• Title/Summary/Keyword: electron microscopy

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Developing a Dental Unit Waterline Model Using General Laboratory Equipments (실험실 일반 장비를 이용한 치과용 유니트 수관 모델 개발)

  • Yoon, Hye Young;Lee, Si Young
    • Journal of dental hygiene science
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    • v.16 no.4
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    • pp.284-292
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    • 2016
  • Water supplied through dental unit waterlines (DUWLs) has been shown to contain high number of bacteria. To reduce the contamination of DUWLs, it is essential to develop effective disinfectants. It is, however, difficulty to obtain proper DUWL samples for studies. The purpose of this study was to establish a simple laboratory model for reproducing DUWL biofilms. The bacteria obtained from DUWLs were cultured in R2A liquid medium for 10 days, and then stored at $-70^{\circ}C$. This stock was inoculated into R2A liquid medium and incubated in batch mode. After 5 days of culturing, it was inoculated into the biofilm formation model developed in this study. Our biofilm formation model comprised of a beaker containing R2A liquid medium and five glass rods attached to DUWL polyurethane tubing. Biofilm was allowed to form on the stir plate and the medium was replaced every 2 days. After 4 days of biofilm formation in the laboratory model, biofilm thickness, morphological characteristics and distribution of the composing bacteria were examined by confocal laser microscopy and scanning electron microscopy. The mean of biofilm accumulation was $4.68{\times}10^4$ colony forming unit/$cm^2$ and its thickness was $10{\sim}14{\mu}m$. In our laboratory model, thick bacterial lumps were observed in some parts of the tubing. To test the suitability of this biofilm model system, the effectiveness of disinfectants such as sodium hypochlorite, hydrogen peroxide, and chlorhexidine, was examined by their application to the biofilm formed in our model. Lower concentrations of disinfectants were less effective in reducing the count of bacteria constituting the biofilm. These results showed that our DUWL biofilm laboratory model was appropriate for comparison of disinfectant effects. Our laboratory model is expected to be useful for various other purposes in further studies.

Establishment of a Dental Unit Biofilm Model Using Well-Plate (Well-Plate를 사용한 치과용 유니트 수관 바이오필름 모델 확립)

  • Yoon, Hye Young;Lee, Si Young
    • Journal of dental hygiene science
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    • v.17 no.4
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    • pp.283-289
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    • 2017
  • The water discharged from dental unit waterlines (DUWLs) is heavily contaminated with bacteria. The development of efficient disinfectants is required to maintain good quality DUWL water. The purpose of this study was to establish a DUWL biofilm model using well-plates to confirm the effectiveness of disinfectants in the laboratory. Bacteria were obtained from the water discharged from DUWLs and incubated in R2A liquid medium for 10 days. The bacterial solution cultured for 10 days was made into stock and these stocks were incubated in R2A broth and batch mode for 5 days. Batch-cultured bacterial culture solution and polyurethane tubing sections were incubated in 12-well plates for 4 days. Biofilm accumulation was confirmed through plating on R2A solid medium. In addition, the thickness of the biofilm and the shape and distribution of the constituent bacteria were confirmed using confocal laser microscopy and scanning electron microscopy. The average accumulation of the cultured biofilm over 4 days amounted to $1.15{\times}10^7CFU/cm^2$. The biofilm was widely distributed on the inner surface of the polyurethane tubing and consisted of cocci, short-length rods and medium-length rods. The biofilm thickness ranged from $2{\mu}m$ to $7{\mu}m$. The DUWL biofilm model produced in this study can be used to develop disinfectants and study DUWL biofilm-forming bacteria.

A Study on Characteristics of Airborne Asbestos Concentrations Using PCM and TEM in Life Environment Surroundings of Seoul (서울지역 생활환경주변의 공기 중 석면분포 특성에 관한 연구)

