• BMB Reports
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• 제47권3호
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• pp.141-148
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• 2014

Bone is an active tissue, in which bone formation by osteoblast is followed by bone resorption by osteoclasts, in a repeating cycle. Proteomics approaches may allow the detection of changes in cell signal transduction, and the regulatory mechanism of cell differentiation. LC-MS/MS-based quantitative methods can be used with labeling strategies, such as SILAC, iTRAQ, TMT and enzymatic labeling. When used in combination with specific protein enrichment strategies, quantitative proteomics methods can identify various signaling molecules and modulators, and their interacting proteins in bone metabolism, to elucidate biological functions for the newly identified proteins in the cellular context. In this article, we will briefly review recent major advances in the application of proteomics for bone biology, especially from the aspect of cellular signaling.

• Bulletin of the Korean Chemical Society
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• 제25권8호
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• pp.1147-1150
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• 2004

Hypericin (1,3,4,6,8,13-hexahydroxy-10,11-dimethylphenanthro[1,10,9,8-opqra]perylene-7,14-dione), an antidepressant which is also known to be a potent protein kinase C (PKC) inhibitor was synthesized as a precursor for radioiodine labeling via two step reactions. Malignant glioma cells express higher PKC activity compared to untransformed glial cell. Here we report the synthesis and radioiodine labeling of hypericin as a potential brain tumor imaging radiopharmaceutical. The reference compound, 2-iodohypericin, and its radiolabelled analogues, 2-[$^{123}I$]iodohypericin and 2-[$^{124}I$]iodohypericin have been prepared by the reaction of hypericin with NaI or [$^{123}I$]NaI or [$^{124}I$]NaI. The labeling yield was 60-65% for each analogue and the optimal reaction time was 10 min. The purification and isolation of the labelled products were achieved by a reversed-phase HPLC.

• Journal of Microbiology and Biotechnology
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• 제19권12호
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• pp.1635-1643
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• 2009

The aim of this study was to provide new insight into the mechanism whereby the housekeeping enzyme glyceraldehyde-3-phosphate dehydrogenase (GAPDH) locates to cell walls of Lactobacillus plantarum 299v. After purification, cytosolic and cell wall GAPDH (cw-GAPDH) forms were characterized and shown to be identical homotetrameric active enzymes. GAPDH concentration on cell walls was growth-time dependent. Free GAPDH was not observed on the culture supernatant at any time during growth, and provoked cell lysis was not concomitant with any reassociation of GAPDH onto the cell surface. Hence, with the possibility of cw-GAPDH resulting from autolysis being unlikely, entrapment of intracellular GAPDH on the cell wall after a passive efflux through altered plasma membrane was investigated. Flow cytometry was used to assess L. plantarum 299v membrane permeabilization after labeling with propidium iodide (PI). By combining PI uptake and cw-GAPDH activity measurements, we demonstrate here that the increase in cw-GAPDH concentration from the early exponential phase to the late stationary phase is closely related to an increase in plasma membrane permeability during growth. Moreover, we observed that increases in both plasma membrane permeability and cw-GAPDH activity were delayed when glucose was added during L. plantarum 299v growth. Using a double labeling of L. plantarum 299v cells with anti-GAPDH antibodies and propidium iodide, we established unambiguously that cells with impaired membrane manifest five times more cw-GAPDH than unaltered cells. Our results show that plasma membrane permeability appears to be closely related to the efflux of GAPDH on the bacterial cell surface, offering new insight into the understanding of the cell wall location of this enzyme.

• Toxicological Research
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• 제34권1호
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• pp.1-6
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• 2018

