Preliminary evaluation of monolithic column high-performance liquid chromatography with tris(2,2’-bipyridyl)ruthenium(II) chemiluminescence detection for the determination of quetiapine in human body fluids

High-performance liquid chromatography (HPLC) with tris(2,2-bipyridyl)ruthenium(II) chemiluminescence detection methodology is reported for the determination of the atypical antipsychotic drug quetiapine and the observation of its major active and inactive metabolites in human urine and serum. The method uses a monolithic chromatographic column allowing high flow rates of 3mL min−1 enabling rapid quantification. Flow injection analysis (FIA) with tris(2,2-bipyridyl)ruthenium(II) chemiluminescence detection and HPLC time of flight mass spectrometry (TOF-MS) were used for the determination of quetiapine in a pharmaceutical preparation to establish its suitability as a calibration standard. The limit of detection achieved with FIA was 2×10−11 mol L−1 in simple aqueous solution. The limits of detection achieved with HPLC were 7×10−8 and 2×10−10 mol L−1 in urine and serum, respectively. The calibration range for FIA was between 5×10−9 and 1×10−6 mol L−1. The calibration ranges for HPLC were between 1×10−7–1×10−4 and 1×10−8–1×10−4 mol L−1 in urine and serum, respectively. The quetiapine concentrations in patient samples were found to be 3×10−6 mol L−1 in urine and 7×10−7 mol L−1 in serum. Without the need for preconcentration, the HPLC detection limits compared favourably with those in previously published methodologies. The metabolites were identified using HPLC-TOF-MS.

Microstructures and bond strengths of the calcium phosphate coatings formed on titanium from different simulated body fluids

Calcium phosphate (Ca-P) coatings were deposited on Ti substrates by a biomimetic method from m-SBF and 10× SBF, respectively. Comparative study of microstructures and bond strengths of the Ca-P coatings deposited from those different SBFs was carried out. Effect of the surface roughness of the substrates on the bond strength of the Ca-P coatings was also studied. Scanning electron microscopy (SEM), X-ray diffractometry (XRD), Fourier transformed infrared spectroscopy (FTIR), inductive coupled plasma spectrometry (ICP) and thermogravimetry (TG) were used to characterize the Ca-P coatings. The bond strengths between the coatings and Ti substrates were measured using an adhesive strength test. Results indicated that the ionic concentrations of the SBFs and the surface roughness of the substrate had a significant influence on the formation, morphology and bond strength of the Ca-P precipitates. The induction period of time to deposit a complete Ca-P layer from the m-SBF is much longer, but the Ca-P coating is denser and has higher bond strength than that formed from the 10× SBF. The Ti with a surface roughness of R_a 0.64 µm and R_z 2.81 µm favoures the formation of a compact Ca-P coating from the m-SBF with the highest bond strength of approximately 15.5 MPa.

Bioassay detects soluble MAdCAM-1 in body fluids

Mucosal addressin cell adhesion molecule (MAdCAM-1) is a key player in mediating the infiltration of leucocytes into chronically inflamed tissues. Five anti-MAdCAM-1 monoclonal antibodies (mAb), designated 17F5, 201F7, 314G8, 377D10 and 355G8, were generated by fusion of P3 × 63Ag8.653 myeloma cells with spleen cells from BALB/c mice immunized with recombinant human MAdCAM-1-Fc. The latter four mAb recognize the ligand-binding first Ig domain, and block T -cell adhesion to MAdCAM-1. The non-blocking mAb 17F5 recognizes the mucin domain. Extensive analysis of a large panel of paraffin-embedded human tissues revealed that the 314G8 mAb detected MAdCAM-1 on venules in the spleen and small intestine. MAdCAM-1 was strongly expressed in the synovium of osteoarthritis patients, predominantly on the endothelial lining of blood vessels, but also within the vessel lumen. An ELISA, based on mAb 377D10 and 355G8, was developed to determine whether soluble MAdCAM-1 was present in body fluids, and to measure the levels present. The assay detected soluble MAdCAM-1 in the serum and urine of healthy donors, at levels similar to those of soluble forms of the related CAM, ICAM-1 and VCAM-1. The anti-MAdCAM-1 antibodies and assay developed here may be useful therapeutically in the treatment of inflammation in humans. Similarly, they may be useful diagnostically to monitor the presence and levels of MAdCAM-1.

