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Eun Jeong Park Yoshikazu Yuki Hiroshi Kiyono Motomu Shimaoka 《Journal of biomedical science》2015,22(1)
Integrins mediate leukocyte accumulation to the sites of inflammation, thereby enhancing their potential as an important therapeutic target for inflammatory disorders. Integrin activation triggered by inflammatory mediators or signaling pathway is a key step to initiate leukocyte migration to inflamed tissues; however, an appropriately regulated integrin deactivation is indispensable for maintaining productive leukocyte migration. While typical integrin antagonists that block integrin activation target the initiation of leukocyte migration, a novel class of experimental compounds has been designed to block integrin deactivation, thereby perturbing the progression of cell migration. Current review discusses the mechanisms by which integrin is activated and subsequently deactivated by focusing on its structure-function relationship. 相似文献
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A continuous single organic phase process for the lipase catalyzed synthesis of peroxy acids increases productivity
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Janine Meyer Angelika E. W. Horst Max Steinhagen Dirk Holtmann Marion B. Ansorge‐Schumacher Matthias Kraume Anja Drews 《Engineering in Life Science》2017,17(7):759-767
The design of an optimal process is particularly crucial when the reactants deactivate the biocatalyst. The reaction cascades of the chemo‐enzymatic epoxidation where the intermediate peroxy acid is produced by an enzyme are still limited by enzyme inhibition and deactivation by hydrogen peroxide. To avoid additional effects caused by interfaces (aq/org) and to reduce the process limiting deactivation by the substrate hydrogen peroxide, a single‐phase concept was applied in a fed‐batch and a continuous process (stirred tank), without the commonly applied addition of a carrier solvent. The synthesis of peroxyoctanoic acid catalyzed by Candida antarctica lipase B was chosen as the model reaction. Here, the feasibility of this biocatalytic reaction in a single‐phase system was shown for the first time. The work shows the economic superiority of the continuous process compared to the fed‐batch process. Employing the fed‐batch process reaction rates up to 36 mmol h?1 per gramcat, and a maximum yield of 96 % was achieved, but activity dropped quickly. In contrast, continuous operation can maintain long‐term enzyme activity. For the first time, the continuous enzymatic reaction could be performed for 55 h without any loss of activity and with a space‐time yield of 154 mmol L?1 h?1, which is three times higher than in the fed‐batch process. The higher catalytic productivity compared to the fed‐batch process (34 vs. 18 gProd g?1cat) shows the increased enzyme stability in the continuous process. 相似文献
4.
Baran J Baj-Krzyworzeka M Weglarczyk K Ruggiero I Zembala M 《Cancer immunology, immunotherapy : CII》2004,53(12):1127-1134
Immunotherapy with Mycobacterium vaccae as an adjuvant to chemotherapy has recently been applied to treatment of patients with cancer. One of the mechanisms of antitumour activity of Mycobacterium bovis bacillus Calmette-Guérin (BCG), the prototype immunomodulator, is associated with activation of monocytes/macrophages. These studies were undertaken to determine how M. vaccae affects monocyte–tumour cell interactions and, in particular, whether it can prevent or reverse deactivation of monocytes that occurrs following their contact with tumour cells during coculture in vitro. Deactivation is characterised by the impaired ability of monocytes to produce tumour necrosis factor (TNF-), interleukin 12 (IL-12), and enhanced IL-10 secretion following their restimulation with tumour cells. To see whether deactivation of monocytes can be either prevented or reversed, three different strains of M. vaccae—B 3805, MB 3683, and SN 920—and BCG were used to stimulate monocytes before or after exposure to tumour cells. Pretreatment of monocytes with M. vaccae MB 3683, SN 920 and BCG before coculture resulted in increased TNF- and decreased IL-10 production. All strains of M. vaccae and BCG used for treatment of deactivated monocytes enhanced depressed TNF- secretion. Strain SN 920 and BCG increased IL-12 release but only BCG treatment inhibited an enhanced IL-10 production by deactivated monocytes. Thus, although some strains of M. vaccae may either prevent or reverse tumour-induced monocyte deactivation, none of them appears to be more effective than BCG. 相似文献
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Krisztián Cser 《BBA》2007,1767(3):233-243
