• 대한방사성의약품학회지
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• 제6권2호
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• pp.92-101
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• 2020

The cyclic Arg-Gly-Asp (cRGD) peptide is well-known as a binding molecule to the integrin α_vβ₃ receptor which is highly expressed on activated endothelial cells and new blood vessels in tumors. Although numerous results have been reported by the usage of cRGD peptide-based ligands for cancer diagnosis and therapy, the distinct mechanisms, and functions of cRGD-integrin binding to cancer cells are still being investigated. In this study, we evaluated the internalization efficacy of different types of cRGD peptides (monomer, dimer and tetramer form) in integrin α_vβ₃ overexpressing cancer cells. Western blot and flow cytometric analysis showed U87MG expresses highly integrin α_vβ₃, whereas CT-26 does not show integrin α_vβ₃ expression. Cytotoxicity assay indicated that all cRGD peptides (0-200 µM) had at least 70-80% of viability in U87MG cells. Fluorescence images showed cRGD dimer peptides have the highest cellular internalization compare to cRGD monomer and cRGD tetramer peptides. Additionally, transmission electron microscope results clearly visualized the endocytic internalization of integrin α_vβ₃ receptors and correlated with confocal microscopic results. These results support the rationale for the use of cRGD dimer peptides for imaging, diagnosis, or therapy of integrin α_vβ₃-rich glioblastoma.

• 치위생과학회지
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• 제23권3호
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• pp.208-215
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• 2023

Background: Dental caries in preschool children can cause early loss of teeth, reduced masticatory function, malocclusion, and speech disorders, making oral care for preschool children essential. However, because children have a poor ability to perform oral hygiene by themselves, educational tools that help establish correct oral health behaviors are required. This study evaluated the effect of toothbrushing application for kids on dental plaque removal and toothbrushing interest in preschool children. Methods: Seven 5-year-old children enrolled in a daycare center participated in the evaluation, and the same children participated in the experiment twice a week. The Pokémon Smile application (App) was used as an auxiliary application for tooth brushing, and the degree of dental plaque removal on the tooth surface was evaluated by quantitative light-induced fluorescence-digital imaging. After the experiment, children's toothbrushing preferences were investigated through interviews. Results: The levels of AREA R30 and AREA R70 measured after the children's toothbrushing as usual decreased compared to those before toothbrushing; however, the change was not significant. The levels of AREA R30 and AREA R70 measured after using the Pokémon Smile App were significantly reduced compared to those before toothbrushing. Children's interest in brushing their teeth increased by 28.59% after using the Pokémon Smile App. Conclusion: The toothbrushing application for kids effectively removes dental plaque by helping preschool children brush their teeth. It also increased preschool children's interest in tooth brushing. Therefore, an oral health education application would be useful for children who need to develop correct oral care methods and habits.

• BMB Reports
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• 제56권12호
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• pp.657-662
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• 2023

Neurodegenerative diseases are characterized by distinct protein aggregates, such as those of α-synuclein and tau. Lysosomal defect is a key contributor to the accumulation and propagation of aberrant protein aggregates in these diseases. The discoveries of common proteinopathies in multiple forms of lysosomal storage diseases (LSDs) and the identification of some LSD genes as susceptible genes for those proteinopathies suggest causative links between LSDs and the proteinopathies. The present study hypothesized that defects in lysosomal genes will differentially affect the propagation of α-synuclein and tau proteins, thereby determining the progression of a specific proteinopathy. We established an imaging-based high-contents screening (HCS) system in Caenorhabditis elegans (C. elegans) model, by which the propagation of α-synuclein or tau is measured by fluorescence intensity. Using this system, we performed RNA interference (RNAi) screening to induce a wide range of lysosomal malfunction through knock down of 79 LSD genes, and to obtain the candidate genes with significant change in protein propagation. While some LSD genes commonly affected both α-synuclein and tau propagation, our study identified the distinct sets of LSD genes that differentially regulate the propagation of either α-synuclein or tau. The specificity and efficacy of these LSD genes were retained in the disease-related phenotypes, such as pharyngeal pumping behavior and life span. This study suggests that distinct lysosomal genes differentially regulate the propagation of α-synuclein and tau, and offer a steppingstone to understanding disease specificity.

