The role of HtrA as a chaperone and protease in bacterial pathogenisis

HtrA (High Temperature Requirement A) is a critical stress response protease and chaperone for many bacteria. HtrA is a multitasking protein which can degrade unfolded proteins, conduct specific proteolysis of some substrates for correct assembly, interact with substrates to ensure correct folding, assembly or localisation, and chaperone unfolded proteins. These functions are critical for the virulence of a number of bacterial pathogens, in some cases not simply due to the broad activities of HtrA in protection against the protein stress conditions which occur during virulence. But also due to the role of HtrA in either specific proteolysis or assembly of key protein substrates which function directly in virulence. Remarkably, these activities are all conducted without any requirement for ATP. The biochemical mechanism of HtrA relies both on the chymotryptic serine protease active site as well as the presence of two PDZ (protein binding) domains. The mechanism is a unique combination of activation by substrate motifs to alter the confirmation of the active site, and assembly into a multimeric complex which has enhanced degradation and may also act as a protective cage for proteins which are not degraded. The role of this protease in the pathogenesis of a number of bacteria and the details of its distinctive biochemical activation and assembly mechanisms are discussed in this chapter.

Vibrational spectroscopy of natural cave mineral monetite CaHPO4 and the synthetic analog

Monetite is a phosphate mineral formed by the reaction of the chemicals in bat guano with calcite substrates and is commonly found in caves. The analog of the mineral monetite CaHPO4 has been synthesized and the Raman and infrared spectra of the natural monetite originating from the Murra-el-elevyn Cave, Eucla, Western Australia, compared. Monetite is characterized by a complex set of phosphate bands that arise because of two sets of pairs of phosphate units in the unit cell. Raman and infrared bands are assigned to HPO4(2-), OH stretching and bending vibrations. Infrared bands at 1346 and 1402 cm−1 are assigned to POH deformation modes. Vibrational spectroscopy confirms the presence of monetite in the cave system.

Laboratory and pilot scale pretreatment of sugarcane bagasse by acidified aqueous glycerol solutions

Pretreatment of sugarcane bagasse with acidified aqueous glycerol solution was evaluated at both laboratory and pilot scales. Laboratory scale pretreatment (4.00 g dry mass in 40.00 g liquid) with glycerol solutions containing ≤ 20 wt% water and 1.2 wt% HCl at 130 °C for 60 min resulted in biomass having glucan digestibilities of ≥ 88%. Comparable glucan enzymatic digestibility of 90% was achieved with bagasse pretreated at pilot scale (10 kg dry mass in 60 kg liquid) using a glycerol solution containing 0.4 wt% HCl and 17 wt% water at 130 °C for 15 min. We attribute more efficient pretreatment at pilot scale (despite shorter reaction time and reduced acid content) to improved mixing and heat transfer in a horizontal reactor. Pretreatment of sugarcane bagasse with acid-catalysed glycerol solutions likely produces glycerol-glycosides, which together with hydrolysed lignin are potential substrates for the production of biopolymers.

Pt/Nanostructured RuO2/SiC Schottky Diode based hydrogen gas sensors

In this paper, we report the development of novel Pt/nanostructured RuO2/SiC Schottky diode based sensors for hydrogen gas applications. The nanostructured ruthenium oxide thin films were deposited on SiC substrates using radio frequency sputtering technique. Scanning electron microscopy revealed the sputtered RuO2 layer consists of nano-cubular structures with dimensions ranging between 10 and 50 nm. X-ray diffraction confirmed the presence of tetragonal ruthenium (IV) oxide, with preferred orientation along the (101) lattice plane. The current-voltage characteristics of the sensors were investigated towards hydrogen gas in synthetic air at different temperatures from 25 °C to 240 °C. The dynamic responses of the sensors were studied at an optimum temperature of 240 °C and a voltage shift of 304 mV was recorded toward 1% hydrogen gas.

Hydrogen gas sensing performance of Pt/SnO2 nanowires/SiC MOS devices

This paper presents material and gas sensing properties of Pt/SnO2 nanowires/SiC metal oxide semiconductor devices towards hydrogen. The SnO2 nanowires were deposited onto the SiC substrates by vapour-liquid-solid growth mechanism. The material properties of the sensors were investigated using scanning electron microscopy, transmission electron microscopy and X-ray photoelectron spectroscopy. The current-voltage characteristics have been analysed. The effective change in the barrier height for 1% hydrogen was found to be 142.91 meV. The dynamic response of the sensors towards hydrogen at different temperatures has also been studied. At 530°C, voltage shift of 310 mV for 1% hydrogen was observed.

