• Biomolecules & Therapeutics
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• 제26권2호
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• pp.182-190
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• 2018

The objective of this work is to evaluate the effect of polyamidoamine (PAMAM) dendrimers on electroosmotic flow (EOF) through skin. The effect of size and concentration of dendrimer was studied, using generation 1, 4 and 7 dendrimer (G1, G4 and G7, respectively). As a marker molecule for the direction and magnitude of EOF, a neutral molecule, acetoaminophen (AAP) was used. The visualization of dendrimer permeation into the current conducting pore (CCP) of skin was made using G4-fluorescein isothiocyanate (FITC) conjugate and confocal microscopy. Without dendrimer, anodal flux of AAP was much higher than cathodal or passive flux. When G1 dendrimer was added, anodal flux decreased, presumably due to the decrease in EOF by the association of G1 dendrimer with net negative charge in CCP. As the generation increased, larger decrease in anodal flux was observed, and the direction of EOF was reversed. Small amount of methanol used for the preparation of dendrimer solution also contributed to the decrease in anodal flux of AAP. Cross-sectional view perpendicular to the skin surface by confocal laser scanning microscope (CLSM) study showed that G4 dendrimer-FITC conjugate (G4-FITC) can penetrate into the viable epidermis and dermis under anodal current. The permeation route seemed to be localized on hair follicle region. These results suggest that PAMAM dendrimers can permeate into CCP and change the magnitude and direction of EOF. Overall, we obtained a better understanding on the mechanistic insights into the electroosmosis phenomena and its role on flux during iontophoresis.

• 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.

• 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.

• 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.

• The Journal of Advanced Prosthodontics
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• 제14권6호
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• pp.335-345
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• 2022

PURPOSE. This in vitro study aimed to evaluate the surface characteristics of a full veneer crown fabricated chairside (CS) from a (Y, Nb)-TZP zirconia block in response to conventional zirconia grinding and polishing. MATERIALS AND METHODS. Zirconia crowns (n = 40) were first prepared and divided into two groups of materials: Labside (LS) and CS, after which each specimen went through a five-step grinding and polishing procedure. Following each surface treatment, surface characteristics were analyzed using confocal laser microscopy (CLSM), average surface roughness (Ra) values were processed from the profile data through Gaussian filtering, and X-ray diffraction pattern analysis was performed to evaluate the monoclinic (M) phase content. Then, a representative specimen was selected for field-emission scanning electron microscopy (FE-SEM), followed by a final analysis of the roughness and X-ray diffraction of the specimens using the independent t-test and repeated measures analysis of variance (RM-ANOVA). RESULTS. In every group, polishing significantly reduced the Ra values (P < .001). There was no significant difference in Ra between the polished state CS and LS. Furthermore, CLSM and FE-SEM investigations revealed that even though grain exposure was visible in CS specimens throughout the as-delivered and ground states, the exposure was reduced after polishing. Moreover, while no phase transformation was visible in the LS, phase transformation was visible in CS after every surface treatment, with the M phase content of the CS group showing a significant reduction after polishing (P < .001). CONCLUSION. Within the limits of this study, clinically acceptable level of surface finishing of (Y, Nb)-TZP can be achieved after conventional zirconia polishing sequence.

• Journal of Microbiology and Biotechnology
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• 제33권1호
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• pp.61-74
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• 2023

