• Title/Summary/Keyword: cell morphology

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THE EFFECTS OF NICOTINE ON HUMAN GINGIVAL FIBROBLAST & PERIODONTAL LIGAMENT CELLS IN VITRO (니코틴이 배양인체 치은섬유모세포 및 치주인대세포의 활성에 미치는 효과)

  • Kong, Young-Hwan;Yoo, Hyung-Keun;Shin, Hyung-Shik
    • Journal of Periodontal and Implant Science
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    • v.25 no.2
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    • pp.181-191
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    • 1995
  • The ability of fibroblasts attach to teeth is of paramount imporance in re-establishing the lost connective tissue attachment after periodontal therapy. Tobacco contains a complex mixture of substances including nicotine. various nitrousamines, trace elements. and a variety of poorly characterized substances. The effects of nicotine on fibroblasts have reported an altered morphology and attachment of fibroblasts to substrates and disturbances in protein synthesis and secretion. This study examined the effect of nicotine, a major component of the particulate phase of tobacco smoke, on human gingival fibroblasts and periodontal ligament cells attachment to tissue culture surfaces and cellular activity of human gingival fibroblasts and periodontal ligament cells. Pooled human gingival fibroblasts made from extraction of 3rd molar were utilized between passage 4 and 5 and plated in 96 well plate at 20,000 cells per well. Cell number were determined using 3-(4,5-dimethylthiazole-2-y)2,5-diphenyltetrazolium bromide(MTI) , which is reflection of mitochondrial dehydrogenase activity. The concentration of nicotine used were 0.025, 0.05, 0.1, 0.2 and $0.4{\mu}M$, the average serum concentration for a smoker being approximately $0.1{\mu}M$. The results were as follows : 1. Attachment effects of nicotine on human gingival fibroblasts and periodontal ligament cells Excepts of $0.4{\mu}M$, the effects on attachment with increasing numbers of cells attaching with increasing nicotine concentrations, compared to control group. But over the 60min, return to control value. 2. The effect of cellular activity on human gingival fibroblasts and periodontal ligament cells. The cellular activity of human gingival fibroblasts and periodontal ligament cells were similar or decrease to control value at 1st incubation day. At 2nd incubation day, 0.05, 0.1, 0.2, $0.4{\mu}M$ concentrations were statistically different from control value on gingival fibroblasts group. But at 3rd incubation day, cellular activities of all experimental group were significantly decrease than control group.

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INTERRELATIONSHIP BETWEEN VIRULENT CLONAL TYPES, SEROTYPES AND LEUKOTOXICITY OF KOREAN STRAINS OF A. ACTINOMYCETEMCOMITANS (한국인 Actinobacillus actinomycetemcomitans 균주의 특이 독성 clone형과 혈청형 및 백혈구독성과의 관계)

  • Ku, Young
    • Journal of Periodontal and Implant Science
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    • v.25 no.3
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    • pp.487-496
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    • 1995
  • Previous studies have demonstrated that not all A. actinomycetemcomitans produced significant level of leukotoxic factor and its leukotoxicity have associated with serotype and genetic variation. Our aim was to investigate on the interrelationship between serotype and leukotoxicity of an A. actinomycetemcomitans consisting of 13 clinically well characterized. Korean isolates and to evaluate if particular virulent clonal types of A. actinomycetemcomitans are associated with periodontal disease. For this study, 13 strains of A. actinomycetemcomitans from 6 patients with periodontal disease were isolated and identified by using a selective medium(tryptic soy agar supplemented with 10% serum, $75{\mu}g$ of bacitracin and $5{\mu}g$ of vancomycin per ml) in 10% C02 incubator for 3days with routine Gram staining, colony morphology and biochemical test..For serotyping, antisera were prepared from reference strains of 5 serotypes. (ATCC 29523,Y4, SUNY aB 67, IDH 781, IDH 1705) and then ammonium sulfate precipitation, immunoabsorption and indirect immunofluoroscent procedures were done. For analysis of leukotoxicity, sonic extract of A. actinomycetemcomitans exposed to PMN, and trypan blue was stained for counting the cell viability. Finally Southern blot analyses of genomic DNA digested with the restriction enzyme Tag I was done and the Southern blots were hybridized with the 530bp fragment, termed delta 530, originating from the ltx promoter of strain 652 and deleted from strain JP2. Also ltxA-3.1 and SC2 probe from strain JP2 were hybridized with genomic DNA fragments. Results reveal that strains isolated showed approximately equal proportions of 3 serotypes(b, d, e) and serotype b was not detected. 2 patients harbored 2 different serotypes in the same disease site. The prevalence of leukotoxic strain was 23% and there was no relationship between serotype, leukotoxicity and clinical observations. Especially virulent clonal types of Actinobacillus actinomycetemcomitan (JP2 strain) could not found. Further studies are necessary on the genetic polymorphism of leukotoxin and its relations to clinical status.

