Increase of proteolytic activity inLactobacillus casei by X-ray irradiation

Summary A study was conducted on the effect of X-ray irradiation on proteolytic activity ofLactobacillus casei (RTS). Substantial increase (82%) in the proteolytic activity was noted among the isolates of X-ray irradiation survivors after 18,000 R.     

GILT is a critical host factor for Listeria monocytogenes infection

Listeria monocytogenes is a gram-positive, intracellular, food-borne pathogen that can cause severe illness in humans and animals. On infection, it is actively phagocytosed by macrophages; it then escapes from the phagosome, replicates in the cytosol, and subsequently spreads from cell to cell by a non-lytic mechanism driven by actin polymerization. Penetration of the phagosomal membrane is initiated by the secreted haemolysin listeriolysin O (LLO), which is essential for vacuolar escape in vitro and for virulence in animal models of infection. Reduction is required to activate the lytic activity of LLO in vitro, and we show here that reduction by the enzyme gamma-interferon-inducible lysosomal thiol reductase (GILT, also called Ifi30) is responsible for the activation of LLO in vivo. GILT is a soluble thiol reductase expressed constitutively within the lysosomes of antigen-presenting cells, and it accumulates in macrophage phagosomes as they mature into phagolysosomes. The enzyme is delivered by a mannose-6-phosphate receptor-dependent mechanism to the endocytic pathway, where amino- and carboxy-terminal pro-peptides are cleaved to generate a 30-kDa mature enzyme. The active site of GILT contains two cysteine residues in a CXXC motif that catalyses the reduction of disulphide bonds. Mice lacking GILT are deficient in generating major histocompatibility complex class-II-restricted CD4(+) T-cell responses to protein antigens that contain disulphide bonds. Here we show that these mice are resistant to L. monocytogenes infection. Replication of the organism in GILT-negative macrophages, or macrophages expressing an enzymatically inactive GILT mutant, is impaired because of delayed escape from the phagosome. GILT activates LLO within the phagosome by the thiol reductase mechanism shared by members of the thioredoxin family. In addition, purified GILT activates recombinant LLO, facilitating membrane permeabilization and red blood cell lysis. The data show that GILT is a critical host factor that facilitates L. monocytogenes infection.     

Kinetic redistribution of native and misfolded RNAs by a DEAD-box chaperone

DExD/H-box proteins are ubiquitously involved in RNA-mediated processes and use ATP to accelerate conformational changes in RNA. However, their mechanisms of action, and what determines which RNA species are targeted, are not well understood. Here we show that the DExD/H-box protein CYT-19, a general RNA chaperone, mediates ATP-dependent unfolding of both the native conformation and a long-lived misfolded conformation of a group I catalytic RNA with efficiencies that depend on the stabilities of the RNA species but not on specific structural features. CYT-19 then allows the RNA to refold, changing the distribution from equilibrium to kinetic control. Because misfolding is favoured kinetically, conditions that allow unfolding of the native RNA yield large increases in the population of misfolded species. Our results suggest that DExD/H-box proteins act with sufficient breadth and efficiency to allow structured RNAs to populate a wider range of conformations than would be present at equilibrium. Thus, RNAs may face selective pressure to stabilize their active conformations relative to inactive ones to avoid significant redistribution by DExD/H-box proteins. Conversely, RNAs whose functions depend on forming multiple conformations may rely on DExD/H-box proteins to increase the populations of less stable conformations, thereby increasing their overall efficiencies.     

Effects of gadolinium addition on the microstructure and mechanical properties of Mg-9Al alloy

This research aims to study the significance of Gd addition (0wt%-2wt%) on the microstructure and mechanical properties of Mg-9Al alloy. The effect of Gd addition on the microstructure was investigated via X-ray diffraction (XRD), optical microscopy, scanning electron microscopy (SEM), and transmission electron microscopy (TEM). The Mg-9Al alloy contained two phases, α-Mg and β-Mg17Al12. Alloying with Gd led to the emergence of a new rectangular-shaped phase, Al₂Gd. The grain size also decreased marginally upon Gd addition. The ultimate tensile strength and microhardness of Mg-9Al alloy increased by 23% and 19%, respectively, upon 1.5wt% Gd addition. We observed that, although Mg-9Al-2.0Gd alloy exhibited the smallest grain size (181 μm) and the highest dislocation density (5.1×1010 m-2) among the investigated compositions, the Mg-9Al-1.5Gd alloy displayed the best mechanical properties. This anomalous behavior was observed because the Al₂Gd phase was uniformly distributed and present in abundance in Mg-9Al-1.5Gd alloy, whereas it was coarsened and asymmetrically conglomerated in Mg-9Al-2.0Gd.     

Microstructures and impact toughness behavior of Al 5083 alloy processed by cryorolling and afterwards annealing

The influence of rolling at liquid nitrogen temperature and annealing on the microstructure and mechanical properties of Al 5083 alloy was studied in this paper. Cryorolled samples of Al 5083 show significant improvements in strength and hardness. The ultimate tensile strength increases up to 340 MPa and 390 MPa for the 30% and 50% cryorolled samples, respectively. The cryorolled samples, with 30% and 50% reduction, were subjected to Charpy impact testing at various temperatures from ?190℃ to 100℃. It is observed that increasing the percentage of reduction of samples during cryorolling has significant effect on decreasing impact toughness at all temperatures by increasing yield strength and decreasing ductility. Annealing of samples after cryorolling shows remarkable increment in impact toughness through recovery and recrystallization. The average grain size of the 50% cryorolled sample (14 μm) after annealing at 350℃ for 1 h is found to be finer than that of the 30% cryorolled sample (25 μm). The scanning electron microscopy (SEM) analysis of fractured surfaces shows a large-size dimpled morphology, resembling the ductile fracture mechanism in the starting material and fibrous structure with very fine dimples in cryorolled samples corresponding to the brittle fracture mechanism.     

Lipid accumulation by a cellulolytic strain of Aspergillus niger.

Lipid accumulation by a cellulolytic mold, Aspergillus niger, was studied. The amount of lipid accumulated ranged from 13.6-16.6% on various carbon sources, namely glucose, xylose, avicel (microcrystalline cellulose) and bagasse (a natural lignocellulosic substrate). Neutral lipids, phospholipids and glycolipids of the mycelia varied from 41.0-46.2%, 34.9-38.4% and 18.7-22.6% of total lipids, respectively. Unsaturated fatty acids comprised around 80% of total fatty materials with linoleic and oleic acid predominating. Of the four nitrogen sources tested, NH4Cl was the best source for lipid synthesis from cellulose (bagasse). Optimum temperature range for growth and lipid synthesis was 25-30 degrees C.