• Nuclear Medicine and Molecular Imaging
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• 제42권4호
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• pp.333-336
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• 2008

A 52-year-old woman with a history of general weakness, fatigue, weight loss, elevated serum levels of liver transaminase enzyme for three months underwent an F-18 FDG PET/CT scan to evaluate a cause of the hepatosplenomegaly found on abdominal ultrasonography. Initial PET/CT revealed markedly enlarged liver and spleen with intense FDG uptake. Otherwise, there were no areas of abnormal FDG uptake in whole body image. Histological evaluation by a hepatic needle biopsy demonstrated diffuse large B cell type lymphoma and final diagnosis for this patient was hepatosplenic B-cell lymphoma. She received five cycles of CHOP chemotherapy, and second PET/CT scan was followed after then. Follow-up PET-CT revealed normal sized liver with disappearance of abnormal FDG uptake. Hepatosplenic B-cell lymphoma is relatively rare and mostly presents as single or multiple nodules.1,2 Diffuse type hepatosplenic lymphoma is extremely rare and poorly recognized entity.3 The diagnosis is very difficult and complicated by the presence of misleading symptoms.4 In this rare hepatosplenic B-cell lymphoma case, F-18 FDG PET/CT scan provided a initial diagnostic clue of hepatosplenic lymphoma and an accurate chemotherapy response.

• Molecules and Cells
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• 제26권2호
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• pp.113-120
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• 2008

Laser light microscopy enables observation of various simultaneously occurring events in living cells. This capability is important for monitoring the spatiotemporal patterns of the molecular interactions underlying such events. Two-photon excited fluorescence microscopy (two-photon microscopy), a technology based on multiphoton excitation, is one of the most promising candidates for such imaging. The advantages of two-photon microscopy have spurred wider adoption of the method, especially in neurological studies. Multicolor excitation capability, one advantage of two-photon microscopy, has enabled the quantification of spatiotemporal patterns of $[Ca^{2+}]_i$ and single episodes of fusion pore openings during exocytosis. In pancreatic acinar cells, we have successfully demonstrated the existence of "sequential compound exocytosis" for the first time, a process which has subsequently been identified in a wide variety of secretory cells including exocrine, endocrine and blood cells. Our newly developed method, the two-photon extracellular polar-tracer imaging-based quantification (TEPIQ) method, can be used for determining fusion pores and the diameters of vesicles smaller than the diffraction-limited resolution. Furthermore, two-photon microscopy has the demonstrated capability of obtaining cross-sectional images from deep layers within nearly intact tissue samples over long observation times with excellent spatial resolution. Recently, we have successfully observed a neuron located deeper than 0.9 mm from the brain cortex surface in an anesthetized mouse. This microscopy also enables the monitoring of long-term changes in neural or glial cells in a living mouse. This minireview describes both the current and anticipated capabilities of two-photon microscopy, based on a discussion of previous publications and recently obtained data.

• 한국환경농학회지
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• 제28권4호
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• pp.435-441
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• 2009

In the present study, we analyzed the defensin protein deduced from Korean radish (Raphanus sativus L.) seeds.To express the genes in E. coli, we constructed a recombinant expression vector with a defensin gene, named rKRs-AFP gene isolated from Korean radish seeds. Over expressed rKRs-AFP proteins was separated by SDS-PAGE to determine the purity, and protein concentration was determined by the Bradford method. Antifungal activity was assessed by disk assay method against the tested fungi. As a result, when 500 mL of cell culture were disrupted by sonicator, 32.5 mg total proteins were obtained. The purified protein showed a single band on SDS-PAGE with estimated molecular weight about 6 KDa, consistent with the molecular mass calculated from the deduced amino acid sequence. The purified rKRs-AFP protein showed remarkable antifungal activities against several fungi including Aspergillus niger, Botrytis cinerea causing the gray mold disease, and Candida albicans. In field tests using the purified rKRs-AFP protein, the protein showed the reducing activity of disease spot and the mitigating effect of spreading of disease like agrichemicals. The immuno-assay of rKRs-AFP protein showed that the purified protein entirely accumulated at B. cinerea cytoplasm through the hyphal septa shown by fluorescence imaging. There was no fluorescence inside the cell, when the hypha was incubated without the protein. These all results indicate that the recombinant rKRs-AFP proteins can be utilized as a potential antifungal drug to control harmful plant fungal pathogens.

