Haemoglobins of the shark, Heterodontus portusjacksoni II. Amino acid sequence of the alpha-chain

The amino acid sequence of the alpha-chain of the principal haemoglobin from the shark, H. portusjacksoni has been determined. The chain has 148 residues and is acetylated at the amino terminal. The soluble peptides obtained by tryptic and chymotryptic digestion of the protein or its cyanogen bromide fragments were isolated by gel filtration, paper ionophoresis and paper chromatography. The amino acid sequences were determined by the dansyl-Edman procedure. The insoluble "core" peptide from the tryptic digestion contained 34 residues and required cleavage by several prosteases before the sequence was established. Compared with human alpha-chain there are 88 amino acid differences including the additional seven residues which appear on the amino terminal of the shark chain. There is also one deletion and one insertion. The chain contains no tryptophan but has four cysteinyl residues which is the highest number of such residues recorded for a vertebrate globin. In the alpha1beta1 contact sites there are four changes in the oxyhaemoglobin form and six deoxy form. Nine of the 16, alpha1beta1 contact sites show variation while three of the haem contact sites have changed in comparison to the residues known to be involved in these interactions in horse haemoglobin alpha-chain. Use of the sequence data to estimate a time of divergence of the shark from the main vertebrate line yielded the value of 410 +/- 46 million years. The data, in general, support the palaeontological view that bony fishes arose before the elasmobranchs.     

Memory for Visuospatial Location Following Selective Hippocampal Sclerosis: The Use of Different Coordinate Systems.

This study addressed the role of the medial temporal lobe regions and, more specifically, the contribution of the human hippocampus in memory for body-centered (egocentric) and environment-centered (allocentric) spatial location. Twenty-one patients with unilateral atrophy of the hippocampus secondary to long-standing epilepsy (left, n = 7; right, n = 14) and 15 normal control participants underwent 3 tasks measuring recall of egocentric or allocentric spatial location. Patients with left hippocampal sclerosis were consistently impaired in the allocentric conditions of all 3 tasks but not in the egocentric conditions. Patients with right hippocampal sclerosis were impaired to a lesser extent and in only 2 of the 3 tasks. It was concluded that hippocampal structures are crucial for allocentric, but not egocentric, spatial memory. (PsycINFO Database Record (c) 2010 APA, all rights reserved)     

Metabolic mapping of the effects of oral alcohol self-administration in rats

A method for the quantitation of protein carbonyls, which have been widely employed as markers of protein oxidative damage, is described. Protein carbonyls were derivatized with tritiated sodium borohydride and the tritiated proteins were separated on SDS-PAGE. Protein bands, visualized by Coomassie blue staining, were then excised and incubated in 30% H2O2 at 60 degrees C for 48 h. Tritium, incorporated into the proteins, was quantitated by liquid scintillation counting after gel solubilization by H2O2. This method can be applied to the measurement of carbonylation of specific proteins as it employs SDS-PAGE and has the advantage that unreacted NaB3H4 in the labeling reaction mixture need not be removed. The present method, when combined with immunochemical detection of protein carbonyls, should be very useful in the quantitation of oxidative damage to individual proteins.     

Effect of Pseudomonas fluorescens on beef discoloration and oxymyoglobin oxidation in vitro

