Soluble Prokaryotic Overexpression and Purification of Human GM-CSF Using the Protein Disulfide Isomerase b′a′ Domain

Granulocyte-macrophage colony-stimulating factor (GM-CSF) is a member of the colony-stimulating factor (CSF) family, which functions to enhance the proliferation and differentiation of hematopoietic stem cells and other hematopoietic lineages such as neutrophils, dendritic cells, or macrophages. These proteins have thus generated considerable interest in clinical therapy research. A current obstacle to the prokaryotic production of human GM-CSF (hGM-CSF) is its low solubility when overexpressed and subsequent complex refolding processes. In our present study, the solubility of hGM-CSF was examined when combined with three N-terminal fusion tags in five E. coli strains at three different expression temperatures. In the five E. coli strains BL21 (DE3), ClearColi BL21 (DE3), LOBSTR, SHuffle T7 and Origami2 (DE3), the hexahistidine-tagged hGM-CSF showed the best expression but was insoluble in all cases at each examined temperature. Tagging with the maltose-binding protein (MBP) and the b′a′ domain of protein disulfide isomerase (PDIb′a′) greatly improved the soluble overexpression of hGM-CSF at 30 °C and 18 °C. The solubility was not improved using the Origami2 (DE3) and SHuffle T7 strains that have been engineered for disulfide bond formation. Two conventional chromatographic steps were used to purify hGM-CSF from the overexpressed PDIb′a′-hGM-CSF produced in ClearColi BL21 (DE3). In the experiment, 0.65 mg of hGM-CSF was isolated from a 0.5 L flask culture of these E. coli and showed a 98% purity by SDS-PAGE analysis and silver staining. The bioactivity of this purified hGM-CSF was measured at an EC₅₀ of 16.4 ± 2 pM by a CCK8 assay in TF-1 human erythroleukemia cells.     

Simulated Microgravity Inhibits the Proliferation of Chang Liver Cells by Attenuation of the Major Cell Cycle Regulators and Cytoskeletal Proteins

Simulated microgravity (SMG) induced the changes in cell proliferation and cytoskeleton organization, which plays an important factor in various cellular processes. The inhibition in cell cycle progression has been considered to be one of the main causes of proliferation inhibition in cells under SMG, but their mechanisms are still not fully understood. This study aimed to evaluate the effects of SMG on the proliferative ability and cytoskeleton changes of Chang Liver Cells (CCL-13). CCL-13 cells were induced SMG by 3D clinostat for 72 h, while the control group were treated in normal gravity at the same time. The results showed that SMG reduced CCL-13 cell proliferation by an increase in the number of CCL-13 cells in G0/G1 phase. This cell cycle phase arrest of CCL-13 cells was due to a downregulation of cell cycle-related proteins, such as cyclin A1 and A2, cyclin D1, and cyclin-dependent kinase 6 (Cdk6). SMG-exposed CCL-13 cells also exhibited a downregulation of α-tubulin 3 and β-actin which induced the cytoskeleton reorganization. These results suggested that the inhibited proliferation of SMG-exposed CCL-13 cells could be associate with the attenuation of major cell cycle regulators and main cytoskeletal proteins.     

A technique for extracting behavioral sequence patterns from GPS recorded data

The mobile wireless market has been attracting many customers. Technically, the paradigm of anytime-anywhere connectivity raises previously unthinkable challenges, including the management of million of mobile customers, their profiles, the profiles-based selective information dissemination, and server-side computing infrastructure design issues to support such a large pool of users automatically and intelligently. In this paper, we propose a data mining technique for discovering frequent behavioral patterns from a collection of trajectories gathered by Global Positioning System. Although the search space for spatiotemporal knowledge is extremely challenging, imposing spatial and temporal constraints on spatiotemporal sequences makes the computation feasible. Specifically, the mined patterns are incorporated with synthetic constraints, namely spatiotemporal sequence length restriction, minimum and maximum timing gap between events, time window of occurrence of the whole pattern, inclusion or exclusion event constraints, and frequent movement patterns predictive of one ore more classes. The algorithm for mining all frequent constrained patterns is named cAllMOP. Moreover, to control the density of pattern regions a clustering algorithm is exploited. The proposed method is efficient and scalable. Its efficiency is better than that of the previous algorithms AllMOP and GSP with respect to the compactness of discovered knowledge, execution time, and memory requirement.     

