The human SHIP gene is differentially expressed in cell lineages of the bone marrow and blood

The macrophage colony-stimulating factor receptor and several other hematopoietic growth factor receptors induce the tyrosine phosphorylation of a 145- to 150-kD protein in murine cells. We have previously cloned a cDNA for the murine 150-kD protein, SHIP, and found that it encodes a unique signaling intermediate that binds the SHC PTB domain through at least one tyrosine phosphorylated (NPXY) site in the carboxyl-terminal region. SHIP also contains several potential SH3 domain-binding sites, an SH2 domain for binding other tyrosine phosphorylated proteins, and an enzymatic activity that removes the phosphate from the 5 position of phosphatidylinositol 3,4,5-phosphate or from inositol 1,3,4,5-phosphate. SHIP has a negative effect on cell growth and therefore loss or modification may have profound effects on hematopoietic cell development. In this study, we have cloned a cDNA for human SHIP and examined mRNA and protein expression of SHIP and related species in bone marrow and blood cells. Flow cytometry indicates that at least 74% of immature CD34+ cells express SHIP cross-reacting protein species, whereas within the more mature population of CD33+ cells, only 10% of cells have similar expression. The majority of T cells react positively with the anti-SHIP antibodies, but significantly fewer B cells are positive. Immunoblotting detects up to seven different cross-reacting SHIP species, with peripheral blood mononuclear cells exhibiting primarily a 100-kD protein and a CD34+ acute myeloblastic leukemia expressing mainly 130-kD and 145-kD forms of SHIP. Overall, these results indicate that there is an enormous diversity in the size of SHIP or SHIP-related mRNA and protein species. Furthermore, the expression of these protein species changes according to both the developmental stage and differentiated lineage of the mature blood cell.     

Toll road crashes of commercial and passenger motor vehicles

Revenue-collection data from toll roads allow for accurate estimates of miles driven by vehicle type and, when combined with crash data, valid estimates of crash involvements per mile driven. Data on vehicle-miles traveled and collisions were obtained from toll road authorities in Florida. Kansas, and New York. In addition, state crash files and published vehicle-miles of travel were obtained for toll roads in Illinois. Indiana, Ohio, and Pennsylvania. Large commercial motor vehicles were significantly underinvolved in single-vehicle crashes on all state toll roads. In five states, commercial motor vehicles were significantly overinvolved in multiple-vehicle crashes relative to passenger vehicles; the exceptions were Kansas, where they had significantly lower multiple-vehicle involvement rates, and Indiana. where there were no significant differences in multiple-vehicle involvements by vehicle type. The risk of commercial motor vehicle involvement in multiple-vehicle crashes resulting in deaths or serious injuries was double that of passenger vehicles in the two states (Ohio and Pennsylvania) that identified serious injuries. Whether crash rates, on toll roads of commercial motor vehicles are higher or lower than those of passenger vehicles appears to depend on the type of crash, specific toll road. and traffic density.     

Effect of two different rubbers as secondary binders on the friction and wear characteristics of non-asbestos organic (NAO) brake friction materials

Binder provides structural integrity by holding all ingredients in the composition of a brake friction material. The modified binders have played a major role in improving the frictional performance and thermal resistance of the friction material. The present research work evaluates the influence of secondary binders (Nitrile Butadiene rubber (NBR) and Styrene Butadiene rubber (SBR)) on the tribological performance of the friction material using a full-scale inertia brake dynamometer as per JASO C406 standard. Three brake pads were developed by varying the type and composition of secondary rubber binder (5%NBR, 5%SBR and 2.5%NBR + 2.5%SBR) with rest of the ingredients kept unaltered. It was found that the quantity of SBR rubber powder present as secondary binder improved dry and wet recovery. Friction coefficient (μ) exhibited better stability during the fade with the inclusion of both the rubber powders. The friction material with the inclusion of both the NBR and SBR rubber powders exhibited overall better performance than compared to the inclusion of only one secondary binder rubber in the composition. The worn-out surface of the developed friction materials and the counter discs were characterised using FESEM.     

