Development of a liquid chromatographic time-of-flight mass spectrometric method for the determination of unlabelled and deuterium-labelled alpha-tocopherol in blood components

A method is described for the analysis of deuterated and undeuterated alpha-tocopherol in blood components using liquid chromatography coupled to an orthogonal acceleration time-of-flight (TOF) mass spectrometer. Optimal ionisation conditions for undeuterated (d0) and tri- and hexadeuterated (d3 or d6) alpha-tocopherol standards were found with negative ion mode electrospray ionisation. Each species produced an isotopically resolved single ion of exact mass. Calibration curves of pure standards were linear in the range tested (0-1.5 muM, 0-15 pmol injected). For quantification of d0 and d6 in blood components following a standard solvent extraction, a stable-isotope-labelled internal standard (d3-alpha-tocopherol) was employed. To counter matrix ion suppression effects, standard response curves were generated following identical solvent extraction procedures to those of the samples. Within-day and between-day precision were determined for quantification of d0- and d6-labelled alpha-tocopherol in each blood component and both averaged 3-10%. Accuracy was assessed by comparison with a standard high-performance liquid chromatography (HPLC) method, achieving good correlation (r(2) = 0.94), and by spiking with known concentrations of alpha-tocopherol (98% accuracy). Limits of detection and quantification were determined to be 5 and 50 fmol injected, respectively. The assay was used to measure the appearance and disappearance of deuterium-labelled alpha-tocopherol in human blood components following deuterium-labelled (d6) RRR-alpha-tocopheryl acetate ingestion. The new LC/TOFMS method was found to be sensitive, required small sample volumes, was reproducible and robust, and was capable of high throughput when large numbers of samples were generated. Copyright (C) 2003 John Wiley Sons, Ltd.

Analysis of the headspace aroma compounds of walnuts (Juglans regia L.)

The aroma volatiles of walnuts from three different geographical locations were studied. Over 110 compounds were identified in the headspace volatiles, many for the first time as walnut components. Walnuts from China and the Ukraine contained high levels of lipid-derived volatiles, in particular hexanal, pentanal, 1-hexanol and 1-pentanol from linoleic acid breakdown, and 1-penten-3-ol from alpha-linolenic acid breakdown. Chilean walnuts, however, contained high levels of alkylbenzenes of molecular weight 120, with the lipid-derived aldehydes and alcohols present at much lower levels than in the other two walnut samples. The relationship between the fatty acid composition of the walnuts and their volatile composition is discussed. Copyright (C) 2005 John Wiley & Sons, Ltd.

Molecular interactions between the penetration enhancer 1,8-cineole and human skin

Penetration enhancers are chemicals that temporarily and reversibly diminish the barrier function of the outermost layer of skin, the stratum corneum, to facilitate drug delivery to and through the tissue. In the current study, the complex mechanisms by which 1,8-cineole, a potent terpene penetration enhancer, disrupts the stratum corneum barrier is investigated using post-mortem skin samples. In order to validate the use of excised tissue for these and related studies, a fibre optical probe coupled to an FT-Raman spectrometer compared spectroscopic information for human skin recorded from in vivo and in vitro sampling arrangements. Spectra from full-thickness (epidermis and dermis) post-mortem skin samples presented to the spectrometer with minimal sample preparation (cold acetone rinse) were compared with the in vivo system (the forearms of human volunteers). No significant differences in the Raman spectra between the in vivo and in vitro samples were observed, endorsing the use of post-mortem or surgical samples for this investigational work. Treating post-mortem samples with the penetration enhancer revealed some unexpected findings: while evidence for enhancer-induced disruption of the barrier lipid packing in the stratum corneum was detected in some samples, spectra from other samples revealed an increase in lipid order on treatment with the permeation promoter. These findings are consistent with phase-separation of the enhancer within the barrier lipid domains as opposed to homogeneous disruption of the lipid lamellae. Copyright (C) 2006 John Wiley & Sons, Ltd.

