Epitope specificity of CD44 for monoclonal antibody-dependent facilitation of marrow engraftment in a canine model

Primary graft rejection after marrow transplantation occurs more frequently in patients receiving HLA-haploidentical compared with HLA-identical sibling transplants. Both human and experimental animal data suggest that the cells responsible for this phenomenon are either host natural killer (NK) cells, T cells, or both. To investigate the mechanisms of graft rejection, we have developed a canine model of marrow transplantation, which uses DLA-nonidentical unrelated donors in the absence of postgrafting immunosuppression. In this model most animals rejected their marrow grafts after a preparative regimen of 9.2 Gy total body irradiation (TBI). However, engraftment of DLA-nonidentical marrow can be facilitated when the recipients are pretreated with monoclonal antibody (MoAb) S5, which recognizes CD44. In this report, we extended these observations by first cloning the canine CD44 and, next, mapping the epitope recognized by S5, which was located in a region conserved among human and canine CD44 and was distinct from the hyaluronan binding domain. However, in vitro binding of S5 caused a conformational change in CD44, which allowed increased hyaluronan binding. Then, we reexamined the in vivo model of marrow transplantation and compared results with MoAb S5 to those with two other anti-CD44 MoAbs, IM7 and S3. Only MoAb S5 significantly increased the engraftment rate of DLA-nonidentical unrelated marrow, whereas the two other anti-CD44 MoAbs were ineffective. The enhanced in vivo effect was not related to differences in the MoAbs' avidities, since both S5 and IM7 had equivalent binding to CD44, but most likely related to the specific epitope that S5 recognizes. Thus, this study shows that the effect of the anti-CD44 MoAb S5 in facilitating engraftment is epitope specific and if one is to use an anti-CD44 to facilitate engraftment of marrow in humans, one cannot assume that any anti-CD44 would work.     

GC/MS characterization of mate tea leaves extracts obtained from high-pressure CO2 extraction

This work presents a study concerning the chemical characteristics and analytical separation process of the essential oil obtained from high-pressure carbon dioxide extraction of Ilex paraguariensis. The experiments were performed in a laboratory-scale unit in the temperature range of 20–40 °C, from 100 to 250 bar. A blend of the I. paraguariensis extracts was percolated through a preparative chromatographic column, containing silica gel, and successively eluted with 150 mL of each of the following solvents: hexane, toluene, dichloromethane, ethyl acetate, acetone and methanol. The raw extract and its fractions were analyzed by gas chromatography coupled to mass spectrometry detection (GC/MS). The fractionation procedure showed to be a good clean up technique due to the isolation of different classes of compounds in each fraction. Chromatographic analyses allowed the identification of caffeine, fatty acids and esters, phytol, squalene, Vitamin E, stigmasterol derivatives and saturated hydrocarbons.     

Oxidation and mineralisation of substituted phenols by Fenton's reagent and catalytic wet oxidation.

Catalytic abatement of solutions of 1,000 mg/L in phenol, ortho and para nitrophenol and ortho and para cresols was acomplished by using two catalytic systems. Fenton's reagent was used at 50 degrees C by adding 10 mg/L of ferrous cation and different dosages of H2O2. The mixture was reacting isothermically in a batch way during 3 hours. Catalytic wet oxidation (CWO) was carried out by using a commercial Activated Carbon, Industrial React FE01606A, CWO runs were carried out in a fixed bed reactor (FBR) with concurrent upflow. Temperature and oxygen pressure of the reactor were set to 160 degrees C and 16 bar, respectively. While phenols are quicky oxidised by the Fenton reagent higher mineralisation was obtained in the CWO process.     

New HPLC Method to Determine Ethyl Carbamate in Alcoholic Beverages Using Fluorescence Detection

ABSTRACT: A new methodology to the quantification of ethyl carbamate (EC) has been developed. This method allows the analysis by HPLC of ethyl carbamate in samples of wine, fortified wine, and wine brandy, by a pre-column derivatization with 9-xanthydrol, and fluorescence detection. This does not require previous sample extraction or concentration. The method presents an average recovery of 96% among samples studied, a detection limit of 4.2μg/L, and an average intermediate precision of 6.3%. The comparison of the results obtained for EC analysis on the same wine brandy samples by GC/MS and HPLC are statistically indistinguishable with 97.5% probability. The results of the analysis of 42 samples are presented.     

Yeast exo-β-glucanases can be used as efficient and readily detectable reporter genes in Saccharomyces cerevisiae

Yeast exo-1,3-β-glucanases are secretable proteins whose function is basically trophic and may also be involved in cell wall glucan hydrolytic processes. Since fluorescein di(β-D -glucopyranoside) is a fluorogenic substrate detectable and quantifiable by flow cytometry, it was used for testing the ability of the EXG1 gene product of Saccharomyces cerevisiae and its homologous gene in Candida albicans to function as reporter genes. These open reading frames were coupled to different promoters in multicopy plasmids, and exoglucanase activity quantified at flow cytometry. Exoglucanases were found to be useful tools for the study of promoter regions in S. cerevisiae. This technique has the advantage over other reporter gene systems—such as β-galactosidase fusions—that it does not require permeabilization of yeast cells and therefore it allows the recovery of viable cells—by sorting—after flow cytometry analysis.     

Analysis of the propagation of leaky magnetostatic modes in normally magnetized microstrip and slot lines

The propagation of leaky forward magnetostatic (MS) volume waves along ferrite-loaded microstrip and slot lines is analyzed. This phenomenon is studied by means of a numerical approach based on the residue calculus technique because of its good numerical convergence and stability. The proposed method allows for a quick and accurate computation of the phase and attenuation constants of the leaky MS modes. A comparative analysis between both microstrip lines and slot lines is carried out, and some new physical effects, such as MS resonances in the radiation loss, are reported. The advantages of the proposed method of analysis over other numerical methods, such as Galerkin's or moment methods, are also discussed.     

Stress Concentration Factors for a Circular Hole in Curved Beams Under Bending Loads

Abstract: As a contribution to the field of stress concentration studies, the case of circular hole in curved bar elements under bending is hereby studied. Trend values of stress concentration factors (SCF) are presented based on selected variables of the problem for two critical hole locations. The results will help to identify an SCF law for practical use by applying the statistical method of regression analysis. Two approximations obtained by means of computational methods are compared with those obtained by photoelastic experimental method.     

Friction Stir Welding of Ceramic Particle Reinforced Aluminium Based Metal Matrix Composites

The mechanical and microstructural properties of 6061+20% Al₂O_3p and 7005+10% Al₂O_3p aluminium based metal matrix composites joined by friction stir welding were analyzed in the present study. The two materials were welded into the form of sheets of 7 mm thickness after T6 treatment and were tested in tension at room temperature. The microstructure of the joints was observed by optical microscopy and the fracture surfaces were analyzed by employing a scanning electron microscope equipped with field emission gun in order to study the micromechanisms involved during the deformation.