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Laboratory passage and characterization of an isolate of Toxoplasma gondii from an ocular patient in Korea

  • Chai, Jong-Yil;Lin, Aifen;Shin, Eun-Hee;Oh, Myoung-Don;Han, Eun-Taek;Nam, Ho-Woo;Lee, Soon-Hyung
    • Parasites, Hosts and Diseases
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    • v.41 no.3
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    • pp.147-154
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    • 2003
  • Toxoplasma gondii tachyzoites were isolated from the blood of an ocular patient, and have been successfully passaged in the laboratory, for over a year, by peritoneal inoculation in mice. The isolated parasite was designated the Korean Isolate-1 (KI-1) and its characteristics were compared with those of the RH strain, a well-known virulent strain originating from a child who suffered from encephalitis. The morphology, pathogenicity, infectivity and cell culture characteristics of the KI-1 were similar to those of the RH strain. Both RH and KI-1 antigens were detected by an anti-T gondii monoclonal antibody (mAb), Tg563, against the major surface protein SAG1 (30 kDa), whereas no reaction was observed against an anti-Neospora caninum mAb, 12B4. The KI-1 was confirmed as an isolate of T. gondii. A long-term laboratory maintenance and characterization of a local T gondii isolate is reported for the first time in the Republic of Korea.

The effect of Fullerene (C60) nanoparticles on the surface of PVDF composite membrane

  • Kim, Kyung Hee;Lee, Ju Sung;Hong, Hyun Pyo;Han, Jun Young;Park, Jin-Won;Min, ByoungRyul
    • Membrane and Water Treatment
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    • v.6 no.5
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    • pp.423-437
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    • 2015
  • Polyvinylidene fluoride/fullerene nanoparticle (PVDF/$C_{60}$) composite microfiltration (MF) membranes were fabricated by a non-solvent induced phase separation (NIPS) using N, N-dimethylacetamide (DMAc) as solvent and deionized water (DI) as coagulation solution. Polyvinylpyrrolidone (PVP) was added to the casting solution to form membrane pores. $C_{60}$ was added in increments of 0.2% from 0.0% to 1.0% to produce six different membrane types: one pristine PVDF membrane type with no $C_{60}$ added as control, and five composite membrane types with varying $C_{60}$ concentrations of 0.2, 0.4, 0.6, 0.8 and 1.0%, respectively. The mechanical strength, morphology, pore size and distribution, hydrophilicity, surface property, permeation performance, and fouling resistance of the six membranes types were characterized using respective analytical methods. The results indicate that membranes containing $C_{60}$ have higher surface porosity and pore density than the pristine membrane. The presence of numerous pores on the membrane caused weaker mechanical strength, but the water flux of the composite membranes increased in spite of their smaller size. Initial flux and surface roughness reached the maximum point among the composite membranes when the $C_{60}$ concentration was 0.6 wt.%.

Protection of spontaneous and glutamate-induced neuronal damages by Soeumin Sibjeundaibo-tang and Soyangin Sibimijihwang-tang in cultured mice cerebrocortical cells

  • Lee, Mi-Young;Ma, Jin-Yeul;Choo, Young-Kug;Jung, Kyu-Yong
    • Advances in Traditional Medicine
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    • v.1 no.1
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    • pp.55-63
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    • 2000
  • Soeumin Sibjeundaibo-tang (SJDBT) and Soyangin Sibimijihwang-tang (SMJHT) have been used traditionally to improve the systemic blood circulation and biological energy production in the patients with circulatory and neuronal diseases. The object of this study is to determine the protective effects of SJDBT and SMJHT extracts on the spontaneous and glutamate-induced neuronal damages in cultured cells derived from mice cerebral cortex. At 14 days after beginning the cultures, the activity of lactate dehydrogenase released into the culture media was significantly decreased by treatment of cerebroneuronal cells with SJDBT and SMJHT (0.1 mg/ml) for 7 days. By comparison with the normal cells, cerebroneuronal morphology was dramatically changed by treatment of glutamate (1 mM) for 12 hrs, and this was conspicuously recovered by pretreatment of cerebroneural cells with SJDBT and SMJHT (0.1-1.0 mg/ml) for 2 days. Moreover, glutamated-induced DNA fragmentation was also protected by pretreatment of cerebroneuronal cells with those extracts. These results suggest that naturally occurring and glutamate-induced degeneration of cultured cerebrocortical cells may be related, in part, to the process of apoptotic cell death. The pharmacological properties of SJDBT and SMJHT extracts to improve cerebroneuronal degeneration may be considered as one of useful medicines that can prevent cerebrocortical impairments resulted from age-dependent and excitotoxicity-induced neuronal degeneration in human brain.

