Peptide-Mediated Gene Transfer into Marine Purple Photosynthetic Bacteria

Use of photosynthetic organisms is one of the sustainable ways to produce high-value products. Marine purple photosynthetic bacteria are one of the research focuses as microbial production hosts. Genetic transformation is indispensable as a biotechnology technique. However, only conjugation has been determined to be an applicable method for the transformation of marine purple photosynthetic bacteria so far. In this study, for the first time, a dual peptide-based transformation method combining cell penetrating peptide (CPP), cationic peptide and Tat-derived peptide (dTat-Sar-EED) (containing D-amino acids of Tat and endosomal escape domain (EED) connected by sarcosine linkers) successfully delivered plasmid DNA into Rhodovulum sulfidophilum, a marine purple photosynthetic bacterium. The plasmid delivery efficiency was greatly improved by dTat-Sar-EED. The concentrations of dTat-Sar-EED, cell growth stage and recovery duration affected the efficiency of plasmid DNA delivery. The delivery was inhibited at 4 °C and by A22, which is an inhibitor of the actin homolog MreB. This suggests that the plasmid DNA delivery occurred via MreB-mediated energy dependent process. Additionally, this peptide-mediated delivery method was also applicable for E. coli cells. Thus, a wide range of bacteria could be genetically transformed by using this novel peptide-based transformation method.     

Influence of post-deposition annealing in oxygen ambient on metal–organic decomposed CeO2 film spin coated on 4H-SiC

Post-deposition annealing (PDA) was performed in oxygen ambient at different temperatures (600, 800, 1,000 and 1,150 °C) onto metal–organic decomposition (MOD) prepared CeO₂ film spin coated on n-type 4H-SiC substrate. Effects of PDA onto the physical and electrical characteristics of MOD-derived CeO₂ were investigated. Four orientations [(111), (200), (220), and (311)] of CeO₂ peaks were revealed by X-ray diffraction analysis in all of the samples with a preferred orientation in (200) direction. However, α-Ce₂O₃ and cerium silicate (Ce₂Si₂O₇) interfacial layers emerged at 1,150 °C. As annealing temperature increased, grain size of films was increased but microstrains were decreased. Electrical results indicated that negative effective oxide charge and slow trap density were decreased as temperature increased. The lowest interface trap density was perceived by sample annealed at 1,000 °C. However, the highest electric field was obtained by sample annealed at 1,150 °C. Reasons that contributed to this observation were discussed.     

Structural studies of aluminium nitride embedded in amorphous carbon

Aluminum nitride containing diamond-like carbon was fabricated with pulsed laser deposition without post processing. The compositions of the targets used were varied at 1, 5, 10, 15 at.% and pure carbon was used as a reference. The films were comprehensively characterized with Atomic force microscope (AFM), X-ray photoelectron spectroscopy (XPS) and Transmission electron microscopy (TEM). Roughness analysis using AFM showed an increasing root-mean-square (RMS) roughness with increasing AIN content in target, while XPS analysis showed that the aluminum-nitrogen bonding was still present in the films after the fabrication process. Microstructural studies and selected area electron diffraction (SAED) pattern confirmed the presence of AIN crystals in DLC matrix. This nanostructured composite material is useful for luminescence applications.     

A novel and wide substrate specific polyhydroxyalkanoate (PHA) synthase from unculturable bacteria found in mangrove soil

