A Method for Isolation of Protoplasts from Dermatophytes

A method has been developed to isolate protoplasts from dermatophytes using Novozym 234. A simple technique of flotation in MgSO, has been adapted to separate protoplasts from incubation mixture. Electron microscopic studies confirmed the absence of cell wall material on these protoplasts. The recovery of DNA from protoplasts was higher than from mycelia.

Separating grazing and rainfall effects at regional scale using remote sensing imagery: A dynamic reference-cover method

Remote detection of management-related trend in the presence of inter-annual climatic variability in the rangelands is difficult. Minimally disturbed reference areas provide a useful guide, but suitable benchmarks are usually difficult to identify. We describe a method that uses a unique conceptual framework to identify reference areas from multitemporal sequences of ground cover derived from Landsat TM and ETM+ imagery. The method does not require ground-based reference sites nor GIS layers about management. We calculate a minimum ground cover image across all years to identify locations of most persistent ground cover in years of lowest rainfall. We then use a moving window approach to calculate the difference between the window's central pixel and its surrounding reference pixels. This difference estimates ground-cover change between successive below-average rainfall years, which provides a seasonally interpreted measure of management effects. We examine the approach's sensitivity to window size and to cover-index percentiles used to define persistence. The method successfully detected management-related change in ground cover in Queensland tropical savanna woodlands in two case studies: (1) a grazing trial where heavy stocking resulted in substantial decline in ground cover in small paddocks, and (2) commercial paddocks where wet-season spelling (destocking) resulted in increased ground cover. At a larger scale, there was broad agreement between our analysis of ground-cover change and ground-based land condition change for commercial beef properties with different a priori ratings of initial condition, but there was also some disagreement where changing condition reflected pasture composition rather than ground cover. We conclude that the method is suitably robust to analyse grazing effects on ground cover across the 1.3 x 10(6) km(2) of Queensland's rangelands. Crown Copyright (c) 2012 Published by Elsevier Inc. All rights reserved.

Focus on opportunities as a boundary condition of the relationship between job control and work engagement: A multi-sample, multi-method study

The concept of focus on opportunities describes how many new goals, options, and possibilities employees believe to have in their personal future at work. In this multi-sample, multi-method study, the authors investigated relationships between focus on opportunities and general and daily work engagement and the moderating role of focus on opportunities on between- and within-person relationships between job control and work engagement. Based on a social cognitive theory framework on the motivating potential of a future temporal focus, it was hypothesized that focus on opportunities is positively related to work engagement. Further, consistent with the notion of compensatory resources, it was expected that job control is not related to work engagement among employees with a high focus on opportunities, whereas job control, as an external resource of the work environment, is positively related to work engagement among employees with a low focus on opportunities. Both a cross-sectional survey study (N=174) and a daily diary study (N=64) supported the hypotheses. The study contributes to research on the job demands-resources model as it emphasizes the role of focus on opportunities as a motivational factor in the relationship between job control and work engagement.

Folates in rye: Determination and enhancement by food processing

The average daily intake of folate, one of the B vitamins, falls below recommendations among the Finnish population. Bread and cereals are the main sources of folate, rye being the most significant single source. Processing is a prerequisite for the consumption of whole grain rye; however, little is known about the effect of processing on folates. Moreover, data on the bioavailability of endogenous cereal folates are scarce. The aim of this study was to examine the variation in as well as the effect of fermentation, germination, and thermal processes on folate contents in rye. Bioavailability of endogenous rye folates was investigated in a four-week human intervention study. One of the objectives throughout the work was to optimise and evaluate analytical methods for determining folate contents in cereals. Affinity chromatographic purification followed by high-performance liquid chromatography (HPLC) was a suitable method for analysing cereal products for folate vitamers, and microbiological assay with Lactobacillus rhamnosus reliably quantified the total folate. However, HPLC gave approximately 30% lower results than the microbiological assay. The folate content of rye was high and could be further increased by targeted processing. The vitamer distribution of whole grain rye was characterised by a large proportion of formylated vitamers followed by 5-methyltetrahydrofolate. In sourdough fermentation of rye, the studied yeasts synthesized and lactic acid bacteria mainly depleted folate. Two endogenous bacteria isolated from rye flour were found to produce folate during fermentation. Inclusion of baker s yeast in sourdough fermentation raised the folate level so that the bread could contain more folate than the flour it was made of. Germination markedly increased the folate content of rye, with particularly high folate concentrations in hypocotylar roots. Thermal treatments caused significant folate losses but the preceding germination compensated well for the losses. In the bioavailability study, moderate amounts of endogenous folates in the form of different rye products and orange juice incorporated in the diet improved the folate status among healthy adults. Endogenous folates from rye and orange juice showed similar bioavailability to folic acid from fortified white bread. In brief, it was shown that the folate content of rye can be enhanced manifold by optimising and combining food processing techniques. This offers some practical means to increase the daily intake of folate in a bioavailable form.

