• Title/Summary/Keyword: Confocal microscopy

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Biomineralization of Calcium Carbonate Polymorphs by the Bacterial Strains Isolated from Calcareous Sites

  • Dhami, Navdeep Kaur;Reddy, M. Sudhakara;Mukherjee, Abhijit
    • Journal of Microbiology and Biotechnology
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    • v.23 no.5
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    • pp.707-714
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    • 2013
  • Microbially induced calcium carbonate precipitation (MICCP) is a naturally occurring biological process that has various applications in remediation and restoration of a range of building materials. In the present investigation, five ureolytic bacterial isolates capable of inducing calcium carbonate precipitation were isolated from calcareous soils on the basis of production of urease, carbonic anhydrase, extrapolymeric substances, and biofilm. Bacterial isolates were identified as Bacillus megaterium, B. cereus, B. thuringiensis, B. subtilis, and Lysinibacillus fusiformis based on 16S rRNA analysis. The calcium carbonate polymorphs produced by various bacterial isolates were analyzed by scanning electron microscopy, confocal laser scanning microscopy, X ray diffraction, and Fourier transmission infra red spectroscopy. A strain-specific precipitation of calcium carbonate forms was observed from different bacterial isolates. Based on the type of polymorph precipitated, the technology of MICCP can be applied for remediation of various building materials.

The Wear Mechanism of Carbon(WC/C) Thin Film in Lubricated Contact (WC/C 박막의 윤활접촉하의 마멸기구)

  • Ahn, Hyo-Sok;Kim, Doo-In
    • Tribology and Lubricants
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    • v.24 no.6
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    • pp.302-307
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    • 2008
  • The running-in behaviour of the metal carbon coating (WC/C) was evaluated with regard to its applicability as wear-resistant coating for key components in engines. Reciprocating wear tests under lubricated condition employing an oscillating friction wear tester were performed to investigate the tribological behaviors of the coatings in ambient environment. Confocal microscopy and scanning electron microscopy were used to observe worn surfaces and the wear scars on the steel balls. Elemental composition of the coating and worn surfaces were characterized using Auger electron spectroscopy. The friction behavior of WC/C coating was comparable to common carbon-based coatings. Thin tribofilm was formed on the worn surface of the steel ball due to material transfer and tribochemical reaction whereas there was no evidence of tribofilm generation on the coating surface. indicating the chemical innertness of WC/C coating.

Morphology of Silk Fibroin/Poly(vinyl alcohol) Blend Film (견피브로인/Poly(vinyl alcohol) 브렌드 필름의 형태학적 구조)

  • 엄인철;박영환
    • Journal of Sericultural and Entomological Science
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    • v.40 no.2
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    • pp.169-175
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    • 1998
  • The morphology of silk fibroin/poly(vinyl alcohol)(PVA)blend films was investigated using optical microscopy and confocal laser scanning microscopy. The effects of blend ratio and molecular weight of silk fibroin and PVA on phase separation were studied. Macro-phase separation occurred for the silk fibroin-rich/poor region whereas micro-phase separation took place for the dispersed/continuous phase, In spite of differences in molecular weight and blend ratio, it is observed that the dispersed phase and continuous one are composed of silk fibroin and PVA component, respectively. As the molecular weight of silk fibroin and silk fibroin content in blend ratio are decreased, the compatibility of blend is increased due to the reduction of micro-phase separation.

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Apoptosis in the craniofacial tissues of irradiated growing rats

  • Heo Min-Suk;Choi Hang-Moon;Lee Sam-Sun;Choi Soon-Chul;Park Tae-Won
    • Imaging Science in Dentistry
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    • v.31 no.4
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    • pp.227-233
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    • 2001
  • Purpose: The purpose of this study was to investigate the apoptosis induction in tissues constituting the craniofacial region of growing rat by irradiation. Materials and Methods: The submandibular gland, brain, articular cartilage of condylar head, and calvarium were extracted from 20-day-old rats irradiated 10 Gy. Apoptosis of each tissue was examined by DNA fragmentation and estimated quantitatively using apoptotic index on TUNEL assay. Apoptotic index of each tissue was calculated by the equation for apoptotic cells/total cells × 1,000 on the images of confocal laser scanning microscopy. Apoptotic index was analyzed statistically according to the time lapse after irradiation on the tissues. Results : In the submandibular gland, apoptotic index was significantly increased from 6 hours after irradiation showing the highest value at 12 hours and decreased to the control level at 3 days after irradiation. In the brain, apoptotic index was abruptly reached to the maximum value at 6 hours after irradiation and decreased to the control level at 4 days after irradiation. Articular cartilage and calvarium showed no or little apoptotic signals. The results obtained by the apoptotic index accorded with that of DNA fragmentation. Conclusion : Radiation was closely related with the apoptosis of submandibular gland and brain but, not related with the apoptosis of the articular cartilage of condylar head and calvarium. The changes induced by radiation of the hard tissues would not be explained by apoptosis.