  • Lee, Jinhyo;Lee, Suhyun;Kim, Jihui;Oh, Seokryul;Shin, Jinho;Eom, Seokwon;Chae, Youngzoo;Lee, Jinsook;Koo, Jayong
    • Journal of Korean Society of Environmental Engineers
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    • v.35 no.9
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    • pp.613-623
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    • 2013
  • This study is purposed to evaluate the airborne asbestos concentrations in life environment surroundings in Seoul. In study, we investigated airborne asbestos concentrations in thirteen subway stations, four monitoring networks and each vicinity roadside, six stream surroundings, four tunnels quarterly and we also investigated relationship between the airborne asbestos concentrations and ambient temperature in monitoring networks and time-based airborne asbestos concentration variability for two typical monitoring networks, two subway stations transferred and used by lots of people through Phase Contrast Microscopy (PCM) and Transmission Electron Microscopy (TEM). The airborne asbestos concentrations by PCM for 4 objects of study were less than the detection limit (7 fiber/$mm^2$) in 111 (50%) out of 223 samples. The highest concentration was 0.0130 f/cc. But additional TEM analysis result for samples exceeding the guideline value for indoor air quality (0.01 f/cc) proposed by the Ministry of Environment (Korea), no asbestos was detected. Similarly TEM analysis result for 124 samples, no asbestos was detected. The average airborne asbestos concentrations by PCM in subway stations, monitoring networks, streams and tunnels were $0.0041{\pm}0.0027$ f/cc, $0.0015{\pm}0.0011$ f/cc, $0.0024{\pm}0.0012$ f/cc and $0.0016{\pm}0.0020$ f/cc. All objects of study were satisfied with the guideline value for indoor air quality. The relationship between the airborne asbestos concentrations and ambient temperature in monitoring networks was generally positive correlation (r = 0.660). The higher ambient temperature was and the more transient population was, the airborne asbestos concentrations by time for two subway stations were increased. While the airborne asbestos concentrations for two monitoring networks showed no variation pattern according to time.

Pathogenicity and Infection Mechanism of Pseudocercospora vitis Causing Leaf Spot Disease on Grapevine in Korea (포도나무 갈색무늬병균(Pseudocercospora vitis (Lếv.) Speg.)의 병원성과 침입기작)

  • Park Jong-Han;Han Kyeong-Suk;Lee Jung-Sup;Seo Sang-Tae;Jang Han-Ik;Kim Heung-Tae
    • Research in Plant Disease
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    • v.12 no.1
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    • pp.15-19
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    • 2006
  • This study was carried out to know pathogenicity of the pathogen isolated in several location of Korea and penetration mechanism of the Pseudocercospora vitis ($(L\acute{e}v)$.) Speg. Inoculation tests at room temperature were performed on both sides of leaves with different isolates of the fungus. The typical symptoms appeared on the abaxial leaf surface, but no symptom was observed on the adaxial leaf surface with all isolates. The average incubation period was nine days, since all symptoms appeared from 8 to 10 days after inoculation. In order to know the mechanism of invasion of P. vitis to grapevine, the behavior of penetration hyphae through stomata were compared in two cultivars having different level of resistance. In order to know the mechanism of resistance of grape vine to P.vitis, two cultivars having different level of resistance were compared by counting the numbers and measuring size of the stomata per leaf. In a susceptible cultivar Campbell Early, the number of stomata was more than that of a resistance cultivar 'Kyoho'. In a susceptible cultivar 'Campbell Early', the fungus entered readily into stomata after inoculation. However, in a resistant cultivar 'Kyoho', the fungus seemed to pass over or surrounded only the guard cells. In comparison of height of guard cells of stomata between the two cultivars, significant differences were observed by scanning electron microscopy and light microscopy. The height of guard cells of 'Campbell Early' showed a little higher than those of 'Kyoho' known to be resistant to the fungus.

Viability of Endothelial Cells in Preserved Human Saphenous Vein Allografts (보존된 사람 동종 복재정맥 이식편혈관 내피세포의 생활성에 관한 연구)