Food and Indoor Air Isolated Bacillus Non-Protein Toxins: : Structures, Physico-Chemical Properties and Mechanisms of Effects on Eukaryotic Cells

We report here the structures and properties of heat-stable, non-protein, and mammalian cell-toxic compounds produced by spore-forming bacilli isolated from indoor air of buildings and from food. Little information is available on the effects and occurrence of heat-stable non-protein toxins produced by bacilli in moisture-damaged buildings. Bacilli emit spores that move in the air and can serve as the carriers of toxins, in a manner similar to that of the spores of toxic fungi found in contaminated indoor air. Bacillus spores in food cause problems because they tolerate the temperatures applied in food manufacture and the spores later initiate growth when food storage conditions are more favorable. Detection of the toxic compounds in Bacillus is based on using the change in mobility of boar spermatozoa as an indicator of toxic exposure. GC, LC, MS, and nuclear magnetic resonance NMR spectroscopy were used for purification, detection, quantitation, and analysis of the properties and structures of the compounds. Toxicity and the mechanisms of toxicity of the compounds were studied using boar spermatozoa, feline lung cells, human neural cells, and mitochondria isolated from rat liver. The ionophoric properties were studied using the BLM (black-lipid membrane) method. One novel toxin, forming ion channels permeant to K+ > Na+ > Ca2+, was found and named amylosin. It is produced by B. amyloliquefaciens isolated from indoor air of moisture-damaged buildings. Amylosin was purified with an RP-HPLC and a monoisotopic mass of 1197 Da was determined with ESI-IT-MS. Furthermore, acid hydrolysis of amylosin followed by analysis of the amino acids with the GS-MS showed that it was a peptide. The presence of a chromophoric polyene group was found using a NMR spectroscopy. The quantification method developed for amylosin based on RP-HPLC-UV, using the macrolactone polyene, amphotericin B (MW 924), as a reference compound. The B. licheniformis strains isolated from a food poisoning case produced a lipopeptide, lichenysin A, that ruptured mammalian cell membranes and was purified with a LC. Lichenysin A was identified by its protonated molecules and sodium- and potassium- cationized molecules with MALDI-TOF-MS. Its protonated forms were observed at m/z 1007, 1021 and 1035. The amino acids of lichenysin A were analyzed with ESI-TQ-MS/MS and, after acid hydrolysis, the stereoisomeric forms of the amino acids with RP-HPLC. The indoor air isolates of the strain of B. amyloliquefaciens produced not only amylosin but also lipopeptides: the cell membrane-damaging surfactin and the fungicidal fengycin. They were identified with ESI-IT-MS observing their protonated molecules, the sodium- and potassium-cationized molecules and analysing the MS/MS spectra. The protonated molecules of surfactin and fengycin showed m/z values of 1009, 1023, and 1037 and 1450, 1463, 1493, and 1506, respectively. Cereulide (MW 1152) was purified with RP-HPLC from a food poisoning strain of B. cereus. Cereulide was identified with ESI-TQ-MS according to the protonated molecule observed at m/z 1154 and the ammonium-, sodium- and potassium-cationized molecules observed at m/z 1171, 1176, and 1192, respectively. The fragment ions of the MS/MS spectrum obtained from the protonated molecule of cereulide at m/z 1154 were also interpreted. We developed a quantification method for cereulide, using RP-HPLC-UV and valinomycin (MW 1110, which structurally resembles cereulide) as the reference compound. Furthermore, we showed empirically, using the BLM method, that the emetic toxin cereulide is a specific and effective potassium ionophore of whose toxicity target is especially the mitochondria.

Application of partition method to vibration problems of plates

The “partition method” or “sub-domain method” consists of expressing the solution of a governing differential equation, partial or ordinary, in terms of functions which satisfy the boundary conditions and setting to zero the error in the differential equation integrated over each of the sub-domains into which the given domain is partitioned. In this paper, the use of this method in eigenvalue problems with particular reference to vibration of plates is investigated. The deflection of the plate is expressed in terms of polynomials satisfying the boundary conditions completely. Setting the integrated error in each of the subdomains to zero results in a set of simultaneous, linear, homogeneous, algebraic equations in the undetermined coefficients of the deflection series. The algebraic eigenvalue problem is then solved for eigenvalues and eigenvectors. Convergence is examined in a few typical cases and is found to be satisfactory. The results obtained are compared with existing results based on other methods and are found to be in very good agreement.

