• Title/Summary/Keyword: Phase contrast microscope

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Cytotoxic Effects of 1, 2-Hexanediol and 1, 2-Hexanediol Galactoside on HaCaT Cell (1, 2-Hexanediol과 1, 2-Hexanediol Galactoside의 HaCaT Cell에 대한 세포독성)

  • Kim, Jun-Sub;Jung, Kyung-Hwan
    • Journal of the Society of Cosmetic Scientists of Korea
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    • v.44 no.3
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    • pp.343-347
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    • 2018
  • We synthesized 1, 2-hexanediol galactoside (HD-Gal) from HD using Escherichia coli (E. coli) ${\beta}-galactosidase$ (${\beta}-gal$), in which the reaction is generally called as transgalactosylation (reverse hydrolysis). In this study, we investigated how much HD-Gal and HD had a cytotoxic effect on HaCaT cell, in order to compare HD-Gal with HD in terms of the cytotoxicity of human skin cell. Cell proliferation assay and phase-contrast microscope observation were used for investigating the cytotoxicity. As a result, HD-Gal had not cytotoxic effect on HaCaT cell in the concentration range from 42.2 to 211 mM. In addition, when we observed the cells using microscopy, there was no change in the cell morphology. Meanwhile, when 42.2 mM and 84.4 mM HD were treated on HaCaT cell, we did not observe the cytotoxicity; however, when 168.8 mM and 211 mM HD were on HaCaT cell, HD had a higher cytotoxic effect on HaCaT cell. In addition, when HD was treated on the cells regardless of the concentration of HD, there were obvious changes in cell morphology and cell number. It was expected hopefully that HD-Gal would be applicable as a substitute for HD as a less toxic preservative in views of safety, health, and well-being.

Assessment of the Fertilizing Capacity of Domestic Animal Spermatozoa by Hamster Test II. Effects of incubation medium and X-ray irradiation on hamster test for boar spermatozoa (Hamster test를 이용한 가축정자의 수정능력 검정 II. 정액배지 및 X-선조사가 돼지정자의 Hamster test에 미치는 영향)

  • Kim Yong-Jun;Ji Dong-Boum
    • Journal of Veterinary Clinics
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    • v.9 no.2
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    • pp.373-390
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    • 1992
  • To assay the fertilizing capacity of domestic animal spermatozoa by hamster test, semen were collected from 13 boars(Duroc. Landrace and Yorkshire) which had been proved to be fertile in the past. then, were preserved in BWW medium or in raw state at 18$^{\circ}C$ or at room temperature. The preserved semen were given each different treatment according to the experimental design and coincubated with zona-free hamster ova for 5 hours. The ova were stained by lacmoid and examined under phase contrast microscope to investigate the rates of ova bound with sperm(sperm binding). ova penetrated by sperm(penetration) and formation of a male pronucleus(pronucleus formation) and also numbers of both bound and penetrated sperm per ovum. Between BWW and TBM medium for boar sperm. no difference in the results of hamster test was obtained. The boar spermatozoa in BWW medium, BWW with caffeine, BWW with heparin, and BWW with both caffeine and heparin showed no difference in the results of hamster test. The boar spermatozoa in BWW medium containing both calcium and RSA showed considerably higher rates of sperm binding, penetration and pronucleus formation as well as higher numbers of both bound and penetrated sperm than those not containing calcium with or without BSA( p<0.01) and also the same results higher than that containing calcium without BSA( p< 0.05). The boar spermatozoa irradiated by X-ray(70 KVP, 20mA) for 3 seconds. then, maintained at 18$^{\circ}C$ for 18 hours showed considerably lower rate of sperm binding than all the other groups including the control and X-ray groups irradiated by smaller dose or maintained for shorter period(p<0.01), and also showed lower number of bound sperm than the other groups(p<0.01, p<0.05). All the control groups of both raw and diluted sperm in BWM medium showed higher rates of sperm binding, penetration and pronucleus formation as well as higher number of penetrated sperm than all the X-ray groups irradiated for 3 seconds(70KVP, 20mA) and maintained for either 3 or 18 hours (p<0.01, p<0.05). At the same time the control groups of diluted sperm showed considerably higher rates of sperm penetration and pronucleus formation than the control group of raw sperm( p<0.01). These results indicates that fertile boar sperm showed considerably lower rates In the results of hamster test, when incubated in the medium without calcium and irradiated by X-ray than when incubated in the medium with calcium and not irradiated by X-ray, respectively, to prove consequently that hamster test would be of great value in assaying the fertilizing capacity of boar spermatozoa.