Novel hypothalamic genes involved in body weight regulation in Psammomys obesus

Focuses on the discovery, characterisation and validation of a gene in the brain, named FIT, that regulates body weight. When suppressed in rats, food intake is inhibited and body weight is reduced. FIT regulates both appetite and metabolic rate, and is therefore a new and exciting target for obesity therapy.

On corrosion behaviour of magnesium alloy AZ31 in simulated body fluids and influence of ionic liquid pretreatments

This paper reports on the corrosion of Mg alloy AZ31 in simulated body fluid (SBF) using static immersion tests and electrochemical impedance spectroscopy. A preliminary study on the effect of flowing SBF on the corrosion behaviour of AZ31 has also been carried out. Low toxicity ionic liquids (ILs) trimethyl(butyl)phosphonium diphenyl phosphate P1444DPP and trihexyl(tetradecyl)-phosphonium bis-2,4,4trimethylpentyl-phosphinate [P66614][ i(C8) 2PO2] have been used to provide corrosion protection for AZ31 in SBF. Time dependent immersion tests indicate that under static conditions, AZ31 suffers severe localised corrosion in SBF, with pits developing predominantly beside the Al-Mn intermetallic phase in the α matrix. At longer immersion times, the corrosion product eventually precipitates and covers the entire specimen surface. When exposed to SBF under flowing conditions with a shear stress of 0·88 Pa, more uniform corrosion was observed. The optical profilometry results and electrochemical impedance spectroscopy analysis suggest that both P
1444DPP and [P66614][i(C8)2PO2] pretreatments can increase the corrosion resistance of AZ31 in SBF, in particular by decreasing the number of deeper pits found on the alloy surface. Cytotoxic test shows that the presence of the ILs P
1444DPP and [P66614][i(C8)2PO2] in cell culture media slightly inhibits the growth of human coronary artery endothelial cells in comparison with the good cell viability around the treated specimen. A pretreatment with IL is used in order to improve the corrosion resistance of this alloy in SBF. © 2012 Institute of Materials, Minerals and Mining.

A potential anti-corrosive ionic liquid coating for MG alloy AZ31 in simulated body fluids

Magnesium alloys are attractive materials for biomedical applications, due to their excellent biocompatibility. However, these alloys show fast corrosion rates in the body that limits their clinical applications. Low-toxic ionic liquid (IL) trimethyl(butyl)phosphonium diphenyl phosphate P1444dpp has been investigated to provide corrosion protection for magnesium alloy AZ31 in simulated body fluids (SBFs). This work reports a preliminary exploration of the influence of different treatment temperatures on the corrosion protection properties of IL films for the magnesium alloy AZ31 in SBFs. Results show that the IL treatment at room temperature did not bring significant improvement in the corrosion performance of the AZ31 in SBF. However, when the treatment temperature was increased to 75°C, the IL treatment resulted in a substantial reduction of the corrosion, in particular the reduction of localized pitting corrosion. The influence of ionic liquid treatment on the corrosion performance of the magnesium alloys AZ31 in SBFs has been investigated by electrochemical impedance spectroscopy (EIS) tests and immersion tests.

