Influence of receptor number on the stimulation by salmeterol of gene transcription in CHO-K1 cells transfected with the human beta2-adrenoceptor

We investigated the behavior of the antioxidant enzymes, superoxide dismutase (SOD), catalase (CAT), and ascorbate peroxidase (APx), in potato tubers during storage at low temperature. SOD activity increased temporarily within 3 weeks and was higher at 1 degree C than at 20 degrees C. APx activity also increased more at low (1 degree C) than at higher temperatures (5 and 20 degrees C). The contents of ascorbic acid (AsA), which is the substrate of APx, decreased immediately within 3 weeks and then gradually decreased until 15 weeks. The activity of CAT, the other enzyme which can scavenge hydrogen peroxide, decreased once in the first six weeks and thereafter increased to 15 weeks. Thus, the enhancement of the active oxygen-scavenging system that was induced by low temperature in potato tubers could result not only in a decrease of AsA but also in combined increases in APx and CAT activity whose manners were different.     

Identification of a novel inhibitor (NSC 665564) of dihydroorotate dehydrogenase with a potency equivalent to brequinar

A novel inhibitor of dihydroorotate dehydrogenase (DHO-DH) has been discovered using data from the National Cancer Institute's in vitro drug screen. Upon analysis of cytotoxicity results from the sixty tumor cell lines used in this screen, the COMPARE program predicted that NSC 665564 was likely to have the same mechanism of inhibition as brequinar, a known potent inhibitor of DHO-DH. We validated this prediction experimentally using MOLT-4 lymphoblast and found the IC50 of brequinar (0.5 microM) and NSC 665564 (0.3 microM) were comparable and that this induced cytotoxicity was reversed by either uridine or cytidine. The enzyme target of NSC 665564 was shown to be identical to that of brequinar when incubation with each drug followed by a 1 h pulse with [14C] sodium bicarbonate resulted in cellular accumulation of [14C]N-carbamyl-L-aspartic acid and [14C]L-dihydroorotic acid, with concurrent marked depletion of CTP and UTP. The Ki's for NSC 665564 and brequinar were 0.14 and 0.24 microM, respectively, when partially purified MOLT-4 mitochondria (the site of DHO-DH) were used. These results show that mechanistic predictions obtained using correlations from the COMPARE algorithm are independent of structure since the structure of NSC 665564 is dissimilar to that of other established DHO-DH inhibitors.     

Screening for diabetic retinopathy: the utility of nonmydriatic retinal photography in Egyptian adults

BACKGROUND AND OBJECTIVE: Drug resistance has become a major cause of treatment failure in patients with acute leukemia. P-glycoprotein (Pgp), which is associated with the multidrug resistance (MDR) phenotype, has been reported to be an important predictor of treatment outcome. The aim of this study was to analyze the value of Pgp expression in bone marrow or peripheral blood as a predictor of the response to remission induction chemotherapy as well as the duration of remission in patients with de novo acute myeloid leukemia (AML). DESIGN AND METHODS: We examined the expression of Pgp in 82 patients with de novo AML using an immunocytochemical assay with the C219 monoclonal antibody. RESULTS: Twenty-seven of the 82 patients (33%) were C219-positive in from 1% to 100% of their cells. Thirteen cases (16%) showed a positive reaction in more than 50% of the leukemic cells. Only hyperleukocytosis was significantly associated with higher expression of Pgp. Although 8 of the 13 cases (62%) with more than 50% of cells having Pgp expression were CD34-positive, this association was not statistically significant. A univariate analysis of resistance to induction therapy showed a significantly higher resistance rate in patients with increased Pgp expression (P = 0.01) as well as in those patients with decreased reactivity to myeloperoxidase. The multivariate analysis revealed the independent prognostic value of Pgp expression. C219 reactivity did not have an influence on remission duration. INTERPRETATION AND CONCLUSIONS: Our data indicate that P-glycoprotein expression is a reliable marker of resistance to induction treatment in patients with de novo AML.     

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.     

Influence of mixing cycle on the degree of mixing of calcite‐filled polyethylene upon stretching

The mixing cycle‐dependent degree of dispersion and degree of mixing of a calcite (calcium carbonate) agglomerate in high‐density polyethylene (HDPE), low‐density polyethylene (LDPE), and linear low‐density polyethylene (LLDPE) matrices upon stretching was investigated using three different techniques: mechanical property, morphological behavior, and image analyzer analyses. The mechanical properties analyzed in terms of the tensile strength and maximum elongation resulted in that the second mixing was the best for giving a better property for all systems except the LDPE system, which exhibited no significant difference between the second and third mixings. The morphological behavior of the three compounds were different, but no distinctive difference was observed to differentiate the degree of mixing from system to system. The number‐, weight‐, and z + 1‐average diameters of the air hole and the aspect ratio upon the stretching and mixing cycle were calculated to analyze the degree of mixing of the calcite‐filled composites. As a consequence, no difference in the average diameter of the air hole was obtained among the three systems, but the aspect ratios of the air hole varied significantly. Thus, the degree of dispersion and the degree of mixing may be influenced by the average calcite agglomerate size, the average diameter of the air hole, and the aspect ratio upon stretching and mixing cycles. Those factors would be formed by the difference in chemical characteristics upon various microstructures of polyethylene and its molecular weight and molecular weight distribution. © 2003 Wiley Periodicals, Inc. J Appl Polym Sci 87: 311–321, 2003     

Cure kinetics and mechanical properties of the blend system of epoxy/diaminodiphenyl sulfone and amine terminated polyetherimide-carboxyl terminated poly(butadiene-co-acrylonitrile) block copolymer

The cure kinetics of blends of epoxy resin (4,4’-tetraglycidyl diaminodiphenyl methane; TGDDM)/curing agent (diaminodiphenyl sulfone; DDS) with ATPEI (amine terminated poly-etherimide) -CTBN (carboxyl terminated poly (butadiene-co-acrylonitrile)) block copolymer (AB type) were studied using differential scanning calorimetry under isothermal conditions to determine the reaction kinetic parameters such as activation energy and reaction constants. Final cure conversion decreased with increasing amount of AB in the blends. A diffusion controlled reaction was observed as the cure conversion increased, and the curing reaction was successfully analyzed by incorporating the diffusion control term in the rate equation for the epoxy/DDS/AB blends. The fracture toughness was improved to about 350% compared to that of the unmodified resin at 30% of AB block copolymer. This is attributed to the formation of co-continuous morphology between the epoxy phase and AB block copolymer phase. By increasing the amount of AB, the modulus of the cured blends decreased, which was due to the presence of CTBN rubbery phases.     

Combined salience based person re-identification

Human eye perceives an object as the entity with global information and local information. Human salience is distinctive local information in matching pedestrians across disjoint camera views, and matching on overall foreground guarantees reliable and robust identification. In this paper, we propose a strategy for the matching of mean salience to identify pedestrians. Also, we consider that person re-identification based on the local single directional matching suffers from the variations of pose, illumination and overlapping, and propose a global bi-directional matching to solve the challenging problems of person re-identification. Furthermore, our matching of mean salience is tightly combined with the global bi-directional matching. Patch matching is utilized to handle the misalignment problem in pedestrian images. We test our feature and matching approaches in person re-identification scenario. Experimental results demonstrate that the mean salience and the global bi-directional matching have promising discriminative capability in comparison with other ones.