The relationship between bacterial growth and oxymyoglobin oxidation in vitro and in meat was studied. In the in vitro study, oxymyoglobin was combined with Pseudomonas fluorescens or sterile nutrient broth (control) in an airtight vessel. P. fluorescens samples showed greater metmyoglobin formation and oxygen consumption than controls. The P. fluorescens population in the reaction vessels was correlated with metmyoglobin formation (r = 0.85, P < 0.05) and oxygen consumption (r = 0.91, P < 0.05). When P. fluorescens and oxymyoglobin were combined in an airtight vessel, reducing the headspace from 13 ml and 9 ml to 3 ml resulted in greater metmyoglobin formation (P < 0.05). In the meat study, beef cores prepared from longissimus lumborum were inoculated with P. fluorescens (10(7) CFU/cm2) or sterile peptone water (control), packaged under 1% O2 (+99% N2), air, or 100% O2 and stored at 4 degrees C. Inoculated beef cores showed higher bacterial loads and metmyoglobin formation than their respective controls during 10 h storage in 1% O2, 3 days in air, and 7 days in 100% O2 (P < 0.05). This finding indicated that P. fluorescens could accelerate beef discoloration. Overall, studies demonstrated that oxygen consumption concomitant with P. fluorescens growth decreased partial oxygen pressure, which accelerated oxymyoglobin oxidation.     

Investigation on microstructure and tensile behaviour of stir cast LM13 aluminium alloy reinforced with copper coated short steel fibers using response surface methodology

Copper coated steel fibers reinforced LM13 aluminium alloy composites have been prepared using stir casting process. Experiments have been designed using response surface methodology by varying wt% of reinforcement (0–10), stirrer speed (350–800 rpm) and pouring temperature (700–800 °C). Microstructure, tensile strength and fracture surface of composites have been investigated. Analysis of variance, significance test and confirmation tests have been performed and regressions models have been developed to predict the tensile strength of composites. Response surface plots reveal that tensile strength of composites increases with increasing wt% of copper coated steel fibers reinforcement up to 6 wt%. Further increase in wt% of steel fibers decreases the tensile strength of composites. However tensile strength of composites increases with increasing stirrer speed due to the uniform and homogeneous dispersion of steel fibers in matrix. Optimum stir cast process parameters for obtaining higher tensile strength are found to be 5.9 wt% of reinforcement, 753 °C pouring temperature and stirrer speed of 633 rpm. Fracture mechanism is dominated by steel fiber pullouts in composites with higher wt% of reinforcement and dimples are observed in the surface of composites containing lower levels of wt% of reinforcement.     

Comparative analysis of 8-oxo-2' -deoxyguanosine in DNA by 32P- and 33P-postlabeling and electrochemical detection

8-Oxo-2'-deoxyguanosine (8-oxo-dG) is emerging as a useful marker for oxidative DNA damage. Reported basal levels determined by 32P-postlabeling (PPL) method were 10-fold or more higher than those obtained with HPLC/electrochemical detection (ECD). This discrepancy was investigated. In commercial calf thymus DNA, levels of 4 +/- 1 and 64 +/- 14 8-oxo-dG per 10(6) 2'-deoxynucleosides (dN) were measured by the standard HPLC/ECD and PPL methods, respectively. DNA digestion by micrococcal nuclease/spleen phosphodiesterase and nuclease P1 (as used in the standard PPL method), followed by ECD analysis resulted in a level of 8 +/- 3. In calf thymus DNA spiked with chemically synthesized 8-oxo-dGp to give an increment of 9 8-oxo-dG/10(6) dN, the added standard produced a significant increase with HPLC/ECD but not PPL. After spiking the DNA with 90 8-oxo-dG/10(6) dN, the added 8-oxo-dGp was detectable also with PPL, with a labeling efficiency of 65%. In order to investigate the role of ionizing radiation from 32P for the higher 8-oxo-dG levels in PPL, incubation times and amounts of radioactivity in the phosphorylation reaction with commercial dGp were increased, and external irradiation of commercial dG with 32P was investigated. All modifications resulted in higher values of 8-oxo-dG measured, but the effect was not large enough to fully explain the discrepancy between PPL and HPLC/ECD. Using [gamma-33P]ATP instead of [gamma-32P]ATP or adding [33P]phosphate to a 32P-PPL assay resulted in even higher levels of 8-oxo-dG measured. The increase in 8-oxo-dG levels during the PPL workup is attributed to the presence and oxidation of unmodified dGp in the reaction mixture. For a determination of true basal levels, the PPL method will have to be modified, including the removal of dGp prior to the phosphorylation reaction.     