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Recombinant Human Epidermal Growth Factor (rhEGF)-loaded Solid Lipid Nanoparticles: Fabrication and Their Skin Accumulation Properties for Topical rhEGF Delivery

  • Hwang, Hee-Jin;Han, Sunhui;Jeon, Sangok;Seo, Joeun;Oh, Dongho;Cho, Seong-Wan;Choi, Young Wook;Lee, Sangkil
    • Bulletin of the Korean Chemical Society
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    • v.35 no.8
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    • pp.2290-2294
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    • 2014
  • For the present study, rhEGF was encapsulated into solid lipid nanoparticles (SLNs). The SLNs were prepared by the $W_1/O/W_2$ double emulsification method combined with the high pressure homogenization method and the physical properties such as particle size, zeta-potential and encapsulation efficiency were measured. The overall particle morphology of SLNs was investigated using a transmission electron microscopy (TEM). The percutaneous skin permeation and accumulation property of rhEGF was evaluated using Franz diffusion cell system along with confocal laser scanning microscopy (CLSM). The mean particle size of rhEGF-loaded SLNs was $104.00{\pm}3.99nm$ and the zeta-potential value was in the range of -$36.99{\pm}0.54mV$, providing a good colloidal stability. The TEM image revealed a spherical shape of SLNs about 100 nm and the encapsulation efficiency was $18.47{\pm}0.22%$. The skin accumulation of rhEGF was enhanced by SLNs. CLSM image analysis provided that the rhEGF rat skin accumulation is facilitated by an entry of SLNs through the pores of skin.

Spermiogenesis and Taxonomical Values of Sperm Ultrastructures in Male Crassostrea ariakensis (Fujita & Wakiya, 1929) (Pteroirmorphia: Ostreidae) in the Estuary of the Seomjin River, Korea

  • Son, Pal Won;Chung, Jae Seung;Kim, Jin Hee;Kim, Sung Han;Chung, Ee-Yung
    • Development and Reproduction
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    • v.18 no.3
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    • pp.179-186
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    • 2014
  • Characteristics of the developmental stages of spermatids during spermiogenesis and phylogenetic classicfication of the species using sperm ultrastructures in male Crassostrea ariakensis were investigated by transmission electron microscope observations. The morphology of the spermatozoon of this species has a primitive type and is similar to those of Ostreidae. Ultrastructures of mature sperms are composed of broad, modified cap-shaped acrosomal vesicle and an axial rod in subacrosomal materials on an oval nucleus, four spherical mitochondria in the sperm midpiece, and satellite fibres which appear near the distal centriole. The axoneme of the sperm tail shows a 9+2 structure. Accordingly, the ultrastructural characteristics of mature sperm of C. ariakensis resemble to those of other investigated ostreids in Ostreidae in the subclass Pteriomorphia. In this study, particularly, two transverse bands (stripes) appear at the anterior region of the acrosomal vesicle of this species, unlike two or three transverse bands (stripes) in C. gigas. It is assumed that differences in this acrosomal substructure are associated with the inability of fertilization between the genus Crassostrea and other genus species in Ostreidae. Therefore, we can use sperm ultrastructures and morphologies in the resolution of taxonomic relationships within the Ostreidae in the subclass Pteriomorphia. These spermatozoa, which contain several ultrastructures such as acrosomal vesicle, an axial rod in the sperm head part and four mitochondria and satellite fibres in the sperm midpiece, belong to the family Ostreidae in the subclass Pteriomorphia.