Hydrogen sensing properties of Pt/Lanthanum oxide- molybdenum oxide nanoplatelet/SiC based Schottky diode

An investigation of the electrical and hydrogen sensing properties of a novel Schottky diode based on a nanostructured lanthanum oxide-molybdenum oxide compound is presented herein. Molybdenum oxide (MoO3) nanoplatelets were grown on SiC substrates via thermal evaporation which was then subsequently coated with lanthanum oxide (La2O3) by RF sputtering. The current-voltage characteristics and hydrogen sensing performance (change in barrier height and sensitivity as well as the dynamic response) were examined from 25 to 300°C. At 180°C, a voltage shift of 2.23V was measured from the sensor while exposed to 1% hydrogen gas under a 100 μA constant reverse bias current. The results indicate that the presence of a La2O3 thin layer substantially improves the hydrogen sensitivity of the MoO3 nanoplatelets.

Pt/TiO2 nanotubes/SiC schottky diodes for hydrogen gas sensing applications

Titanium oxide nanotubes Schottky diodes were fabricated for hydrogen gas sensing applications. The TiO2 nanotubes were synthesized via anodization of RF sputtered titanium films on SiC substrates. Two anodization potentials of 5 V and 20 V were used. Scanning electron microscopy of the synthesized films revealed nanotubes with avarage diameters of 20 nm and 75 nm. X-ray diffraction analysis revealed that the composition of the oxide varied with the anodization potential. TiO2 (anatase) being formed preferentially at 5 V and TiO (no anatase) at 20 V. Current-voltage characteristics of the diodes were studied towards hydrogen at temperatures from 25°C to 250°C. At constant current bias of −500 μA and 250°C, the lateral voltage shifts of 800 mV and 520 mV were recorded towards 1% hydrogen for the 5 V and 20 V anodized nanotubes, respectively.

Pt/MoO3 nano-flower/SiC Schottky diode based hydrogen gas sensor

In this paper, we report the development of a novel Pt/MoO3 nano-flower/SiC Schottky diode based device for hydrogen gas sensing applications. The MoO3 nanostructured thin films were deposited on SiC substrates via thermal evaporation. Morphological characterization of the nanostructured MoO3 by scanning electron microscopy revealed randomly orientated thin nanoplatelets in a densely packed formation of nano-flowers with dimensions ranging from 250 nm to 1 μm. Current-voltage characteristics of the sensor were measured at temperatures from 25°C to 250°C. The sensor showed greater sensitivity in a reverse bias condition than in forward bias. Dynamic response of the sensor was investigated towards different concentrations of hydrogen gas in a synthetic air mixture at 250°C and a large voltage shift of 5.7 V was recorded upon exposure to 1% hydrogen.

The feral horse foot. Part A : observational study of the effect of environment on the morphometrics of the feet of 100 Australian feral horses

Aim To better understand the morphology of, and the effect of different travel patterns and varying substrate environments on, the feral horse foot to better manage the feet of domestic horses. Methods The left forefeet of 20 adult feral horses from each of five geographically separated populations in Australia (n = 100) were investigated. Populations were selected on the basis of substrate hardness under foot and the amount of travel typical for the population. Feet were radiographed and photographed and 40 morphometric measurements of each foot were obtained. Results Of the 40 parameters, 37 differed significantly (P < 0.05) among the populations, which suggested that substrate hardness and travel distance have an effect on foot morphology. Harder substrates and longer travel distances were associated with short hoof walls and minimal hoof wall flaring. Softer substrates and moderate travel distances were associated with long flared walls, similar to that of typical untrimmed feet of domestic horses. Conclusions The morphology of the feral horse foot appeared to be affected by the distance travelled and by the abrasive qualities and mechanical properties of the substrate under foot. There were marked differences in some conformation parameters between the feral horses in the current study and domestic horses in previous studies. Although the conformation of the feral horse foot may have some prescriptive value, concerns regarding abnormal foot anatomy warrant further investigation.

