Dilute oxalic acid pretreatment for biorefining giant reed (Arundo donax L.)

Biomass pretreatment is essential to overcome recalcitrance of lignocellulose for ethanol production. In the present study we pretreated giant reed (Arundo donax L.), a perennial, rhizomatous lignocellulosic grass with dilute oxalic acid. The effects of temperature (170-190 °C), acid loading (2-10% w/w) and reaction time (15-40 min) were handled as a single parameter, combined severity. We explored the change in hemicellulose, cellulose and lignin composition following pretreatment and glucan conversion after enzymatic hydrolysis of the solid residue. Two different yeast strains, Scheffersomyces (Pichia) stipitis CBS 6054, which is a native xylose and cellobiose fermenter, and Saccharomyces carlsbergensis FPL-450, which does not ferment xylose or cellobiose, were used along with commercial cellulolytic enzymes in simultaneous saccharification and fermentation (SSF). S. carlsbergensis attained a maximum ethanol concentration of 15.9 g/l after 48 h at pH 5.0, while S. stipitis, at the same condition, took 96 h to reach a similar ethanol value; increasing the pH to 6.0 reduced the S. stipitis lag phase and attained 18.0 g/l of ethanol within 72 h.     

Light and electron microscopy study of the spermatheca of Eupholidoptera chabrieri bimucronata (Ramme, 1927) and Uromenus brevicollis trinacriae La Greca 1964 (Orthoptera: Tettigoniidae)

A study by both optical and electron microscopy has been carried out on the spermatheca of Eupholidoptera chabrieri bimucronata and Uromenus brevicollis trinacriae (Orthoptera: Tettigoniidae). In both the examined species, the spermatheca consists of a sac/kidney‐shaped seminal receptacle and a more or less tortuous spermathecal duct that opens into the common oviduct. The wall of both the organs consists of a pseudostratified epithelium surmounted by a cuticular intima; the latter is made up of a thicker endocuticle and an epicuticle. The epithelium shows two different cell types, irregularly arranged and with well differentiated functions: cuticle‐forming and gland cells. In both the species, the cuticle‐forming cells perform other functions, in addition to producing the cuticular intima. The gland cells never come in contact with the cuticular intima, have inside the reservoir a secretion whose appearance can diversify also in contiguous zones of the seminal receptacle. Based on our findings in both the species, the functions of the seminal receptacle would differ from those of the spermathecal duct. In the latter, some areas of the wall of the connecting tract show an activity of lysis, by contiguous epithelial cells, that could play a role in control and selection of spermatozoa. As for the feather‐shaped spermatodesms, similar in both the species, freeze‐fracture observations have shown that the acrosome of each spermatozoon regularly covers three‐quarters of the extension of the acrosome of the following spermatozoon. Finally, the significance of our findings, compared with what is known in literature, is discussed. Microsc. Res. Tech. 78:577–586, 2015. © 2015 Wiley Periodicals, Inc.     

New High-Permittivity AgNb1-xTaxO3 Microwave Ceramics: Part II, Dielectric Characteristics

Ceramic samples with the AgNb_{1− x} Ta _x O₃ (0.46 < x < 0.54) composition were synthesized and analyzed, with respect to their dielectric properties, in a microwave-frequency range and at radio frequencies. The permittivity over the compositional range 0.46 lessthan equal to x lessthan equal to 0.52 was >410, whereas the Q × f value, measured at 2 GHz, reaches values of 640–900 GHz. Dependence of the resonant frequency on temperature was not linear but did display a minimum at a particular temperature, which was dependent on the composition of the ceramic. Samples that were measured at radio frequencies displayed low dissipation factors ( tg δ= 7 ×10⁻⁴−18 × 10⁻⁴), a low aging rate (0.2% per time decade), and good dielectric strength (8 V/μm at a thickness of 0.6 mm). Microstructural investigations revealed the presence of precipitates and transformation twins that should be eliminated during additional processing optimization.     

Different response to acetic acid stress in Saccharomyces cerevisiae wild‐type and l‐ascorbic acid‐producing strains

Biotechnological processes are of increasing significance for industrial production of fine and bulk chemicals, including biofuels. Unfortunately, under operative conditions microorganisms meet multiple stresses, such as non‐optimal pH, temperature, oxygenation and osmotic stress. Moreover, they have to face inhibitory compounds released during the pretreatment of lignocellulosic biomasses, which constitute the preferential substrate for second‐generation processes. Inhibitors include furan derivatives, phenolic compounds and weak organic acids, among which acetic acid is one of the most abundant and detrimental for cells. They impair cellular metabolism and growth, reducing the productivity of the process: therefore, the development of robust cell factories with improved production rates and resistance is of crucial importance. Here we show that a yeast strain engineered to endogenously produce vitamin C exhibits an increased tolerance compared to the parental strain when exposed to acetic acid at moderately toxic concentrations, measured as viability on plates. Starting from this evidence, we investigated more deeply: (a) the nature and levels of reactive oxygen species (ROS); (b) the activation of enzymes that act directly as detoxifiers of reactive oxygen species, such as superoxide dismutase (SOD) and catalase, in parental and engineered strains during acetic acid stress. The data indicate that the engineered strain can better recover from stress by limiting ROS accumulation, independently from SOD activation. The engineered yeast can be proposed as a model for further investigating direct and indirect mechanism(s) by which an antioxidant can rescue cells from organic acid damage; moreover, these studies will possibly provide additional targets for further strain improvements. Copyright © 2013 John Wiley & Sons, Ltd.     