The purpose of this study was to detect cell death in the liver of mice treated with thioacetamide (TAA) using fluorescence bioimaging and compare this outcome with that using conventional histopathological examination. At 6 weeks of age, 24 mice were randomly divided into three groups: group 1 (G1), control group; group 2 (G2), fluorescence probe control group; group 3 (G3), TAA-treated group. G3 mice were treated with TAA. Twenty-two hours after TAA treatment, G2 and G3 mice were treated with Annexin-Vivo 750. Fluorescence in vivo bioimaging was performed by fluorescence molecular tomography at two hours after Annexin-Vivo 750 treatment, and fluorescence ex vivo bioimaging of the liver was performed. Liver damage was validated by histopathological examination. In vivo bioimaging showed that the fluorescence intensity was increased in the right upper part of G3 mice compared with that in G2 mice, whereas G1 mice showed no signal. Additionally ex vivo bioimaging showed that the fluorescence intensity was significantly increased in the livers of G3 mice compared with those in G1 or G2 mice (p < 0.05). Histopathological examination of the liver showed no cell death in G1 and G2 mice. However, in G3 mice, there was destruction of hepatocytes and increased cell death. Terminal deoxynucleotidyl transferase dUTP nick end labeling staining confirmed many cell death features in the liver of G3 mice, whereas no pathological findings were observed in the liver of G1 and G2 mice. Taken together, fluorescence bioimaging in this study showed the detection of cell death and made it possible to quantify the level of cell death in male mice. The outcome was correlated with conventional biomedical examination. As it was difficult to differentiate histological location by fluorescent bioimaging, it is necessary to develop specific fluorescent dyes for monitoring hepatic disease progression and to exploit new bioimaging techniques without dye-labeling.

• Journal of Nutrition and Health
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• 제31권4호
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• pp.697-707
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• 1998

This study was designed to observe the effect of dietary fiber and fat on colon tumor incidence and cell proliferation. Male Sqraue Dawley rats(n=225) at 7 weeks of age, were divided into 3 groups depending on the type of fat b(beef tallow, corn oil and DHA-rich fish oil) and each group was again divided into 3 groups depending on type of fiber(fiber-free, perctin and cellulose) . The experimental diet containing dietary fat at 15%(w/w) and fiber at 6%(w/w) levels was fed for 25 weeks. At the same time, each rats was intramuscularly injected with DMH two times a week for 6 weeks to geive total dose of 180mg/kg body weight. Cell proliferation was measured by in vivo incroporation of bromodeoxyuridine (BrdU) into DNA. Fish oil decreased the tumor incidence (9.67%) compared with beef talow (33.39%) and corn oil (21.21%). Tumor incidence was decreased in all groups that fed cellulose (11.67%) compared with those of fiber-free(21.74%) and pectic(19.70%). Most of tumors was distributed at the site of the distal colon. The rats fed both fish oil and cellulose significantly decreased th enumber of tumors and tumor incidence compared to other groups. Fish oil was more effective in preventing cell prolofieration by decreasing crypt length and labeling index(LI) compared with beef tallow(p<0.05). Cell proliferation in distal colon was more developed to the upper part of the crypt compared to proximal colon. Overall tumor incidence and cell proliferation were more affected by dietary fat. But the effect of dietary fiber was different depending on type of fat in the experimental diet. These results suggest that a DHA -rich fish oil may has more decisive effect in inhibiting the cell proliferation in colon.

• ALGAE
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• 제33권2호
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• pp.191-203
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• 2018

Fragilariopsis cylindrus is one of the most successful psychrophiles in the Southern Ocean. To investigate the molecular mechanism of biomineralization in this species, we attempted to synchronize F. cylindrus growth, since new cell wall formation is tightly coupled to the cell division process. Nutrient limitation analysis showed that F. cylindrus cultures rapidly stopped growing when deprived of silicate or light, while growth continued to a certain extent in the absence of nitrate. Flow cytometry analysis indicated that deprivation of either silicate or light could effectively arrest the cell cycle of this diatom species at the G1 phase, suggesting that synchrony can be established using either factor. Fluorescence labeling of new cell walls was faintly detectable as early as approximately 6 h after silicon repletion or light irradiation, and labeling was markedly intensified by 18 h. It is revealed that the synthesis of girdle bands begins before valve synthesis in this species, with active valve synthesis occurring during the G2 / M phase. Expression profiling revealed that selective member(s) of the F. cylindrus SIT genes (FcSIT) respond to silicate and light, with a different set of genes being responsive to each factor. The Si / light double depletion experiments demonstrated that expression of one FcSIT gene is possibly correlated to transition to G2 / M phase of the cell cycle, when the valve is actively formed.

