Challenges in mass spectrometry-based quantification of bioactive peptides: A case study exploring the neuropeptide Y family

The study of biologically active peptides is critical to the understanding of physiological pathways, especially those involved in the development of disease. Historically, the measurement of biologically active endogenous peptides has been undertaken by radioimmunoassay, a highly sensitive and robust technique that permits the detection of physiological concentrations in different biofluid and tissue extracts. Over recent years, a range of mass spectrometric approaches have been applied to peptide quantification with limited degrees of success. Neuropeptide Y (NPY), peptide YY (PYY), and pancreatic polypeptide (PP) belong to the NPY family exhibiting regulatory effects on appetite and feeding behavior. The physiological significance of these peptides depends on their molecular forms and in vivo concentrations systemically and at local sites within tissues. In this report, we describe an approach for quantification of individual peptides within mixtures using high-performance liquid chromatography electrospray ionization tandem mass spectrometry analysis of the NPY family peptides. Aspects of quantification including sample preparation, the use of matrix-matched calibration curves, and internal standards will be discussed. This method for the simultaneous determination of NPY, PYY, and PP was accurate and reproducible but lacks the sensitivity required for measurement of their endogenous concentration in plasma. The advantages of mass spectrometric quantification will be discussed alongside the current obstacles and challenges. © 2012 Wiley Periodicals, Inc. Biopolymers (Pept Sci) 98: 357–366, 2012.

Preliminary validation of a novel high-resolution melt-based typing method based on the multilocus sequence typing scheme of Streptococcus pyogenes

The major limitation of current typing methods for Streptococcus pyogenes, such as emm sequence typing and T typing, is that these are based on regions subject to considerable selective pressure. Multilocus sequence typing (MLST) is a better indicator of the genetic backbone of a strain but is not widely used due to high costs. The objective of this study was to develop a robust and cost-effective alternative to S. pyogenes MLST. A 10-member single nucleotide polymorphism (SNP) set that provides a Simpson’s Index of Diversity (D) of 0.99 with respect to the S. pyogenes MLST database was derived. A typing format involving high-resolution melting (HRM) analysis of small fragments nucleated by each of the resolution-optimized SNPs was developed. The fragments were 59–119 bp in size and, based on differences in G+C content, were predicted to generate three to six resolvable HRM curves. The combination of curves across each of the 10 fragments can be used to generate a melt type (MelT) for each sequence type (ST). The 525 STs currently in the S. pyogenes MLST database are predicted to resolve into 298 distinct MelTs and the method is calculated to provide a D of 0.996 against the MLST database. The MelTs are concordant with the S. pyogenes population structure. To validate the method we examined clinical isolates of S. pyogenes of 70 STs. Curves were generated as predicted by G+C content discriminating the 70 STs into 65 distinct MelTs.

Secondary curve behavior in Lenke Type 1C adolescent idiopathic scoliosis after thoracoscopic selective anterior thoracic fusion

Study Design. Analysis of a case series of 24 Lenke 1C adolescent idiopathic scoliosis (AIS) patients receiving selective thoracoscopic anterior scoliosis correction. Objective. To report the behaviour of the compensatory lumbar curve in a group of Lenke IC AIS patients following thoracoscopic anterior scoliosis correction, and to compare the results of this study with previously published data. Summary of Background Data. Several prior studies have reported spontaneous lumbar curve correction for both anterior and posterior selective fusion in Lenke 1C/King-Moe II patients; however to our knowledge no previous studies have reported outcomes of thoracoscopic anterior correction for this curve type. Methods. All AIS patients with a curve classification of Lenke 1C and a minimum of 24 months follow-up were retrieved from a consecutive series of 190 AIS patients who underwent thoracoscopic anterior instrumented fusion. Cobb angles of the major curve, instrumented levels, compensatory lumbar curve, and T5-T12 kyphosis were recorded, as well as coronal spinal balance, T1 tilt angle and shoulder balance. All radiographic parameters were measured before surgery and at 2, 6, 12 and 24 months after surgery. Results. Twenty-four female patients with right thoracic curves had a mean thoracic Cobb angle of 53.0° before surgery, decreasing to 24.9° two years after surgery. The mean lumbar compensatory Cobb angle was 43.5° before surgery, spontaneously correcting to 25.4° two years after surgery, indicating balance between the thoracic and lumbar scoliotic curves. The lumbar correction achieved (41.8%) compares favourably to previous studies. Conclusions. Selective thoracoscopic anterior fusion allows spontaneous lumbar curve correction and achieves coronal balance of main thoracic and compensatory lumbar curves, good cosmesis and patient satisfaction. Correction and balance are maintained 24 months after surgery.