Small amphiphilic organics, coordination extended solids, and constant curvature structures

In this Account, we explore the role constant curvature structures play in amphiphilic small molecule crystals and extended coordination solids. A constant curvature structure is one in which there is a surface or interface that has the same curvature throughout its surface. Simple examples of such structures contain spheres (micelles), columns, or layers. Yet another family are cubic as is found in the gyroid topology. For amphiphilic systems, there are two domains, one generally hydrophobic and the other hydrophilic. We find that the interfaces between these two domains in small molecule structures resemble those in larger scale systems and adopt topologies related to constant curvature structures. The hydrophobic-to-total volume ratio, a molecular parameter, can be used to predict which type of constant curvature structure is adopted. In the case of three coordinate extended solids, constant curvature plays a role both in the extended net topology and in the hydrophobic-to-hydrophilic interface.     

Darstellung,Struktur und Reaktivität quartärer Ammoniumcellulosate

Tetraalkylammonium cellulosates were prepared by means of an exchange reaction between cellulose and tetraalkylammonium methoxides in anhydrous methanol and dimethylsulfoxide up to a degree of substitution of about 0,7 without chain degradation. The course of this reaction and the structure of the cellulosates in comparison to lithium and sodium cellulosate has been studied. The reaction rate and the equilibrium substitution decreased with increasing size of the cation in the methoxide. The distribution of the alcoholate groups in equilibrium at C-2, C-3 and C-6 of the glucopyranosyl unit in the cellulosates, which was investigated by methylation, hydrolysis of the methylcelluloses, and analysis of the methylglucoses by gas chromatography, was independent of the nature of the cation. Under the given reaction conditions the hydroxyl group at C-2 was more acidic than those at C-3 and C-6, while for the latter practically no difference was found. The tetraalkylammonium cellulosates are highly reactive intermediates for the preparation of cellulose derivatives. The reactivity of the cellulosates increased with increasing size of the cation, Li^⊕ < Na^⊕ < N(CH₃)₄^⊕ <[(CH₃)₃N? CH₂? C₆H₅]^⊕.     

Hepatic Δ9 and Δ6 desaturase activities during the recovery period following carbon tetrachloride poisoningdesaturase activities during the recovery period following carbon tetrachloride poisoning

The liver microsomal Δ⁹ and Δ⁶ desaturase activities have been studied in rats with carbon tetrachloride-induced hepatitis. Immediately after poisoning, significant decreases were observed for both types of desaturase activity. However, recovery kinetics were slower for the Δ⁶ desaturase than for the Δ⁹ desaturase. The activities of NADH-ferricyanide and NADH-cytochrome C reductases, proteins involved in the electron transfers associated with microsomal desaturation, were also measured. There was a fall in both activities after poisoning, but this decrease was less than that of the desaturase activities.     

Efficacy of iron fortification compared to iron supplementation among Vietnamese schoolchildren

Background  

Parents of six children are facing a trial on charges of aggravated manslaughter in the care a 5 1/2 month old infant who died suddenly and neglect of their four older children for causing them to be malnourished by feeding them all an exclusively raw foods vegan diet. Both parents declined plea bargains and plan to defend themselves in court.     

Page segmentation using minimum homogeneity algorithm and adaptive mathematical morphology

Document layout analysis or page segmentation is the task of decomposing document images into many different regions such as texts, images, separators, and tables. It is still a challenging problem due to the variety of document layouts. In this paper, we propose a novel hybrid method, which includes three main stages to deal with this problem. In the first stage, the text and non-text elements are classified by using minimum homogeneity algorithm. This method is the combination of connected component analysis and multilevel homogeneity structure. Then, in the second stage, a new homogeneity structure is combined with an adaptive mathematical morphology in the text document to get a set of text regions. Besides, on the non-text document, further classification of non-text elements is applied to get separator regions, table regions, image regions, etc. The final stage, in refinement region and noise detection process, all regions both in the text document and non-text document are refined to eliminate noises and get the geometric layout of each region. The proposed method has been tested with the dataset of ICDAR2009 page segmentation competition and many other databases with different languages. The results of these tests showed that our proposed method achieves a higher accuracy compared to other methods. This proves the effectiveness and superiority of our method.