Compensatory UTE/T2W Imaging of Inflammatory Vascular Wall in Hyperlipidemic Rabbits

ObjectivesTo obtain compensatory ultra-short echo time (UTE) imaging and T2-weighted (T2W) imaging of Watanabe heritable hyperlipidemic (WHHL) rabbits following dextran-coated magnetic nanocluster (DMNC) injection for the effective in vivo detection of inflammatory vascular wall.MethodsMagnetic nanoparticle was synthesized by thermal decomposition and encapsulated with dextran to prepare DMNC. The contrast enhancement efficiency of DMNC was investigated using UTE (repetition time [TR] = 5.58 and TE = 0.07 ms) and T2W (TR = 4000 and TE = 60 ms) imaging sequences. To confirm the internalization of DMNC into macrophages, DMNC-treated macrophages were visualized by cellular transmission electron microscope (TEM) and magnetic resonance (MR) imaging. WHHL rabbits expressing macrophage-rich plaques were subjected to UTE and T2W imaging before and after intravenous DMNC (120 μmol Fe/kg) treatment. Ex vivo MR imaging of plaques and immunostaining studies were also performed.ResultsPositive and negative contrast enhancement of DMNC solutions with increasing Fe concentrations were observed in UTE and T2W imaging, respectively. The relative signal intensities of the DMNC solution containing 2.9 mM Fe were calculated as 3.53 and 0.99 in UTE and T2W imaging, respectively. DMNC uptake into the macrophage cytoplasm was visualized by electron microscopy. Cellular MR imaging of DMNC-treated macrophages revealed relative signals of 3.00 in UTE imaging and 0.98 in T2W imaging. In vivo MR images revealed significant brightening and darkening of plaque areas in the WHHL rabbit 24 h after DMNC injection in UTE and T2W imaging, respectively. Ex vivo MR imaging results agreed with these in vivo MR imaging results. Histological analysis showed that DMNCs were localized to areas of inflammatory vascular wall.ConclusionsUsing compensatory UTE and T2W imaging in conjunction with DMNC is an effective approach for the noninvasive in vivo imaging of atherosclerotic plaque.     

Oroxylin A,a Flavonoid,Stimulates Adult Neurogenesis in the Hippocampal Dentate Gyrus Region of Mice

Previously, we reported the cognitive enhancing effects of oroxylin A in unimpaired mice and its memory ameliorating activity in various memory impaired mice. To elucidate the mechanism mediating the cognitive effects of oroxylin A, this study examined the consequences of oroxylin A administration on neurogenesis in the hippocampal dentate gyrus using immunostaining for 5-bromo-2-deoxyuridine (BrdU) incorporation. In addition, we determined whether the new cells adopted a neuronal or glial fate by examining the co-localization of BrdU staining with neuronal or glial markers. Administration of oroxylin A in a dose-dependent and time-dependent manner increased the number of BrdU-incorporating cells. Moreover, the percentage of BrdU-incorporating cells co-localized with neuronal markers, neuronal nuclei, was significantly increased by the oroxylin A administration. These results suggest that the increased neurogenesis induced by the administration of oroxylin A could be, at least in part, associated with its positive effects on cognitive processing.     

Regenerative potential of mouse embryonic stem cell-derived PDGFRα+ cardiac lineage committed cells in infarcted myocardium

BACKGROUND Pluripotent stem cell-derived cardiomyocytes(CMs) have become one of the most attractive cellular resources for cell-based therapy to rescue damaged cardiac tissue.AIM We investigated the regenerative potential of mouse embryonic stem cell(ESC)-derived platelet-derived growth factor receptor-α(PDGFRα)+ cardiac lineagecommitted cells(CLCs), which have a proliferative capacity but are in a morphologically and functionally immature state compared with differentiated CMs.METHODSWe induced mouse ESCs into PDGFRα+ CLCs and αMHC+ CMs using a combination of the small molecule cyclosporin A, the rho-associated coiled-coil kinase inhibitor Y27632, the antioxidant Trolox, and the ALK5 inhibitor EW7197.We implanted PDGFRα+ CLCs and differentiated αMHC+ CMs into a myocardial infarction(MI) murine model and performed functional analysis using transthoracic echocardiography(TTE) and histologic analysis.RESULTS Compared with the untreated MI hearts, the anterior and septal regional wall motion and systolic functional parameters were notably and similarly improved in the MI hearts implanted with PDGFRα+ CLCs and αMHC+ CMs based on TTE.In histologic analysis, the untreated MI hearts contained a thinner ventricular wall than did the controls, while the ventricular walls of MI hearts implanted with PDGFRα+ CLCs and αMHC+ CMs were similarly thicker compared with that of the untreated MI hearts. Furthermore, implanted PDGFRα+ CLCs aligned and integrated with host CMs and were mostly differentiated into α-actinin+ CMs,and they did not convert into CD31+ endothelial cells or αSMA+ mural cells.CONCLUSION PDGFRα+ CLCs from mouse ESCs exhibiting proliferative capacity showed a regenerative effect in infarcted myocardium. Therefore, mouse ESC-derived PDGFRα+ CLCs may represent a potential cellular resource for cardiac regeneration.     

