Elucidation of protein aggregation in frozen cod and haddock by transmission electron microscopy/immunocytochemistry,light microscopy and atomic force microscopy

Polyclonal antibodies raised to both native cod myosin and actin as well as to aggregated proteins obtained from frozen cod stored for 11 months at ?10 °C were used to investigate disposition of muscle proteins in frozen cod and haddock fillets by transmission electron microscopy. Specimens from cod and haddock fillets, stored at ?10 °C, treated with anti‐aggregate antibody as the primary antibody, showed significantly more gold particles, especially around the protein aggregates and muscle fibres compared with fish stored at ?30 °C. Samples that were treated with anti‐myosin or anti‐actin antibody showed opposite results. Similar binding properties were observed in ELISA experiments involving the reaction of actin and myosin to both native and aggregate antibodies; thus immunological tests can be used for monitoring aggregate and texture changes in frozen stored fish. In addition, atomic force microscopy images obtained from cod muscle also indicated structural changes in frozen cod muscle proteins. The mica surface was covered with a continuous layer of muscle proteins comprising mainly small globular particles and a few large particles for the control cod sample stored at ?30 °C for 11 months. In contrast, cod fillets stored at ?10 °C showed a thin layer of proteins with small holes and an increased number of large particles denoting aggregates. Formation of ice crystals between the muscle fibres of frozen cod and haddock muscle was monitored without thawing by light microscopy at ?20 °C. The micrographs showed a greater proportion of large ice crystals and extensive protein fibre changes in fillets stored at ?10 °C compared with the control at ?30 °C. Copyright © 2004 Society of Chemical Industry     

Comparison of different extraction methods for the extraction of major bioactive flavonoid compounds from spearmint (Mentha spicata L.) leaves

Different bioactive flavonoid compounds including catechin, epicatechin, rutin, myricetin, luteolin, apigenin and naringenin were obtained from spearmint (Mentha spicata L.) leaves by using conventional soxhlet extraction (CSE) and supercritical carbon dioxide (SC-CO₂) extraction at different extraction schemes and parameters. The effect of different parameters such as temperature (40, 50 and 60 °C), pressure (100, 200 and 300 bar) and dynamic extraction time (30, 60 and 90 min) on the supercritical carbon dioxide (SC-CO₂) extraction of spearmint flavonoids was investigated using full factorial arrangement in a completely randomized design (CRD). The extracts of spearmint leaves obtained by CSE and optimal SC-CO₂ extraction conditions were further analyzed by high performance liquid chromatography (HPLC) to identify and quantify major bioactive flavonoid compounds profile. Comparable results were obtained by optimum SC-CO₂ extraction condition (60 °C, 200 bar, 60 min) and 70% ethanol soxhlet extraction. As revealed by the results, soxhlet extraction had a higher crude extract yield (257.67 mg/g) comparing to the SC-CO₂ extraction (60.57 mg/g). Supercritical carbon dioxide extract (optimum condition) was found to have more main flavonoid compounds (seven bioactive flavonoids) with high concentration comparing to the 70% ethanol soxhlet extraction (five bioactive flavonoids). Therefore, SC-CO₂ extraction is considered as an alternative process compared to the CSE for obtaining the bioactive flavonoid compounds with high concentration from spearmint leaves.     

Surface-Induced Interphases During Curing Processes Between Bi- and Pentafunctional Components: Reactive Coarse-Grained Molecular Dynamics Simulations

The present reactive molecular dynamics (RMD) simulations discuss the formation of interphase regions in cured polymer adhesives. The latter are obtained from the curing of reactive liquid mixtures composed of pentafunctional linkers and bifunctional monomers in contact with idealized surfaces. The present reactive scheme mimics the one of epoxies with amine linkers, i.e., processes investigated experimentally by Possart and co-workers. Generic RMD simulations are performed in a coarse-grained (CG) resolution to evaluate basic principles in curing characterized by preferential interactions. The creation of linker-rich domains is promoted by preferential surface-linker as well as linker-linker interactions in the reactive mixtures. The dimension of the interphase both in the starting mixture and the cured network depends on these preferential interactions which lead to a retardation of the curing velocity. This retardation behavior is mapped by conversion curves as a function of the number of reactive steps and by the spatially resolved profiles of the connected linkers. Although derived by generic potentials, the simulated reduction of the curing velocity is in agreement with experimental results in epoxies. The chosen interactions also imply a smaller number of linker bonds in the interphase than in the bulk region. The present RMD approach offers insight into key parameters of curing processes under the influence of preferential surface interactions coupled to selective attractions in the liquid starting mixture.     

4-D x 3-D ultrasound: real-time scan conversion, filtering, and display of displacement vectors with a motorized curvilinear transducer

Recent research in the field of elastography has sought to expand displacement tracking to three dimensions. Once the 3-D volumes of displacement data have been obtained, they must be scan converted so that further processing, such as inversion methods to obtain tissue elasticity, can take place in Cartesian coordinates. This paper details an efficient and geometrically accurate algorithm to scan convert 3-D volumes of displacement vectors obtained from a motorized sector transducer. The proposed algorithm utilizes the physical scan geometry to convert the 3-D volumes of displacement data to both Cartesian coordinates and Cartesian displacements. Spatially varying filters are also proposed to prevent aliasing while minimizing data loss. Validation of the system has shown the algorithm to be correct to floating point precision, and the 3-D scan conversion and filtering can be performed faster than the native rate of data acquisition for the motorized transducer.     