Integrated manufacturing,employee and business performance: Australian and New Zealand evidence

This article reports on a large field study of Integrated Manufacturing (IM), consisting of Total Quality Management (TQM), Just in Time (JIT) and Advanced Manufacturing Technology (AMT). The relationship between these facets of Integrated Manufacturing and performance was examined empirically, including measures of employee performance and manufacturing performance. There was a significant correlation between employee performance and manufacturing performance. While all three facets of Integrated Manufacturing were relatively strongly connected with employee performance, manufacturing performance was significantly but less strongly connected to the intensity of TQM and JIT than employee performance, and not connected with the intensity of Advanced Manufacturing Technology. Some second-order interactive effects between the facets of Integrated Manufacturing and performance were found. Further, by partitioning the large data set into three using the AMT factor as a cutting variable, it was found that the stronger the AMT environment, the stronger the explanatory power of TQM and JIT on both employee and manufacturing performance. The interactive impact of AMT and TQM was significantly positively related to both employee and manufacturing performance.     

Quantitative detection method for Roundup Ready soybean in food using duplex real‐time PCR MGB chemistry

BACKGROUND: Methodologies that enable the detection of genetically modified organisms (GMOs) (authorized and non‐authorized) in food and feed strongly influence the potential for adequate updating and implementation of legislation together with labeling requirements. Quantitative polymerase chain reaction (qPCR) systems were designed to boost the sensitivity and specificity on the identification of GMOs in highly degraded DNA samples; however, such testing will become economically difficult to cope with due to increasing numbers of approved genetically modified (GM) lines. Multiplexing approaches are therefore in development to provide cost‐efficient solution. RESULTS: Construct‐specific primers and probe were developed for quantitative analysis of Roundup Ready ^® soybean (RRS) event glyphosate‐tolerant soybean (GTS) 40‐3‐2. The lectin gene (Le1) was used as a reference gene, and its specificity was verified. RRS‐ and Le1‐specific quantitative real‐time PCR (qRTPCR) were optimized in a duplex platform that has been validated with respect to limit of detection (LOD) and limit of quantification (LOQ), as well as accuracy. The analysis of model processed food samples showed that the degradation of DNA has no adverse or little effects on the performance of quantification assay. CONCLUSION: In this study, a duplex qRTPCR using TaqMan minor groove binder‐non‐fluorescent quencher (MGB‐NFQ) chemistry was developed for specific detection and quantification of RRS event GTS 40‐3‐2 that can be used for practical monitoring in processed food products. Copyright © 2010 Society of Chemical Industry     

rGO Sheets/ZnFe2O4 Nanocomposities as an Efficient Electro Catalyst Material for I3−/I− Reaction for High Performance DSSCs

In this paper, Reduced Graphene Oxide (rGO)/ZnFe₂O₄ (rZnF) nanocomposite is synthesized by a simple hydrothermal method and employed as a counter electrode (CE) material for tri-iodide redox reactions in Dye sensitized solar cells (DSSC) to replace the traditional high cost platinum (Pt) CE. X-ray diffraction analysis and High resolution Transmission electron microscopy, clearly indicated the formation of rZnF nanocomposite and also amorphous rGO sheets were smoothly distributed on the surface of ZnFe₂O₄ (ZnF) nanostructure. The rZnF-50 CE shows excellent electro catalytic activity toward I₃^? reduction, which has simultaneously been confirmed by cyclic voltammetry, electrochemical impedance spectroscopy and Tafel polarization measurements. A DSSC developed by rZnF-50 CE (η?=?8.71%) obtained quite higher than the Pt (η?=?8.53%) based CE under the same condition. The superior performances of rZnF-50 CE due to addition of graphene in to Spinel (ZnF) nanostructure results in creation of highly active electrochemical sites, fast electron transport linkage between CE and electrolyte. Thus it’s a promising low cost CE material for DSSCs.