Finite element modelling of load bearing steel stud walls under real building fires

Fire safety has become an important part in structural design due to the ever increasing loss of properties and lives during fires. Conventionally the fire rating of load bearing wall systems made of Light gauge Steel Frames (LSF) is determined using fire tests based on the standard time-temperature curve given in ISO 834 (ISO, 1999). The standard time-temperature curve given in ISO 834 (ISO, 1999) originated from the application of wood burning furnaces in the early 1900s. However, modern commercial and residential buildings make use of thermoplastic materials, which mean considerably high fuel loads. Hence a detailed fire research study into the performance of LSF walls was undertaken using the developed real fire curves based on Eurocode parametric curves (ECS, 2002) and Barnett’s BFD curves (Barnett, 2002) using both full scale fire tests and numerical studies. It included LSF walls without any insulation, and the recently developed externally insulated composite panel system. This paper presents the details of the numerical studies and the results. It also includes brief details of the development of real building fire curves and experimental studies.

Fire tests of load bearing steel stud walls exposed to real building fires

Abstract. Fire resistance has become an important part in structural design due to the ever increasing loss of properties and lives every year. Conventionally the fire rating of load bearing Light gauge Steel Frame (LSF) walls is determined using standard fire tests based on the time-temperature curve given in ISO 834 [1]. Full scale fire testing based on this standard time-temperature curve originated from the application of wood burning furnaces in the early 1900s and it is questionable whether it truly represents the fuel loads in modern buildings. Hence a detailed fire research study into the performance of LSF walls was undertaken using real design fires based on Eurocode parametric curves [2] and Barnett’s ‘BFD’ curves [3]. This paper presents the development of these real fire curves and the results of full scale experimental study into the structural and fire behaviour of load bearing LSF stud wall systems.

The persistence of phase-separation in LiFePO4 with two-dimensional Li+ transport : the Cahn-Hilliard-reaction equation and the role of defects

We examine the solution of the two-dimensional Cahn-Hilliard-reaction (CHR) equation in the xy plane as a model of Li+ intercalation into LiFePO4 material. We validate our numerical solution against the solution of the depth-averaged equation, which has been used to model intercalation in the limit of highly orthotropic diffusivity and gradient penalty tensors. We then examine the phase-change behaviour in the full CHR system as these parameters become more isotropic, and find that as the Li+ diffusivity is increased in the x direction, phase separation persists at high currents, even in small crystals with averaged coherency strain included. The resulting voltage curves decrease monotonically, which has previously been considered a hallmark of crystals that fill homogeneously.

Influence of atmospheric sublimation and evaporation on the heat pump fluid bed drying of bovine intestines

Experiments on atmospheric two-stage fluidized bed drying of bovine intestines with heat pump were carried out. The investigation covers innovative fluidized bed heat pump drying of bovine intestines. The two-stage drying consists of atmospheric moisture sublimation immediately followed by evaporation. Studies were done to establish the influence of the drying condition on the drying characteristics and product quality of bovine intestines and properties focusing on kinetics, diffusion, and color. The investigation of the drying characteristics has been conducted during moisture removal by evaporation and combined sublimation and evaporation. The effect of drying temperature on the drying constants was determined by fitting the experimental data using regression analysis techniques. The investigation revealed that the drying kinetics is most significantly affected by temperature. Correlations expressing the drying constants and effective moisture diffusivity dependence on the drying conditions are reported.