The Morphology Control of BaTiO3 Particles Synthesized in Water and a Water/Ethanol Solvent

The experimental conditions for the growth of shape‐controlled BaTiO₃ particles in NaOH and Ba(NO₃)₂ aqueous and water/ethanol solutions using various TiO₂‐containing precursors were studied at 80°C–100°C. The different chemistries and physical characteristics of the precursors resulted in different BaTiO₃ formation rates and morphologies_. Nanocrystalline anatase led to irregularly shaped BaTiO₃ particles, whereas star‐like, single‐crystalline BaTiO₃ particles grew from aerogel TiO₂ and sodium titanate (NT) belts in alkaline aqueous solutions. With the addition of ethanol, the star‐like BaTiO₃ particles changed to square‐like, the size of which decreased with an increase in the ethanol content. The electron microscopy observations supported a dissolution–precipitation mechanism as the primary reaction mechanism for the formation of BaTiO₃ nanocrystals, which further aggregated into single‐crystalline star‐ or square‐like particles by oriented attachment. The modification in the water solution with ethanol is believed to influence both the nucleation and aggregation process and consequently influence the particle shape and size.     

Visualizing and exploring profiles with calling context ring charts

Calling context profiling is an important technique for analyzing the performance of object‐oriented software with complex inter‐procedural control flow. The Calling Context Tree (CCT) is a common data structure that stores dynamic metrics, such as CPU time, separately for each calling context. As CCTs may comprise millions of nodes, there is a need for a condensed visualization that eases the localization of performance bottlenecks. In this article, we discuss Calling Context Ring Charts (CCRCs), a compact visualization for CCTs, where callee methods are represented in ring segments surrounding the caller's ring segment. In order to reveal hot methods, their callers, and callees, the ring segments can be sized according to a chosen dynamic metric. We describe two case studies where CCRCs help us to detect and fix performance problems in applications. A performance evaluation also confirms that our implementation can efficiently handle large CCTs. Copyright © 2010 John Wiley & Sons, Ltd.     

Molecular Mechanism of Action of 2‐Ferrocenyl‐1,1‐diphenylbut‐1‐ene on HL‐60 Leukemia Cells

The aim of this work was to investigate the mechanism of action of 2‐ferrocenyl‐1,1‐diphenylbut‐1‐ene ( 1 ) on HL‐60 human leukemia cells. While inactive against noncancerous cells, 1 provoked a concentration‐dependent decrease in viable tumor cells, primarily via apoptosis, as evidenced by analysis of cell morphology, activation of caspases 3 and 7, increased DNA fragmentation, and externalization of phosphatidylserine. Necrosis was observed only at the highest tested concentration (4 μM ). Compound 1 interfered with the cell cycle, causing an accumulation of cells in the G₁/G₀ phase. Interaction of 1 with dsDNA and ssDNA was observed by differential pulse voltammetry and confirmed by hyperchromicity in the UV/Vis spectra of dsDNA, with an interaction constant of 2×10⁴ M ^?1. Both the organic analogue 1,1,2‐triphenylbut‐1‐ene ( 2 ) and ferrocene were inactive against cancer and noncancer cell lines and did not react with DNA. These results reinforce the idea that the hybrid strategy of conjugating ferrocene to the structure of tamoxifen derivatives is advantageous in finding new substances with antineoplastic activity.     

The Mitochondrial Amidoxime Reducing Component (mARC): Involvement in Metabolic Reduction of N‐Oxides,Oximes and N‐Hydroxyamidinohydrazones

The mitochondrial amidoxime reducing component (mARC) is a molybdenum‐containing enzyme and capable of reducing N‐hydroxylated structures such as amidoxime prodrugs. In this study, we tested the involvement of mARC in the reduction of N‐oxides (amitriptyline‐N‐oxide, nicotinamide‐N‐oxide), oximes ((E)‐/(Z)‐2,4,6‐trimethylacetophenonoxime) and a N‐hydroxyamidinohydrazone (guanoxabenz). All groups are reduced by mARC proteins, and the enzymes are therefore involved in the interconversion of N‐oxygenated metabolites originating from cytochrome P450s and flavin‐containing monooxygenases. In addition, these structures open up further options for serving as prodrugs. Thus, with respect to these reactions, testing of candidates with N‐oxygenated structures should not solely be carried out in microsomal enzyme sources but as well in mitochondria. However, differences in the reduction of oximes and N‐oxides between the two isoforms, namely mARC1 and mARC2, were detectable; N‐oxides are exclusively reduced by mARC1. We therefore assume differences between the so far unknown 3D structures of the two proteins.