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Ultrastructures of Oocyte Development and Electrophoretic Patterns of the Yolk Protein Following HCG Treatment in Korean Native Catfish (Silurus asotus)

  • Yoon, J.M.;Chung, E.Y.;Kim, G.W.
    • Asian-Australasian Journal of Animal Sciences
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    • v.14 no.2
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    • pp.174-183
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    • 2001
  • During the rapid phase of gonadal development of the freshwater teleost, the catfish (Silurus asotus), the influence of hCG upon the inducement of final oocyte maturation and spawning was investigated electrophoretically and ultrastructurally. The electrophoretic patterns obtained were different in the presence and absence of some of the major or minor zones, because of the hormone level in catfish. The vitellogenin of hormone-treated fish was stained more intensively than that of sham-treated fish. These proteins showed some minor or main bands of egg extracts which migrated at positions corresponding to molecular weights of approximately 90,000. However, the thickness of electrophoretic band in molecular weight for hCG-treated fish was slightly lower than that for saline control. It seemed the plasma protein with molecular weight of approximately 45,000 in hCG-treated fish disappeared. In contrast to the control fish, the ovaries in the catfish treated with hCG shows a marked ultrastructural change under the electron microscope. No dilated profiles were seen in the granulosa cells of the mature oocyte before ovulation. After germinal vesicle breakdown (GVBD), the zona radiata interna (ZRI) becomes more compact, and there is a loss of all the processes from the pore canals. There is a wide space between the vitelline membrane and zona radiata. Also, during final maturation, the microvillar processes from the oocyte are seen no longer to penetrate deeply into the extracellular spaces of the overlying granulosa cells, and the reticulate patterns of the zona radiata interna becomes occluded, giving the zona radiata a more solid appearance. It has been possible to initiate 100% oocyte maturation in yolk granules and follicles in vivo by treatment with hCG and a high water temperature ($27^{\circ}C$). In hCG-treated fish, the percentages of successful artificial fertilization and hatching were maximal at 15 h after a single injection. It seems clear that a long acting preparation containing hCG can be successfully used in prespawning fish to advance the final events of gonadal maturation and initiate spawning. Further studies are necessary to evaluate the potential of hCG to either stimulate or inhibit the reproductive development of fish at other stages of the seasonal reproductive cycle.

Study of Apoptotic Effect on Hydrogen Peroxide-induced Rat PC-12 Cells by Aster tataricus Water Extract (자완 수추출물(水抽出物)이 $H_2O_2$에 의해 유도(誘導)된 PC-12 세포주(細胞株)의 세포사(細胞死)에 미치는 영향(影響))

  • Shin, Yoo-Jung;Kim, Seung-Mo;Park, Chi-Sang;Shin, Oh-Chul
    • The Journal of Korean Medicine
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    • v.28 no.2 s.70
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    • pp.213-223
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    • 2007
  • Objective : Alzheimer's disease (AD) is a geriatric dementia that is widespread in old age. With an aging populace, AD is a looming problem in public health service. Alzheimer's disease is characterized by specific neuronal degeneration in certain areas of the brain. Mutations and abnormal expression of several genes are associated with ${\beta}-amyloid$ deposits and Alzheimer's disease; among them APP, PS1, and PS2, SOD, free radical, ROS. Methods:We studied herbal medicines that have a relationship to brain degeneration. From pre-modern times, although a variety of oriental prescriptions of Aster tataricus have been traditionally utilized for the treatment of AD, their pharmacological effects and action mechanisms have not yet been fully elucidated. Result : Based on morphological observations by phase-contrast microscope, TUNEL assay and MTT in the culture media, $H_20_2-induced$ cell death was significantly inhibited by Aticus. We examined by ROS formation, catalase activity and GSH activity. We studied the protective effect and inhibitory effects of neurotoxicity in $H_20_2-induced$ PC-12 cells by Aticus. Findings from our experiments show that Aticus inhibits apoptosis, which has neurotoxicities and cell damage in PC-12 cells. In addition, treatment with Aticus ($>25{\mu}g/ml$ for 6hrs) partially prevented $H_20_2-induced$ cytotoxicity in PC-12 cells, and induced a protective effect. Conclusion : As the result of this study, in the Aticus group, the apoptosis in the nervous system was inhibited, protected against the degeneration of PC-12 cells by $H_20_2$. Taken together, Aticus exhibited inhibition of $H_20_2-induced$ apoptotic cell death. Aticus was found to induce protective effect on GSH and catalase in PC-12 cells. Based on these findings, Aticus may be beneficial for the treatment of AD.