• 대한방사성의약품학회지
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• 제9권1호
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• pp.9-16
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• 2023

Liposomes as drug delivery system have proved useful carrier for various disease, including cancer. In addition, perfluorocarbon cored microbubbles are utilized in conjunction with high-intensity focused-ultrasound (HIFU) to enable simultaneous diagnosis and treatment. However, microbubbles generally exhibit lower drug loading efficiency, so the need for the development of a novel liposome-based drug delivery material that can efficiently load and deliver drugs to targeted areas via HIFU. This study aims to develop a liposome-based drug delivery material by introducing a substance that can burst liposomes using ultrasound energy and confirm the ability to target tumors using PET imaging. Liposomes (Lipo-DOX, Lipo-DOX-Au, Lipo-DOX-Au-RGD) were synthesized with gold nanoparticles using an avidin-biotin bond, and doxorubicin was mounted inside by pH gradient method. The size distribution was measured by DLS, and encapsulation efficiency of doxorubicin was analyzed by UV-vis spectrometer. The target specificity and cytotoxicity of liposomes were assessed in vitro by glioblastoma U87mg cells to HIFU treatment and analyzed using CCK-8 assay, and fluorescence microscopy at 6-hour intervals for up to 24 hours. For the in vivo study, U87mg model mouse were injected intravenously with 1.48 MBq of ⁶⁴Cu-labeled Lipo-DOX-Au and Lipo-DOX-Au-RGD, and PET images were taken at 0, 2, 4, 8, and 24 hours. As a result, the size of liposomes was 108.3 ± 5.0 nm at Lipo-DOX-Au and 94.1 ± 12.2 nm at Lipo-DOX-Au-RGD, and it was observed that doxorubicin was mounted inside the liposome up to 52%. After 6 hours of HIFU treatment, the viability of U87mg cells treated with Lipo-DOX-Au decreased by around 20% compared to Lipo-DOX, and Lipo-DOX-Au-RGD had a higher uptake rate than Lipo-DOX. In vivo study using PET images, it was confirmed that ⁶⁴Cu-Lipo-DOX-Au-RGD was taken up into the tumor immediately after injection and maintained for up to 4 hours. In this study, drugs released from liposomes-gold nanoparticles via ultrasound and RGD targeting were confirmed by non-invasive imaging. In cell-level experiments, HIFU treatment of gold nanoparticle-coupled liposomes significantly decreased tumor survival, while RGD-liposomes exhibited high tumor targeting and rapid release in vivo imaging. It is expected that the combination of these models with ultrasound is served as an effective drug delivery material with therapeutic outcomes.

• 대한핵의학회지
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• 제39권1호
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• pp.26-33
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• 2005

목적 : 이 연구는 $^{99m}Tc$-HMPAO에 표지된 미성숙 또는 성숙 수지상 세포의 마우스 생체 내 분포와 이동 양상에 대해 알아보고자 하였다. 대상 및 방법: 마우스의 대퇴골과 경골의 골수로부터 수지상 세포를 배양하고 미성숙, 성숙 수지상세포를 $^{99m}Tc$-HMPAO로 표지하였다. 방사성 표지 전후에 수지상 세포의 기능 및 표현형의 변화 유무를 알기 위해 동종 혼합 림프구 반응 (allogeneic mixed lymphocyte reaction)과 형광 활성 세포 선별 (fluorescence-activated cell sorting)을 시행하였다. 정맥 주사된 수지상 세포의 생체 내 이동은 감마 카메라 영상과 생체 분포 실험을 통하여 평가하였고, 피하 종양 마우스 모델과 대조군에서 비교하였다. 폐, 간, 비장, 신장, 종양 등 조직에서 그램 당 주사량의 백분율(%ID/g)을 계산하였다. 결과: 미성숙, 성숙 수지상 세포의 표지 효율은 각각 $60.4{\pm}5.4%$와 $61.8{\pm}6.7%$ 였다. 수지상 세포의 정맥주사 후 방사능은 폐에서 가장 먼저 관찰되었고, 이후 간과 비장에 분포되었다. 성숙 수지상 세포가 미성숙 수지상 세포에 비해 비장으로 더 많이 이동하였다(대조군; $38.3{\pm}4.0%\;vs.\;32.2{\pm}4.1%$, 종양이식 군: $40.4{\pm}4.1%\;vs.\;35.9{\pm}3.8%$, p<0.05). 종양으로의 이동 역시 성숙 수지상 세포가 미성숙 수지상 세포에 비해 더 많은 비율을 보였다($2.4{\pm}0.3%\;vs\;1.7{\pm}0.2%$; p=0.034). 결론: $^{99m}Tc$-HMPAO 에 표지된 수지상 세포를 이용하여 마우스 생체 내 이동을 실시간 영상화 할 수 있었다. 마우스 정맥에 주사되었을 때, 더 많은 비율의 성숙 수지상 세포가 미성숙 수지상 세포에 비해서 비장이나 종양으로 이동함을 알 수 있었다.