The mechanism of charge recombination was studied in Photosystem II by using flash induced chlorophyll fluorescence and thermoluminescence measurements. The experiments were performed in intact cells of the cyanobacterium Synechocystis 6803 in which the redox properties of the primary pheophytin electron acceptor, Phe, the primary electron donor, P680, and the first quinone electron acceptor, QA, were modified. In the D1Gln130Glu or D1His198Ala mutants, which shift the free energy of the primary radical pair to more positive values, charge recombination from the S2QA− and S2QB− states was accelerated relative to the wild type as shown by the faster decay of chlorophyll fluorescence yield, and the downshifted peak temperature of the thermoluminescence Q and B bands. The opposite effect, i.e. strong stabilization of charge recombination from both the S2QA− and S2QB− states was observed in the D1Gln130Leu or D1His198Lys mutants, which shift the free energy level of the primary radical pair to more negative values, as shown by the retarded decay of flash induced chlorophyll fluorescence and upshifted thermoluminescence peak temperatures. Importantly, these mutations caused a drastic change in the intensity of thermoluminescence, manifested by 8- and 22-fold increase in the D1Gln130Leu and D1His198Lys mutants, respectively, as well as by a 4- and 2.5-fold decrease in the D1Gln130Glu and D1His198Ala mutants, relative to the wild type, respectively. In the presence of the electron transport inhibitor bromoxynil, which decreases the redox potential of QA/QA− relative to that observed in the presence of DCMU, charge recombination from the S2QA− state was accelerated in the wild type and all mutant strains. Our data confirm that in PSII the dominant pathway of charge recombination goes through the P680+Phe− radical pair. This indirect recombination is branched into radiative and non-radiative pathways, which proceed via repopulation of P680* from 1[P680+Ph−] and direct recombination of the 3[P680+Ph−] and 1[P680+Ph−] radical states, respectively. An additional non-radiative pathway involves direct recombination of P680+QA−. The yield of these charge recombination pathways is affected by the free energy gaps between the Photosystem II electron transfer components in a complex way: Increase of ΔG(P680* ↔ P680+Phe−) decreases the yield of the indirect radiative pathway (in the 22-0.2% range). On the other hand, increase of ΔG(P680+Phe− ↔ P680+QA−) increases the yield of the direct pathway (in the 2-50% range) and decreases the yield of the indirect non-radiative pathway (in the 97-37% range). 相似文献
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《Biocatalysis and Biotransformation》2013,31(5-6):413-425
In developing enzyme-catalysed processes, a highly active, selective and especially stable biocatalyst forms the basis for an economic industrial technology. Enzymatic stability is one of the most important properties, but there are no good methods for determining it quickly and efficiently. In order to solve this problem, a new method for the “accelerated measurement of activity and stability of enzymes (AMASE)” has been developed. Compared to conventional methods, the new approach differs in two main points. First, the process of ageing is accelerated by a permanent increase in temperature during a continuous experiment, which generates a non-stable response of the enzyme-reactor system. Second, the evaluation procedure is strongly based on a mathematical-mechanistic approach consisting of four steps: assume a deactivation mechanism, formulate the mathematical model, determine the model parameters and finally calculate the biocatalyst characteristics. In this paper, the application of the method for the Lipozyme IM-catalysed hydrolysis of triglycerides in different media is presented. 相似文献
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Sakatani K Yamashita D Yamanaka T Oda M Yamashita Y Hoshino T Fujiwara N Murata Y Katayama Y 《Life sciences》2006,78(23):2734-2741
To determine the alterations in optical characteristics and cerebral blood oxygenation (CBO) during activation and deactivation, we evaluated the changes in mean optical pathlength (MOP) and CBO induced by a verbal fluency task (VFT) and driving simulation in the right and left prefrontal cortex (PFC), employing a newly developed time-resolved near infrared spectroscopy, which allows quantitative measurements of the evoked-CBO changes by determining the MOP with a sampling time of 1 s. The results demonstrated differences in MOP in the foreheads with the subjects and wavelength; however, there was no significant difference between the right and left foreheads (p > 0.05). Also, both the VFT and driving simulation task did not affect the MOP significantly as compared to that before the tasks (p > 0.05). In the bilateral PFCs, the VFT caused increases of oxyhemoglobin and total hemoglobin associated with a decrease of deoxyhemoglobin, while the driving simulation task caused decreases of oxyhemoglobin and total hemoglobin associated with an increase of deoxyhemoglobin; there were no significant differences in evoked-CBO changes between the right and left PFC. The present results will be useful for quantitative measurement of hemodynamic changes during activation and deactivation in the adults by near infrared spectroscopy. 相似文献