  • 지현근;김용진
    • Journal of Chest Surgery
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    • v.36 no.4
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    • pp.229-241
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    • 2003
  • Background:Autogenous vein is the preferred vascular graft for patients who require coronary artery bypass surgery or peripheral arterial bypass surgery. When an autogenous vein is not available, an allograft saphenous vein can be used as an alternative conduit. Although arterial homograft has been under investigation since the beginning of this century, the viability of endothelial cells and the optimum mode of storage for the venous and arterial allografts is controversial. In addition, with the recently gained knowledge of vascular endothelial functions, such as the production of nitric oxide or thrombomodulin, the viability and antigenicity of endothelial cells are being studied again. The purpose of this study was to evaluate the viability of endothelial cells in the preserved human saphenous veins. Material and Method: The veins were stored in a $4^{\circ}C$ RPMI (Roswell Park Memorial Institute) 1640 solution including 10% fetal calf serum, for one, three, five, seven or fourteen days. After the completion of the storage period, the veins were divided into two groups: Group I: studied immediately at $4^{\circ}C$ (cold) storage (I-1, I-3, I-5, I-7, I-14), and Group II: studied after storage at $-196^{\circ}C$ liquid nitrogen tank (cryopreservation) in an RPMI 1640 solution containing 10% DMSO for two weeks (II-1, II-3, II-5, II-7, II-14). Light microscopy and scanning electron microscopy (SEM), frypan blue exclusion testing, and thrombomodulin immunohistochemistry were performed. Result: In a morphometric study using SEM, there was statistically significant increase in Gundry Score in Groups I-7, I-14, II-5, II-7, and II-14 and showed cellular destruction (p<0.05). In the thrombomodulin immunohistochemistry study, there was reactivity in Groups I-1, I-3, and I-5, but the cryopreserved group revealed decreased reactivity (p<0.05). The trypan blue exclusion testing also showed superior viability in cold storage Group I. Conclusion: Venous allografts preserved in a $4^{\circ}C$ RPMI 1640 solution showed well preserved endothelial cellular integrity and thrombomodulin expression at up to seven days of preservation. Although cryopreservation of venous allografts stored in 10% DMSO -RPMI 1640 solution maintained the endothelial cellular structure on SEM, immunohistochemistry from the thrombomodulin and trypan blue exclusion testing showed decreased viability, It remains to be seen whether the decreased thrombomodulin reactivity could be restored, and what the nature to the relationship is between thrombomodulin and long-term patency of allografts.

Geochemical Characteristics of the Gyeongju LILW Repository II. Rock and Mineral (중.저준위 방사성폐기물 처분부지의 지구화학 특성 II. 암석 및 광물)

  • Kim, Geon-Young;Koh, Yong-Kwon;Choi, Byoung-Young;Shin, Seon-Ho;Kim, Doo-Haeng
    • Journal of Nuclear Fuel Cycle and Waste Technology(JNFCWT)
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    • v.6 no.4
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    • pp.307-327
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    • 2008
  • Geochemical study on the rocks and minerals of the Gyeongju low and intermediate level waste repository was carried out in order to provide geochemical data for the safety assessment and geochemical modeling. Polarized microscopy, X-ray diffraction method, chemical analysis for the major and trace elements, scanning electron microscopy(SEM), and stable isotope analysis were applied. Fracture zones are locally developed with various degrees of alteration in the study area. The study area is mainly composed of granodiorite and diorite and their relation is gradational in the field. However, they could be easily distinguished by their chemical property. The granodiorite showed higher $SiO_2$ content and lower MgO and $Fe_2O_3$ contents than the diorite. Variation trends of the major elements of the granodiorite and diorite were plotted on the same line according to the increase of $SiO_2$ content suggesting that they were differentiated from the same magma. Spatial distribution of the various elements showed that the diorite region had lower $SiO_2,\;Al_2O_3,\;Na_2O\;and\;K_2O$ contents, and higher CaO, $Fe_2O_3$ contents than the granodiorite region. Especially, because the differences in the CaO and $Na_2O$ distribution were most distinct and their trends were reciprocal, the chemical variation of the plagioclase of the granitic rocks was the main parameter of the chemical variation of the host rocks in the study area. Identified fracture-filling minerals from the drill core were montmorillonite, zeolite minerals, chlorite, illite, calcite and pyrite. Especially pyrite and laumontite, which are known as indicating minerals of hydrothermal alteration, were widely distributed in the study area indicating that the study area was affected by mineralization and/or hydrothermal alteration. Sulfur isotope analysis for the pyrite and oxygen-hydrogen stable isotope analysis for the clay minerals indicated that they were originated from the magma. Therefore, it is considered that the fracture-filling minerals from the study area were affected by the hydrothermal solution as well as the simply water-rock interaction.

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Ultrastructure of Degenerating Axon Terminals in the Basal Forebrain Nuclei of the Rat following Prefrontal Decortication (이마앞겉질을 제거시킨 흰쥐 앞뇌의 바닥핵무리에서 변성축삭종말의 미세구조연구)