• 미생물학회지
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• 제51권3호
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• pp.241-248
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• 2015

프로바이오틱스에 대한 연구는 주로 생균의 효과가 많이 알려져 있지만 가열 처리된 유산균인 사균체의 기능에 대한 연구도 활발히 이루어지고 있다. 본 연구에서는 락토바실러스 아시도필러스 CBT LA1의 사균체의 장관장벽 기능에 대하여 in vitro, in vivo에서 실험하였다. 이를 위하여, 세포표면 소수성 상호작용력(cell surface hydrophobicity), 자가응집력(autoaggregation), 세포에 부착하는 능력(cell adhesion)과 자가응집력(autoaggregation), LPS와의 결합력을 조사하였다. 또한 HT-29 장상피세포에서 LPS로 유도되는 IL-8의 발현을 억제하는 효과를 조사하였다. CBT LA1을 80도에서 121도까지 10분 동안 열을 처리하였을 때, 80도에서 열을 처리한 CBT LA1 사균체가 가장 높은 세포에 부착하는 능력을 보여 주었다. CBT LA1 생균과 비교했을 때, 80도에서 열을 처리한 CBT LA1 사균체는 높은 LPS와의 결합력, 소수성 상호작용력, 자가응집력, HT-29 세포에 부착하는 능력과 IL-8의 발현을 억제하는 능력을 보여주었다. In vivo 실험에서 FITC로 label된 LPS를 투여하였을 때 16시간 후, CBT LA1 사균체를 섭취한 동물의 장관 내 LPS가 가장 많이 제거되었다. 이러한 연구 결과들은 CBT LA1 생균처럼 CBT LA1 사균체도 장관장벽 기능을 가지며 이는, 파마바이오틱스로서 그 가능성을 시사한다.

• 생명과학회지
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• 제30권6호
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• pp.563-569
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• 2020

유방암의 수술적 치료에 있어서 종양성형학적 수술이란 종양학적 안정성과 성형적 만족도를 모두 충족시키는 수술을 일컫는 용어이다. 처음 시작한 서양에서는 적은 절개나 절제, 미세침습수술을 지향하기 보다 유방축소술을 기본으로 제거될 조직에 병변을 포함하도록 계획한다. 충분한 절제가 가능하기 때문에 수술 전 검사, 절제연에 대한 고민, 수술 중 병리 검사의 필요성 등도 줄면서 종양학적 안전성에도 도움이 된다고 알려져 왔다. 개개인의 체형에 대한 병변의 종양학적 안정성을 충족시키는 최소 절제 비율에 따라 전위 또는 보충으로 나뉘게 된다. 환자의 기본 체형은 인종적인 영향을 받는 유방암의 특징으로, 유럽, 미국이나 영국의 경우 평균 크기가 대개 36D(600 ㎤) 이상으로, 유방고정술이나 유방축소술이 주로 사용되고, 성형 측면의 최대 목표인 대칭을 위해서는 반대측 유방의 축소도 흔히 이루어지고 있다. 부산대학교병원 유방암 환자 671명의 평균 분포를 분석해 본 결과, 평균은 33A (75%, <35B)로, 대략 체적은 300 ㎤ (75%, <400 ㎤)라고 생각할 수 있다. 따라서 서양과는 다른 수술법이 선택될 수 밖에 없다. 수술 전 MRI를 이용한 범위의 확인을 통한 대상 선정과 함께 초음파와 피부 표식을 이용한 정확한 절제 범위의 계획도 중요하다. 병리적 판독 기준도 절제연 음성이면 종양학적 안정성에 충분하다는 것으로 관점이 바뀐 것은 보존 수술에 유리한 측면이 된다. 동결절편검사는 여전히 고급 전문 인력, 시간이 지나치게 소요되고 있어 편리하고 안전한 병변의 절제에 도움이 될 만한 새로운 방법들이 필요하여 연구되고 있다. 대표적인 최근 연구로는 무선주파수를 이용하는 margin probe, 형광 색소를 결합한 Aqueous Quantum-Dot-Molecular Probes 등을 들 수 있다. 합병증 측면에서 박리 면적이 늘어나는 종양성형학적 수술의 단점을 보강할 수 있는 최근 기술로 초음파 에너지 기구들이 유용하고, 수술 범위가 늘어남에 따라 생길 수 있는 혈종이나 장액 저류의 감소에 도움이 될 수 있다.

