• International journal of thyroidology
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• 제11권2호
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• pp.160-166
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• 2018

Background and objectives: Salivary hypofunction is one of the common side effects after radioiodine therapy, and its pathophysiology is salivary ductal stenosis resulting from ductal cell injury. This study aimed to develop the functional culture environment of human parotid gland ductal cells in in vitro three-dimensional perfusion culture system. Materials and Methods: We compared plastic dish culture method and three-dimensional culture system containing Matrigel and nanofiber. Morphogenesis of reconstituted salivary structures was assessed by histomorphometry. Functional characteristics were assessed by immunohistochemistry and reverse transcription polymerase chain reaction (aquaporin 5, CK7, CK18, connexin 43, and p21). In addition, we designed the media perfusion culture system and identified higher rate of cell proliferation and expression of connexin 43 in perfusion system comparing to dish. Results: Human parotid ductal cells were well proliferated with the ductal cell characters under environment with Matrigel. In the presence of Matrigel, aquaporin 5, CK18 and connexin 43 were more expressed than 2D dish and 3D nanofiber setting. In the media perfusion culture system, ductal cells in 3D culture media showed higher cells count and connexin 43 expression compared to 2D dish. Conclusion: This in vitro ductal cell perfusion culture system using Matrigel could be used to study for radioiodine induced sialadenitis model in vivo.

• Journal of Microbiology and Biotechnology
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• 제18권2호
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• pp.295-298
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• 2008

Creatine kinase (CK; E.C. 2.7.3.2) is an important enzyme that catalyzes the reversible transfer of a phosphoryl group from ATP to creatine in energy homeostasis. The brain-type cytosolic isoform of creatine kinase (BB-CK), which is found mainly in the brain and retina, is a key enzyme in brain energy metabolism, because high-energy phosphates are transfered through the creatine kinase/phosphocreatine shuttle system. The recombinant human BB-CK protein was overexpressed as a soluble form in Escherichia coli and crystallized at $22^{\circ}C$ using PEG 4000 as a precipitant. Native X-ray diffraction data were collected to $2.2{\AA}$ resolution using synchrotron radiation. The crystals belonged to the tetragonal space group $P4_32_12$, with cell parameters of a=b=97.963, $c=164.312{\AA},\;and\;{\alpha}={\beta}={\gamma}=90^{\circ}$. The asymmetric unit contained two molecules of CK, giving a crystal volume per protein mass $(V_m)$ of $1.80{\AA}^3\;Da^{-1}$ and a solvent content of 31.6%.

• Journal of Microbiology and Biotechnology
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• 제16권8호
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• pp.1201-1209
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• 2006

The purpose of this study was to isolate probiotic lactic acid bacteria (LAB) that produce bile salt hydrolase (BSH), and to evaluate its effects on serum cholesterol level. One-hundred-twenty bacterial colonies were initially isolated from human feces, and five strains were selected after screening based on their resistance to acids, tolerance against bile salts, and inhibitory activity on Escherichia coli. The Lactobacillus plantarum strain with the highest level of BSH activity was identified using 16S rRNA sequences, and was named L. plantarum CK 102. L. plantarum CK 102 at a level of 1.36$\times$10$^8$cfu/ml survived in pH 2 buffer for 6 h and exhibited excellent tolerance for bile salt. Coculturing the strain with E. coli in MRS broth resulted in strong inhibition against growth of E. coli at 18 h. Furthermore, the potential effect of CK 102 on serum cholesterol level was evaluated in rats. Thirty-two rats [Sprague-Dawley (SD) male, 129$\pm$l g, 5 weeks old] were divided into four groups of eight each. For six weeks, Group 1 was fed a normal diet (negative control); Group 2 was fed a cholesterol-enriched diet (positive control); Group 3 was fed a cholesterol-enriched diet plus L. plantarum CK 102 at 1.0$\times$10$^7$cfu/ml; and Group 4 was fed a cholesterol-enriched diet plus L. plantarum CK 102 at 5.0$\times$10$^7$cfu/ml. Blood samples were collected, serum lipids were analyzed, and weights of the organs were measured. Total blood cholesterol level, triglyceride, LDL-cholesterol, and free-cholesterol values were lower in rats that were fed 1. plantarum CK 102 than in those not fed L. plantarum CK 102. This cholesterol lowering effect implies that L. plantarum CK 102 could be utilized as an additive for health-assistance foods. In conclusion, these results suggest that the 1. plantarum CK 102 isolated could be used commercially as a probiotic.