The global increase in multidrug-resistant (MDR) bacteria has inspired researchers to develop new strategies to overcome this problem. In this study, 23 morphologically different, soil-isolated actinomycete cultures were screened for their antibacterial ability against MDR isolates of ESKAPE pathogens. Among them, isolate BOGE18 exhibited a broad antibacterial spectrum, so it was selected and identified based on cultural, morphological, physiological, and biochemical characteristics. Chemotaxonomic analysis was also performed together with nucleotide sequencing of the 16S rRNA gene, which showed this strain to have identity with Streptomyces lienomycini. The ethyl acetate extract of the cell-free filtrate (CFF) of strain BOGE18 was evaluated for its antibacterial spectrum, and the minimum inhibitory concentration (MIC) ranged from 62.5 to 250 ㎍/ml. The recorded results from the in vitro anti-biofilm microtiter assay and confocal laser scanning microscopy (CLSM) of sub-MIC concentrations revealed a significant reduction in biofilm formation in a concentration-dependent manner. The extract also displayed significant scavenging activity, reaching 91.61 ± 4.1% and 85.06 ± 3.14% of 2,2-diphenyl-1-picrylhydrazyl (DPPH) and 2,2'-azinobis(3-ethylbenzothiazoline-6-sulfonic acid) (ABTS), respectively. A promising cytotoxic ability against breast (MCF-7) and hepatocellular (HePG2) cancer cell lines was obtained from the extract with IC₅₀ values of 47.15 ± 13.10 and 122.69 ± 9.12 ㎍/ml, respectively. Moreover, based on gas chromatography-mass spectrometry (GC-MS) analysis, nine known compounds were detected in the BOGE18 extract, suggesting their contribution to the multitude of biological activities recorded in this study. Overall, Streptomyces lienomycini BOGE18-derived extract is a good candidate for use in a natural combating strategy to prevent bacterial infection, especially by MDR pathogens.

• 한국식품영양과학회지
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• 제46권9호
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• pp.1071-1080
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• 2017

본 연구에서는 공초점 주사현미경(CLSM)을 활용하여 장내 유익균과 국산 통곡물(whole grain) 소재들과의 부착을 통해 생육환경에 있어 도움이 될 수 있는 서식공간을 제공받는 것을 시각화하고, 소재들의 프리바이오틱스 효과를 in vitro 배양을 통해 확인하여 통곡물 소재들로 제조된 프리바이오틱스 팽화식품을 개발하는 것을 목표하였고, 그 결과 장 속에서 장내 유익균 균총을 향상시켜 인체의 건강증진에 유익한 팽화과자 제조에 필요한 기초 자료를 확보하였다. 선정곡물들의 일반성분 함량은 수분이 8.99~12.19%, 조회분 1.34~2.39%, 조단백질 7.34~15.38%, 조지방 1.84~8.12%로 함량이 분석되었고, 불용성 식이섬유 3.08~18.62%, 수용성 식이섬유 0.10~3.13%, 그리고 총 식이섬유 함량 5.26~21.36%로 확인되었다. 곡물분말의 프리바이오틱스 부착여부를 CLSM으로 관찰한 결과 곡물분말과 혼합된 Lactobacillus acidophilus는 곳곳에 산재되어 있었지만, 주변 유동액 층보다 상대적으로 곡물분말 주변에 부착되어 존재하였다. 선정곡물들의 장내 유익균의 증진에 활성 효과가 있음을 검증한 결과 L. acidophilus에서는 찰보리 추출물에서 가장 높은 흡광도 값을 보였다. Lactobacillus delbrueckii subsp. lactis에서는 귀리 추출물에서 가장 높은 효과를 보였다. Streptococcus salivarius subsp. thermophilus에서는 control보다 높은 수치를 보였지만, 두드러진 차이는 보이지 않았다. Bifidobacterium bifidum은 12시간 이후 흡광도 값은 일정하게 유지되었다. 곡물 소재 동결건조 추출물 간의 장내 유익균 활성 정도를 비교한 실험 결과 L. acidophilus에서는 찰보리와 율무에서 가장 높은 생육증진 효과를 보였고 현미를 제외한 다른 곡물 모두 유의한 증진 효과를 보였다. L. delbrueckii subsp. lactis에서는 율무에서 가장 높은 증진 효과를 보였고, S. salivarius subsp. thermophilus에서는 찰보리와 찰수수에서 가장 높은 증진 효과를 보였다. 그리고 B. bifidum에서는 찰보리와 찰기장에서 높은 증진 효과를 보였다. 팽화식품 제조 시 배합에 사용될 6종의 혼합곡물의 장내 유익균 활성 효과를 알아본 결과, 24시간 배양 시 L. acidophilus는 control에 비해 1.72배이상 상승하였고, L. delbrueckii subsp. lactis는 1.89배, S. salivarius subsp. thermophilus는 1.66배 이상, B. bifidum는 1.47배가량 상승한 것으로 분석되었다. 제조된 팽화과자의 색도 측정결과 L값은 치즈맛의 시즈닝 첨가군이 대조군보다 높게 나왔다. a값은 대조군이 적색에 가깝게, 시즈닝 첨가군은 녹색 쪽으로 나타났다. b값은 두 제품에서 크게 유의한 차이를 보이진 않았다. 팽화과자의 경도는 첨가군이 대조군보다 높았고, 바삭거림은 첨가군이 대조군보다 약한 것으로 확인되었다.

