• Title/Summary/Keyword: confocal microscopy

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Adhesion control of Campylobacter jejuni in chicken skin using emulsifiers (유화제를 이용한 계육 표면에서 Campylobacter jejuni의 부착 제어)

  • Oh, Do Geon;Kim, Kwang Yup
    • Korean Journal of Food Science and Technology
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    • v.52 no.6
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    • pp.670-677
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    • 2020
  • To prevent contamination by Campylobacter jejuni during chicken carcass processing, the effect of emulsifiers on C. jejuni inoculated on chicken skin was investigated using confocal laser scanning microscopy. Among the 8 emulsifiers (SWA-10D, L-7D, M-7D, S-1670, L-1695, P-1670, polysorbate 20, polysorbate 80) tested for antimicrobial activity by the paper disk method, 4 emulsifiers (L-7D, L-1695, polysorbate 20, polysorbate 80) were screened further. Emulsifier L-1695 showed the largest clear zone at a concentration of 200 mg/mL. The 4 emulsifiers subjected to primary screening were screened for heat and pH stability. In the contact surface test, emulsifier L-1695 showed the lowest log CFU/㎠ value on both stainless steel and ceramic surfaces. When emulsifier L-1695 was applied via general and electrostatic spray methods, the number of C. jejuni entrapped inside chicken skin follicles was significantly reduced in both methods. In conclusion, the emulsifier L-1695 could be employed as a microbial detachment agent in the chicken carcass processing industry.

Effect of Potassium Iodide on Erythrosine-Mediated Photodynamic Therapy on Streptococcus Mutans Biofilms (Streptococcus mutans 바이오필름에 대한 에리스로신 매개 광역동 치료 시 potassium iodide의 효과)

  • Yongsoon, Kim;Howon, Park;Juhyun, Lee;Haeni, Kim;Siyoung, Lee
    • Journal of the korean academy of Pediatric Dentistry
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    • v.49 no.3
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    • pp.321-328
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    • 2022
  • The aim of this in-vitro study is to evaluate the effect of potassium iodide (KI) on erythrosine-mediated photodynamic therapy (PDT) against Streptococcus mutans biofilms. S. mutans ATCC 25175 was cultured to form a biofilm on a hydroxyapatite disk. After diluting erythrosine to 20 μM and KI to 10, 50, and 100 mM, respectively, PDT was performed. The number of surviving bacteria was calculated as colony forming units (CFU)/mL and the statistical significance of the difference between groups was confirmed by Bonferroni post-hoc analysis. Cell viability was visually evaluated using confocal laser scanning microscopy (CLSM). As a result of the experiment, a significant decrease (p < 0.05) in CFU was observed in the experimental groups in which PDT was performed after applying KI regardless of the concentration of KI. In addition, a significant reduction (p < 0.05) in CFU was observed in the experimental group to which 100 mM KI was applied compared to 10 mM KI. The same results were confirmed when observing CLSM. KI significantly improved the efficacy of erythrosine-mediated PDT on S. mutans biofilms at all concentrations. This may compensate for the low sensitivity of PDT to biofilm-state bacteria strains, but it is necessary to establish an optimal clinical protocol through further research.

EDTA Surface Capped Water-Dispersible ZnSe and ZnS:Mn Nanocrystals

  • Lee, Jae-Woog;Lee, Sang-Min;Huh, Young-Duk;Hwang, Cheong-Soo
    • Bulletin of the Korean Chemical Society
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    • v.31 no.7
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    • pp.1997-2002
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    • 2010
  • ZnSe and ZnS:Mn nanocrystals were synthesized via the thermal decomposition of their corresponding organometallic precursors in a hot coordinating solvent (TOP/TOPO) mixture. The organic surface capping agents were substituted with EDTA molecules to impart hydrophilic surface properties to the resulting nanocrystals. The optical properties of the water-dispersible nanocrystals were analyzed by UV-visible and room temperature solution photoluminescence (PL) spectroscopy. The powders were characterized by X-ray diffraction (XRD), high resolution transmission electron microscopy (HR-TEM), and confocal laser scanning microscopy (CLSM). The solution PL spectra revealed emission peaks at 390 (ZnSe-EDTA) and 597 (ZnS:Mn-EDTA) nm with PL efficiencies of 4.0 (former) and 2.4% (latter), respectively. Two-photon spectra were obtained by fixing the excitation light source wavelengths at 616 nm (ZnSe-EDTA) and 560 nm (ZnS:Mn-EDTA). The emission peaks appeared at the same positions to that of the PL spectra but with lower peak intensity. In addition, the morphology and sizes of the nanocrystals were estimated from the corresponding HR-TEM images. The measured average particle sizes were 5.4 nm (ZnSe-EDTA) with a standard deviation of 1.2 nm, and 4.7 nm (ZnS:Mn-EDTA) with a standard deviation of 0.8 nm, respectively.