• 한국식품영양과학회지
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• 제35권4호
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• pp.410-415
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• 2006

청국장 분말이 고지방식이를 섭취한 흰쥐의 지질대사에 미치는 영향을 조사하고자, 성숙한(생후 20주령) 숫쥐를 4주간 고지방식이(0.5% 콜레스테롤, 10% 지방)로 고지혈증을 유도한 후 대조군(고지방식이), 청국장군(고지방식이에 단백질원으로 청국장 분말을 첨가한 식이) 및 상황버섯 청국장군(고지방식이에 단백질원으로 상황버섯 청국장 분말을 첨가한 식이 ) 등 3군으로 나누어 5주간 사육한 결과는 다음과 같다. 실험동물의 체중은 대조군이 41.8 g증가함에 비하여 청국장군 및 상황버섯 청국장군은 각 -3.7 g 및 -0.8 g으로 유의하게 감소되었고, 식이섭취량 및 식이효율은 대조군에 비하여 청국장군들이 유의하게 감소되었다. 간과 부고환지방의 무게 및 간조직의 콜레스테롤과 중성지질 농도는 대조군에 비하여 청국장군 및 상황버섯 청국장군이 유의하게 감소시키는 효과가 나타났다. 혈청의 중성지질, 총 콜레스테롤, LDL-콜레스테롤, VLDL-콜레스테롤 농도 및 동맥경화지수는 대조군에 비해 청국장군 및 상황버섯 청국장군이 유의하게 감소되었다. 총 콜레스테롤에 대한 HDL-콜레스테롤의 비율은 대조군에 비해 청국장군들이 유의하게 증가되었다. 변의 무게, 총 지질, 중성지질 및 콜레스테롤 배설량은 청국장군들이 대조군보다 유의하게 증가된 것으로 나타났다. 이상의 결과로 보아 고지방식이에 청국장 및 상황버섯청국장 분말을 첨가 섭취시 혈청과 간조직의 중성지질, 총콜레스테롤과 LDL-콜레스테롤 농도 및 동맥 경화지수를 낮추고, 총 콜레스테롤에 대한 HDL-콜레스테롤의 비율을 증가시키는 지질대사의 개선 효과가 나타났다.

• Asian Pacific Journal of Cancer Prevention
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• 제15권3호
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• pp.1187-1192
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• 2014

Background: To correlate breast cancer subtypes with prognostic factors, microvessel density (MVD), vascular endothelial growth factor (VEGF) expression and clinical features. Materials and Methods: One hundred cases of primary breast carcinoma were classified using biomarkers on tissue microarray as: luminal A [estrogen receptor (ER)+, HER2-, $Ki-67{\leq}14%$], luminal B [ER+, HER2+ or ER+, HER2-, Ki-67>14%], HER2, triple negative basal-like (TNB) [any basal cytokeratins (CKs, 5, 14, 17) and/or endothelial growth factor receptor (EGFR) expression], and TN without such markers [TNN, null], and assessed for p53, vimentin, VEGF and CD31 immunoperoxidase. Results: Of the 100 cases (mean age, 51 years; mean tumor size, 3.2cm; 56% with nodal metastasis; 89 invasive ductal carcinomas, not otherwise specified, 4 invasive lobular carcinomas, 3 metaplastic carcinomas, and 4 other types) there were 39 luminal A, 18 luminal B, 18 HER2, 15 TNB and 10 TNN. The positivities of basal-like markers in the basal-like subtype were 78.3% for CK5, 40% for CK14, 20% for CK17, 46.7% for EGFR. There was no significant difference in age distribution, tumor size, degree of tubular formation, pleomorphism, lymphovascular invasion, nodal metastasis, MVD, VEGF expression and survival among subgroups. TNs demonstrated significantly higher tumor grade, mitotic count, Ki-67 index, p53 and vimentin and decreased overall survival compared with nonTN. Conclusions: The distribution of breast cancer subtypes in this study was similar to other Asian countries with a high prevalence of TN. The high grade character of TN was confirmed and CK5 expression was found to be common in our basal-like subtype. No significant elevation of MVD or VEGF expression was apparent.