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Isolation and Characterization of Endosome Subpopulation in Chinese Hamster Ovarian Cells

  • Suh, Duk-Joon;Park, Mi-Yeon;Jung, Dong-Keun;Bae, Hae-Rahn
    • The Korean Journal of Physiology
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    • v.30 no.2
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    • pp.197-208
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    • 1996
  • Endosomes lower their internal pH by an ATP-driven proton pump, which is critical to dissociation of many receptor-ligand complexes, the first step in the intracellular sorting of internalized receptors and ligands. Endosomes are known to exhibit n great range of pH values that can vary between 5.0 and 7.0 within a single cell although the factors that regulate endosomal pH remain uncertain. To evaluate the morphological and topological differences of endosomes in the different stages, confocal microscopy was used. The early endosomes labeled with fluorescein isothiocyanate-dextran for 10 min at $37^{\circ}C$ were identifiable at the peripheral and tubule-vesicular endosome compartment. In contrast, the late endosomes formed by 10 min pulse and 20 min trace were located deeper in the cytoplasm and showed more vesicular features than early endosomes. For the purpose of determining whether ATP-dependent acidification was heterogeneous and whether the differences in acidification were attributed to differences in the activity of $Na^{+}-K^{+}$-ATPase and/or $Cl^{-}$ channel, endocytic compartments were fractionated into subpopulation using percoll gradient and measured ATP-dependent acidification. While all fractions exhibited ATP-dependent acidification activity, both the initial rate of acidification and extent of proton translocation were lower in early endosomes and gradually increased in late endosomes. Phosphorylation by PKA and ATP enhanced ATP-dependent acidification in both early and late endosomes, hut there was no difference in the degree of enhancement by phosphorylation between two subpopulations. When ATP-dependent acidification was determined in the presence or absence of vanadate ($Na_{3}VO_{4}$) or ouabain, only early endosomes exhibited the vanadate or ouabain dependent stimulation of acidification activity, suggesting the inhibition of $Na^{+}-K^{+}$-ATPase. Therefore, it seems probable that the inhibition of early endosome acidification by $Na^{+}-K^{+}$-ATPase observed in vitro at least in part plays a physiological role in controlling the acidification of early endosomes in vivo.

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Generation and characterization of a monoclonal antibody with high species-specificity to Schistosoma japonicum glutathione S-transferase

  • Kim, Jung-Hwan;Park, Jung-Hyun;Ju, Sung-Kyu;Lee, Myung-Kyu;Kim, Kil Lyong
    • IMMUNE NETWORK
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    • v.1 no.3
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    • pp.187-195
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    • 2001
  • The expression of recombinant proteins fused to 26 kDa glutathione S-transferase (GST) extracted from Schistosoma japonicum represents an attractive system for purifiying proteins of interest in a single step using GST-affinity chromatography. In addition, the GST-tag is used conveniently for detecting fused proteins since its high solubility as well as its relatively small size rarely interferes with the biological activity of the fused protein. In this regard, the GST system is frequently applied for tracing fusion proteins in both prokaryotic and eukaryotic cells to elucidate the physiological interactions and functional compartments of proteins. To provide a further tool in analyzing GST-fusion proteins, a new monoclonal antibody, with a high specificity to the S. japonicum GST was produced. Methods: BALB/c mice were immunized both with recombinant S. japonicum GST proteins, and by the fusion of splenocytes from these mice with myeloma cells. From this, a new anti -GST monoclonal antibody, termed SARAH, was generated. The specificity and reactivity of this antibody was confirmed by ELISA and by Western blot analysis. Results: SARAH showed a high reactivity to recombinant GST and GST fusion protein but not with native mammalian GST proteins as derived from other species including humans, cows, rabbits and rats. The applicability of SARAH was further demonstrated by confocal laser scanning microscopy, where GST proteins that were expressed transiently in mouse fibroblast cells, were specifically detected without interference of endogenous GST. Conclusion: SARAH is new monoclonal antibody with a high specificity to recombinant GST proteins but not to endogenous GST in mammalian cells.