• Imaging Science in Dentistry
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• 제34권2호
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• pp.99-106
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• 2004

Purpose: To investigate the effects of irradiation on the phenotypic expression of the MC3T3-El osteoblastic cell line, especially on the osteonectin and bone sialoprotein. Materials and Methods: Cells were irradiated with a single dose of 0.5, 1, 4 and 8 Gy at a dose rate of 5.38 Gy/min using Cs-I37 irradiator. After specimens were harvested, total RNA was extracted on the 3rd, 7th, 14th, 21st day after irradiation. The total RNA was reverse-transcribed and the resulting cDNAs were subjected to amplification by PCR with a pair of primers. Results: The irradiated cells showed a dose-dependent increase in osteonectin mRNA expression when compared with the unirradiated control group. The irradiated cells showed no difference in bone sialoprotein mRNA expression when compared with the unirradiated control group. In accordance with the duration of culture period after irradiation, the level of osteonectin mRNA expression showed no difference, but it increased a little at the 21st day in the 4 and 8 Gy exposure groups. In the case of bone sialoprotein, however, the level of mRNA expression increased significantly at the 3rd and 7th day after irradiation, but it showed no difference at the 14th and 21st day when compared with the control group. Conclusion: These results showed that each single dose of 0.5, 1, 4 and 8 Gy influenced the mRNA expression of osteonectin and bone sialoprotein at the calcification stage of osteoblastic cells, suggesting that single dose of irradiation affected the osteoblastic bone formation at the cell level.

• Nuclear Medicine and Molecular Imaging
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• 제43권5호
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• pp.487-494
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• 2009

목적: 작은 크기의 재조합 단일사슬 항체는 빠른 혈중 제거율과 종양의 항체 집적율이 증가되는 등의 장점을 가지고 있다. 반면에 항체의 작은 크기는 방사성 또는 형광물질의 표지를 위한 킬레이터 결합에 중요한 아미노산 그룹의 감소를 의미하기도 한다. 본 연구에서는 단일사슬 lym-1 염기서열 C-말단에 lysine 아미노산 태그를 삽입하여 형광 물질의 직접표지 및 그 표지수율 증가를 확인하고자 하였다. 대상 및 방법: 대장균 pET-22b (+) 벡터에 재조합 된 lysine 삽입 단일사슬 lym-1유전자는 대장균 BL21 (DE3)에 형질전환하여 발현하였다. 생산된 lysine lym-1 항체는 Ni-NTA 컬럽과 분자량 컬럼을 사용해 정제하였고. 단백질 전기 영동과 western blot을 통해 확인하였다. lysine lym-1 항체에 방사성 동위원소인 I-124, I-125, I-131 과 Tc-99m를 표지하여 그 수율을 확인하였으며 유세포계측기를 사용해 형광물질인 FITC가 직접표지된 라이신 lym-1 항체의 면역반응성을 사람의 버킷 림프종 세포주인 Raji 세포주에서 면역반응성을 확인하였다. 결과 Lysine도입 단일사슬 lym-1 항체는 두 과정의 정제를 통하여 획득하였으며 그 크기는 약 48 KDa이었고, 방사성동위원소인 I-124, I-125, I-131과 Tc-99m의 표지수율은 각각 >99%, >99%, >95%, >99%로 확인되었다. 유세포계측을 통한 lysine 도입 단일사슬 lym-1항체의 면역반응성은 기존의 단일사슬 lym-1항체와 유사함을 확인하였다. 결론: 재조합 lym-1 항체에 형광 물질을 직접 표지하기 위한 lysine 아미노산의 도입은 항체의 면역반응성 감소를 최소화 시키면서 직접표지 수율을 증가시킬 수 있는 유용한 방법임을 확인하였다.

