• 제목/요약/키워드: confocal microscopy

검색결과 437건 처리시간 0.035초

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

  • 오도건;김광엽
    • 한국식품과학회지
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    • 제52권6호
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    • pp.670-677
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    • 2020
  • 본 연구에서는 가장 빈번하게 발생하는 세균성 식중독의 원인균인 C. jejuni의 주 감염원인 계육에 대하여, 식품공전에 등재된 식품첨가물 중 유화제를 이용하여 계육에서 C. jejuni의 부착을 제어 할 수 있는 기술을 확보하고자 하였다. 8종의 유화제를 200 mg/mL의 농도에서 paper disc agar diffusion method로 C. jejuni에 대한 항균활성을 검색한 결과 L-7D, L-1695, polysorbate 20, polysorbate 80 등 4종의 유화제에서 생육억제환을 생성하였다. L-7D, L-1695, polysorbate 20, polysorbate 80 4종의 유화제를 25, 50, 100, 200 mg/mL의 농도에서 항균활성을 검색한 결과, 농도가 작아질수록 생육억제환의 크기도 줄어들었으며, 유화제 중 L-1695 샘플이 200 mg/mL에서 가장 큰 생육억제환을 생성하였다. pH 및 열에 대해 안정성을 측정한 결과 L-7D, L-1695, polysorbate 20, polysorbate 80 4종의 유화제 모두 pH 및 열에 안정성을 가지고 있다는 것을 확인하였다. 최소저해농도를 측정한 결과 다른 유화제 L-7D, polysorbate 20, polysorbate 80 샘플과 비교하였을때 L-1695 샘플이 1.56 mg/mL에서 가장 좋은 최소저해농도를 나타냈다. 최소살균 효과는 L-7D, L-1695, polysorbate 20, polysorbate 80 4종의 유화제 모두 나타나지 않았다. 접촉표면의 부착제어능력을 확인하기 위해서 stainless steel과 ceramic에서 실험한 결과, 두 접촉 표면 모두에서 L-1695 샘플 처리 시 가장 적은 생균수를 나타냈다. 앞선 실험의 종합적인 결과에 따라 L-1695 유화제를 최종적으로 선별하고, 계육 피부에 부착된 C. jejuni에 영향을 주는지 CLSM으로 분석한 결과 대조구에 비하여 conventional spray 및 electrostatic spray를 처리하였을 때 모공에 부착된 균이 대다 수 떨어졌음을 확인하였다. 그러나 생균수를 확인해 본 결과 conventional spray와 electrostatic spray 처리 시에는 유의한 차이가 나타나지 않았다. 위 결과들을 종합한 결과 C. jejuni KCTC 5327에 대하여 L-1695 유화제는 생육을 억제시키지만, 살균효과는 없는 것을 확인하였다. 그러나 L-1695 유화제는 식품접촉표면에서 캠필로박터균의 부착을 저해하는 특성을 갖고 있으며, 계육 피부에 인위접종된 C. jejuni를 효과적으로 감소시키는 것을 볼 때, 실제 도계공정에서 세척수에 포함시켜 C. jejuni의 제어에 사용될 수 있을 것으로 기대된다.

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

  • 김용순;박호원;이주현;김해니;이시영
    • 대한소아치과학회지
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    • 제49권3호
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    • pp.321-328
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    • 2022
  • 이 연구는 Streptococcus mutans 바이오필름에 대한 에리스로신 매개 광역동 치료에서 potassium iodide (KI)의 효과를 평가하는 In vitro 실험이다. S. mutans ATCC 25175를 배양하여 hydroxyapatite disk에 바이오필름을 형성하였다. 광감각제인 에리스로신을 20 μM, KI를 각각 10, 50, 100 mM로 희석하여 적용한 뒤 광역동 치료를 시행하였다. 생존한 세균의 수는 colony forming units (CFU)/mL로 산정하였으며 Bonferroni 사후 분석을 통해 그룹 간 차이의 유의성을 확인하였다. Confocal laser scanning microscopy (CLSM)을 이용하여 세포 생존율을 시각적으로 평가하였다. 실험 결과 KI를 적용한 후 광역동 치료를 시행한 실험군에서 KI의 농도의 관계 없이 현저한 CFU의 감소가 관찰되었다(p < 0.05). 또한 10 mM KI에 비해 100 mM KI를 적용한 실험군에서 유의한 CFU의 감소가 관찰되었다(p < 0.05). CLSM 관찰 시에도 동일한 결과를 확인하였다. KI는 모든 농도에서 S. mutans 바이오필름에 대한 에리스로신 매개 광역동 치료의 효과를 유의하게 향상시켰다. 이는 바이오필름 상태의 균주에 대한 광역동 치료의 낮은 감수성을 보완할 수 있지만, 추가적인 연구를 통한 최적의 임상 프로토콜 확립이 필요하다.

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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    • 제31권7호
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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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    • 제30권3호
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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.

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

  • 장정우;최소영;권대근;배용철;김진수;이상한
    • Journal of the Korean Association of Oral and Maxillofacial Surgeons
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    • 제36권5호
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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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    • 제41권2호
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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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    • 제27권8호
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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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    • 제1권4호
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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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    • 제35권8호
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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
    • 한국가금학회:학술대회논문집
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    • 한국가금학회 2000년도 제17차 정기총회 및 학술발표
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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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