Characterization of sky-region morphological-temporal airborne collision detection

Automated airborne collision-detection systems are a key enabling technology for facilitat- ing the integration of unmanned aerial vehicles (UAVs) into the national airspace. These safety-critical systems must be sensitive enough to provide timely warnings of genuine air- borne collision threats, but not so sensitive as to cause excessive false-alarms. Hence, an accurate characterisation of detection and false alarm sensitivity is essential for understand- ing performance trade-offs, and system designers can exploit this characterisation to help achieve a desired balance in system performance. In this paper we experimentally evaluate a sky-region, image based, aircraft collision detection system that is based on morphologi- cal and temporal processing techniques. (Note that the examined detection approaches are not suitable for the detection of potential collision threats against a ground clutter back- ground). A novel collection methodology for collecting realistic airborne collision-course target footage in both head-on and tail-chase engagement geometries is described. Under (hazy) blue sky conditions, our proposed system achieved detection ranges greater than 1540m in 3 flight test cases with no false alarm events in 14.14 hours of non-target data (under cloudy conditions, the system achieved detection ranges greater than 1170m in 4 flight test cases with no false alarm events in 6.63 hours of non-target data). Importantly, this paper is the first documented presentation of detection range versus false alarm curves generated from airborne target and non-target image data.

Melt-textured YBa2Cu3O7-x wires having plate-like grains

YBCO wires which consist of well oriented plate-like fine grains are fabricated using a moving furnace to achieve higher mechanical strength. Melt-texturing experiments have been undertaken on YBCO wires with two different compositions: YBa1.5Cu2.9O7-x, and YBa1.8Cu3.0O7-x. Wires are extruded from a mixture of precursor powders (formed by a coprecipitation process) then textured by firing in a moving furnace. Size of secondary phases such as barium cuprate and copper oxide, and overall composition of the sample affect the orientation of the fine grains. At zero magnetic field, the YBa1.5Cu2.9O7-x wire shows the highest critical current density of 1,450 Acm-2 and 8,770 Acm-2 at 77K and 4.2K, respectively. At 1 T, critical current densities of 30 Acm-2 and 200 Acm-2, respectively, are obtained at 77K and 4.2K. Magnetisation curves are also obtained for one sample to evaluate critical current density using the Bean model. Analysis of the microstructure indicates that the starting composition of the green body significantly affects the achievement of grain alignment via melt-texturing processes.

Evaluation of the "reconceptualising early mathematics learning" project

The Pattern and Structure Mathematics Awareness Project (PASMAP) has investigated the development of patterning and early algebraic reasoning among 4 to 8 year olds over a series of related studies. We assert that an awareness of mathematical pattern and structure enables mathematical thinking and simple forms of generalisation from an early age. The project aims to promote a strong foundation for mathematical development by focusing on critical, underlying features of mathematics learning. This paper provides an overview of key aspects of the assessment and intervention, and analyses of the impact of PASMAP on students’ representation, abstraction and generalisation of mathematical ideas. A purposive sample of four large primary schools, two in Sydney and two in Brisbane, representing 316 students from diverse socio-economic and cultural contexts, participated in the evaluation throughout the 2009 school year and a follow-up assessment in 2010. Two different mathematics programs were implemented: in each school, two Kindergarten teachers implemented the PASMAP and another two implemented their regular program. The study shows that both groups of students made substantial gains on the ‘I Can Do Maths’ assessment and a Pattern and Structure Assessment (PASA) interview, but highly significant differences were found on the latter with PASMAP students outperforming the regular group on PASA scores. Qualitative analysis of students’ responses for structural development showed increased levels for the PASMAP students; those categorised as low ability developed improved structural responses over a relatively short period of time.

Numerical modelling and full scale testing of load bearing steel walls under real fires

Fire safety has become an important part in structural design due to the ever increasing loss of properties and lives during fires. Fire rating of load bearing wall systems made of Light gauge Steel Frames (LSF) is determined using fire tests based on the standard time-temperature curve given in ISO 834. However, modern residential buildings make use of thermoplastic materials, which mean considerably high fuel loads. Hence a detailed fire research study into the performance of load bearing LSF walls was undertaken using a series of realistic design fire curves developed based on Eurocode parametric curves and Barnett’s BFD curves. It included both full scale fire tests and numerical studies of LSF walls without any insulation, and the recently developed externally insulated composite panels. This paper presents the details of fire tests first, and then the numerical models of tested LSF wall studs. It shows that suitable finite element models can be developed to predict the fire rating of load bearing walls under real fire conditions. The paper also describes the structural and fire performances of externally insulated LSF walls in comparison to the non-insulated walls under real fires, and highlights the effects of standard and real fire curves on fire performance of LSF walls.

