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A selective molecularly imprinted polymer for immobilization of acetylcholinesterase (AChE): an active enzyme targeted and efficient method
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Gökhan Demirci Yasemin İspirli Doğaç Mustafa Teke 《Journal of molecular recognition : JMR》2015,28(11):645-650
In the present study, we immobilized acetylcholinesterase (AChE) enzyme onto acetylcholine removed imprinted polymer and acetylcholine containing polymer. First, the polymers were produced with acetylcholine, substrate of AChE, by dispersion polymerization. Then, the enzyme was immobilized onto the polymers by using two different methods: In the first method (method A), acetylcholine was removed from the polymer, and then AChE was immobilized onto this polymer (acetylcholine removed imprinted polymer). In the second method (method B), AChE was immobilized onto acetylcholine containing polymer by affinity. In method A, enzyme‐specific species (binding sites) occurred by removing acetylcholine from the polymer. The immobilized AChE reached 240% relative specific activity comparison with free AChE because the active enzyme molecules bounded onto the polymer. Transmission electron microscopy results were taken before and after immobilization of AChE for the assessment of morphological structure of polymer. Also, the experiments, which include optimum temperature (25–65°C), optimum pH (3–10), thermal stability (4–70°C), kinetic parameters, operational stability and reusability, were performed to determine the characteristic of the immobilized AChE. Copyright © 2015 John Wiley & Sons, Ltd. 相似文献
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Guanmei Wen Cheng Zhang Qishan Chen Le Anh Luong Arif Mustafa Shu Ye Qingzhong Xiao 《The Journal of biological chemistry》2015,290(31):19158-19172
Matrix metalloproteinase-8 (MMP8) has been shown to influence various cellular functions. As monocytes and macrophages (Mφ) express MMP8, we investigated if MMP8 played a role in macrophage differentiation and polarization. MMP8 expression was significantly increased during monocyte differentiation into Mφ. Monocyte-derived Mφ from MMP8-deficient mice expressed higher levels of M1-Mφ markers but lower levels of M2-Mφ markers than monocyte-derived Mφ from wild-type mice. Although Mφ from either MMP8-deficient or wild-type mice were inducible by interferon-γ into M1-Mφ, only wild-type Mφ but not MMP8-deficient Mφ could be induced into M2-Mφ by interleukin-4. However, MMP8-deficient Mφ exposed to conditioned culture media of wild-type Mφ developed a M2-Mφ phenotype. Compared with conditioned culture media of wild-type Mφ, conditioned culture media of MMP8-deficient Mφ contained a lower concentration of active transforming growth factor-β (TGF-β), an M2-Mφ inducer. Moreover, evidence also showed that the degradation of the TGF-β sequester, fibromodulin, was modulated by MMP8. The data indicate a previously unknown role of MMP8 in M2-Mφ polarization by cleaving fibromodulin and therefore increasing the bioavailability of the M2-Mφ inducer TGF-β. 相似文献
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G?k?e Güllü Irem Peker Aptullah Haholu Fatih Eren Zafer Kü?ükodaci Bülent Güle? Hüseyin Baloglu Can Erzik Ayse ?zer Mustafa Akkiprik 《Genetics and molecular biology》2015,38(1):21-29
The functional role of IGFBP5 in breast cancer is complicated. Experimental and
bioinformatics studies have shown that IGFBP5 is targeted by miR-140-5p and miR-193b,
although this has not yet been proven in clinical samples. The aim of this study was
to evaluate the expression of miR-140-5p and miR-193b in breast cancer and adjacent
normal tissue and assess its correlation with IGFBP5 and the clinicopathological
characteristics of the tumors. IGFBP5 protein expression was analyzed
immunohistochemically and IGFBP5, miR-140 and miR-193b mRNA expression levels were
analyzed with real-time RT-PCR. Tumor tissue had higher miR-140-5p expression than
adjacent normal tissue (p = 0.015). Samples with no immunohistochemical staining for
IGFBP5 showed increased miR-140-5p expression (p = 0.009). miR-140-5p expression was
elevated in invasive ductal carcinomas (p = 0.002), whereas basal-like tumors had
decreased expression of miR-140-5p compared to other tumors (p = 0.008). Lymph
node-positive samples showed an approximately 13-fold increase in miR-140-5p
expression compared to lymph node-negative tissue (p = 0.049). These findings suggest
that miR-140-5p, but not miR-193b, could be an important determinant of IGFBP5
expression and clinical phenotype in breast cancer patients. Further studies are
needed to clarify the expressional regulation of IGFBP5 by miR-140-5p. 相似文献