• IMMUNE NETWORK
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• 제7권3호
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• pp.117-123
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• 2007

Background: This study was designed to investigate the role of the hepatocyte growth factor (HGF) with regards to differentiation of somatic stem cells originating from the human umbilical cord blood (UCB) into hepatic lineage cells in vitro culture system. Methods: Mononuclear cells from UCB were cultured with and without HGF based on the fibroblast growth factor (FGF)-1, FGF-2, and stem cell factor. The cultured cells were confirmed by immunofluorescent staining analysis with albumin (ALB), cytokeratin-19 (CK-19), and proliferating cell nuclear antigen (PCNA) MoAb. ALB and CK-18 mRNA were also evaluated by reverse transcription-polymerase chain reaction. In order to observe changes in proliferating capacity with respect to the cultured period, CFSE with affinity to proliferating cells were tagged and later underwent flow cytometry. Results: In the HGF-treated group, cultured cells had a large oval shaped appearance with adherent, but easily detachable characteristics. In the HGF-non treated group, these cells were spindle-shaped with strong adherent characteristics. Expressions of ALB and CK-19 were evident in HGF-treated group compared to non-expression of those in to HGF-non treated group. Dual immunostaining analysis of the ALB producing cells showed presence of PCNA in their nuclei, and ALB and CK-18 mRNA were detected on the 21st day of cultured cells in the HGF-treated group. Conclusion: Our findings suggest that HGF has a pivotal role in differentiating somatic stem cells of human UCB into hepatic lineage cells in vitro.

• 응용통계연구
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• 제18권2호
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• pp.329-341
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• 2005

군집방법의 비교시 사용되는 Rand(1971)의 $C_k$, k = 2, 3, . . ., N-1 통계량에 대한 점근 결과를 이용하여 자료에 존재하는 군집수를 예측하는 방법을 제안하였다. 제안된 방법과 $C_k$ 통계량의 변화 형태에 따라 군집수를 예측하는 Chae와 Warde(1991)와 허명회와 이용구(2004)의 방법을 비교하기 위하여 모의실험을 하였다. 현실적인 문제를 고려하여 실제자료에 대해서는 계속적인 재표본의 형성을 위하여 붓스트랩방법을 사용하였다.

• Journal of Nutrition and Health
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• 제39권4호
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• pp.331-337
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• 2006

청국장 분말이 고지방식이를 섭취한 흰쥐의 지질대사에 미치는 영향을 조사하고자, 성숙한 암쥐에 4주간 고지방식이(0.1% 콜레스테롤, 10% 지방)로 고지혈증을 유도한 후 대조군 (고지방식이), 청국장군 (고지방식이에 단백질원으로 청국장 분말을 첨가한 식이) 및 신령버섯 청국장군 (고지방식이에 단백질원으로 신령버섯 청국장 분말을 첨가한 식이) 등 3군으로 나누어 8주간 사육한 결과는 다음과 같다. 실험동물의 체중변화는 대조군은 16.1 g증가에 비하여 청국장군 및 신령버섯 청국장군은 각 1.0 g 및 5.9 g 증가로 유의하게 감소되었고, 식이섭취량 및 식이효율도 대조군에 비하여 청국장군들이 감소되었다. 자궁주위 지방의 무게 및 간조직의 콜레스테롤과 중성지질 농도는 대조군에 비하여 청국장군 및 신령버섯 청국장군이 유의하게 감소되었다. 혈청의 중성지질. 총 콜레스테롤, LDL-콜레스테롤 농도 및 동맥경화지수는 대조군에 비해 청국장군 및 신려버섯 청국장군이 유의하게 감소되었고, 총 콜레스테롤에 대한 HDL-콜레스테롤의 비율은 대조군에 비해 청국장군들이 유의하게 증가되었다. 변의 무게, 수분함량 및 총 지질 배설량은 청국장군들이 대조군 보다 유의하게 증가된 것으로 나타났다. 혈청 단백질 및 혈당 농도는 청국장 및 신령버섯 청국장 섭취에 의한 영향은 나타나지 않았으나, 혈색소 농도는 청국장군들이 증가되는 것으로 나타타났다. 이상의 결과로 보아 고지방식이에 청국장 및 신령버섯 청국장 분말 첨가 섭취시 흰쥐의 체중 감소와 혈청과 간조직의 중성지질, 총 콜레스테롤과 LDL-콜레스테롤 농도 및 동맥경화지수를 낮추고, 총 콜레스테롤에 대한 HDL-콜레스테롤의 비율이 증가되어 지질대사의 개선 효과가 있는 것으로 나타났다.

