• Restorative Dentistry and Endodontics
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• 제43권1호
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• pp.2.1-2.10
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• 2018

Objectives: To determine the effect of size and insertion depth of irrigation needle on the amount of apical extruded debris and the amount of penetration depth of sealer using a confocal laser scanning microscope (CLSM). Materials and Methods: Twenty maxillary premolars were assigned to 2 groups (n = 10), according to the size of needle tip, 28 G or 30 G. Buccal roots of samples were irrigated with respective needle type inserted 1 mm short of the working length (WL), while palatal roots were irrigated with respective needle type inserted 3 mm short of the WL. Prepared teeth were removed from the pre-weighed Eppendorf tubes. Canals were filled with F3 gutta-percha cone and rhodamine B dye-labeled AH 26 sealer. Teeth were transversally sectioned at 1 and 3 mm levels from the apex and observed under a CLSM. Eppendorf tubes were incubated to evaporate the irrigant and were weighed again. The difference between pre- and post-weights was calculated, and statistical evaluation was performed. Results: Inserting needles closer to the apex and using needles with wider diameters were associated with significantly more debris extrusion (p < 0.05). The position of needles and level of sections had statistically significant effects on sealer penetration depth (p < 0.05 for both). Conclusions: Following preparation, inserting narrower needles compatible with the final apical diameter of the prepared root canal at 3 mm short of WL during final irrigation might prevent debris extrusion and improve sealer penetration in the apical third.

• Journal of Microbiology and Biotechnology
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• 제23권12호
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• pp.1673-1682
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• 2013

Citrobacter sp. is a cause of significant opportunistic nosocomial infection and is frequently found in human and animal feces, soil, and sewage water, and even in industrial waste or putrefaction. Biofilm formation is an important virulence trait of Citrobacter sp. pathogens but the process and characteristics of this formation are unclear. Therefore, we employed in vitro assays to study the nutritional and environmental parameters that might influence biofilm formation of C. werkmanii BF-6 using 96-well microtiter plates. In addition, we detected the relative transcript levels of biofilm formation genes by RT-PCR. Our results indicated that the capacity of C. werkmanii BF-6 to form biofilms was affected by culture temperature, media, time, pH, and the osmotic agents glucose, sucrose, NaCl, and KCl. Confocal laser scanning microscopy results illustrated that the structure of biofilms and extracellular polysaccharide was influenced by 100 mM NaCl or 100 mM KCl. In addition, nine biofilm formation genes (bsmA, bssR, bssS, csgD, csgE, csgF, mrkA, mrkB, and mrkE) were found to contribute to planktonic and biofilm growth. Our data suggest that biofilm formation by C. werkmanii BF-6 is affected by nutritional and environmental factors, which could pave the way to the prevention and elimination of biofilm formation using proper strategies.

• Applied Microscopy
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• 제44권1호
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• pp.21-29
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• 2014

Retinal glial responses to hypertensive glaucomatous injury were spatiotemporally surveyed. Retinas as a whole or vertical sections were processed for anti-glial fibrillary acidic protein (GFAP), anti-Iba1, anti-nerve growth factor (NGF), and anti-tumor necrosis factor (TNF)-${\alpha}$ immunohistochemistry for confocal microscopic analyses. The optic nerve head of paired controls was processed for electron microscopy. GFAP positive astrocytes appeared in the nerve fiber layer in the glaucomatous and control retinas, changing from fine protoplasmic to stout fibrous parallel to glaucomatous duration. Iba1 positive microglia appeared in both retinas, and enormous reaction appeared at the latest glaucomatous. M$\ddot{u}$ller reaction detected by GFAP reactivity expanded from the end feet to whole profile following to duration in the glaucomatous. NGF reactivity expended from the end feet to the proximal radial processes of the M$\ddot{u}$ller cells in both retinas according to glaucomatous duration. TNF-${\alpha}$ immunoreactivity in the nerve fiber layer was stronger in both the glaucomatous and controls than in the normal, and exceptionally at the latest glaucomatous was even lower than the normal. The astrocytes in the optic nerve head are interconnected with each other via gap junction. These results demonstrate that astrocyte reaction propagates to the contralateral via physical links, and TNF-${\alpha}$ is correlated with NGF production for neuroprotection in response to hypertensive glaucomatous injury.

