DNA and a CpG oligonucleotide derived from Babesia bovis are mitogenic for bovine B cells

DNAs from bacteria and variety of nonvertebrate organisms, including nematodes, mollusks, yeasts, and insects, cause polyclonal activation of murine B lymphocytes. Similar studies have not been reported for bovine B cells, and to date no studies have reported mitogenic properties of protozoal DNA for any species. However, we and others have observed that protozoal parasite antigens can induce the proliferation of lymphocytes from nonexposed donors. Extending these studies, we now show that the mitogenic property of protozoal antigen preparations is in part attributable to parasite DNA and that Babesia bovis DNA is directly mitogenic for bovine B cells. DNase treatment of B. bovis extracts abrogated B. bovis-induced proliferation of peripheral blood mononuclear cells from nonexposed cattle. Like DNAs from other organisms that were mitogenic for murine B cells, B. bovis DNA is largely nonmethylated and induced a dose-dependent proliferation of bovine B cells, which was reduced upon methylation. Furthermore, B. bovis and E. coli DNAs enhanced immunoglobulin secretion by cultured B cells, inducing moderate increases in immunoglobulin G1 and stronger increases in immunoglobulin G2. Because certain nonmethylated CpG motifs present in bacterial DNA are known to stimulate proliferation of murine and human B cells, an 11-kb fragment of B. bovis DNA was analyzed for CG dinucleotide content and for the presence of known immunostimulatory sequences (ISS) centered on a CG motif. The frequency of CG dinucleotides was approximately one-half of the expected frequency, and several CpG hexameric sequences with known activity for murine B cells were identified. An oligodeoxynucleotide containing one of these ISS (AACGTT), which is present within the rhoptry-associated protein-1 (rap-1) open reading frame, was shown to stimulate B-cell proliferation. These ISS may be involved in host immune modulation during protozoal infection and may be useful as vaccine adjuvants.     

beta-Amyloid precursor protein isoforms show correlations with neurones but not with glia of demented subjects

Post-mortem cerebral cortex from 15 demented patients was specially collected to minimise autolysis and two membrane fractions and one soluble fraction were quantitatively examined for the major species of beta-amyloid precursor protein (APP) of high apparent molecular mass (> or = 80 kDa) together with the major mRNA species encoding APP isoforms. The number of pyramidal neurones and astrocytes, putative biochemical indices of interneurones and pyramidal neurones, and choline acetyl transferase activity were also determined. Multiple regression analysis has been used to investigate intercorrelations of APP species with biochemical and morphometric measures, free of any effects of confounding demographic variables. Subjects with Alzheimer's disease showed a loss of cholinergic activity and D-aspartate uptake compared with patients with other causes of dementia. The major finding of the study is that measures of neurones rather than astrocytes most closely correlate with the concentration of APP. Pyramidal cell numbers were positively correlated with mRNA for APP695. APP in the soluble fraction showed a negative correlation with pyramidal cell numbers and cholinergic activity. These results indicate that neurones within the cerebral cortex are the major source of APP, and that secretion of APP is dependent upon cortical pyramidal neuronal activity and cholinergic activity.     

Molecular analysis of the rfaD gene, for heptose synthesis, and the rfaF gene, for heptose transfer, in lipopolysaccharide synthesis in Salmonella typhimurium

We report the analysis of three open reading frames of Salmonella typhimurium LT2 which we identified as rfaF, the structural gene for ADP-heptose:LPS heptosyltransferase II; rfaD, the structural gene for ADP-L-glycero-D-manno-heptose-6-epimerase; and part of kbl, the structural gene for 2-amino-3-ketobutyrate CoA ligase. A plasmid carrying rfaF complements an rfaF mutant of S. typhimurium; rfaD and kbl are homologous to and in the same location as the equivalent genes in Escherichia coli K-12. The RfaF (heptosyl transferase II) protein shares regions of amino acid homology with RfaC (heptosyltransferase I), RfaQ (postulated to be heptosyltransferase III), and KdtA (ketodeoxyoctonate transferase), suggesting that these regions function in heptose binding. E. coli contains a block of DNA of about 1,200 bp between kbl and rfaD which is missing from S. typhimurium. This DNA includes yibB, which is an open reading frame of unknown function, and two promoters upstream of rfaD (P3, a heat-shock promoter, and P2). Both S. typhimurium and E. coli rfaD genes share a normal consensus promoter (P1). We postulate that the yibB segment is an insertion into the line leading to E. coli from the common ancestor of the two genera, though it could be a deletion from the line leading to S. typhimurium. The G+C content of the rfaLKZYJI genes of both S. typhimurium LT2 and E. coli K-12 is about 35%, much lower than the average of enteric bacteria; if this low G+C content is due to lateral transfer from a source of low G+C content, it must have occurred prior to evolutionary divergence of the two genera.     