A role for skeletal muscle stearoyl-CoA desaturase 1 in control of thermogenesis

An enhanced metabolic efficiency for accelerating the recovery of fat mass (or catch-up fat) is a characteristic feature of body weight regulation after weight loss or growth retardation and is the outcome of an "adipose-specific" suppression of thermogenesis, i.e., a feedback control system in which signals from the depleted adipose tissue fat stores exert a suppressive effect on thermogenesis. Using a previously described rat model of semistarvation-refeeding in which catch-up fat results from suppressed thermogenesis per se, we report here that the gene expression of stearoyl-coenzyme A desaturase 1 (SCD1) is elevated in skeletal muscle after 2 wk of semistarvation and remains elevated in parallel to the phase of suppressed thermogenesis favoring catch-up fat during refeeding. These elevations in the SCD1 transcript are skeletal muscle specific and are associated with elevations in microsomal ^9 desaturase enzyme activity, in the ^9 desaturation index, and in the relative content of SCD1-derived monounsaturates in several lipid fractions extracted from skeletal muscle. An elevated skeletal muscle SCD1, by desaturating the products of de novo lipogenesis and diverting them away from mitochondrial oxidation, would inhibit substrate cycling between de novo lipogenesis and lipid oxidation, thereby leading to a state of suppressed thermogenesis that regulates the body’s fat stores.—Mainieri, D., Summermatter, S., Seydoux, J., Montani, J. P., Rusconi, S., Russell, A. P., Boss, O., Buchala, A. J., Dulloo, A. G. A role for skeletal muscle stearoyl-CoA desaturase 1 in control of thermogenesis.

Effect of Jersey type on thermoregulatory responses during exercise in a warm humid environment

The thermoregulatory responses of subjects wearing two different forms of rugby league jersey, one with plastic sponsorship recognition and numbering (trial Gl) and one without (trial G2), and a lightweight alternative (trial G3), were compared with a trial without any form of upper body garment (trial GO). Ten male volunteers, mean age 20.9 (±2.3) years, height 179.8 (±4.7) cm, weight 80.2 (±8.9) kg, and body surface area 1.99 (±0.13) m2, participated in this study. Subjects had a mean maximal oxygen uptake capacity of 56.0 (±6.3) ml.kg.min-1 and a sum of 8 skinfolds of 80.6 (±23.8) mm. Subjects were exercised at approximately 50% of maximal oxygen uptake in a warm humid environment for 50 minutes. Mean ambient temperature was 27.6°C (±0.32) with a relative humidity of 64.7% (±1.44). Measurements of core and skin (7 sites) temperature, heart rate, oxygen uptake, plasma volume, peak lactate concentration, and pre- and post-trial body weight, hematocrit and garment weight were recorded. The statistical results showed that all subjects experienced significant (p ≤.0001) decreases in body weight representing a percentage decrease ranging from 1.2-1.3%. No significant difference was found between trials with respect to body weight change. No significant effect of garment type was found on pre- and post-trial hematocrit, plasma volume changes or peak blood lactic acid concentration. However, mean peak lactate was highest for trial Gl (5.6 mmol.L-1 ±2.2) and lowest for trial G3 (4.6 mmol.L-1 ±1.27). Post-trial core temperature was significantly (p≤ .0001) higher than the resting value; no significant difference was found between trials. The mean absolute increase for all experimental trials was 0.9°C. A significant (p≤.005) difference between mean total (7 sites) skin temperature was found with a post-hoc test revealing that trials Gl and G2 were significantly higher than trial GO; no significant difference was found when comparing trial G3 with trial GO or when comparing the garments between each other. Mean skin temperature under the garment (4 sites) was found to be significantly (p≤.05) higher for all trials involving a garment when compared with mean skin temperature outside (3 sites) the garment; no significant difference was found between trials. Mean oxygen uptake was significantly different between trials (p≤.005), with trial Gl and G3 found to be significantly lower than trial GO; no difference was found when comparing the garments with each other. Post-trial garment weights were significantly (p≤.001) heavier than pre-trial and were significantly (p≤.0001) different when compared with each other. There was no significant effect on heart rate, haematocrit, plasma volume changes, peak blood lactic acid concentration, or core temperature due to garment type. However, differences in skin temperature suggest that the garment used in trial G3 may have a benefit. Further research should consider the impact of increased exercise intensity and/or environmental temperature and humidity on the measured parameters while wearing the garments described in this study.