Information, risk and trust in the food chain : Ex-ante valuation of consumer willingness to pay for beef quality information using the contingent valuation method

The safety of food has become an increasingly interesting issue to consumers and the media. It has also become a source of concern, as the amount of information on the risks related to food safety continues to expand. Today, risk and safety are permanent elements within the concept of food quality. Safety, in particular, is the attribute that consumers find very difficult to assess. The literature in this study consists of three main themes: traceability; consumer behaviour related to both quality and safety issues and perception of risk; and valuation methods. The empirical scope of the study was restricted to beef, because the beef labelling system enables reliable tracing of the origin of beef, as well as attributes related to safety, environmental friendliness and animal welfare. The purpose of this study was to examine what kind of information flows are required to ensure quality and safety in the food chain for beef, and who should produce that information. Studying the willingness to pay of consumers makes it possible to determine whether the consumers consider the quantity of information available on the safety and quality of beef sufficient. One of the main findings of this study was that the majority of Finnish consumers (73%) regard increased quality information as beneficial. These benefits were assessed using the contingent valuation method. The results showed that those who were willing to pay for increased information on the quality and safety of beef would accept an average price increase of 24% per kilogram. The results showed that certain risk factors impact consumer willingness to pay. If the respondents considered genetic modification of food or foodborne zoonotic diseases as harmful or extremely harmful risk factors in food, they were more likely to be willing to pay for quality information. The results produced by the models thus confirmed the premise that certain food-related risks affect willingness to pay for beef quality information. The results also showed that safety-related quality cues are significant to the consumers. In the first place, the consumers would like to receive information on the control of zoonotic diseases that are contagious to humans. Similarly, other process-control related information ranked high among the top responses. Information on any potential genetic modification was also considered important, even though genetic modification was not regarded as a high risk factor.

A Method of Characteristics for Solving Two-dimensional Unsteady Flow Problems

The aim of this investigation is to evolve a method of solving two-dimensional unsteady flow problems by the method of characteristics. This involves the reduction of the given system of equations to an equivalent system where only interior derivatives occur on a characteristic surface. From this system, four special bicharacteristic directional derivatives are chosen. A finite difference scheme is prescribed for solving the equations. General rectangular lattices are also considered. As an example, we investigate the propagation of an initial pressure distribution in a medium at rest.

Intrusion warning and assessment method for site safety enhancement

Intrusion (unauthorized stepping-into/staying-in a hazardous area), as a common type of near-miss, is the prime cause of the majority of incidents on construction sites including fall from heights, and striking against or being struck by moving objects. Accidents often occur because workers take shortcuts moving about the site without fully perceiving the potential dangers. A number of researches have been devoted to developing methods to prevent such behaviors mainly based on the theory of Behavior-Based Safety (BBS), which aims to cultivate safety behaviors among workers in accordance with safety regulations. In current BBS practice, trained observers and safety supervisors are responsible for safety behavior inspections following safety plans and operation regulations. The observation process is time-consuming and its effectiveness depends largely on the observer’s safety knowledge and experience, which often results in omissions or bias. This paper presents a reformed safety behavior modification approach by integrating a location-based technology with BBS. Firstly, a detailed background is provided, covering current intrusion problems on site, existing use of BBS for behavior improvement, difficulties in achieving widespread adoption and potential technologies for location tracking and in-time feedback. Then, a conceptual framework of positioning technology-enhanced BBS is developed, followed by details of the corresponding on-line supporting system, Real Time Location System (RTLS) and Virtual Construction System (VCS). The application of the system is then demonstrated and tested in a construction site in Hong Kong. Final comments are made concerning further research direction and prospects for wider adoption.

New nondestructive method for three-dimensional photoelasticity

A new set of equations describing completely the optical phenomena in a model involving continuous rotation of secondary axes and secondary principal-stress differences are obtained. These are solved by Peano-Baker method using experimentally determined characteristic parameters for several wavelengths of light. Experimental verifications are obtained for a rectangular bar subjected to combined torsion and tension. Paper was presented at Third SESA International Congress on Experimental Mechanics held in Los Angeles, CA on May 13–18, 1973.