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Antimicrobial Effect of Photodynamic Therapy Using Plaque Disclosing Agent (치면착색제를 이용한 광역동 치료의 항균 효과)

  • Kho, Junhee;Park, Howon;Lee, Juhyun;Seo, Hyunwoo;Lee, Siyoung
    • Journal of the korean academy of Pediatric Dentistry
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    • v.47 no.2
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    • pp.120-127
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    • 2020
  • The aim of this study was to evaluate the antimicrobial effect of photodynamic therapy (PDT) using plaque disclosing agent, 10 - 20 mM erythrosine, as a photosensitizer. Multispecies cariogenic biofilms containing Streptococcus mutans, Lactobacillus casei and Candida albicans were formed on hydroxyapatite disc. 20 μM, 10 mM and 20 mM erythrosine were applied as a photosensitizer for 3 minutes, and then light-emitting diode (LED) irradiated for 24 seconds. Colony-forming unit (CFU) were measured and biofilms were observed using confocal laser scanning microscopy (CLSM). CFU were significantly decreased in the PDT groups using 10 - 20 mM erythrosine (10 mM, 20mM) and the results were also confirmed by CLSM. This study confirms the high antimicrobial effect of photodynamic therapy using plaque disclosing agent as a photosensitizer.

Comparison of Microtubule Distributions between Somatic Cell Nuclear Transfer and Parthenogenetic Porcine Embryos

  • Park, Joo-Hee;Kwon, Dae-JinK;Lee, Beom-Ki;Hwang, In-Sun;Park, Choon-Keun;Yang, Boo-Keun;Cheong, Hee-Tae
    • Reproductive and Developmental Biology
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    • v.33 no.1
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    • pp.13-18
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    • 2009
  • The aim of this study was to examine the microtubule distributions of somatic cell nuclear transfer (SCNT) and parthenogenetic porcine embryos. Porcine SCNT embryos were produced by fusion of serum-starved fetal fibroblast cells with enucleated oocytes. Reconstituted and mature oocytes were activated by electric pulses combined with 6-dimethlyaminopurine treatment. SCNT and parthenogenetic embryos were cultured in vitro for 6 days. Microtubule assembly of embryos was examined by confocal microscopy 1 hr and 20 hr after fusion or activation, respectively. The proportions of embryos developed to the blastocyst stage were 25.7% and 30.4% in SCNT and parthenogenetic embryos, respectively. The frequency of embryos showing $\beta$-tubulins was 81.8% in parthenogenetic embryos, whereas 31.3% in SCNT embryos 1 hr after activation or fusion. The frequency of the embryos underwent normal mitotic phase was low in SCNT embryos (40.6%) compared to that of parthenogenetic ones (59.7%) 20 hr after fusion or activation (p<0.05). The rate of SCNT embryos with an abnormal mitosis pattern is about twice compared to that of parthenogenetic ones. The spindle assembly and its distribution of SCNT embryos in the first mitotic phase were not different from those of parthenogenetic ones. The result shows that although microtubule distribution of porcine SCNT embryos shortly after fusion is different from parthenogenetic embryos, and the frequency of abnormal mitosis 20 hr after fusion or activation is slightly increased in SCNT embryos, microtubule distributions at the first mitotic phase are similar in both SCNT and parthenogenetic embryos.

Antifungal Mechanism of Antifungal Peptide Derived from Cecropin A(1-8)- Melittin(1-12) Hybrid against Aspergillus fumigatus

  • Lee, Dong-Gun;Jin, Zhe-Zhu;Maeng, Cheol-Young;Shin, Song-Yub;Seo, Moo-Yeol;Kim, Kil-Lyong;Hahm, Kyung-Soo
    • Journal of Microbiology and Biotechnology
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    • v.9 no.2
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    • pp.168-172
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    • 1999
  • The antifungal mechanism of the antifungal peptide against Aspergillus fumigatus, $K^{18,19}$-CA(l-8)-ME(l-12), derived from cecropin A(l-8)-melittin(l-12) was investigated by confocal laser scanning microscopy, cell wall regeneration, ATPase activity inhibition, and released potassium ion. By confocal laser scanning microscopy, $K^{18,19}$-CA(l-8)-ME(l-12) was detected on the surface of A. fumigatus, while cecropin A used as a negative control peptide was not detected. The protoplast of A. fumigatus treated with$K^{18,19}$-CA(1-8)-ME(1-12) failed to regenerate the fungal cell walls. Compared with cecropin A, the amount of potassium ion released by $K^{18,19}$-CA(l-8)-ME(l-12) was increased. Furthermore, $K^{18,19}$-CA(l-8)-ME(l-12) inhibited the ATPase activity on the plasma membrane. These results suggested that $K^{18,19}$-CA(l-8)-ME(1-12) acts on the plasma membrane of A. fumigatus and its antifungal action is due to the ion channel or pore formation on the plasma membrane.

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