• KSBB Journal
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• 제22권4호
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• pp.239-243
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• 2007

본 연구에서는 미네랄오일에 유화제를 사용하여 한천 현탁액을 제조하고 여기에 피브로인을 첨가하여 마이크로캡슐을 제조하는 방법을 개발하였다. 먼저 한천 유화액에 첨가될 유화제의 농도를 10%로 결정하였고 피브로인을 투입하는 속도를 조절하여 피브로인으로 피막이 형성된 마이크로캡슐을 감압건조를 통하여 수분을 제거하고 에탄올 침전물로 회수하였다. 공초점 현미경으로 피브로인 피막이 형성되어 있음과 마이크로캡슐의 90%가 $1.32{\mu}m$에서 $6.0{\mu}m$사이에 존재함을 확인하였다. 에탄올 침전물 형태의 마이크로캡슐과 이들이 분산되는 것을 전자현미경으로 확인하였으며 열중량 분석을 통하여 마이크로캡슐은 중량비로 한천이 51.2%, 피브로인이 13.8%, Span 80이 1.4%로 구성되어있음을 확인하였다.

• 대한화장품학회지
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• 제40권3호
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• pp.247-257
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• 2014

새로운 베지클인 glyceryl citrate/ lactate/ linoleate/ oleate를 이용한 수중유형 형태의 아스타잔틴 나노에멀젼에 대해 항산화 효과, 세포 생존력, 단백질과 관련한 효소의 영향, 피부 침투도 그리고 피부에 대한 보습 및 탄력 등의 약용화장품적인 측면에서의 전반적 연구를 실시하였다. 항산화력 및 세포 생존력에 대해선 각각 DPPH법과 MMT assay를 이용하여 측정하였다. 아스타잔틴 나노에멀젼에 대한 또 다른 성질은 2D-Page를 이용한 단백질 분석 및 컨포칼, in-vivo 테스트를 통해 측정하였다. 본 연구를 통해, 아스타잔틴을 포함하는 나노에멀젼은 MMP발현에 관련한 단백질 억제 및 세포외 기질의 분해를 막고 라디칼의 소거에 매우 우수한 결과를 보였다. 종전의 레시친을 이용한 나노에멀젼 보다는 새로운 베지클을 이용한 아스타잔틴 나노에멀젼의 피부 침투가 매우 효과적임을 CLSM을 통해 측정하였다. 또한 28일 동안의 한국 성인 여성 11명을 통한 보습 및 탄력 인비보 테스트에서 우수한 효과를 확인할 수 있었다.

• Molecules and Cells
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• 제26권1호
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• pp.100-105
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• 2008

Glycogen synthase kinase $3{\beta}$ (GSK $3{\beta}$) is a serine/threonine kinase that phosphorylates substrates such as ${\beta}$-catenin and is involved in a variety of biological processes, including embryonic development, metabolism, tumorigenesis, and cell death. Here, we present evidence that human GSK $3{\beta}$ is associated with Fe65, which has the characteristics of an adaptor protein, possessing a WW domain, and two phosphotyrosine interaction domains, PID1 and PID2. The GSK $3{\beta}$ catalytic domain also contains a putative WW domain binding motif ($^{371}PPLA^{374}$), and we observed, using a pull down approach and co-immunoprecipitation, that it interacts physically with Fe65 via this motif. In addition, we detected co-localization of GSK $3{\beta}$ and Fe65 by confocal microscopy, and this co-localization was disrupted by mutation of the putative WW domain binding motif of GSK $3{\beta}$. Finally, in transient transfection assays interaction of GSK $3{\beta}$ (wt) with Fe65 induced substantial cell apoptosis, whereas interaction with the GSK $3{\beta}$ AALA mutant ($^{371}AALA^{374}$) did not, and we noted that phosphorylation of the Tyr 216 residue of the GSK $3{\beta}$ AALA mutant was significantly reduced compared to that of GSK $3{\beta}$ wild type. Thus, our observations indicate that GSK $3{\beta}$ binds to Fe65 through its $^{371}PPLA^{374}$ motif and that this interaction regulates apoptosis and phosphorylation of Tyr 216 of GSK $3{\beta}$.

