• Title/Summary/Keyword: glycosylated

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Purification and Characterization of Recombinant Human Follicle Stimulating Hormone Produced by Chinese Hamster Ovary Cells

  • NA KYU HEUM;KIM SEUNG CHUL;SEO KWANG SEOK;LEE SUNG HEE;KIM WON BAE;LEE KANG CHOON
    • Journal of Microbiology and Biotechnology
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    • v.15 no.2
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    • pp.395-402
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    • 2005
  • Biologically active recombinant human follicle stimulating hormone (rhFSH) was produced in Chinese hamster ovary cells and purified by a series of chromatographic steps. The chromatographic steps included anion-exchange chromatography (DEAE Sepharose F/F, Q Sepharose F/F), hydrophobic interaction chromatography (Source 15 PHE), and hydroxyapatite chromatography (Macro-Prep ceramic hydroxyapatite type I). A distinctive step of the purification process developed was the use of ZnCl$_2$ for the removal of non-glycosylated or lowly-glycosylated FSH and impurities through co-precipitation with Zn$^{2+}$. Purified rhFSH was identified and characterized by several physicochemical and biological methods such as gel electrophoresis, high-performance liquid chromatography, amino acid analysis, carbohydrate analysis, and biological activity. The overall yield of the purification was ~$30\%$. The rhFSH preparation obtained showed high purity (>$99\%$) and high in vivo potency (>16,000 IU/mg). Carbohydrate analysis suggested that the purified rhFSH contained approximately $40\%$ (w/w) carbohydrate with di­or tri-antennary structure on average, which is somewhat more heavily sialylated than commercially available rhFSH. In conclusion, the results of these analyses established an identity of the purified rhFSH with natural FSH from human pituitary glands, and furthermore, the purified rhFSH preparation showed higher in vivo potency and was slightly more heavily sialylated than commercially available rhFSH.

The Anti-melanogenic Effect of Whitening Agent Containing Arbutin and ${\beta}-Glucosidase$ (알부틴과 베타-글루코시다제를 함유하는 미백 화장품의 항멜라닌 효과)

  • Lew Bark-Lin;Ryou Ji-Ho;Lee Mu-Hyoung
    • Journal of the Society of Cosmetic Scientists of Korea
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    • v.30 no.3 s.47
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    • pp.415-418
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    • 2004
  • Arbutin is a glycosylated hydroquinone found at high concentration in certain plants capable of surviving extreme and sustained dehydration. It has been reported to have an inhibitory effect of melanogenesis and to be hydrolyzed easily to yield glucose and hydroquinone by ${\beta}-glucosidase.$ While hydroquinone also has an anti-melanogenic effect, however, is carefully used as a topical whitening agent because of side effects. The present study was undertaken to examine the inhibitory effect of an whitening agent containing arbutin and ${\beta}-glucosidase$ on UV radiation induced pigmentation in human skin. Experimental subjects were UVB-irradiated on the back. UVB-irradiated areas were assigned to three groups: arbutin and ${\beta}-glucosidase$ treated group, vehicle control, and no-application control. Arbutin and ${\beta}-glucosidase$ treatment inhibited pigmentation by 50.17 percent, compared with the controls (N : 10: P<0.05). These results suggest that the whitening agent containing arbutin and ${\beta}-glucosidase$may be used as an agent to inhibit melanin formation induced by UV radiation.

Hybridization of Quercus aliena Blume and Q. serrata Murray in Korea - Analyses of Morphological variation and Flavonoid chemistry -

  • Park, Jin Hee;Park, Chong-Wook
    • Korean Journal of Environment and Ecology
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    • v.29 no.2
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    • pp.145-161
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    • 2015
  • This research was conducted in order to understand the hybridization between Quercus aliena Blume and Q. serrata Murray in Korea which show wide range of morphological variations within species and interspecific variations of diverse overlapping characteristics caused by hybridization. Morphological analysis (principal components analysis; PCA) of 116 individuals representing two species and their intermediates were performed. As a result, two species were clearly distinguished in terms of morphology, and intermediate morpho-types assumed to be hybrids between the two species were mostly located in the middle of each parent species in the plot of the principal components analysis. There was a clear distinction between two species in trichome distribution pattern which is an important diagnostic character in taxonomy of genus Quercus, whereas intermediate morpho-types showed intermediate state between two species' trichome distributions. Forty-two individuals representing two species and their intermediates were examined for leaf flavonoid constituents. Twenty-three flavonoid compounds were isolated and identified: They were glycosylated derivatives of flavonols, kaempferol, quercetin, isorhamnetin and myricetin. The flavonoid constituents of Q. aliena were five glycosylated derivatives: kaempferol 3-O-galactoside, kaempferol 3-O-glucoside, quercetin 3-O-galactoside, quercetin 3-O-glucoside, and Isorhamnetin 3-O-glucoside. The flavonoid constituents of Q. serrata had 20 diverse flavonol compounds including five flavonoid compounds found in Q. aliena. It was found that there is a clear difference in flavonoid constituents of Q. aliena and Q. serrata. Flavonoid chemistry is very useful in recognizing each species and putative hybrids. The flavonoid constituents of intermediates were a mixture of the two species' constituents and they generally showed similar characteristics to morpho-types. The hybrids between Q. aliena and Q. serrata showed morphologically and chemically diverse characteristics and it is assumed that there are frequent interspecific hybridization and introgression.