Pt/Graphene Nano-sheet Based Hydrogen Gas Sensor

In this paper, we present gas sensing properties of Pt/graphene-like nano-sheets towards hydrogen gas. The graphene-like nano-sheets were produced via the reduction of spray-coated graphite oxide deposited on SiC substrates by hydrazine vapor. Structural and morphological characterizations of the graphene sheets were analyzed by scanning electron and atomic force microscopy. Current-voltage and dynamic responses of the sensors were investigated towards different concentrations of hydrogen gas in a synthetic air mixture at 100°C. A voltage shift of 100 mV was recorded at 1 mA reverse bias current.

Pt/ZnO/SiC thin film for hydrogen gas sensing

Zinc oxide (ZnO) is one of the most promising electronic and photonic materials to date. In this work, we present an enhanced ZnO Schottky gas sensor deposited on SiC substrates in comparison to those reported previously in literature. The performance of ZnO/SiC based Schottky thin film gas sensors produced a forward lateral voltage shift of 12.99mV and 111.87mV in response to concentrations of hydrogen gas at 0.06% and 1% in air at optimum temperature of 330 ºC. The maximum change in barrier height was calculated as 37.9 meV for 1% H2 sensing operation at the optimum temperature.

Pt/anodized TiO2/SiC-based MOS device for hydrocarbon sensing

Pt/anodized TiO2/SiC based metal-oxide-semiconductor (MOS) devices were fabricated and characterized for their sensitivity towards propene (C3H6). Titanium (Ti) thin films were deposited onto the SiC substrates using a filtered cathodic vacuum arc (FCVA) method. Fluoride ions containing neutral electrolyte (0.5 wt% NH4F in ethylene glycol)were used to anodize the Ti films. The anodized films were subsequently annealed at 600 °C for 4 hrs in an oxygen rich environment to obtain TiO2. The current-voltage(I-V) characteristics of the Pt/TiO2/SiC devices were measured in different concentrations of propene. Exposure to the analyte gas caused a change in the Schottky barrier height and hence a lateral shift in the I-V characteristics. The effective change in the barrier height for 1% propene was calculated as 32.8 meV at 620°C. The dynamic response of the sensors was also investigated and a voltage shift of 157 mV was measured at 620°C during exposure to 1% propene.

Pt/SnO2 nanowires/SiC based hydrogen gas sensor

Pt/SnO2 nanowires/SiC based metal-oxidesemiconductor (MOS) devices were fabricated and tested for their gas sensitivity towards hydrogen. Tin oxide (SnO2) nanowires were grown on SiC substrates by the vapour liquid solid growth process. The material properties of the SnO2 nanowires such as its formation and dimensions were analyzed using scanning electron microscopy (SEM). The currentvoltage (I-V) characteristics at different hydrogen concentrations are presented. The effective change in the barrier height for 0.06 and 1% hydrogen were found to be 20.78 and 131.59 meV, respectively. A voltage shift of 310 mV at 530°C for 1% hydrogen was measured.

Metabolic and ecological study of environmental pentose utilizing bacteria (E-PUB)