Role of leptin receptors in the development of obesity

The focus of this dissertation was leptin and the leptin receptor, and the role of these genes (OB and OB-R) in the development of obesity and type 2 diabetes in humans and Psammomys obesus, a polygenic rodent model of obesity and type 2 diabetes. Studies in humans showed that circulating leptin concentrations were positively associated with adiposity, and independently associated with circulating insulin and triglyceride concentrations. Analysis of two leptin receptor sequence polymorphisms in a Caucasian Australian population and a population of Nauruan males, with very high prevalence rates of obesity, showed no associations between sequence variation within the OB-R gene and obesity- or diabetes-related phenotypic measures. In addition, these two OB-R polymorphisms were not associated with longitudinal changes in body mass or composition in either of the populations examined. A unique analysis of the effects of multiple gene defects in the Nauruan population, demonstrated that the presence of sequence alterations in both the OB and OB-R genes were associated with insulin resistance. Psammomys obesus is regarded as an excellent rodent model in which to study the development of obesity and type 2 diabetes in humans. Examination of circulating leptin concentrations in Psammomys revealed that, as in humans, leptin concentrations were associated with adiposity, and independently associated with circulating insulin concentrations. This animal model was utilised to examine expression of OB-R, and the regulation of expression of this gene after dietary manipulation. OB-R is known to have several isoforms, and in particular, OB-RA and OB-RB gene expression were examined. OB-RB is the main signalling isoform of the leptin receptors. It has a long intracellular domain and has previously been shown to play an important role in energy balance and body weight regulation in rodents and humans. OB-RA is a much shorter isoform of OB-R, and although it lacks the long intracellular domain necessary to activate the JAK/STAT pathway, OB-RA is also capable of signalling, although to a lesser degree than OB-RB. OB-RA is found to be expressed almost ubiquitously throughout the body, and this isoform may be involved in transport of leptin into the cell, although its role remains unclear. OB-RA and OB-RB were both found to be expressed in a large number of tissues in Psammomys obesus. Interestingly, obese Psammomys were found to have lower levels of expression of OB-RA and OB-RB in the hypothalamus, compared to lean animals. This finding raises the possibility that decreased leptin signalling in the brain of obese, hyperleptinemic Psammomys obesus may contribute to the leptin resistance previously described in this animal model. However, the primary defect is unclear, as alternatively, increased circulating leptin concentrations may lead to down-regulation of leptin receptors. The effect of fasting on leptin concentrations and gene expression of OB-RA and OB-RB was also examined. A 24-hour fast resulted in no change in body weight, but a reduction in circulating leptin concentrations, and an increase in hypothalamic OB-RB gene expression in lean Psammomys. In obese animals, fasting again did not alter body weight, but resulted in an increase in both circulating leptin concentrations and hypothalamic OB-RB gene expression. In the liver, fasting resulted in a large increase in OB-RA gene expression in both lean and obese animals. These results highlighted the fact that regulation of leptin receptor gene expression in polygenic models of obesity and type 2 diabetes is complex, and not solely under the control of circulating leptin concentrations. Sucrose-feeding is an established method of inducing obesity and type 2 diabetes in rodents, and this experimental paradigm was utilised to examine the effects of longer term perturbations of energy balance on the leptin signalling pathway in Psammomys obesus. Addition of a 5% sucrose solution to the diet of lean and obese Psammomys resulted in increased body weight in both groups of animals, however only obese Psammomys showed increased fat mass and the development of type 2 diabetes. The changes in body mass and composition with sucrose-feeding were accompanied by decreased circulating leptin concentrations in both groups of animals, as well as a range of changes in leptin receptor gene expression. Sucrose-feeding increased hypothalamic OB-RB gene expression in obese Psammomys only, while in the liver there was evidence of a reduction in OB-RA and OB-RB gene expression in both lean and obese animals. The direct effects of sucrose on the leptin signalling pathway are unclear, however it is possible to speculate that the effect of sucrose to decrease leptin concentrations may have been involved in the exacerbation of obesity and the development of type 2 diabetes in obese Psammomys, From these studies, it appears that sequence variation in the OB and OB-R genes is unlikely to be a major factor in the etiology of obesity in human populations. The ability to examine regulation of expression of these genes in Psammomys obesus, however, has demonstrated that the effects of nutritional modifications on leptin receptor gene expression need closer attention. The role of the OB and OB-R genes in metabolism and the development of type 2 diabetes also warrants further examination, with particular attention on the differential effects of dietary modifications on leptin receptor gene expression across a range of tissues.