• Journal of Microbiology and Biotechnology
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• 제10권6호
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• pp.844-850
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• 2000

The specific binding and internalization of viral particles is an essential step for the successful infection of viral pathogens. In the case of the hepatitis B virus (HBV), virions bind to the host cell via the preS domain of the viral surface antigen and are subsequently internalized by endocytosis. HBV-preS specific receptors are primarily expressed on hepatocytes, however, viral DNA and proteins have also been detected in extrahepatic sites, suggsting that celluar recepators for HBV may also exist on extrahepatic cells. Recently, an EBV-transformed B-cell line was identified onto which the preS region binds in a receptor-ligand specific manner. In this study, this specific interaction was further characterized, and the binding region within the preS protein was locaized. Also the internalization after host cell attachment was visualized and analyzed by fluorescence-labeled HBV-preS1 proteins using confocal microscopy. Energy depletion by sodium azide treatment effectively inhibited the internalization of the membrane-bound preS1 ligands, thereby indicating an energy-dependent receptor-mediated endocytotic pathway. Accordingly, the interaction of HBV-pres! with this specific B-cell line may serve as an effective model for an infection pathway in extrahepatic cells.

• 생명과학회지
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• 제22권10호
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• pp.1415-1419
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• 2012

Thymosin ${\beta}4$ 는 대장암에서 암 줄기세포 마커인 CD133을 지닌 세포에서 지니지 않은 세포에 비해 강하게 발현된다고 보고되어 있다. 본 연구에서는 thymosin ${\beta}4$와 줄기세포 마커인 CD133의 상관관계를 정상 위 조직에서 관찰하였다. Thymosin ${\beta}4$와 CD133의 발현 양상은 tissue microarray 조직상에서 면역화학적 방법으로 관찰하였으며 thymosin ${\beta}4$와 CD133의 존재 위치는 면역형광 염색법 및 confocal microscope를 사용하여 조사하였다. Thymosin ${\beta}4$와 CD133은 동일한 양상으로 발현되었으며 모두 위의 선조직에서 강하게 발현되었다. 면역 형광 염색법으로 두가지 단백질을 동시에 염색한 결과 두 단백질이 동일한 위치에서 함께 존재하는 것으로 규명되었다. 이러한 결과는 thymosin ${\beta}4$와 CD133은 정상위의 선조직에서 발현되며 두 단백질의 발현 양상 및 위치가 동일하여 서로 긴밀한 상호작용을 할 가능성을 제시하고 있다.

• 한국정보과학회논문지:소프트웨어및응용
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• 제31권7호
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• pp.922-928
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• 2004

본 논문에서는 볼륨데이타에서의 레이블링(labeling)을 위한 알고리즘을 제안하고자한다. 3차원 볼륨은 2차원 슬라이스 데이타의 연속으로 보고 각 슬라이스의 레이블링 정보를 바탕으로 하는 SIL(Slice Information based Labeling)방법을 제안한다. 이는 기존의 알고리즘에 비해 효율적인 메모리 사용이 가능하고 분석하고자 하는 데이타의 특성에 맞는 2차원 레이블링과의 조합이 가능한 장점이 있다. 기존 알고리즘과 제안하는 방법을 3차원 세포영상에서 비교하여 보았으며, SIL을 2차원 레이블링 CCCL(Contour based Connected Component Labeling)과 함께 볼륨데이타에 적용하여 본 결과 기존의 알고리즘 보다 약 2배 빠른 성능을 보였다. 다양한 3차원 레이블링 방법 중 적용되는 영상에 따라 각기 다른 결과를 얻었지만, 3차원 세포영상의 분석에서는 SIL 방법이 우수하다는 결론을 얻었다.

• Journal of Microbiology and Biotechnology
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• 제17권2호
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• pp.202-206
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• 2007

The adhesion of Campylobacter jejuni to chicken skin, along with the associated morphological changes under aerobic conditions at 4, 25, and $37^{\circ}C$ and microaerobic ($O_2\;5%,\;CO_2\;10%,\;N_2\;85%$) conditions, were investigated using confocal laser scanning microscopy (CLSM), flow cytometry, and plate counting. The morphological change of C. jejuni from a spiral shape to a coccoid form or VBNC form (viable but nonculturable form) progressed rapidly under aerobic conditions at 25, 37, and $4^{\circ}C$. As regards adhesion, the C. jejuni cells were mostly located in the crevices and feather follicles of the chicken skin, where the cells in the feather follicles floated freely in the entrapped water, even after the skin was rinsed quite thoroughly. CLSM also revealed the penetration of some spiral-shaped C. jejuni cells into the chicken skin. Even after changing their shape at various temperatures, coccoid-form C. jejuni cells were still found in the crevices and feather follicles of the chicken skin.