This study reports the discovery of a polyhydroxyalkanoate (PHA) synthase (PhaC) possessing very wide substrate specificity from a mangrove soil metagenome. For the first time, putative PhaCs were identified from a metagenome using next-generation sequencing (NGS) and bioinformatic approaches. High-throughput shotgun metagenomic sequencing was conducted using the Illumina HiSeq 2000 platform. Sequence annotation and bioinformatic analyses were performed using the MG-RAST metagenomic pipeline. Reads annotated as PhaC against the NCBI RefSeq database were retrieved using the MG-RAST RESTful API (Application Programming Interface). PhaC gene sequence assembly was accomplished using the SPAdes assembler. A total of two de novo assembled contigs were subjected to sequence verification. A putative PhaC sequence, “BP-M-CPF4”, was selected for functional assessment by in vivo PHA biosynthesis in a PHA-negative mutant. An artificial stop codon was added at the 3′-end of the incomplete coding gene sequence. This novel PhaC showed very broad substrate specificity with the ability to incorporate six types of PHA monomers, 3-hydroxybutyrate (3HB), 3-hydroxyvalerate (3HV), 4-hydroxybutyrate (4HB), 3-hydroxy-4-methylvalerate (3H4MV), 5-hydroxyvalerate (5HV) and 3-hydroxyhexanoate (3HHx) in the presence of suitable precursors. This PHA synthase is suitable for the biosynthesis of PHAs that can be used in various biomedical applications due to its ability to incorporate the lipase-degradable monomer sequences of 4HB and 5HV. This study demonstrates that a functional metagenomic approach using next-generation sequencing can be used to mine novel PHA synthases with interesting substrate specificities from unculturable microorganisms.     

An integrated case-based reasoning approach for intelligent help desk fault management

As modern business functions become more complex and knowledge-intensive, with increasing demands for quality services, there is an emerging trend for organisations to develop and deploy intelligent knowledge-based systems for mission-critical operations. Some of the challenges in successfully implementing this breed of systems depend on how well the intelligent system is integrated with conventional existing information systems and workflow, and the quality of the intelligent system itself. Developing quality expert systems lies in the effective modelling of cognitive processes of human experts and representation of various forms of related knowledge in a domain. An integrated intelligent system called the Intelligent Help Desk Facilitator (IHDF), has been developed for computer and network fault management. The system, which comprises various modules including an expert system, is successfully deployed in a problem response help desk environment of a local bank. This paper describes a cognitive-driven approach to the development of the expert system based on a hybrid knowledge representation and reasoning strategy. The approach incorporates a hybrid case-based reasoning (CBR) framework of techniques which include case memory organisation structures (discrimination networks and shared-featured networks), case indexing and retrieval schemes (fuzzy character-matching, nearest-neighbour similarity matching and knowledge-guided indexing); and an interactive and incremental style of reasoning. The paper discusses the design and implementation of the expert system component of IHDF and illustrates the appropriateness of the hybrid architecture for problem resolution and diagnostic types of applications.     

Coal gasification in a spouted bed

Low Btu gas has been produced by gasifying coal in a spouted bed reactor. Coal of size 0.8-3.6 mm is fed continuously to a 0.15 m diameter spouted bed of inerts and gasified using mixtures of steam and air. Tests of the process with Western Canadian coals of free swelling index 0, 4 and 7 are reported. Gases of heating value to 3.61 MJ/m³ were produced at coal throughputs of 0.188 kg/m²s with the reactor operating at atmospheric pressure and temperatures to 930°C. Characteristics of the spouted bed gasifier are presented and results compared to commercial moving and fluidized bed systems. A simple mathematical model based on the two-region spouted bed model of Mathur and Lim is used to predict the effect on steam utilization of bed composition, bed height and diameter, and particle size.     

Lateral Optical Sensor With Slip Detection for Locating Live Products on Moving Conveyor

This paper presents a method to determine the 2-D profile and motion of a live product (such as chicken for poultry meat processing) on a moving conveyor from a lateral optical sensor that consists of an orthogonal pair of line array (LA) scanners. Unlike most line array (LA) scanners designed to provide a 2-D image of a static object, the lateral optical sensor presented here offers a practical means to detect object slippage on the conveyor in real time. Three examples are given to illustrate the effectiveness of this sensing method. The first simulates the 2-D boundary of a geometrically well-defined object on an accelerating conveyor, which offers intuitive insights on the effects of conveyor dynamics and object slippage on the accuracy of the 2-D boundary measurement. The second experimentally demonstrates the extendibility of LA sensors to detect both engineering and natural objects. The final example illustrates the application of the lateral optical sensor as a real time feedback sensor for active singulation of natural objects.