• Journal of Korean Biological Nursing Science
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• 제1권1호
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• pp.25-41
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• 1999

Physiological activity of osteoblast including bone formation is known to be closely related to the increase of intracellular $Ca^{2+}$ activity($[Ca^{2+}]_i$) in osteoblast. $Ca^{2+}$ is an important intracellular messenger in diverse cellular functions, and regulation of its level is mediated by the transmembrane $Ca^{2+}$ movement via $Ca^{2+}$ channels, $Na^+-Ca^{2+}$ exchange, and by intracellular $Ca^{2+}$ movement through the intracellular stores. The purpose of this study is to investigate how the intracellular $Ca^{2+}$ is regulated in osteoblast-like cells(OLCs) by measuring $Ca^{2+}$ activity with cell imaging technique. OLCs were isolated from femur and tibia of neonatal rats, and cultured for 7 days. Cultured OLCs were loaded with a $Ca^{2+}$-sensitive fluorescent dye, Fura-2, and fluorescence images were monitored with a cooled CCD camera. The images were processed and analyzed with an image analyzing software. The results were as follows. (1) $[Ca^{2+}]_i$ of OLC decreased as the $Ca^{2+}$ concentration in the superfusing Tyrode solution was lowered. When $Na^+$ concentration in the superfusing solution was decreased, $[Ca^{2+}]_i$ increased.. These suggest that $Ca^{2+}$ flux occurs via the $Na^+-Ca^{2+}$ exchange mechanism. (2) When $Na^+$ in the superfusing solution was removed. a transient $Ca^{2+}$, increase($Ca^{2+}$ spike) was occasionally observed. However, $Ca^{2+}$ spike was not observed after adding 1 ${\mu}M$ thapsigargin. This implies that the generation of $Ca^{2+}$ spike is mediated by the release of $Ca^{2+}$ from endoplasmic reticulum(ER). (3) As the $Ca^{2+}$ concentration in the superfusing solution was raised, the frequency of 0mM $Na^+$-induced $Ca^{2+}$ spike increased, suggesting that $Ca^{2+}$-induced $Ca^{2+}$ release(CICR) mechanism exists. (4) After $[Ca^{2+}]_i$ was decreased with the superfusion of $Ca^{2+}$-free solution containing thapsigargin, the recovery of $[Ca^{2+}]_i$ with reperfusion of 2.5mM $Ca^{2+}$ solution transiently exceeded the control level, suggesting that the depletion of $Ca^{2+}$ in ER induces $Ca^{2+}$ influx from extracellular medium via store-operated $Ca^{2+}$ influx(SOCI) mechanism. (5) $[Ca^{2+}]_i$ was not affected by the superfusion of 25mM $K^+$ Tyrode solution. These results suggest that intracellular $Ca^{2+}$ activity in osteoblast is regulated by transmembrane $Ca^{2+}$ flux via $Na^+-Ca^{2+}$ exchange, $Ca^{2+}$ release from the internal store (ER) via $Ca^{2+}$-induced $Ca^{2+}$ release, and store-operated $Ca^{2+}$ influx across the cell membrane.