Novel sphene coatings on Ti–6Al–4V for orthopedic implants using sol–gel method

Hydroxyapatite (HAp) is commonly used to coat titanium alloys (Ti–6Al–4V) for orthopedic implants. However, their poor adhesion strength and insufficient long-term stability limit their application. Novel sphene (CaTiSiO₅) ceramics possess excellent chemical stability and cytocompatibility. The aim of this study is to use the novel sphene ceramics as coatings for Ti–6Al–4V. The sol–gel method was used to produce the coatings and the thermal properties, phase composition, microstructure, thickness, surface roughness and adhesion strength of sphene coatings were analyzed by differential thermal analysis–thermal gravity (DTA–TG), X-ray diffraction (XRD), scanning electron microscopy (SEM), atom force microscopy (AFM) and scratch test, respectively. DTA analysis confirmed that the temperature of the sphene phase formation is 875 °C and XRD analysis indicated pure sphene coatings were obtained. A uniform structure of the sphene coating was found across the Ti–6Al–4V surface, with a thickness and surface roughness of the coating of about 0.5–1 μm and 0.38 μm, respectively. Sphene-coated Ti–6Al–4V possessed a significantly improved adhesion strength compared to that for HAp coating and their chemical stability was evaluated by testing the profile element distribution and the dissolution kinetics of calcium (Ca) ions after soaking the sphene-coated Ti–6Al–4V in Tris–HCl solution. Sphene coatings had a significantly improved chemical stability compared to the HAp coatings. A layer of apatite formed on the sphene-coated Ti–6Al–4V after they were soaked in simulated body fluids (SBF). Our results indicate that sol–gel coating of novel sphene onto Ti–6Al–4V possessed improved adhesion strength and chemical stability, compared to HAp-coated Ti–6Al–4V prepared under the same conditions, suggesting their potential application as coatings for orthopedic implants.

Two-dimensional correlation localized surface plasmon resonance spectroscopy for analysis of the interaction between metal nanoparticles and bovine serum albumin

Time-resolved extinction spectra assisted with two-dimensional correlation spectroscopy (2DCOS) analysis and principal component analysis (PCA) were employed to investigate the interaction between bovine serum albumin (BSA) and metal nanoparticles (NPs). A series of localized surface plasmon resonance (LSPR) spectra of metal NPs were measured just after a small amount of BSA was added into metal colloids. Through 2DCOS analysis, remarkable changes in the intensities of the LSPR were observed. The interaction process was totally divided into three periods according to the PCA. Transmission electron microscopy, dynamic light scattering, and ζ-potential measurements were also employed to characterize the interaction between BSA and metal NPs. The addition of BSA brings silver NPs to aggregate through the electrostatic interaction between them, but it has less effect on gold NPs. In a gold and silver mixed system, gold NPs can affect the interaction of silver NPs and BSA, leading it to weaken. The combination of 2DCOS analysis and LSPR spectroscopy is powerful for exploring the LSPR spectra of the metal NP involved systems. This combined technique holds great potential in LSPR sensing through analysis of slight, slim spectral changes of metal colloids

The effect of treatment temperature on corrosion resistance and hydrophilicity of an ionic liquid coating for mg-based stents