The synthesis and cytotoxic evaluation of a series of benzodioxole substituted titanocenes

Using 6-benzo[1,3]dioxolefulvene (1a), a series of benzodioxole substituted titanocenes was synthesized. The benzyl-substituted titanocene bis[(benzo[1,3]dioxole)-5-methylcyclopentadienyl] titanium (IV) dichloride (2a) was synthesized from the reaction of Super Hydride with 1a. An X-ray determined crystal structure was obtained for 2a. The ansa-titanocene (1,2-di(cyclopentadienyl)1,2-di-(benzo[1,3]dioxole)-ethanediyl) titanium(IV) dichloride (2b) was synthesized by reductive dimerisation of la with titanium dichloride. The diarylmethyl substituted titanocene bis(di(benzo[1,3]dioxole)-S-methylcyclopentadienyl) titanium(IV) dichloride (20 was synthesized by reacting la with the para-lithiated benzodioxole followed by transmetallation with titanium tetrachloride. When titanocenes 2a-c were tested against pig kidney (LLC-PK) cells inhibitory concentrations (IC50) of 2.8 X 10(-4), 1.6 x 10(-4) and 7.6 x 10(-5) m, respectively, were observed. These values represent improved cytotoxicity against LLC-PK, when compared with unsubstituted titanocene dichloride, but are not as impressive as values obtained for titanocenes previously synthesized using the above methods. Copyright (c) 2006 John Wiley & Sons, Ltd.

Isopimaric acid from Pinus nigra shows activity against multidrug-resistant and EMRSA strains of Staphylococcus aureus

The diterpene isopimaric acid was extracted from the immature cones of Pinus nigra (Arnold) using bioassay. guided fractionation of a crude hexane extract. Isopimaric acid was assayed against multidrug-resistant (MDR) and methicillin-resistant Staphylococcus aureus (MRSA). The minimum inhibitory concentrations (MIC) were 32-64 mu g/mL and compared with a commercially obtained resin acid, abietic acid, with MICs of 64 mu g/mL. Resin acids are known to have antibacterial activity and are valued in traditional medicine for their antiseptic properties: These results show that isopimaric acid is active against MDR an MRSA strains of S. aureus which are becoming, increasingly resistant to antibiotics. Both compounds were evaluated for modulation activity in combination with antibiotics, but did not potentiate the activity of the antibiotics tested. However, the compounds were also assayed in combination with the efflux pump inhibitor reserpine, to ice if inhibition of the TetK or NorA efflux pump increased their activity. Interestingly, rather than a potentiation of activity by a reduction in MIC, a two to four-fold increase in MIC was seen. It may he that isopimaric acid and abietic acid are not substrates for these efflux pumps, but it is also possible that an antagonistic interaction with reserpine may render the antibiotics inactive. H-1-NMR of abietic acid and reserpine taken individually and in combination, revealed a shift in resonance of some peaks for both compounds when mixed together compared with the spectra of the compounds on their own. It is proposed that this may he due to complex formation between abietic acid and reserpine and that this complex formation is responsible for a reduction in activity and elevation of MIC. Copyright (c) 2005 John Wiley & Sons, Ltd.

Natural products as alternative treatments for metabolic bone disorders and for maintenance of bone health

Bone metabolism involves a complex balance between the deposition of matrix and mineralization and resorption. There is now good evidence that dietary components and herbal products can influence these processes, particularly by inhibiting bone resorption, thus having beneficial effects on the skeleton. For example, it has been reported that a number of common vegetables, including onion, garlic and parsley, can inhibit bone resorption in ovariectomized rats. Essential oils derived from sage, rosemary, thyme and other herbs inhibit osteoclast activity in vitro and in vitro and leading to an increase in bone mineral density. Soya, a rich source of isoflavones, has shown promising results and epidemiological evidence to support a use in maintaining bone health, and various traditional herbal formulae in Chinese and Ayurvedic medicine also have demonstrable effects in pharmacological models of osteoporosis. Recently, cannabinoids have been described as having positive effects on osteoblast differentiation, and the presence of cannabinoid receptors in bone tissue indicates a more complex role in bone metabolism than previously thought. The first part of this review briefly discusses normal bone metabolism and disorders caused by its disruption, with particular reference to osteoporosis and current pharmacological treatments. The effects of natural products on bone and connective tissue are then discussed, to include items of diet, herbal extracts and food supplements, with evidence for their efficacy outlined. Copyright (c) 2006 John Wiley & Sons, Ltd.

Comparison of the antidiabetic activity of Berberis lyceum root extract and berberine in alloxan-induced diabetic rats