• 한국생물공학회:학술대회논문집
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• 한국생물공학회 2001년도 추계학술발표대회
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• pp.282-285
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• 2001

질산화 생물여과 시스템 내 생물막 안에 존재하는 ammonia oxidizers 및 nitrite oxidizers의 군집 구조 및 공간적 분포를 조사하였다. FISH 분석 결과 생물막 내 숫적으로 우점종을 이루는 미생물은 ammonia oxidizer인 Nitrosomonas spp.로 나타났으며 nitrite oxidizer 인 Nilrospira spp.에 비해 3 ${\sim}$ 5 정도 더 많이 존재하였다. 이는 실협 기간동안 완전한 질산화를 보였지만 반응기가 2 년 이상 nitrite 축적을 위해 높은 free ammonia 농도 빛 낮은 용존 산소 상태에서 운선되어 nitrite oxidizers에 저해를 주었기 때문인 것으로 사료된다. FISH와 결합된 CLSM 관찰 결과 생물막 전체에 걸쳐 ammonia oxidizer가 분포하는 반면 안쪽으로 갈수록 nitrite oxidizers가 분포함을 보였다. 이는 폐수의 ammonium 을 생물막 내 ammon ia oxidizer가 먼저 nitrite로 산화시키고 이를 nitrite oxidizers가 곧바로 nitrate로 산화시키기 때문이다.

• Journal of Microbiology and Biotechnology
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• 제20권8호
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• pp.1185-1188
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• 2010

Previously, papiliocin was isolated from the swallowtail butterfly Papilio xuthus and its antimicrobial activity was suggested. In this study, the antifungal mechanism of papiliocin against Candida albicans was investigated. Confocal laser scanning microscopy (CLSM) and 1,6-diphenyl-1,3,5-hexatriene (DPH) fluorescence analysis indicated that papiliocin disturbed the fungal plasma membrane. Moreover, the assessment of the release of FITC-dextran (FD) from liposomes further demonstrated that the antifungal mechanism of papiliocin could have originated from the pore-forming action and that the radius of the pores was presumed to be anywhere from 2.3 to 3.3 nm.

• Advances in environmental research
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• 제7권1호
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• pp.39-52
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• 2018

Direct membrane filtration (DMF) of wastewater has many advantages over conventional biological wastewater treatment processes. DMF is not only compact, but potentially energy efficient due to the lack of biological aeration. It also produces more biosolids that can be used to produce methane gas through anaerobic digestion. Most of ammoniacal nitrogen in wastewater is preserved in effluent and is used as fertilizer when effluent is recycled for irrigation. In this study, a technical feasibility of DMF was explored. Organic and nitrogen removal efficiencies were compared between DMF and membrane bioreactor (MBR). Despite the extremely high F/V ratio, e.g., $14.4kg\;COD/m^3/d$, DMF provided very high COD removal efficiencies at ~93%. Soluble microbial products (SMP) and extracellular polymeric substances (EPS) were less in DMF sludge, but membrane fouling rate was far greater than in MBR. The diversity of microbial community in DMF appeared very narrow based on the morphological observation using optical microscope. On the contrary, highly diverse microbial community was observed in the MBR. Microorganisms tended to form jelly globs and attach on reactor wall in DMF. FT-IR study revealed that the biological globs were structurally supported by feather-like materials made of secondary amines. Confocal laser scanning microscopy (CLSM) study showed microorganisms mainly resided on the external surface of microbial globs rather than the internal spaces.