Enzymatic esterification of beta-methylglucoside with acrylic/methacrylic acid in organic solvents

The enzymatic esterifications of beta-methylglucoside with acrylic acid/methacrylic acid were carried out using Novozym 435. t-Butanol indicating the highest conversion value was determined as an optimal solvent. The molar ratio (beta-methylglucoside:acids) of 1:15 was most favorable to the esterification. The enzyme concentration of 5% (w/v), and the temperature (50 degrees C for beta-methylglucoside:acrylic acid, 45 degrees C for beta-methylglucoside:methacrylic acid) resulted in the highest final conversion. Beta-methylglucoside of 60gl(-1) was found to be most effective in terms of short reaction time as well as product concentrations. Under these conditions, the maximum conversions for the esterification of beta-methylglucoside with acrylic acid and beta-methylglucoside with methacrylic acid were 59.3% after 12h and 71.3% after 72h, respectively. The structural analysis of the products was performed by FT-IR spectroscopy and (1)H NMR.     

Enhanced secretion of<Emphasis Type="Italic"> Bacillus stearothermophilus</Emphasis> L1 lipase in<Emphasis Type="Italic"> Saccharomyces cerevisiae</Emphasis> by translational fusion to cellulose-binding domain

The secretion of Bacillus stearothermophilus L1 lipase in Saccharomyces cerevisiae was investigated by employing a fusion partner, a cellulose-binding domain (CBD) from Trichoderma harzianum endoglucanase II (THEG). The CBD was connected to the N-terminal of L1 lipase through an endogenous linker peptide from THEG. The expression cassette for the fusion protein in S. cerevisiae was constructed using the -amylase signal peptide and the galactose-inducible GAL10 promoter. Secretion of CBD-linker-L1 lipase by this fusion construct was dramatically 7-fold enhanced, compared with that of the mature L1 lipase without CBD-fusion. The fusion protein was secreted into the culture medium, reaching levels of approximately 1.3 g/l in high-cell-density fed-batch cultures. Insertion of a KEX2 cleavage site into the junction between CBD-linker and L1 lipase resulted in the same level of enhanced secretion, indicating that the CBD-linker fusion probably plays a critical role in secretion from endoplasmic reticulum to Golgi apparatus. Therefore, the CBD from THEG can be used both as an affinity tag and as a secretion enhancer for the secretory production of heterologous proteins in S. cerevisiae, since in vivo breakage at the linker was almost negligible.     

Enzymatic synthesis of alkylglucosides by amphiphilic phase enzyme reaction

Alkyl--glucosides were synthesized with -glucosidase, from almond, in an amphiphilic phase reaction system using different water-miscible cosolvents. By means of stepwise addition of glucose, we obtained about 1.5 times higher concentration of hexylglucoside than that of normal reaction. Using the same way, octyl-, decyl- and dodecylglucoside were synthesized and their concentrations were 6.64, 5.3 and 4.48 g l^–1, respectively.     

A modular control design method for a flexible manufacturing cell including error handling

This paper introduces a new modular control design method for a cell controller with integrated error handling. To make the complexity of the cell controller manageable, its control logic is separated into two parts: resource allocation control and operation control. To create the operation control not only quickly but correctly, modular operation blocks integrated with error handling are developed. An algorithm automatically generates the operation control. The operation control created by the proposed method is proved to have desired control behaviors. The method is applied to an example system.