Lignocellulosic materials, such as sugar cane bagasse, a waste product of the sugarcane processing industry, agricultural residues and herbaceous crops, may serve as an abundant and comparatively cheap feedstock for largescale industrial fermentation, resulting in the production of marketable end-products. However, the complex structure of lignocellulosic materials, the presence of various hexose and pentose sugars in the hemicellulose component, and the presence of various compounds that inhibit the organisms selected for the fermentation process, all constitute barriers that add to the production costs and make full scale industrial production economically less feasible. The work presented in this thesis was conducted in order to screen microorganisms for ability to utilize pentose sugars derived from the sugar mill industrial waste. A large number of individual bacterial strains were investigated from hemi-cellulose rich material collected at the Proserpine and Maryborough sugar mills, notably soil samples from the mill sites. The research conducted to isolation of six pentose-capable Gram-positive organisms from the actinomycetes group by using pentose as a sole carbon source in the cultivation process. The isolates were identified as Corynebacterium glutamicum, Actinomyces odontolyticus, Nocardia elegans, and Propionibacterium freudenreichii all of which were isolated from the hemicellulose-enriched soil. Pentose degrading microbes are very rare in the environment, so this was a significant discovery. Previous research indicated that microbes could degrade pentose after genetic modification but the microbes discovered in this research were able to naturally utilize pentose. Six isolates, identified as four different genera, were investigated for their ability to utilize single sugars as substrates (glucose, xylose, arabinose or ribose), and also dual sugars as substrates (a hexose plus a pentose). The results demonstrated that C. glutamicum, A. odontolyticus, N. elegans, and P. freudenreichii were pentose-capable (able to grow using xylose or other pentose sugar), and also showed diauxie growth characteristics during the dual-sugar (glucose, in combination with xylose, arabinose or ribose) carbon source tests. In addition, it was shown that the isolates displayed very small differences in growth rates when grown on dual sugars as compared to single sugars, whether pentose or hexose in nature. The anabolic characteristics of C. glutamicum, A. odontolyticus, N. elegans and P. freudenreichii were subsequently investigated by qualitative analysis of their end-products, using high performance liquid chromatography (HPLC). All of the organisms produced arginine and cysteine after utilization of the pentose substrates alone. In addition, P. freudenreichii produced alanine and glycine. The end-product profile arising from culture with dual carbon sources was also tested. Interestingly, this time the product was different. All of them produced the amino acid glycine, when grown on a combination substrate-mix of glucose with xylose, and also glucose with arabinose. Only N. elegans was able to break down ribose, either singly or in combination with glucose, and the end-product of metabolism of the glucose plus ribose substrate combination was glutamic acid. The ecological analysis of microbial abundance in sugar mill waste was performed using denaturing gradient gel electrophoresis (DGGE) and also the metagenomic microarray PhyloChip method. Eleven solid samples and seven liquid samples were investigated. A very complex bacterial ecosystem was demonstrated in the seven liquid samples after testing with the PhyloChip method. It was also shown that bagasse leachate was the most different, compared to all of the other samples, by virtue of its richness in variety of taxa and the complexity of its bacterial community. The bacterial community in solid samples from Proserpine, Mackay and Maryborough sugar mills showed huge diversity. The information found from 16S rDNA sequencing results was that the bacterial genera Brevibacillus, Rhodospirillaceae, Bacillus, Vibrio and Pseudomonas were present in greatest abundance. In addition, Corynebacterium was also found in the soil samples. The metagenomic studies of the sugar mill samples demonstrate two important outcomes: firstly that the bagasse leachate, as potentially the most pentose-rich sample tested, had the most complex and diverse bacterial community; and secondly that the pentose-capable isolates that were initially discovered at the beginning of this study, were not amongst the most abundant taxonomic groups discovered in the sugar mill samples, and in fact were, as suspected, very rare. As a bioprospecting exercise, therefore, the study has discovered organisms that are naturally present, but in very small numbers, in the appropriate natural environment. This has implications for the industrial application of E-PUB, in that a seeding process using a starter culture will be necessary for industrial purposes, rather than simply assuming that natural fermentation might occur.

trans-Polyacetylene on sodium and cesium mordenites: a resonance Raman spectroscopic study

Sodium and cesium mordenite (denoted NaM and CsM, respectively) were investigated as potential catalysts for the synthesis of polyacetylene ((CH) x). Both were successful in initiating polymerization of purified gaseous acetylene at room temperature as evidenced by Raman spectroscopic studies. The polyacetylene synthesised in this way exhibited resonance enhancement of the polyene skeletal vibrations. trans-Polyacetylene, but no cis-(CH) x, was detected. As no apparent coloration of the NaM and CsM substrates accompanied the formation of trans-(CH) x it was concluded that only small quantities of the polymer were present. The number of conjugated double bonds was estimated from the frequencies of the Raman active C-C and C=C stretching vibrations, and it was shown that the trans-(CH) x formed on CsM has a distribution of conjugation lengths ranging from less than 6 to at least 30 double bonds. The polyacetylene formed on NaM was significantly shorter and was produced in lower yields than that synthesized on CsM. "Sliced" resonance excitation profiles of polyacetylene formed on CsM were obtained using nearly 40 different excitation wavelengths and these confirmed that the adsorbed trans-(CH) x was composed of segments having a distribution of conjugated lengths. The architecture of the mordenite pore system permitted only a single polymer molecule per channel, thereby preventing cross-linking. Raman spectroscopic studies of the effects of exposure to air revealed that progressive oxidative degradation occurred with a reduction in the number of conjugated double bond