• Imaging Science in Dentistry
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• 제30권1호
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• pp.71-79
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• 2000

Purpose : This study was aimed to evaluate the cell cycle arrest and apoptosis induction after irradiation and epidermal growth factor (EGF) treatment in three human epithelial tumor cell lines (A431, Siha, KB). Materials and Methods: Single irradiation of 2, 5 and 10 Gy was done on three cell lines with 5.38 Gy/min dose rate using Cs-137 irradiator at room temperature. Also, EGF of 10 ng/ml was added immediately after 10 Gy irradiation. Cell growth was evaluated by counting the living cell number using a hemocytometer at 1 day, 2 days, 3 days, 4 days and 5 days after irradiation. Cell cycle arrest and apoptosis induction were assayed with the flow cytometry at 8 hours, 12 hours, 1 day, 2 days, 3 days, 4 days and 5 days after irradiation. Results : Growth of irradiated three cell lines were inhibited in proportion to radiation dose. EGF treatment after irradiation showed various results according to cell lines. On all cell lines, G2 arrest was detected after 8 hours and maximized after 12 hours or 1 day. Amount of G2 arrest was positively dose dependent. However, EGF showed no significant change on G2 arrest. G2 arrest was recovered with time at 2 Gy and 5 Gy irradiation. However, at 10 Gy irradiation, G2 arrest was continued. Apoptosis was detected at 10 Gy irradiation. On EGF treated group after irradiation, A431 and Siha cell lines showed slightly increased apoptosis but there was no statistically significant difference. KB cell line showed no marked change of apoptosis induction. Conclusion : Irradiation effects on cell cycle arrest and apoptosis induction in three human epithelial tumor cell lines, however epidermal growth factor doesn't effect on.

• Imaging Science in Dentistry
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• 제30권3호
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• pp.169-181
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• 2000

Purpose: To investigate the expression of bone morphogenetic protein (BMP)-2/4 during eary tooth development after irradiation and calcium-deficient diet. Materials and Methods: The pregnant three-week-old Sprague-Dawley rats were used for the study. The control group was non-irradiation/normal diet group (Group 1), and the experimental groups were irradiation/normal diet group (Group 2) and irradiation/calcium-deficient diet group (Group 3). The abdomen of the rats at the 9th day of pregnancy were irradiated with single dose of 350 cGy. The rat pups were sacrificed at embryonic 18 days, 3 days and 14 days after delivery and the maxillae tooth germs were taken. The tissue sections of specimen were stained immunohisto-chemically with anti-BMP-2/4 antibody. Results: At embryo-18 days, immunoreacivity for BMP-2/4 of the Group 1 was modetate in stratum intermedium of dental organ and weak in dental papilla and dental follicle, but that of Group 2 was weak in cell layer of dental organ, and no immunoreacivity was shown in dental papilla and dental follice of Group 2 and in all tissue components of the Group 3. At postnatal-3 days, immunoreacivity for BMP-2/4 of the Group 1 was strong in cell layer of dental organ, odontoblasts and developing alveolar bone, but that of Group of 2 and Group 3 was weak in odontoblasts and developing alveolar bone. At postnatal-14 days, immunoreacivity for BMP-2/4 of the Group 1 was strong in newly formed cementum, alveolar bone and odontoblasts, but that of Group 2 was weaker than that of Group 1. In the Group 3, tooth forming cell layer showed weak immunoreactivity, but other cell layers showed no immunoreactivity. Couclusion : The expression of bone morphogenetic protein (BMP)-2/4 during early tooth development was disturbed after irradiation and calcium-deficient diet.

• The Korean Journal of Physiology and Pharmacology
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• 제28권1호
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• pp.93-103
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• 2024

Satellite glial cells (SGCs), a major type of glial cell in the autonomic ganglia, closely envelop the cell body and even the synaptic regions of a single neuron with a very narrow gap. This structurally unique organization suggests that autonomic neurons and SGCs may communicate reciprocally. Glial Ca²⁺ signaling is critical for controlling neural activity. Here, for the first time we identified the machinery of store-operated Ca²⁺ entry (SOCE) which is critical for cellular Ca²⁺ homeostasis in rat sympathetic ganglia under normal and pathological states. Quantitative realtime PCR and immunostaining analyses showed that Orai1 and stromal interaction molecules 1 (STIM1) proteins are the primary components of SOCE machinery in the sympathetic ganglia. When the internal Ca²⁺ stores were depleted in the absence of extracellular Ca²⁺, the number of plasmalemmal Orai1 puncta was increased in neurons and SGCs, suggesting activation of the Ca²⁺ entry channels. Intracellular Ca²⁺ imaging revealed that SOCE was present in SGCs and neurons; however, the magnitude of SOCE was much larger in the SGCs than in the neurons. The SOCE was significantly suppressed by GSK7975A, a selective Orai1 blocker, and Pyr6, a SOCE blocker. Lipopolysaccharide (LPS) upregulated the glial fibrillary acidic protein and Toll-like receptor 4 in the sympathetic ganglia. Importantly, LPS attenuated SOCE via downregulating Orai1 and STIM1 expression. In conclusion, sympathetic SGCs functionally express the SOCE machinery, which is indispensable for intracellular Ca²⁺ signaling. The SOCE is highly susceptible to inflammation, which may affect sympathetic neuronal activity and thereby autonomic output.