Effects of Luteolin on Fetal Bovine Serum-induced Events in Cultured Rat Vascular Smooth Muscle Cells (소태아혈청으로 유도된 흰쥐 혈관평활근세포의 luteolin 효과)

  • Lim, Yong
    • Journal of Life Science
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    • v.22 no.12
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    • pp.1595-1599
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    • 2012
  • Cell cycle activation and progression in vascular proliferative disease represent potent therapeutic targets. Luteolin, which occurs as glycosylated forms in celery, green pepper, perilla leaf, and camomile tea, has demonstrated antimutagenic, antitumorigenic, antioxidant, and antiinflammatory properties. In this study, we investigated the effect of luteolin on the proliferation of primary cultured rat aortic vascular smooth muscle cells induced by 5% fetal bovine serum. Luteolin at concentrations of 5, 20, and $50{\mu}M$ significantly inhibited this proliferation by 29.6, 50.8, and 83.1%, respectively. The incorporation of $[^3H]$-thymidine into DNA was also inhibited by 25.8, 57.6, and 81.0%, respectively. Flow cytometry analysis of DNA content revealed that FBS-inducible cell cycle progression was blocked by luteolin. Luteolin showed no cytotoxicity in VSMCs in this experimental condition according to WST-1 assays. Luteolin may represent a potential anti-proliferative agent for treatment of angioplasty restenosis and atherosclerosis.

Treatment Costs and Factors Associated with Glycemic Control among Patients with Diabetes in the United Arab Emirates

  • Lee, Seung-Mi;Song, Inmyung;Suh, David;Chang, Chongwon;Suh, Dong-Churl
    • Journal of Obesity & Metabolic Syndrome
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    • v.27 no.4
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    • pp.238-247
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    • 2018
  • Background: We aimed to estimate the proportion of patients with diabetes who achieved target glycemic control, to estimate diabetes-related costs attributable to poor control, and to identify factors associated with them in the United Arab Emirates. Methods: This retrospective cohort study used administrative claims data handled by Abu Dhabi Health Authority (January 2010 to June 2012) to determine glycemic control and diabetes-related treatment costs. A total of 4,058 patients were matched using propensity scores to eliminate selection bias between patients with glycosylated hemoglobin (HbA1c) <7% and HbA1c ${\geq}7%$. Diabetes-related costs attributable to poor control were estimated using a recycled prediction method. Factors associated with glycemic control were investigated using logistic regression and factors associated with these costs were identified using a generalized linear model. Results: During the 1-year follow-up period, 46.6% of the patients achieved HbA1c <7%. Older age, female sex, better insurance coverage, non-use of insulin in the index diagnosis month, and non-use of antidiabetic medications during the follow-up period were significantly associated with improved glycemic control. The mean diabetes-related annual costs were $2,282 and $2,667 for patients with and without glycemic control, respectively, and the cost attributable to poor glycemic control was $172 (95% confidence interval [CI], $164-180). The diabetes-related costs were lower with mean HbA1c levels <7% (cost ratio, 0.94; 95% CI, 0.88-0.99). The costs were significantly higher in patients aged ${\geq}65$ years than those aged ${\leq}44$ years (cost ratio, 1.45; 95% CI, 1.25-1.70). Conclusion: More than 50% of patients with diabetes had poorly controlled HbA1c. Poor glycemic control may increase diabetes-related costs.

Synthesis and evaluation of 64Cu-labeled avidin for lymph node imaging

  • Kang, Choong Mo;Kim, Hyunjung;Lee, Yong Jin;Choe, Yearn Seong
    • Journal of Radiopharmaceuticals and Molecular Probes
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    • v.5 no.1
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    • pp.54-60
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    • 2019
  • Sentinel lymph node (SLN) imaging plays an important role in surgery of patients with breast cancer and melanoma. In this study, avidin (Av), a tetrameric protein glycosylated with mannose and N-acetylglucosamine molecules, was labeled with $^{64}Cu$ and then evaluated for LN imaging. $^{64}Cu$-Labeled $NeutrAvidin^{TM}$ (NAv), a non-glycosylated form of Av, was used for comparison. 1,4,7,10-Tetraazacyclododecane-N,N',N'',N'''-tetraacetic acid (DOTA)-conjugated Av and NAv were prepared from the corresponding proteins and DOTA-NHS ester, which were then labeled with copper-64 and purified using PD-10 columns. The numbers of DOTA molecules conjugated to Av and NAv were 4.9 and 3.3, respectively. [$^{64}Cu$]Cu-DOTA-conjugated Av and NAv were prepared in 93% and 73% radiochemical yields, respectively. In vitro serum stability study showed that copper-64 remained stable on all radiotracers for 24 h (>97%). MicroPET/CT images showed that high radioactivity was accumulated in LNs within 15 min after footpad-injection of radiotracers. Tissue distribution data of mice demonstrated significantly higher uptake in the popliteal (PO) LN than lumbar (LU) LN for $^{64}Cu$-labeled Av (relative % ID/g excluding the injection sites: 66.2% and 26.0%, respectively) compared with those of $^{64}Cu$-labeled NAv (43.0% and 49.2%, respectively). The results of this study suggest that mannose molecules on Av enabled the radiotracer to retain in the first LN after mouse footpad-injection.