Complete Genome and Calcium Carbonate Precipitation of Alkaliphilic Bacillus sp. AK13 for Self-Healing Concrete

  • Jung, Yoonhee;Kim, Wonjae;Kim, Wook;Park, Woojun
    • Journal of Microbiology and Biotechnology
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    • v.30 no.3
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    • pp.404-416
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    • 2020
  • Bacteria that are resistant to high temperatures and alkaline environments are essential for the biological repair of damaged concrete. Alkaliphilic and halotolerant Bacillus sp. AK13 was isolated from the rhizosphere of Miscanthus sacchariflorus. Unlike other tested Bacillus species, the AK13 strain grows at pH 13 and withstands 11% (w/v) NaCl. Growth of the AK13 strain at elevated pH without urea promoted calcium carbonate (CaCO3) formation. Irregular vaterite-like CaCO3 minerals that were tightly attached to cells were observed using field-emission scanning electron microscopy. Energy-dispersive X-ray spectrometry, confocal laser scanning microscopy, and X-ray diffraction analyses confirmed the presence of CaCO3 around the cell. Isotope ration mass spectrometry analysis confirmed that the majority of CO32- ions in the CaCO3 were produced by cellular respiration rather than being derived from atmospheric carbon dioxide. The minerals produced from calcium acetate-added growth medium formed smaller crystals than those formed in calcium lactate-added medium. Strain AK13 appears to heal cracks on mortar specimens when applied as a pelletized spore powder. Alkaliphilic Bacillus sp. AK13 is a promising candidate for self-healing agents in concrete.

Demyelination of neurofilament protein 200 immune positive never fibers in human pulp (사람 치수 내에서 neurofilament protein 200 면역양성반응을 나타내는 신경섬유의 탈말이집 현상에 대한 연구)

  • Jang, Jung-Woo;Choi, So-Young;Kwon, Dae-Geon;Bae, Yong-Chul;Kim, Chin-Soo;Lee, Sang-Han
    • Journal of the Korean Association of Oral and Maxillofacial Surgeons
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    • v.36 no.5
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    • pp.360-365
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    • 2010
  • Introduction: Mammalian tooth pulp is densely innervated by sensory nerves that are mostly C fibers and A delta fibers. However, there is evidence suggesting that many unmyelinated axons in the pulp are in fact parent meylinated axons. Immunohistochemical staining for neurofilament protein 200 kDa (NFP200) was performed to identify the demyelinated but parent myelinated axons. Materials and Methods: The pulp was removed from healthy premolars and 3rd molars extracted from juveniles and adults undergoing orthodontic treatment, and immunohistochemical staining were applied with NPF200 antibodies, which specifically dye myelinated axons. The specimens underwent an electron microscopy examination with diaminobenzidine (DAB) immunostaining after observation and analysis by fluorescence and confocal laser scanning microscopy. Results: The NPF200 immuno-positive axons in the radicular pulp areas were observed as bundles of many nerve fibers. Many small bundles were formed with fewer axons when firing to the coronal pulp areas and then reachrd a different direction. In the radicular pulp, unmyelinated axons and myelinated axons were present together. However, in the coronal pulp, unmyelinated axons were most common and NFP200 immuno-positive unmyelinated axons with a larger diameter than those in the radicular pulp were observed more frequently. On the other hand, most of the immuno-positive unmyelinated fibers were similar in size to that of typically well-known unmyelinated fibers. Conclusion: Myelinated fibers innervated to the dental pulp maintain their myelins in the radicular portion, but these fibers lost myelins in the coronal portion. After the loss of myelin, the size of the axoplasm also decreased.

Initial adhesion of bone marrow stromal cells to various bone graft substitutes

  • Jo, Young-Jae;Kim, Kyoung-Hwa;Koo, Ki-Tae;Kim, Tae-Il;Seol, Yang-Jo;Lee, Yong-Moo;Ku, Young;Chung, Chong-Pyoung;Rhyu, In-Chul
    • Journal of Periodontal and Implant Science
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    • v.41 no.2
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    • pp.67-72
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    • 2011
  • Purpose: The aim of this study is to determine whether certain biomaterials have the potential to support cell attachment. After seeding bone marrow stromal cells onto the biomaterials, we investigated their responses to each material in vitro. Methods: Rat bone marrow derived stromal cells were used. The biomaterials were deproteinized bovine bone mineral (DBBM), DBBM coated with fibronectin (FN), synthetic hydroxyapatite (HA), HA coated with FN, HA coated with $\beta$-tricalcium phosphate (TCP), and pure $\beta$-TCP. With confocal laser scanning microscopy, actin filaments and vinculin were observed after 6, 12, and 24 hours of cell seeding. The morphological features of cells on each biomaterial were observed using scanning electron microscopy at day 1 and 7. Results: The cells on HA/FN and HA spread widely and showed better defined actin cytoskeletons than those on the other biomaterials. At the initial phase, FN seemed to have a favorable effect on cell adhesion. In DBBM, very few cells adhered to the surface. Conclusions: Within the limitations of this study, we can conclude that in contrast with DBBM not supporting cell attachment, HA provided a more favorable environment with respect to cell attachment.