Calphostin C triggers calcium-dependent apoptosis in human acute lymphoblastic leukemia cells

Recent studies have demonstrated that the naturally occurring perylenequinone antibiotic calphostin C is a potent inhibitor of protein kinase C and can induce apoptosis in some tumor cell lines by an as yet unknown mechanism. Here we demonstrate that calphostin C induces dose-dependent apoptosis in DT40 chicken lymphoma B-cells, and targeted disruption of lyn, syk, btk, PLCgamma2, or IP3R genes does not prevent or attenuate its cytotoxicity. In our study, calphostin C also induced rapid apoptosis in human acute lymphoblastic leukemia (ALL) cell lines ALL-1 (BCR-ABL+ pre-pre-B ALL), RS4;11 (MLL-AF4+ pro-B ALL), NALM-6 (pre-B ALL), DAUDI (Burkitt's/B-cell ALL), MOLT-3 (T-ALL), and JURKAT (T-ALL), whereas other potent PKC inhibitors did not. In biochemical studies, calphostin C was discovered to induce rapid calcium mobilization from intracellular stores of ALL cell lines, and its cytotoxicity against ALL cell lines was well correlated with the magnitude of this calcium signal. Calphostin C-induced apoptosis was markedly suppressed by BAPTA/AM, a cell-permeable Ca2+ chelator as well as NiCl2, an inhibitor of Ca2+/Mg2+-dependent endonucleases. Inhibition of the Ca2+/calmodulin-dependent phosphatase calcineurin with perfluoreperazine dimadeate (a calmodulin antagonist) or cyclosporin A (a specific inhibitor of calcineurin) also reduced the magnitude of calphostin C-induced apoptosis in ALL cell lines. Calphostin C was capable of inducing calcium mobilization and apoptosis in freshly obtained primary leukemic cells from children with ALL. Taken together, our results provide unprecedented evidence that calphostin C triggers a Ca2+-dependent apoptotic signal in human ALL cells.     

Corrosion and oxidation of alloys of the Mg-Y-Zr-REM system

The corrosion resistance of an alloy of the Mg-Y-Zr-REM system obtained by sand casting and subjected to different modes of heat treatment is studied with the use of potentiodynamic polarization, immersion tests, scanning electron microscope, and Auger electron spectroscope. The microstructure of the alloy is described.     

Pilot-scale two-stage process: a combination of acidogenic hydrogenesis and methanogenesis.

This study was performed to optimize both acidogenic hydrogenesis and methanogenesis, and then to develop a pilot-scale two-stage process producing not only CH4 but also H2. Firstly, acidogenic hydrogenesis of food waste was examined in pilot-scale leaching-bed reactors using dilution rate (D) as a tool to improve the environmental conditions. The maximum efficiency of 71.4% was obtained by adjusting D from 4.5 to 2.5 d(-1) depending on the state of degradation. Secondly, the wastewater from acidogenic hydrogenesis was converted to CH4 in a pilot-scale UASB reactor. The COD removal efficiency exceeded 95% up to the loading rates of 13.1 g COD/Ld, which corresponded to HRT of 0.25 d (6 h). Lastly, a pilot-scale two-stage process was devised based on a combination of acidogenic hydrogenesis and methanogenesis. Over 120 days, the pilot-scale process resulted in large VS reduction of 70.9% at the high loading rate of 12.5 kg VS/m3/d in a short SRT of 8 days.     

Plasma enhanced machining of Inconel 718: modeling of workpiece temperature with plasma heating and experimental results

A numerical and experimental analysis of plasma enhanced machining (PEM) of Inconel 718 is presented in this paper. Surface temperatures due to plasma heating are systematically characterized through numerical modeling and experimental investigation using infrared radiation thermometry. A three-dimensional finite difference model is established to determine the temperature distribution in a cylindrical workpiece subjected to intense localized heating. The results are compared with experimental results obtained with a radiation pyrometer. A sensitivity analysis is presented to examine the effects of machining parameters on the temperature distribution. Benefits of PEM are also demonstrated through the reduction of cutting forces and improved surface roughness over a wide range of cutting conditions. In addition, improvement of productivity in machining Inconel with PEM is illustrated.     

A computational approach for prediction of the damage evolution and crushing behavior of chopped random fiber composites

A computational model is developed, by implementing the damage models previously proposed by authors into a finite element code, for simulating the damage evolution and crushing behavior of chopped random fiber composites. Material damages induced by fiber debonding and crack nucleation and growth are considered. Systematic computational algorithms are developed to combine the damage models into the constitutive relation. Based on the implemented computational model, a range of simulations are carried out to probe the behavior of the composites and to validate the proposed methodology. Numerical examples show that the present computational model is capable of modeling progressive deterioration of effective stiffness and softening behavior after the peak load. Crushing behavior of composite tube is also simulated, which shows the applicability of the proposed computational model for crashworthiness simulations.     

Test for measuring cut resistance of yarns

A test procedure for evaluating the cut resistance of yarns under tension-shear loading conditions is described and demonstrated. A knife blade is pressed transversely at a constant rate against a yarn gripped at its ends, the load-deflection relation is measured, and the energy required to cut through the yarn is computed. Results for Zylon (polybenzobisoxazole or PBO) are presented. The cut energy and strain to initiate cutting depend on the sharpness of the blade, the slicing angle, and the pre-tension in the yarn. The dependencies are explained by changes in failure mode of the fibers within the yarn. The test provides information needed to extend a computational model of ballistic response of fabrics to sharp fragments and to design a cutting tool.