Mg alloys are attractive candidate materials for biodegradable stents. However, there are few commercially available Mg-based stents in clinical use because Mg alloys generally undergo rapid localized corrosion in the body. In this study, we report a new surface coating for Mg alloy AZ31 based on a low-toxicity ionic liquid (IL), tributyl(methyl)phosphonium diphenyl phosphate (P1,4,4,4 dpp), to control its corrosion rate. Emphasis is placed on the effect of treatment temperature. We showed that enhancing the treatment temperature provided remarkable improvements in the performances of both corrosion resistance and biocompatibility. Increasing treatment temperature resulted in a thicker (although still nanometer scale) and more homogeneous IL film on the surface. Scanning electron microscopy and optical profilometry observations showed that there were many large, deep pits formed on the surface of bare AZ31 after 2 h of immersion in simulated body fluid (SBF). The IL coating (particularly when formed at 100 °C for 1 h) significantly suppressed the formation of these pits on the surface, making corrosion occur more uniformly. The P1,4,4,4 dpp IL film formed at 100 °C was more hydrophilic than the bare AZ31 surface, which was believed to be beneficial for avoiding the deposition of the proteins and cells on the surface and therefore improving the biocompatibility of AZ31 in blood. The interaction mechanism between this IL and AZ31 was also investigated using ATR-FTIR, which showed that both anion and cation of this IL were present in the film, and there was a chemical interaction between dpp(-) anion and the surface of AZ31 during the film formation.

The evolution of the avian bill as a thermoregulatory organ.

The avian bill is a textbook example of how evolution shapes morphology in response to changing environments. Bills of seed-specialist finches in particular have been the focus of intense study demonstrating how climatic fluctuations acting on food availability drive bill size and shape. The avian bill also plays an important but under-appreciated role in body temperature regulation, and therefore in energetics. Birds are endothermic and rely on numerous mechanisms for balancing internal heat production with biophysical constraints of the environment. The bill is highly vascularised and heat exchange with the environment can vary substantially, ranging from around 2% to as high as 400% of basal heat production in certain species. This heat exchange may impact how birds respond to heat stress, substitute for evaporative water loss at elevated temperatures or environments of altered water availability, or be an energetic liability at low environmental temperatures. As a result, in numerous taxa, there is evidence for a positive association between bill size and environmental temperatures, both within and among species. Therefore, bill size is both developmentally flexible and evolutionarily adaptive in response to temperature. Understanding the evolution of variation in bill size however, requires explanations of all potential mechanisms. The purpose of this review, therefore, is to promote a greater understanding of the role of temperature on shaping bill size over spatial gradients as well as developmental, seasonal, and evolutionary timescales.

Effects of approach velocity and foot-target characteristics on the visual regulation of step length

Two questions emerge from the literature concerning the perceptual-motor processes underlying the visual regulation of step length. The first concerns the effects of velocity on the onset of visual control (VCO), when visual regulation of step length begins during goal-directed locomotion. The second concerns the effects of different obstacles such as a target or raised surface on step length regulation. In two separate experiments, participants (Experiment 1 & 2: n=12, 6 female, 6 male) walked, jogged, or sprinted towards an obstacle along a 10 m walkway, consisting of two marker-strips with alternating black and white 0.50 m markings. Each experiment consisted of three targeting or obstacle tasks with the requirement to both negotiate and continue moving (run-through) through the target. Five trials were conducted for each task and approach speed, with trials block randomised between the six participants of each gender. One 50 Hz video camera panned and filmed each trial from an elevated position, adjacent to the walkway. Video footage was digitized to deduce the gait characteristics. Results for the targeting tasks indicate a linear relationship between approach velocity and accuracy of final foot placement (r=0.89). When foot placement was highly constrained by the obstacle step length shortened during the entire approach. VCO was found to occur at an earlier tau-margin for lower approach velocities for both experiments, indicating that the optical variable ‘tau' is affected by approach velocity. A three-phase kinematic profile was found for all tasks, except for the take-off board condition when sprinting. Further research is needed to determine whether this velocity affect on VCO is due to ‘whole-body' approach velocity or whether it is a function of the differences between gait modes.