Shall we dance? How a multicopper oxidase chooses its electron transfer partner

Multicopper oxidases (MCOs) are encoded in the genomes of Eukarya, Bacteria, and Archea. These proteins are unique in that they contain at least four Cu atom prosthetic groups organized into one each of the three spectral classifications of copper sites in biology: type 1 (T1), type 2 (T2), and binuclear type 3 (T3), where the T2 and T3 sites form a trinuclear Cu cluster. With these four redox-active copper sites, the multicopper oxidases catalyze the four-electron (4e(-)) reduction of dioxygen to 2H2O, an activity that they alone share with the terminal heme-containing oxidases. Most MCOs exhibit broad specificity towards organic reductants, while a relatively small number of family members exhibit equally robust activity towards metal ions like Fe(II), Cu(I), and Mn(II) and, thus, are considered metallo-oxidases. This Account analyzes the structure-activity features of multicopper oxidases that determine their relative substrate specificity. Since the substrate oxidation step involves an outer-sphere electron transfer from the reductant to the T1Cu site in the protein, the concepts of Marcus theory are applied to unravel the origin of the substrate specificity of the multicopper ferroxidases.     

Rapid detection of Salmonella from hydrodynamic pressure-treated poultry using molecular beacon real-time PCR

A real-time polymerase chain reaction (PCR) assay was evaluated to detect Salmonella in hydrodynamic pressure (HDP)-treated chicken using molecular beacon probes available as a commercial kit (iQ-Check, Bio-Rad Laboratories). The sensitivity and accuracy of the assay were compared with the conventional USDA microbiological procedure using artificially contaminated minced chicken. Chicken fillets were irradiated at 10 kGy to completely destroy any naturally occurring Salmonella. These fillets were minced and inoculated with as low as 2+/-1 cfu of S. typhimurium per 25 g chicken. The minced chicken samples were vacuum packed in multi-layer barrier bags, heat shrunk, and treated with HDP. Results showed that all inoculated samples (n=36) were detected by the PCR assay and conventional USDA procedure. Similarly, all uninoculated controls (n=11) were negative by both PCR assay and USDA procedure. As few as 2+/-1 cfu could be detected from 25 g HDP-treated chicken following 16-18 h enrichment in buffered peptone water. Real-time PCR proved to be an effective method for Salmonella detection in HDP-treated chicken with high sensitivity and more importantly, a rapid and high-throughput detection in 18 h, compared to 3-8 days for the conventional microbiological methods. HDP treatment, which has been reported to reduce spoilage bacteria in various meats, was unable to kill pathogenic Salmonella in minced chicken.     

Phosphorus speciation in manure and manure-amended soils using XANES spectroscopy

Previous studies suggested an increase in the proportion of calcium phosphates (CaP) of the total phosphorus (P) pool in soils with a long-term poultry manure application history versus those with no or limited application histories. To understand and predict long-term P accumulation and release dynamics in these highly amended soils, it is important to understand what specific P species are being formed. We assessed forms of CaP formed in poultry manure and originally acidic soil in response to different lengths of mostly poultry manure applications using P K-edge X-ray absorption near-edge structure (XANES) spectroscopy. Phosphorus K-edge XANES spectra of poultry manure showed no evidences of crystalline P minerals but dominance of soluble CaP species and free and weakly bound phosphates (aqueous phosphate and phosphate adsorbed on soil minerals). Phosphate in an unamended neighboring forest soil (pH 4.3) was mainly associated with iron (Fe) compounds such as strengite and Fe-oxides. Soils with a short-term manure history contained both Fe-associated phosphates and soluble CaP species such as dibasic calcium phosphate (DCP) and amorphous calcium phosphate (ACP). Long-term manure application resulted in a dominance of CaP forms confirming our earlier results obtained with sequential extractions, and a transformation from soluble to more stable CaP species such as beta-tricalcium calcium phosphate (TCP). Even after long-term manure application (> 25 yr and total P in soil up to 13,307 mg kg(-1)), however, none of the manure-amended soils showed the presence of crystalline CaP. With a reduction or elimination of poultry manure application to naturally acidic soils, the pH of the soil is likely to decrease, thereby increasing the solubility of Ca-bonded inorganic P minerals. Maintaining a high pH is therefore an important strategy to minimize P leaching in these soils.