  • Ahn, Byung-June;Ko, Jeong-Sik;Ahn, E-Tay
    • Applied Microscopy
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    • v.35 no.3
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    • pp.135-152
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    • 2005
  • Prefrontal cortex is a psychological and metaphysical cortex, which deals with feeling, memory, planning, attention, personality, etc. And it also integrates above-mentioned events with motor control and locomotor activities. Prefrontal cortex works as a highest CNS center, since the above mentioned functions are very important for one's successful life, and further more they are upgraded every moments through memory and learning. Many of these highest functions are supposed to be generated via forebrain basal nuclei (caudate nucleus, fundus striati nucleus, accumbens septi nucleus, septal nucleus, etc.). In this experiment, prefrontal efferent terminals within basal forebrain nuclei were ultrastructurally studied. Spraque Dawley rats, weighing $250{\sim}300g$ each, were anesthetized and their heads were fixed on the stereotaxic apparatus (experimental model, David Kopf Co.). Rats were incised their scalp, perforated a 3mm-wide hole on the right side of skull at the 11mm anterior point from the frontal O point (Ref. 13, Fig. 1), suctioned out the prefrontal cortex including cortex of the frontal pole, with suction instrument. Two days following the operations, small tissue blocks of basal forebrain nuclei were punched out, fixed in 1% glutaraldehyde-1% paraformaldehyde solution followed by 2% osmium tetroxide solutions. Ultrathin sections were stained with 1% borax-toluidin blue solution, and the stained sections were obserbed with an electron microscope. Degenerating axon terminals were found within all the basal forbrain nuclei. Numbers of degenerated terminals were largest in the caudate nucleus, next in order, in the fundus striati nucleus, in the accumbens septi nucleus, and the least in the septal nucleus. Only axospinous terminals were degenerated within the caudate nucleus and the fundus striati nucleus, and they showed the characters of striatal motor control system. Axodendritic and axospinous terminals were degenerated within the accumbens septi nucleus and the lateral septal nucleus, and they showed the characters of visceral limbic system. Prefrontal role in integrating the limbic system with the striatal system, en route basal forebrain nuclei, was discussed.

Distribution and Differentiation of the Choline Acetyltransferase-immunoreactive Nerve Cells in the Basal Nucleus of Meynert of the Rat Forebrains During the Postnatal Development (흰쥐 출생후 발생에 따른 전뇌 기저부 Meynert기저핵에서 Choline Acetyltranseferase 면역반응 신경세포의 분포 및 분화)

  • Lee, Nam-Seob;Chung, Young-Wha
    • Applied Microscopy
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    • v.29 no.4
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    • pp.479-491
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    • 1999
  • This study was performed to investigate the distribution and differentiation of choline acetyltransferase (ChAT)-immunoreactive cells in the basal nucleus of Meynert of the postnatal and adult rat forebrains, utilizing techniques of immunocytochemistry. According to the cell shape and the ratio of long axis vs short axis of cell soma, the ChAT-immunoreactive nerve cells in the basal nucleus of Meynert of the adult rat were classified into six types. In the adult rat, the frequency distributions (FD) of round, oval, elongated, fusiform, triangular and polygonal cells were 9.4%, 35.5%, 32.1%, 5.9%, 9.1% and 8.0%, respectively. The FD of oval and round nerve cells on the postnatal day (PND) 14 were observed to be 18.7% and 51.5%, respectively. Those were shown to be progressively decreased during developmental process to the adult. Also, those of elongated and triangular nerve cells on the PND 21 were observed to be 30.4% and 10.1%, respectively. Those were shown to be same phenomenon a,1 those in the round and oval cells. Meanwhile, those of the triangular and polygonal nerve cells were progressively increased from the early postnatal stage to the adult. The total mean volumes of ChAT-immunoreactive cell somata in the PND 7 rat were the lowest $(1,083{\mu}m^3)$ and those in the PND 21 rat were shown to be the highest $(5,045{\mu}m^3)$. But in the adult, those were decreased to $(2,731{\mu}m^3)$. Those in the PND 21 rat were shown to be about 84.7% larger than those in the adult. On the electron micrography, the cell organelles such as ribosomes, polysomes, rough endoplasmic reticula (RER) and mitochondria were well developed in the PND 21 rat forebrains, but Golgi complexes were shown to be proliferating phase. Especially, ribosomes, polysomes and RER were immunoreactive in the tissues treated with 0.05% triton X-100. According to the observations in the present study, it is considered that the ChAT-immunoreactive nerve cells in the basal nucleus of Meynert of the rat forebrains are differentiated throughout the following processes of changes during the postnatal development: 1) increase of cell soma volumes with the differentiation of tell organelles and neurites, 2) increase in the FD of differentiated tell types and 3) cell schrinkage without cell loss. The ribosomes, polysomes and RER are considered to be closely related to the intracellular localization and biosynthesis of the ChAT but not Colgi complex.