• Applied Microscopy
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• 제32권2호
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• pp.175-183
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• 2002

한국 연근해산 살오징어와 서해낙지의 시엽에서 신경전달물질을 분비하는 neuron의 특성 및 기능을 확인하기 위해 rabbit anti-dopamine (TH)과 rabbit anti-calbindin-$D_{28K}$ 등의 항체를 이용한 면역염색과 면역금지법을 시행한 결과는 다음과 같았다. 항-dopamine을 사용한 면역염색 결과 살오징어에서는 외과립세포층 상단에 위치한 대형 무축삭세포와 수질부의 섬을 이루는 세포들 중 $2{\sim}3$개의 비교적 소량의 세포만이 양성반응을 보인 반면, 서해낙지에서는 외과립세포층 상단 및 중간 부위의 $2{\sim}3$개의 세포와 수질부섬을 이루는 5개 이상의 세포에서 강한 양성반응을 보였다. 항-calbindin-D28k인 경우는 살오징어의 외과립세포층 상단에 위치한 $2{\sim}3$개의 소형 무축삭세포와 내과립세포층 하단에 위치한 $1{\sim}2$개의 세포에서 양성반응을 보인 반면, 서해낙지인 경우는 외과립세포층에서 4개 정도의 세포에서 양성 반응을 보였으나, 내과립세포층에서는 반응을 확인할 수 없었다. 면역금지법을 시행한 결과 살오징어에서는 항-dopamine과 항-calbindin에 면역반응을 보인 세포들의 세포질 $0.5{\mu}m^2$당 $17{\sim}26$개 정도의 비교적 많은 수의 금입자가 표지된 반면, 서해낙지에서는 세포질 $0.5{\mu}m^2$당 평균 10개 정도인 소량의 금입자만이 표지되어, 살오징어가 서해낙지에 비해 표지된 금입자가 거의 2배 정도 많았다.

• 대한한의학회지
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• 제24권4호
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• pp.127-135
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• 2003

Objectives : Intracerebral hemorrhage (ICH) is one of the most devastating types of stroke. The effect of acupuncture on the intrastriatal hemorrhage-induced neuronal cell death and cell proliferation in rats is examined. Methods : Cell death and cell proliferation in rats was investigated via terminal deoxynucleotidyl transferase-mediated dUTP nick end labeling (TUNEL) assay and immunohistochemistry for caspase-3 and 5-bromo-2'-deoxyuridine (BrdU). Results : Results showed that apoptotic cell death in the striatum and cell proliferation in the hippocampal dentate gyrus significantly increased following intrastriatal hemorrhage in rats, and that acupunctural treatment at the Zusanli acupoint suppressed the hemorrhage-induced increase in apoptosis in the striatum and cell proliferation in the dentate gyrus. Conclusions : It is suggested that acupunctural treatment, especially at the Zusanli acupoint, may aid recovery following central nervous system sequelae following ICH.

• 센서학회지
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• 제21권5호
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• pp.324-328
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• 2012

In the past a few decades, various kinds of cells have been examined in laboratories all over the world, and their interesting results have been expressed through various methods in journal publications. For a representative example, the increment or reduction of cell numbers during a bio-related experimental process has been demonstrated using the hazard ratio in survival analysis or in the form of a graph. In addition, the condition of cells such as their normality or abnormality would be indicated by the images of the cell nuclei or membranes treated with proper fluorescent labeling. However, the above methods seem to not be quantitative but rather qualitative assessments, which might be difficult to provide people with the eidetic understanding through parameters or numerical data. With adequate suggestions on any indices enabling the explanation for cell conditions, some analyses may be underestimated due to the lack of objectiveness caused by merely linguistic evaluation for the cell conditions, not numerally scientific interpretation. Therefore, in this study, we would suggest some indices enabling quantitative analysis on the cellular conditions.

• 한국멀티미디어학회논문지
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• 제20권5호
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• pp.740-747
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• 2017

A fast cell contour extraction method using CUDA parallel processing technique is presented. The cell contour extraction is one of important processes to analyze cell information in pathology. However, conventional sequential contour extraction methods are slow for a huge high-resolution medical image, so they are not adequate to use in the field. We developed a parallel morphology operation algorithm to extract cell contour more quickly. The algorithm can create an inner contour and fail to extract the contour from the concave part of the cell. We solved these problems by subdividing the contour extraction process into four steps: morphology operation, labeling, positioning and contour extraction. Experimental results show that the proposed method is four times faster than the conventional one.