2-D high-frame-rate dynamic elastography using delay compensated and angularly compounded motion vectors: preliminary results

This paper describes a new ultrasound-based system for high-frame-rate measurement of periodic motion in 2-D for tissue elasticity imaging. Similarly to conventional 2-D flow vector imaging, the system acquires the RF signals from the region of interest at multiple steering angles. A custom sector subdivision technique is used to increase the temporal resolution while keeping the total acquisition time within the range suitable for real-time applications. Within each sector, 1-D motion is estimated along the beam direction. The intra- and inter-sector delays are compensated using our recently introduced delay compensation algorithm. In-plane 2-D motion vectors are then reconstructed from these delay-compensated 1-D motions. We show that Young's modulus images can be reconstructed from these 2-D motion vectors using local inversion algorithms. The performance of the system is validated quantitatively using a commercial flow phantom and a commercial elasticity phantom. At the frame rate of 1667 Hz, the estimated flow velocities with the system are in agreement with the velocity measured with a pulsed-wave Doppler imaging mode of a commercial ultrasound machine with manual angle correction. At the frame rate of 1250 Hz, phantom Young's moduli of 29, 6, and 54 kPa for the background, the soft inclusion, and the hard inclusion, are estimated to be 30, 11, and 53 kPa, respectively.     

Heat treatment in high Cr white cast iron Nb alloy

Wear resistance of high Cr white cast irons can be improved by means of heat treatment. This type of cast iron alloy may present a microstructure with retained austenite. The amount of retained austenite changes with the applied heat treatment, which will have an influence on wear properties. The purpose of this work was to study the influence of several parameters such as quenching and tempering temperatures and subzero treatment in the wear performance of the high Cr white cast iron Nb alloy. In this way, the performance was evaluated using pin-on-disc abrasion test. The worn surface was examined by scanning electron microscopy, and the main wear mechanisms were identified. The microstructural characterization was also performed with carbide identification. This Fe alloy has proven to be good for applications in mining and alcohol-sugar industries.     

Steady state and transient photoconductivity measurements in a-Se90 − xSb10Inx thin films

Amorphous thin films of Se_90 − xSb₁₀In_x (0 ≤ x ≤ 15) have been prepared by electron beam evaporation method. The steady state and transient photoconductivity measurements on the thin films of Se_90 − xSb₁₀In_x (0 ≤ x ≤ 15) were carried out at different levels of light intensities (500 lx-5000 lx) at room temperature (301 K). The plot of photocurrent (I_ph) versus light intensity (F) follows a power law I_ph ∝ F^γ. The value of exponent γ lies between 0.5 and 1.0, which indicates there exists a continuous distribution of localized states in the mobility gap of Se_90 − xSb₁₀In_x (0 ≤ x ≤ 15) thin films. For transient photoconductivity, when the samples were illuminated with light, the photocurrent reaches the maximum value during the first 5 s of exposure time and thereafter, it starts decreasing and becomes stable after 15 min of exposure. This kind of phenomenon is termed as photo-degradation of photocurrent. The results have been explained on the basis of charged defect model and the intercluster interaction model. The magnitude of photocurrent of the system a-Se₇₅Sb₁₀In₁₅ is higher than the parent system a-Se₉₀Sb₁₀. The photosensitivity shows a minimum value at 5 atomic percentage of indium (In) concentration, which is explained based on chemically ordered network model and the topological model.     

Production and Characterization of Peptide Antibodies to the C-Terminal of Frameshifted Calreticulin Associated with Myeloproliferative Diseases

Myeloproliferative Neoplasms (MPNs) constitute a group of rare blood cancers that are characterized by mutations in bone marrow stem cells leading to the overproduction of erythrocytes, leukocytes, and thrombocytes. Mutations in calreticulin (CRT) genes may initiate MPNs, causing a novel variable polybasic stretch terminating in a common C-terminal sequence in the frameshifted CRT (CRTfs) proteins. Peptide antibodies to the mutated C-terminal are important reagents for research in the molecular mechanisms of MPNs and for the development of new diagnostic assays and therapies. In this study, eight peptide antibodies targeting the C-terminal of CRTfs were produced and characterised by modified enzyme-linked immunosorbent assays using resin-bound peptides. The antibodies reacted to two epitopes: CREACLQGWTE for SSI-HYB 385-01, 385-02, 385-03, 385-04, 385-07, 385-08, and 385-09 and CLQGWT for SSI-HYB 385-06. For the majority of antibodies, the residues Cys¹, Trp⁹, and Glu¹¹ were essential for reactivity. SSI-HYB 385-06, with the highest affinity, recognised recombinant CRTfs produced in yeast and the MARIMO cell line expressing CRTfs when examined in Western immunoblotting. Moreover, SSI-HYB 385-06 occasionally reacted to CRTfs from MPN patients when analysed by flow cytometry. The characterized antibodies may be used to understand the role of CRTfs in the pathogenesis of MPNs and to design and develop new diagnostic assays and therapeutic targets.     

Temporary Tasks Assignment Resolved

Among all basic on-line load balancing problems, the only unresolved problem was load balancing of temporary tasks on unrelated machines. This open problem existed for almost a decade, see [12]. We resolve this problem by providing an inapproximability result. In addition, a newer open question is to identify the dependency of the competitive ratio on the durations of jobs in the case where durations are known. We resolve this problem by characterizing this dependency. Finally, we provide a PTAS for the off-line problem with a fixed number of machines and show a 2 -inapproximability result for the general case.