• 한국컴퓨터그래픽스학회논문지
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• 제24권1호
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• pp.9-16
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• 2018

본 논문에서는 광 회절 단층 촬영 (Optical Diffraction Tomography, ODT) 기법을 사용해 얻어진 세포 영상을 3차원 가상 공간에 시각적으로 표현하고 기존의 세포 영상들과의 일치감을 주는 색상 매핑 기술을 포함한 가시화 프레임 워크를 소개한다. 전체 볼륨을 구성하는 내부 구조에 대한 정보가 잘 알려져 있거나 명확하게 구분 가능한 인체의 장기 또는 산업 기기와 같은 기존의 볼륨 데이터와는 달리 세포 영상 데이터는 세포소기관들 간의 경계가 모호하거나 상황에 따라 형상의 변화가 다양하다는 특징을 가지고 있어, 세포의 형상에 대한 일관적인 시각 표현이 상대적으로 어렵다는 문제가 있다. 본 논문에서는 이를 해소하기 위해 세포의 3차원 형상을 실시간으로 렌더링 할 수 있는 가시화 기법을 제안한다. 제안하는 기법에서는 우선 세포의 3차원 형상을 나타내기 위해 볼륨 데이터의 가시화에서 널리 활용되고 있는 볼륨 렌더링 기법을 ODT 영상에 맞게 적용했으며, 빈 공간 교란 기법을 통한 렌더링 결과의 개선으로 세포내 구조의 연속성을 나타낼 수 있게 했다. 또한 다중 전이 함수에 대해 레이어 기반 독립 렌더링을 적용하는 것을 통해 다수의 세포내 구조를 하나의 화면에 표현하는 복합 가시화 기법을 제안했다. ODT 영상 및 염색 영상을 동시에 촬영 가능한 현미경으로부터 얻어진 세포 영상을 가시화 하는 것을 통해 제시된 가시화 기법의 유용성을 확인했다.

• International Journal of Oral Biology
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• 제31권4호
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• pp.141-148
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• 2006

The effects of adenosine triphosphate(ATP) on salivary glands have been recognized since 1982. The presence of purinergic recepetors(P2Rs) that mediate the effects of ATP in various tissues, including parotid and submandibular salivary gland, has been supported by the cloning of receptor cDNAs and the expression of the receptor proteins. P2Rs have many subtypes, and the activation of these receptor subtypes increase intracellular $Ca^{2+}$, a key ion in the regulation of the secretion in the salivary gland. The apical pores of taste buds in circumvallate and foliate papillae are surrounded by the saliva from von Ebner salivary gland(vEG). Thus, it is important how the secretion of vEG is controlled. This study was designed to elucidate the roles of P2Rs on salivary secretion of vEG. Male Sprague-Dawley rats (about 200 g) were used for this experiment. vEG-rich tissues were obtained from dissecting $500-1,000\;{\mu}m$ thick posterior tongue slices under stereomicroscope view. P2Rs mRNA in vEG acinar cells were identified with RT-PCR. To observe the change in intracellular $Ca^{2+}$ activity, we employed $Ca^{2+}-ion$ specific fluorescence analysis with fura-2. Single acinar cells and cell clusters were isolated by a sequential trypsin/collagenase treatment and were loaded with $10\;{\mu}M$ fura -2 AM for 60 minutes at room temperature. Several agonists and antagonists were used to test a receptor specificity. RT-PCR revealed that the mRNAs of $P2X_4$, $P2Y_1$, $P2Y_2$ and $P2Y_3$ are expressed in vEG acinar cells. The intracellular calcium activity was increased in response to $10\;{\mu}M$ ATP, a P2Rs agonist, and 2-MeSATP, a $P2Y_1$ and $P2Y_2R$ agonist. However, $300\;{\mu}M\;{\alpha}{\beta}-MeATP$, a $P2X_1$ and $P2X_3R$ agonist, did not elicit the response. The responses elicited by $10\;{\mu}M$ ATP and UTP, a $P2Y_2R$ agonists, were maintained when extracellular calcium was removed. $10\;{\mu}M$ suramin, a P2XR antagonist, and reactive blue 2, a P2YR antagonist, partially blocked ATP-induced response. However, when extracellular calciums were removed, suramin did not abolish the responses elicited by ATP. These results suggest that P2Rs play an important role in salivary secretion of vEG acinar cells and the effects of ATP on vEG salivary secretion may be mediated by $P2X_4$, $P2Y_1$, $P2Y_2$, and/or $P2Y_3$.