• Imaging Science in Dentistry
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• 제38권3호
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• pp.125-132
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• 2008

Purpose : To investigate the effects of irradiation on transforming growth factor ${\beta}_1$ (TGF-${\beta}_1$) mRNA expression and calcific nodule formation in MC3T3-E1 osteoblastic cell line. Materials and Methods : Cells were cultured in alpha-minimum essential medium ($\alpha$-MEM) supplemented with 10% fetal bovine serum and antibiotics. When the cells reached the level of 70-80% confluence, culture media were changed with $\alpha$-MEM supplemented with 10% FBS, 5 mM $\beta$-glycerol phosphate, and $50\;{\mu}g/mL$ ascorbic acid. Thereafter the cells were irradiated with a single dose of 2, 4, 6, 8 Gy at a dose rate of 1.5 Gy/min. The expression pattern of TGF-${\beta}_1$ mRNA, calcium content and calcific nodule formation were examined on day 3, 7, 14, 21, 28, respectively, after the irradiation. Results : The amount of TGF-${\beta}_1$ mRNA expression decreased significantly on day 7 after irradiation of 4, 6, 8 Gy. It also decreased on day 14 after irradiation of 6, 8 Gy. and decreased on day 21 after irradiation of 8 Gy. The amount of calcium deposition decreased significantly on day 7 after irradiation of 4, 8 Gy (P < 0.01) and showed a decreased tendency on day 14, 21 after irradiation of 4, 6, 8 Gy. The number of calcific nodules was decreased on day 7 after irradiation of 4, 8 Gy. Conclusion: Irradiation with a single dose of 4, 6, 8 Gy influences negatively the bone formation at the molecular level by affecting the TGF-${\beta}_1$ mRNA expression that was associated with proliferation and the production of extracellular matrix in MC3T3-E1 osteoblastic cell line.

• Imaging Science in Dentistry
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• 제35권1호
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• pp.1-8
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• 2005

Purpose : To investigate the effects of irradiation on the calcium content and calcific nodule formation in the MC3T3-E1 osteoblastic cell line. Materials and Methods : Cells were irradiated with a single dose of 2,4 and 8 Gy at a dose rate of 5.38 Gy/min using a Cs-137 irradiator. After irradiation, the calcium content and calcific nodule formation were examined on the 1 st, 2nd, 3rd and 4th week. Results : A decreasing dose-dependent tendency of the cell proliferation rate was found in all irradiated groups of this experiment when compared with the unirradiated control group. In accordance with the duration of culture, there was no significant difference in the cell proliferation rate after irradiation of 2 Gy when compared with the unirradiated group, however a decreasing tendency was found in 4 Gy- and 8 Gy-irradiated groups. While an increase in total calcium content after irradiation of 2 Gy was found at week 1, week 2, and week 4, there was a decrease in calcium content at week 1 through 4 in the 8 Gy- irradiated group. Calcific nodule formation was increased in irradiated experimental groups when compared with the unirradiated control group in the 2 Gy-irradiated group, but decreased in the 4 Gy- and 8 Gy-irradiated groups at the same stage. Conclusion : The results showed a mild increasing tendency of the calcific nodule formation after irradiation of 2 Gy. However, a decreased calcific nodule formation in 4 Gy- and 8 Gy-irradiated groups was found. Taken together, the irradiation of 2 Gy mildly activated bone formation, however 4 Gy or 8 Gy suppressed bone formation by decreasing cell numbers in the MC3T3-El osteoblastic cell line.