Thermal performance of composite panels under fire conditions using numerical studies : plasterboards, rockwool, glass fibre and cellulose insulations

In recent times, light gauge steel frame (LSF) wall systems are increasingly used in the building industry. They are usually made of cold-formed and thin-walled steel studs that are fire-protected by two layers of plasterboard on both sides. A composite LSF wall panel system was developed recently, where an insulation layer was used externally between the two plasterboards to improve the fire performance of LSF wall panels. In this research, finite element thermal models of the new composite panels were developed using a finite element program, SAFIR, to simulate their thermal performance under both standard and Eurocode design fire curves. Suitable apparent thermal properties of both the gypsum plasterboard and insulation materials were proposed and used in the numerical models. The developed models were then validated by comparing their results with available standard fire test results of composite panels. This paper presents the details of the finite element models of composite panels, the thermal analysis results in the form of time-temperature profiles under standard and Eurocode design fire curves and their comparisons with fire test results. Effects of using rockwool, glass fibre and cellulose fibre insulations with varying thickness and density were also investigated, and the results are presented in this paper. The results show that the use of composite panels in LSF wall systems will improve their fire rating, and that Eurocode design fires are likely to cause severe damage to LSF walls than standard fires.

Exponential asymptotics of free surface flow due to a line source

The steady problem of free surface flow due to a submerged line source is revisited for the case in which the fluid depth is finite and there is a stagnation point on the free surface directly above the source. Both the strength of the source and the fluid speed in the far field are measured by a dimensionless parameter, the Froude number. By applying techniques in exponential asymptotics, it is shown that there is a train of periodic waves on the surface of the fluid with an amplitude which is exponentially small in the limit that the Froude number vanishes. This study clarifies that periodic waves do form for flows due to a source, contrary to a suggestion by Chapman & Vanden-Broeck (2006, J. Fluid Mech., 567, 299--326). The exponentially small nature of the waves means they appear beyond all orders of the original power series expansion; this result explains why attempts at describing these flows using a finite number of terms in an algebraic power series incorrectly predict a flat free surface in the far field.

Kinetic study of goethite dehydration and the effect of aluminium substitution on the dehydrate

Goethite and Al-substituted goethite were synthesized and were characterized using XRD and XRF. The kinetic study of goethite dehydrate was investigated by TG and DTG at different heating rates (2, 5, 10, 15, 20 ◦C/min) and the effect of Al substitution for Fe on dehydrate was studied. The results showed that two types of absorbed water with the same Ed values of 3.4, 6.2 kJ/mol were confirmed on goethite and Alsubstituted goethite. Three types of hydroxyl units were proved, one being on the surface and the other two being in the structure of goethite. The substitution of Al for Fe in the structure of goethite decreases the desorption rate of hydroxyl, increases the dehydroxylation temperature, broadens the desorption peaks in DTG curves, and improves the Ed values from 19.4, 20.4, 26.1 kJ/mol to 21.6, 30, 33.6 kJ/mol when Al substitution comes to 9.1%.

Principles of motor learning in ecological dynamics : a comment on functions of learning and the acquisition of motor skills (with reference to sport)

This paper provides a commentary on the contribution by Dr Chow who questioned whether the functions of learning are general across all categories of tasks or whether there are some task-particular aspects to the functions of learning in relation to task type. Specifically, they queried whether principles and practice for the acquisition of sport skills are different than what they are for musical, industrial, military and human factors skills. In this commentary we argue that ecological dynamics contains general principles of motor learning that can be instantiated in specific performance contexts to underpin learning design. In this proposal, we highlight the importance of conducting skill acquisition research in sport, rather than relying on empirical outcomes of research from a variety of different performance contexts. Here we discuss how task constraints of different performance contexts (sport, industry, military, music) provide different specific information sources that individuals use to couple their actions when performing and acquiring skills. We conclude by suggesting that his relationship between performance task constraints and learning processes might help explain the traditional emphasis on performance curves and performance outcomes to infer motor learning.