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Semen treatment to enhance the use of hamster test in the dog (개에서 Hamster test의 이용을 높이기 위한 정액처리조건)

  • Kim, Yong-jun;Lee, Hae-iee
    • Korean Journal of Veterinary Research
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    • v.33 no.2
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    • pp.337-343
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    • 1993
  • To determine the test conditions to enhance the use of hamster test in dogs, semen were collected from four dogs which had been proven to be fertile in the past and then preserved in BWW (Biggers, Whitten, Whittingham) medium for about 20 hours. The semen were given each different treatment according to the experimental design and coincubated with zona-free hamster ova for 5 hours. The ova were stained by lacmoid and examined under phase contrast microscope to investigate the rates of ova bound with sperm(sperm binding) and ova penetrated by sperm (penetration), and also numbers of both bound and penetrated sperm per ovum. In comparison of different concentrations of canine sperm, the rate of sperm binding was higher in $1.5{\times}10^8$, $1{\times}10^8$, and $5{\times}10^7$ sperm concentrations than $5{\times}10^5$ concentration(p<0.01), and also than $5{\times}10^6$ concentration(p<0.05), respectively. The number of bound sperm per ovum was considerably higher in $1.5{\times}10^8$ sperm concentration than $5{\times}10^7$, $1.5{\times}10^6$, and $5{\times}10^5$ concentrations(p<0.01). The rate of penetration was considerably higher in $1.5{\times}10^8$ and $1{\times}10^8$ sperm concentrations than $5{\times}10^5$ concentration,(p<0.01), and also the higher result of penetration was shown in $5{\times}10^7$ than $5{\times}10^5$ (p<0.05). The number of penetrated sperm per ovum was considerably higher in $1.5{\times}10^8$ sperm concentrations than $5{\times}10^5$(p<0.01), and also the higher number was shown in $1{\times}10^8$ than $5{\times}10^5$ (p<0.05). In comparison of the different preincubation period of canine spermatozoa, no difference was obtained in the results of hamster test among the preincubation periods of 4 hours, 18~24 hours and 48 hours. The canine spermatozoa in BWW medium with $Ca^{2+}$(1.3mM) and without FCS(fetal calf serum), with both $Ca^{2+}$(1.3mM) and FCS, with $Ca^{2+}$(2.6mM) and without FCS, and with both $Ca^{2+}$(2.6mM) and FCS showed no difference in the results of hamster test.These results indicated that the appropriate concentration of sperm should be given in hamster test for dog sperm.

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Clients Satisfaction with Oral Hygiene Care Services System Provided by the Dental Hygiene Clinics in the Department of Dental Hygiene, Yonsei University (연세대학교 치위생학과 구강위생교육실을 방문한 대상자의 구강위생관리 서비스체계에 대한 만족도 조사)