Antidiabetic Effect of Herbal Formula Containing Mori Folium, Euonymi Lignum Suberalatum and Ginseng Radix in db/db Mice (db/db 마우스에서 상엽, 귀전우, 인삼 복합처방의 항당뇨 활성)

  • Park, Keum-Ju;Han, Eun-Jung;Choi, Yun-Sook;Han, Gi-Cheol;Park, Jong-Seok;Chung, Sung-Hyun
    • Korean Journal of Pharmacognosy
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    • v.38 no.1
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    • pp.10-14
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    • 2007
  • Type 2 diabetes mellitus relavant to insulin resistance is a chronic and hard to control. In order to develop an antidiabetic agent from natural products, anti-hyperglycemic effect of herbal formula containing Mori Follium, Euonymi Lignum Suberalatum and Ginseng Radix(MEG) was investigated in db/db mice. Treatment group was administered orally with MEG formula at a dose of 300 mg/kg for 5 weeks, and blood glucose, insulin and lipid levels were determined. MEG treatment group showed a marked decrease in fasting blood glucose level and insulin resistance index(IRI) compared to those in diabetic control. Improvement of insulin resistance(60.6%) was indicative of reducing lipid levels in plasma and triglyceride contents in muscle and adipose tissue. In addition, expressions of an insulin responsive gene, glucose transporter 4(Glut4), in muscle and adipose tissue were upregulated in MEG treatment group. Compared islet morphology between groups, MEG formula prevented the ${\beta}$-cell destruction caused by high blood glucose. Taken together, MEG formula can act as an anti-hyperglycemic agent with insulin sensitizing effect, and thus deserves a clinical trial in the future.

Morphological Classification of Trichomes Associated with Possible Biotic Stress Resistance in the Genus Capsicum

  • Kim, Hyun-Jung;Seo, Eun-Young;Kim, Ji-Hyun;Cheong, Hee-Jin;Kang, Byoung-Cheorl;Choi, Do-Il
    • The Plant Pathology Journal
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    • v.28 no.1
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    • pp.107-113
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    • 2012
  • Trichomes are specialized epidermal structure having the functions of physical and chemical block against biotic and abiotic stresses. Several studies on $Capsicum$ species revealed that virus and herbivore resistance is associated with trichome-formation. However, there is no research on the structural characterization of trichomes developed on the epidermis of $Capsicum$ spp. Thus, this study attempts to charaterize the trichome morphologies in 5 species of $Capsicum$ using a Field Emission Scanning Electron Microscopy (FESEM). Six main trichome types were identified by their morphology under FESEM. Both glandular and non-glandular types of trichomes were developed on the epidermal tissues of $Capsicum$ spp. The glandular trichome were further classified into type I, IV and VII according to their base, stalk length, and stalk. Non-glandular trichomes were also classified into type II, III, and V based on stalk cell number and norphology. Almost all the species in $C.$ $chinense$ and $C.$ $pubescens$ had glandular trichomes. To our knowledge, this is the first study on classification of trichomes in the genus $Capsicum$ and, our results could provide basic informations for understanding the structure and function of trichomes on the epidermal differentiation and association with biotic stress tolerance.

Altered Gene Expression and Intracellular Changes of the Viable But Nonculturable State in Ralstonia solanacearum by Copper Treatment

  • Um, Hae Young;Kong, Hyun Gi;Lee, Hyoung Ju;Choi, Hye Kyung;Park, Eun Jin;Kim, Sun Tae;Murugiyan, Senthilkumar;Chung, Eunsook;Kang, Kyu Young;Lee, Seon-Woo
    • The Plant Pathology Journal
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    • v.29 no.4
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    • pp.374-385
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    • 2013
  • Environmental stresses induce several plant pathogenic bacteria into a viable but nonculturable (VBNC) state, but the basis for VBNC is largely uncharacterized. We investigated the physiology and morphology of the copper-induced VBNC state in the plant pathogen Ralstonia solanacearum in liquid microcosm. Supplementation of $200{\mu}M$ copper sulfate to the liquid microcosm completely suppressed bacterial colony formation on culture media; however, LIVE/DEAD BacLight bacterial viability staining showed that the bacterial cells maintained viability, and that the viable cells contain higher level of DNA. Based on electron microscopic observations, the bacterial cells in the VBNC state were unchanged in size, but heavily aggregated and surrounded by an unknown extracellular material. Cellular ribosome contents, however, were less, resulting in a reduction of the total RNA in VBNC cells. Proteome comparison and reverse transcription PCR analysis showed that the Dps protein production was up-regulated at the transcriptional level and that 2 catalases/peroxidases were present at lower level in VBNC cells. Cell aggregation and elevated levels of Dps protein are typical oxidative stress responses. $H_2O_2$ levels also increased in VBNC cells, which could result if catalase/peroxidase levels are reduced. Some of phenotypic changes in VBNC cells of R. solanacearum could be an oxidative stress response due to $H_2O_2$ accumulation. This report is the first of the distinct phenotypic changes in cells of R. solanacearum in the VBNC state.