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Histochemical Detection of Ionic Zinc in the Rat Olfactory Mucosa: Zinc Selenium Autometallography ($ZnSe^{AMG}$) (랫드 후각점막내 Zinc 이온의 조직화학적 동정)

  • Nam, Dong-Woo;Sun, Yuan-Jie;Kim, Sung-Joo;Kim, Yong-Kuk;Kim, Soo-Jin;Yu, Yun-Cho;Jeong, Young-Gil;Jo, Seung-Mook
    • Applied Microscopy
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    • v.33 no.2
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    • pp.145-154
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    • 2003
  • The present study was designed to demonstrate ionic zinc in the rat nasal mucosa by means of zinc selenium autometallography ($ZnSe^{AMG}$). Rats were given sodium selenide either intraperitoneally (i.p) or intranasally (i.n). Prior to the i.n. administration the rats were anesthetized with pentobarbital sodium (30 mg/kg, i.p.). A thin plastic tube coupled to a Hamilton syringe was then inserted into the right nostril and $10{\mu}l$ of the solution was instilled. For the i.p. administration non-anesthetized rats were given $100{\mu}l$ of the sodium selenide solution (10 mg/kg). Control rats were instilled with saline. After 2 hrs survival, the rats were anaesthetized and transcardially perfused with 3% glutaraldehyde. The olfactory area was removed and put into same fixative. The nose was then sectioned ($30{\mu}m$) horizontally, autometallography (AMG) was performed according to Danscher et al. (1997). After silver enhancement, fine AMG grains were scattered in the whole length of the olfactory epithelium containing olfactory receptor neurons, sustentacular and basal cells. However, much higher concentration of the AMG grains occupied near the surface and in the basal region of the olfactory epithelium. Both groups of i.p. and i.n. administration showed almost same level in the concentration of the AMG grains. In i.n. group, few AMG grains were also found in olfactory nerves of the lamina propria, suggesting zinc transport into the olfactory bulb via olfactory axons. At the electron microscopic level, the AMG grains were most entirely found in the supporting cells of the olfactory epithelium, and they were mostly localized in lysosome-like organelles. The i.n. group showed various signs of tissue damage of the olfactory mucosa, where dense concentration of AMG grains were localized at crystalloid structures. The present study demonstrated dense population of ionic zinc in the rat olfactory epithelium. zinc may play a role in the olfactory functioin and in the pathogenesis of the neurodegerative disorders affecting nose.

Increased Protein of the Secretory Leukocyte Pretense Inhibitor (SLPI) and the Expression of Growth Factors in NIH3T3 Cells by LPS Stimulation (NIH3T3 세포주에서 LPS자극에 의한 분비백혈구단백분해효소억제제 (SLPI)의 단백질증가와 성장인자들의 발현)

  • Lee, Sang-Hwa;Choi, Baik-Dong;Jeong, Soon-Jeong;Jang, Hyun-Seon;Kim, Byung-Ock;Lim, Do-Seon;Park, Joo-Cheol;Wang, Guan-Lin;Jeong, Moon-Jin
    • Applied Microscopy
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    • v.36 no.3
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    • pp.165-172
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    • 2006
  • Secretory leukocyte protease inhibitor (SLPI) involves tissue protection against the destructive action of neutrophil elastase at the site of inflammation. Several studies on new functions of SLPI have demonstrated that SLPI may play a primary role in innate immunity than protease inhibitor, To identify the function of SLPI by lipopolysaccharide (LPS) stimulation in the embryonic fibroblast (NIH3T3) cells. we studied the expression of SLPI compared to other growth factors involving the LPS treatment. To address this, we performed the reverse transcriptase polymerase chain reaction (RT-PCR) and Western blots for the detection of mRNA and protein expression of the SLPI and some growth factors such as VEGF. bFGF, and PDGF-BB after LPS stimulation. NIH3T3 cells were exposed 100 ng/mL Escherichia coli LPS for 30min, 60min, 90min, 24h, and 48h, respectively. The result of RT-PCR showed that SLPI and VEGF mRNA was expressed strongly in NIH3T3 without related to LPS stimulation. mRNA of bFGF was weakly expressed such as the expression of the control. PDGF mRNA expression gradually increased follows at time course. However, SLPI protein level was increased in lysates and culture medium by LPS stimulation. Phase contrast microscopic and scanning electron microscopic observation showed that the increased cell number and cytoplasmic enlargement of the NIH3T3 cells. Therefore, it suggests that the LPS upregulates SLPI expression in NIH3T3 cells. Moreover, secreted SLPI may stimulate cell proliferation and migration.