  • Kim, Nam-Hee;Kwon, Hye-Ri;Kim, Da-Hye;Kim, Da-Hee;Kim, Min-Hee;Yoo, Seung-Hee;Choi, Jin-Ah;Chung, Won-Gyun
    • Journal of Korean society of Dental Hygiene
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    • v.7 no.4
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    • pp.419-431
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    • 2007
  • The setting of dental hygiene clinics is very important to dental hygiene education, which is the place not only to educate students but also to care clients. The purpose of this study is to provide basic research material for improvement of dental hygiene care system in dental hygiene clinics by analyzing the client satisfaction. A questionnaire survey by means of self-entry method was conducted to find out satisfaction of the client, who was visited to the dental hygiene clinics in the department of dental hygiene, Yonsei University. An analysis of frequency, one way ANOVA and T-Test were performed through SPSS 12.0K program. 1. Most clients were mainly composed of students in Wonju College of Medicine. 2. The clients visited for scaling(85.8%) and oral examination(9.73%) were much than treatment(4.42%). 3. Clients aged 21 to 25 were relatively lower in satisfaction with the facilities, system, attitude than any other ages. 4. The dental hygiene students are the lowest group in satisfaction with the facility, system, attitude than medical and nursing students. 5. The clients satisfaction with dental hygiene clinics was decreased in reverse proportion to visiting frequency. 6. Most of the clients pointed out the problems of appointment system(54.0%) and fee(23.0%), which should be improved than any other operation conditions. 7. Most of the clients were not satisfied with chair time and pain during care. 8. Most clients recognized to receive the better care service than other dental offices(81.3%). Especially, they paid attention to oral health education using phase-contrast microscope. 9. Many clients were dissatisfied with facilities of the dental hygiene clinics(71.7%). The problems of appointment system(54.0%) and chair time of dental hygiene care services(63.6%) had also inconvenienced to clients. The dental hygiene clinics in school play a crucial role in dental hygiene education to foster the student to be competent as a professional dental hygienist in the future. Therefore, well-organized dental hygiene care program based on dental hygiene process is essential. It is also required to improve the environment of dental hygiene clinics including facilities, appointment system and fee etc.

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Preparation of Chitosan-coated Magnetite Nanoparticles (키토산이 피복된 나노 크기의 자성체 분말 제조)

  • Cho, Jun-Hee;Ko, Sang-Gil;Ahn, Yang-Kyu;Song, Ki-Chang;Choi, Eun-Jung
    • Journal of the Korean Magnetics Society
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    • v.16 no.1
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    • pp.102-106
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    • 2006
  • Magnetic nanoparticles can be used for a variety of biomedical applications. They can be used in the targeted delivery of therapeutic agents in vivo, in the hyperthermic treatment of cancers. in magnetic resonance (MR) imaging as contrast agents and in the biomagnetic separations of biomolecules. We have synthesized magnetite $(Fe_3O_4)$ nanoparticles using chemical coprecipitation technique with sodium oleate as surfactant. Nanoparticle size can be varied from 2 to 8nm by controlling the sodium oleate concentration. Magnetite phase nanoparticles could be observed from X-ray diffraction. Magnetic colloid suspensions containing particles with sodium oleate and chitosan have been prepared. Nanoparticles, both oleate-coated and chitosan-coated, have been characterized by several techniques. Atomic farce microscope (AFM) was used to image the coated nanoparticles. Magnetic hysteresis measurement were performed using a superconducting quantum interference device (SQUID) magnetometer at room temperature to investigate the magnetic properties of the magnetite nanoparticles. The SQUID measurements revealed superparamagnetism of nanoparticles.

Marine Blue-green Algae in Korea (II) (한국 해산 남조류 (II))

  • Yu, Sun-Ae;Chae, Seung-Mun;Lee, Gi-Wan
    • The Journal of Natural Sciences
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    • v.6 no.1
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    • pp.5-39
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    • 1993
  • The Blue-green algae collected from whole coasts of Korea were investigated morphotaxonomically in order to list up Korean marine Cyanophyta and clarify their taxnomic position. As a result, 36 species, 20 genus, 6 families belonging to 3 orders were identified. Among these, 14 species were recorded for the first time in Korea. They are Chroococcus minutus (K$\"{u}$tzing) N$\"{a}$geli, Merismopedia punctata Meyen, Microcystis ichtyoblabe K$\"{u}$zing, Dermocarpa leibleiniae (Reinsch) Born. et Thur., Hydrocoleum confluens (Setchell et Gardner) Drouet, Lyngbya sordida (Zanard.) Gomont, Phormidium forveolarum (Mont.) Gomont, Phormidium hansgieri Schmidle, Skujaella hildebrandtii (Gomont.) de Toni, Sphaeronema lithophila (Ercegovic) Umezaki, Spirulina tenerrima K$\"{u}$tzing, Hormothamnion enteromorphoides Grunow, Michrochaete aeruginea Batters, Michrochaete grisea Thuret ex Born. et flah.. Using the phase contrast microscope and the Nomarski interference micrope, we made photomicrographs of minute blue green algae. The cellular inclusions especially PHB(poly-$\SS$-hydroxy-butyrate) granules of the blue-green algae identified were investigated. The species clearly characteriged to have PHB granule were Lyngbya confervoides, L. semiplena, Phormidium corium, Sirocoleum kurzii, Hormothamnion enteromorphoides and Calothrix crustacea. These result would be fundamental data for estabilishing phylogenetic system of blue-green algae based on physio-biochemical characteristics in future.