Large-chamber methane and nitrous oxide measurements are comparable to the backward Lagrangian stochastic method

Measurement of individual emission sources (e.g., animals or pen manure) within intensive livestock enterprises is necessary to test emission calculation protocols and to identify targets for decreased emissions. In this study, a vented, fabric-covered large chamber (4.5 × 4.5 m, 1.5 m high; encompassing greater spatial variability than a smaller chamber) in combination with on-line analysis (nitrous oxide [N2O] and methane [CH4] via Fourier Transform Infrared Spectroscopy; 1 analysis min-1) was tested as a means to isolate and measure emissions from beef feedlot pen manure sources. An exponential model relating chamber concentrations to ambient gas concentrations, air exchange (e.g., due to poor sealing with the surface; model linear when ≈ 0 m3 s-1), and chamber dimensions allowed data to be fitted with high confidence. Alternating manure source emission measurements using the large-chamber and the backward Lagrangian stochastic (bLS) technique (5-mo period; bLS validated via tracer gas release, recovery 94-104%) produced comparable N2O and CH4 emission values (no significant difference at P < 0.05). Greater precision of individual measurements was achieved via the large chamber than for the bLS (mean ± standard error of variance components: bLS half-hour measurements, 99.5 ± 325 mg CH4 s-1 and 9.26 ± 20.6 mg N2O s-1; large-chamber measurements, 99.6 ± 64.2 mg CH4 s-1 and 8.18 ± 0.3 mg N2O s-1). The large-chamber design is suitable for measurement of emissions from manure on pen surfaces, isolating these emissions from surrounding emission sources, including enteric emissions. © © American Society of Agronomy, Crop Science Society of America, and Soil Science Society of America.

Quick abnormal-bid-detection method for construction contract auctions

Non-competitive bids have recently become a major concern in both Public and Private sector construction contract auctions. Consequently, several models have been developed to help identify bidders potentially involved in collusive practices. However, most of these models require complex calculations and extensive information that is difficult to obtain. The aim of this paper is to utilize recent developments for detecting abnormal bids in capped auctions (auctions with an upper bid limit set by the auctioner) and extend them to the more conventional uncapped auctions (where no such limits are set). To accomplish this, a new method is developed for estimating the values of bid distribution supports by using the solution to what has become known as the German tank problem. The model is then demonstrated and tested on a sample of real construction bid data and shown to detect cover bids with high accuracy. This work contributes to an improved understanding of abnormal bid behavior as an aid to detecting and monitoring potential collusive bid practices.

Scientists in Control: a progressive approach to preparing work ready graduates in QC, EQA and method evaluation

Introduction QC, EQA and method evaluation are integral to delivery of quality patient results. To ensure QUT graduates have a solid grounding in these key areas of practice, a theory-to-practice approach is used to progressively develop and consolidate these skills. Methods Using a BCG assay for serum albumin, each student undertakes an eight week project analysing two levels of QC alongside ‘patient’ samples. Results are assessed using both single rules and Multirules. Concomitantly with the QC analyses, an EQA project is undertaken; students analyse two EQA samples, twice in the semester. Results are submitted using cloud software and data for the full ‘peer group’ returned to students in spreadsheets and incomplete Youden plots. Youden plots are completed with target values and calculated ALP values and analysed for ‘lab’ and method performance. The method has a low-level positive bias, which leads to the need to investigate an alternative method. Building directly on the EQA of the first project and using the scenario of a lab that services renal patients, students undertake a method validation comparing BCP and BCG assays in another eight-week project. Precision and patient comparison studies allow students to assess whether the BCP method addresses the proportional bias of the BCG method and overall is a ‘better’ alternative method for analysing serum albumin, accounting for pragmatic factors, such as cost, as well as performance characteristics. Results Students develop understanding of the purpose and importance of QC and EQA in delivering quality results, the need to optimise testing to deliver quality results and importantly, a working knowledge of the analyses that go into ensuring this quality. In parallel to developing these key workplace competencies, students become confident, competent practitioners, able to pipette accurately and precisely and organise themselves in a busy, time pressured work environment.

Parents as co-researchers at home: Using an observational method to document young children’s use of technology

This paper discusses the use of observational video recordings to document young children’s use of technology in their homes. Although observational research practices have been used for decades, often with video-based techniques, the participant group in this study (i.e., very young children) and the setting (i.e., private homes), provide a rich space for exploring the benefits and limitations of qualitative observation. The data gathered in this study point to a number of key decisions and issues that researchers must face in designing observational research, particularly where non-researchers (in this case, parents) act as surrogates for the researcher at the data collection stage. The involvement of parents and children as research videographers in the home resulted in very rich and detailed data about children’s use of technology in their daily lives. However, limitations noted in the dataset (e.g., image quality) provide important guidance for researchers developing projects using similar methods in future. The paper provides recommendations for future observational designs in similar settings and/or with similar participant groups.