Molecular Cloning and Characterization of a Lipocalin in the Bumblebee Bombus Ignitus

  • Hu, Zhigang;Yoon, Hyung-Joo;Sohn, Hung-Dae;Jin, Byung-Rae
    • International Journal of Industrial Entomology and Biomaterials
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    • v.19 no.2
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    • pp.229-235
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    • 2009
  • We have cloned and characterized a lipocalin from the bumblebee Bombus ignitus (Bi-lipocalin). The Bi-lipocalin gene spans 2284 bp and consists of four exons coding for 270 amino acid residues. Sequence analysis revealed that Bi-lipocalin possesses three structurally conserved regions (SCTs) that characterize lipocalins. Recombinant Bi-lipocalin, expressed as a 37 kDa protein in baculovirus-infected insect cells, was N-glycosylated, indicating that the carbohydrate moieties are necessary for secretion. Tissue distribution analysis revealed ubiquitous expression of Bi-lipocalin in all tissues examined. Bi-lipocalin transcripts were upregulated by stress, such as wounding, $H_2O_2$ exposure, and external temperature shock. These results indicate that Bi-lipocalin is a stress-inducible protein that acts on wounding, $H_2O_2$ overexposure and temperature stimulation.

in vitro 에서 Human lactadherin gene의 발현 및 동정

  • 전길수;염행철
    • Proceedings of the Korean Society of Developmental Biology Conference
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    • 2003.10a
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    • pp.93-93
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    • 2003
  • 모유의 뮤신 복합체는 rotavirus에 특이적으로 결합하여 항 바이러스활동을 보여주는 것으로 나타났다. 자연 상태에서의 뮤신은 몇몇 작은 분자들과 복합적으로 연합되어 있는데 70kda의 glycoprotein, butyrophilin, 그리고 glycosylated component, lactadherin을 포함하고 있다. 그중 rotavirus에 가장 높은 결합력과 항바이러스 활동을 나타내는 Lactadherin은 모유의 유단백질의 하나인 뮤신과 결합되어 분비되는 당단백질의 하나로 분자량이 46kda이고, 지방구막 속에 연합되어있다. 이러한 배경에서 본 연구에서는 한국 여성의 breast tissue로부터 lactadherin 유전자의 cloning 및 in vitro에서의 발현을 유도하여 lactadherin band만을 purify하였고 여기서 얻은 lactadherin을 항체 생산을 위한 항원으로 사용하여 anti-lactadherin antibody를 확보하였다. 이 항체는 human milk에서의 lactadherin을 동정하는데 사용하였는데 western blot결과 lactadherin을 포함하여 몇 몇 단백질들이 확인되었다. Human milk내 mucin은 몇몇 작은 분자들과 복합체를 형성하는 것으로 확인되어졌는데 70kda의 glycoprotein, butyrophilin 그리고 46kda의 glyxosylated component, lactadherin을 포함하고 있는 milk mucin은 associated molecular임을 확인할 수 있었다.

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Luteolin Induces the Differentiation of Osteoblasts

  • Ko, Seon-Yle
    • International Journal of Oral Biology
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    • v.35 no.3
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    • pp.99-106
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    • 2010
  • Luteolin is a flavonoid that exists in a glycosylated form in celery and green pepper. Flavonoids possess antioxidant and anti-inflammatory properties and can reduce the expression of key inflammatory molecules in macrophages and monocytes. It has been reported also that some flavonoids have effects on bone metabolism. The effects of luteolin on the function of osteoblasts were investigated by measuring cell viability, alkaline phosphatase activity, type I collagen production, osteoprotegerin secretion, Wnt promoter activity, BMP-2 and Runx2 expression and calcified nodule formation. Luteolin has no effects upon osteoblast viability but induced an increase in alkaline phosphatase activity, type I collagen production and a decrease in osteoprotegerin secretion in these cells. Luteolin treatment also upregulated BMP-2 mRNA expression. These results suggest that luteolin may be a regulatory molecule that facilitates the differentiation of osteoblasts.