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Antifungal Activity of Rheum undulatum on Candida albicans by the Changes in Membrane Permeability (막투과성 변화로 인한 대황의 Candida albicans에 대한 항진균 활성)

  • Lee, Heung-Shick;Kim, Younhee
    • Korean Journal of Microbiology
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    • v.50 no.4
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    • pp.360-367
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    • 2014
  • Candida albicans is an opportunistic and the most prevalent fungal pathogen that can cause superficial and systemic infections in immunocompromised patients. C. albicans can promote the transition from budding yeast to filamentous form, generating biofilms. Infections associated with C. albicans biofilms are frequently resistant to conventional antifungal therapy. Therefore, the development of more effective antifungal drugs related with biofilm formation is required urgently. The roots of Rheum undulatum have been used for medicinal purposes in Korea and China traditionally. The aim of present study was to evaluate the effect of R. undulatum extract upon preformed biofilms of 12 clinical C. albicans isolates and the antifungal activities. Its effect on preformed biofilms was evaluated using XTT reduction assay, and metabolic activity of all tested strains was reduced significantly ($49.4{\pm}6.0%$) at 0.098 mg/ml R. undulatum. The R. undulatum extract blocked the adhesion of C. albicans biofilms to polystyrene surfaces, and damaged the cell membrane integrity of C. albicans which was analyzed by CFDA, AM, and propidium iodide double staining. It caused cell lysis which was observed by Confocal laser scanning and phase contrast microscope after propidium iodide and neutral red staining, respectively. Membrane permeability was changed as evidenced by crystal violet uptake. The data suggest that R. undulatum inhibits biofilm formation by C. albicans, which can be associated with the damage of the cell membrane integrity, the changes in the membrane permeability and the cell lysis of C. albicans.

THE MORPHOLOGICAL OBSERVATION OF HUMAN PERIODONTAL LIGAMENT CELLS ATTACHMENT AND SPREADING ON THE SURFACE OF SLIDE GLASS (치주인대세포의 부착과 전개에 관한 형태학적 관찰)

  • Lee, Jin-Mi;Suh, Jo-Young;Park, Joon-Bong
    • Journal of Periodontal and Implant Science
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    • v.23 no.1
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    • pp.97-108
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    • 1993
  • One of the important initial events required for periodontal regeneration is the attachment and subsequent spreading of periodontal ligament cells on the root surface. The purposes of this study is to investigate the attachment and spreading pattern of human periodontal ligament cell on the surface of glass slides. After establishment of a cell line of the primary cell culture from the periodontal ligament of 1st premolar teeth which were extracted for the purpose of orthodontic treatment, author dispersed the cells at $5{\times}10^3\;cells/ml$ into the each 35mm culture petri-dish containing 2 glass slides. To observe the morphological changes of the cells which attached to the surfaces of glasses at every designed time schedule, author used the inverted phase contrast microscope and scanning electron microscope. During the whole experiment culture condition was at $37^{\circ}C$, 100% Humidity, 5% $CO_2$ gas incubator. The following results were obtained. Periodontal ligament cells showed spherical outline and started to attach to glass surface by basal sytoplasmic extension after 10min in culture. After 30min in culture, periodontal ligament cells were attached to glass surface by well - developed filopodia which protruded from the lamellipodia. The cell surface is covered with bubble-like structures and occasional microvillus can be seen with diffculty among these structures. After 1.5hr in culture, peridontal ligament cells shhowed radially well-spread cytoplasm and the nucleus was centered on its cytoplasm. Unspread central region of the cell was covered with numerous microvilli. The change of cell attachment and spreading pattern was manifest at 6hr in culture. At this time, periodontal ligament cell showed elongated outline and an oval-shaped nucleus. After 12hr in culture, periodontal ligament cells showed more stretched fibroblast-like appearance with polarity. Two long lamellipodia can be seen around the both terminal ends of cells. After 24hr in culture, periodontal ligament cells showed spindle shapes and an oval-shaped nucleus was slanted toward one side of the cell.

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