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Jiyeon Lee Hye-Jin Kim Jooho Roh Youngsil Seo Minjae Kim Hye-Ryeong Jun Chuong D. Pham Myung-Hee Kwon 《The Journal of biological chemistry》2013,288(50):35877-35885
Many murine monoclonal anti-DNA antibodies (Abs) derived from mice models for systemic lupus erythematosus have additional cell-penetration and/or nucleic acid-hydrolysis properties. Here, we examined the influence of deactivating each complementarity-determining region (CDR) within a multifunctional anti-nucleic acid antibody (Ab) that possesses these activities, the catalytic 3D8 single chain variable fragment (scFv). CDR-deactivated 3D8 scFv variants were generated by replacing all of the amino acids within each CDR with Gly/Ser residues. The structure of 3D8 scFv accommodated single complete CDR deactivations. Different functional activities of 3D8 scFv were affected differently depending on which CDR was deactivated. The only exception was CDR1, located within the light chain (LCDR1); deactivation of LCDR1 abolished all of the functional activities of 3D8 scFv. A hybrid Ab, HW6/3D8L1, in which the LCDR1 from an unrelated Ab (HW6) was replaced with the LCDR1 from 3D8, acquired all activities associated with the 3D8 scFv. These results suggest that the activity of a multifunctional 3D8 scFv Ab can be modulated by single complete CDR deactivation and that the LCDR1 plays a crucial role in maintaining Ab properties. This study presents a new approach for determining the role of individual CDRs in multifunctional Abs with important implications for the future of Ab engineering. 相似文献
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Recent studies have suggested a role for a chloride current in the modulation of pacemaker potentials generated by interstitial
cells of Cajal. Patch-clamp recordings were made from inside–out patches of cultured interstitial cells of Cajal from the
murine small intestine. The majority of patches were quiescent immediately after excision, but in some patches currents activated
spontaneously after a period of 10 min to 1 h. Currents could also be activated by strongly polarizing the patch. It was found
that the currents activated in both cases included a chloride channel. This channel could also be activated by ATP and the
catalytic subunit of protein kinase A. The channel had conductance states (±SD) of 53 ± 25.35, 126 ± 21.44, 180 ± 12.57 and
211 ± 8.86 pS. It was outwardly rectifying (as a function of open probability) and deactivated (i.e., gave a tail current)
but showed no inactivation. The permeability sequence of the channel was I−>>Br−≥Cl−>Asp−. It was unaffected in magnitude or rectification by changing the free Ca2+ concentration of the bath between <10 nm, 100 nm (control) and 2 mm. 相似文献
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Chuanyi Yao Shaokun Tang Jinhong Zhang Yaoting Yu 《Journal of Molecular Catalysis .B, Enzymatic》2002,18(4-6):279-284
The stability of lipase in AOT/isooctane reversed micellar solution was investigated. It was found that the lipase deactivated to a stable state that was not completely inactivated. The lipase residual activity after achieving the stable state in AOT/isooctane reversed micelles at 30 °C, pH 7.0, W0=8.0 was found to be 0.15, and the first-order deactivation rate coefficient of lipase at the same conditions was regressed to be 0.75 h−1. The stability of lipase was increased while oleic acid was added. Assuming the protection of oleic acid to lipase stability is due to the lipase–oleic acid complex does not decay, the kinetic model of lipase deactivation in AOT/isooctane reversed micellar solution including the influence of oleic acid was established. It was shown with the model equation that the increase in stability of the enzyme by oleic acid could be quantitatively estimated by the dissociation constant of lipase–oleic acid complex which was determined by product inhibition experiments. The model equation fit the experimental data well with an average relative deviation of 3.40%. 相似文献