• Journal of Microbiology and Biotechnology
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• 제18권6호
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• pp.1081-1089
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• 2008

A novel ginsenoside-hydrolyzing ${\beta}$-glucosidase was purified from Paecilomyces Bainier sp. 229 by a combination of Q-Sepharose FF, phenyl-Sepharose CL-4B, and CHT ceramic hydroxyapatite column chromatography. The purified enzyme was a monomeric protein with a molecular mass estimated to be 115 kDa. The optimal enzyme activity was observed at pH 3.5 and $60^{\circ}C$. It was highly stable within pH 3-9 and at temperatures lower than $55^{\circ}C$. The enzyme was specific to ${\beta}$-glucoside. The order of enzyme activities against different types of ${\beta}$-glucosidic linkages was ${\beta}$-(1-6)>${\beta}$-(1-2)>${\beta}$-(1-4). The enzyme converted ginsenoside Rb1 to CK specifically and efficiently. An 84.3% amount of ginsenoside Rb1, with an initial concentration of 2 mM, was converted into CK in 24 h by the enzyme at $45^{\circ}C$ and pH 3.5. The hydrolysis pathway of ginsenoside Rb1 by the enzyme was $Rb1{\to}Rd{\to}F2{\to}CK$. Five tryptic peptide fragments of the enzyme were identified by a newly developed de novo sequencing method of post-source decay (PSD) matrix-assisted laser desorption/ionization (MALDI) mass spectrometry. By comparing the five identified peptide sequences with the NCBI database, this purified ${\beta}$-glucosidase proves to be a new protein that has not been reported before.

• Asian Pacific Journal of Cancer Prevention
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• 제13권10호
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• pp.5233-5236
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• 2012

CK2 is a serine threonine kinase that participates in a variety of cellular processes with more than 300 defined substrates. This critical enzyme is known to be upregulated in cancers, but the role of this upregulation in carcinogenesis is not yet fully understood but c-myc, one of the defined CK2 substrates, is a well-known proto-oncogene that is normally essential in developmental process but is also involved in tumor development. We evaluated the optimal enzyme and substrate concentrations for CK2 activity in both neoplastic and non-neoplastic human lung tissues using the c-$myc^{424-434}$ peptide (EQKLISEEDL) as a substrate. The activities measured for the neoplastic tissue were 600-750 U/mg protein while those for the control tissue was in the range of 650-800 U/mg. $K_m$ value for c-myc peptide was determined as $0.33{\mu}M$ in non-neoplastic tissue and $0.18{\mu}M$ in neoplastic tissue. In this study, we did not observe an increased activity in the neoplastic tissue when compared with the non-neoplastic lung tissue, but we recorded two times higher affinity for c-$myc^{424-434}$ in cancer tissue. Considering the metabolic position of c-$myc^{424-434}$, our results suggest that phosphorylation by CK2 may be important in dimerization and thus it might affect the regulation of c-myc in cancer tissues.