Enhanced Internalization of Macromolecular Drugs into Mycobacterium smegmatis with the Assistance of Silver Nanoparticles

  • Sun, Fangfang;Oh, Sangjin;Kim, Jeonghyo;Kato, Tatsuya;Kim, Hwa-Jung;Lee, Jaebeom;Park, Enoch Y.
    • Journal of Microbiology and Biotechnology
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    • v.27 no.8
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    • pp.1483-1490
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    • 2017
  • In this study, silver nanoparticles (AgNPs) were synthesized by the citrate reduction process and, with the assistance of n-hydroxysuccinimide and 1-ethyl-3-(3-dimethylaminopropyl) carbodiimide, were successfully loaded with the macromolecular drug vancomycin (VAM) to form AgNP-VAM bioconjugates. The synthesized AgNPs, VAM, and AgNP-VAM conjugate were characterized by UV-visible spectroscopy, zeta potential analysis, confocal microscopy, and transmission electron microscopy. The effect of loading VAM onto AgNPs was investigated by testing the internalization of the bioconjugate into Mycobacterium smegmatis. After treatment with the AgNP-VAM conjugate, the bacterial cells showed a significant decrease in UV absorption, indicating that loading of the VAM on AgNPs had vastly improved the drug's internalization compared with that of AgNPs. All the experimental assessments showed that, compared with free AgNPs and VAM, enhanced internalization had been successfully achieved with the AgNP-VAM conjugate, thus leading to significantly better delivery of the macromolecular drug into the M. smegmatis cell. The current research provides a new potential drug delivery system for the treatment of mycobacterial infections.

Bactericidal and wound disinfection efficacy of nanostructured titania

  • Azad, Abdul-Majeed;Aboelzahab, Asem;Goel, Vijay
    • Advances in materials Research
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    • v.1 no.4
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    • pp.311-347
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    • 2012
  • Infections are caused due to the infiltration of tissue or organ space by infectious bacterial agents, among which Staphylococcus aureus bacteria are clinically most relevant. While current treatment modalities are in general quite effective, several bacterial strains exhibit high resistance to them, leading to complications and additional surgeries, thereby increasing the patient morbidity rates. Titanium dioxide is a celebrated photoactive material and has been utilized extensively in antibacterial functions, making it a leading infection mitigating agent. In view of the property amelioration in materials via nanofication, free-standing titania nanofibers (pure and nominally doped) and nanocoatings (on Ti and Ti6Al4V implants) were fabricated and evaluated to assess their efficacy to mitigate the viability and growth of S. aureus upon brief (30 s) activation by a portable hand-held infrared laser. In order to gauge the effect of exposure and its correlation with the antibacterial activities, both isolated (only titania substrate) and simultaneous (substrate submerged in the bacterial suspension) activations were performed. The bactericidal efficacy of the IR-activated $TiO_2$ nanocoatings was also tested against E. coli biofilms. Toxicity study was conducted to assess any potential harm to the tissue cells in the presence of photoactivated materials. These investigations showed that the photoactivated titania nanofibers caused greater than 97% bacterial necrosis of S. aureus. In the case of titania-coated Ti-implant surrogates, the bactericidal efficacy exceeded 90% in the case of pre-activation and was 100% in the case of simultaneous-activation. In addition to their high bactericidal efficacy against S. aureus, the benignity of titania nanofibers and nanocoatings towards tissue cells during in-vivo exposure was also demonstrated, making them safe for use in implant devices.

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.

Examination Of The Migratory Ability Of Primordial Germ Cells From Embryonic Gonads At Different Developmental Stages In Quail

  • Kim, Duk-Kyung;Park, Tae ub;Lee, Yong-Mok;Kim, Mi-Ah;Kim, Gwi-Sook;Kim, Ki-Dong;Han, Jae-Yong
    • Proceedings of the Korea Society of Poultry Science Conference
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    • 2000.11a
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    • pp.75-77
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    • 2000
  • Retaining migratory activity is a prerequisite for the manipulation and use of PGCs. This study was conducted to examine whether migratory activity is retained in the primordial germ cells(PGCs) from gonads at the later embryonic developmental stage. In the present study, gonads were dissected from 5-, 6- and 10-day-old quail embryos and treated with trypsin-EDTA for the degradation of gonadal tissue. Gonadal PGCs (gPGCs) were purified by Ficoll density gradient centrifugation and labeled with PKH26 fluorescent dye. The PKH26-labeled gPGCs were microinjected into the blood vessels of recipient quail embryo. After further incubation of 3 days, the manipulated recipients were embedded in paraffin and sectioned. The gPGCs were detected by their fluorescence under the fluorescent microscopy and the confocal laser microscopy. As a result, 10-day-old quail gPGCs as well as 5-and 6-day-old gPGCs, could migrate to recipient embryonic gonads and settle down. These results suggest that the 10-day-old gPGCs have the properties of circulating PGCs at early stage. Therefore the PGCs from 10-day old embryonic gonads can be used for the tools of genetic manipulation.

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