다른 종류의 식중독균들에 대한 적용 가능 연구가 필요할 것으로 사료된다.