Berberine has been shown to have hypoglycaemic activity in several in vitro and in vivo models, although the mechanism of action is not fully known. Berberis lyceum Royle root produces high concentrations of berberine, and in traditional medicine, the whole extract of this plant is used widely to treat diabetes. The antidiabetic activity of the ethanol root extract of Berberis lyceum was compared with pure berberine in normal and alloxan-diabetic rats using similar doses of each. The concentration of berberine in the extract was determined to be 80% dry weight with only trace amounts of other alkaloids present. The purpose of the study was to investigate the effects of berberine and a whole extract of Berberis lyceum on blood glucose and other parameters associated with diabetes, to compare the effects of the crude extract with those of pure berberine and thus validate its use as a therapeutic agent, and finally to identify any contribution of the other components of the extract to these effects. Oral administration of 50 mg/kg of Berberis extract and berberine to normal and experimental diabetic rats produced a significant (p < 0.05) reduction in blood glucose levels from days 3-7 days of treatment. Significant effects were also observed on the glucose tolerance, glycosylated haemoglobin, serum lipid profiles and body weight of experimental animals. Berberis extract and berberine demonstrated similar effects on all parameters measured, and although the extract was comparable in efficacy to berberine, it did not produce any effects additional to those shown by pure berberine. The results support the use of the extract in traditional medicine, and demonstrate that apart from being a highly cost-effective means of treating with berberine, the total extract does not appear to confer any additional benefits or disadvantages compared with the pure compound. Copyright (c) 2008 John Wiley & Sons, Ltd.

The multidimensional measurement of the positive symptoms of psychosis

The measures most frequently used to assess psychotic symptoms fail to reflect important dimensions. The Psychotic Symptom Rating Scale (PSYRATS) aims to capture the multidimensional nature of auditory hallucinations and delusions. Individuals (N = 276) who had recently relapsed with positive symptoms completed the auditory hallucinations and delusions PSYRATS scales. These scores were compared with the relevant items from the SAPS and PANSS, and with measures of current mood. Total scores and distribution of items of the PSYRATS scales are presented and correlated with other measures. Positive symptom items from the SAPS and PANSS reflected the more objective aspects of PSYRATS ratings of auditory hallucinations and delusions (frequency and conviction) but were relatively poor at measuring distress. A major strength of the PSYRATS scales is the specific measurement of the distress dimension of symptoms, which is a key target of psychological intervention. It is advised that the PSYRATS should not be used as a total score alone, whilst further research is needed to clarify the best use of potential subscales. Copyright (c) 2007 John Wiley & Sons, Ltd.

Effects of active information processing on the understanding of risk information

Two experiments investigated effects of active processing of risk information on participants' understanding and judgments. It was hypothesized that more active processing would lead to better understanding and differences in affective judgments (e.g. increased satisfaction and reduced perceived risk to health). In both experiments participants were given a written scenario about their being prescribed a fictitious medication. This medication was said to cause side effects in 2% of people who took it. Before answering a series of written questions, participants in the active conditions of both experiments were asked to carry out a reflective task (portraying the size of risk on a bar chart in Experiment 1 and answering a reflective question in Experiment 2). The results showed that active participants rated the likelihood of experiencing possible side effects significantly lower than passive participants (Experiment 1), and that active participants were significantly more satisfied with the information and judged perceived risk to health from taking the medication significantly lower than passive participants (Experiment 2). In both experiments, active participants were significantly more correct in their probability and frequency estimates. The studies demonstrate that active processing of risk information leads to improved understanding of the information given. This has important implications for risk communication. In the context of health, better understanding should lead to improved decision-making and health outcomes. Copyright (C) 2004 John Wiley Sons, Ltd.

Training conditional and cumulative risk judgements: the role of frequencies, problem-structure and Einstellung

Two experiments implement and evaluate a training scheme for learning to apply frequency formats to probability judgements couched in terms of percentages. Results indicate that both conditional and cumulative probability judgements can be improved in this manner, however the scheme is insufficient to promote any deeper understanding of the problem structure. In both experiments, training on one problem type only (either conditional or cumulative risk judgements) resulted in an inappropriate transfer of a learned method at test. The obstacles facing a frequency-based training programme for teaching appropriate use of probability data are discussed. Copyright (c) 2006 John Wiley & Sons, Ltd.

Distinguishing novelty and familiarity effects in infant preference procedures

This paper considers possible problems researchers might face when interpreting the results of studies that employ variants of the preference procedure. Infants show a tendency to shift their preference from familiar to novel stimuli with increasing exposure to the familiar stimulus, a behaviour that is exploited by the habituation paradigm. This change in attentional preference with exposure leads us to suggest that researchers interested in infants' pre-experimental or spontaneous preferences should beware of the potentially confounding effects of exposing infants to familiarization trials prior to employing the preference procedure. The notion that infant attentional preference is dynamic also calls into question the use of the direction of post-familiarization preference per se when interpreting the knowledge or strategies available to infants. We look into the results of a cross-modal word learning study to show how the interpretation of results may be difficult when infants exhibit a significant preference in an unexpected direction. As a possible solution to this problem we propose that significant preferences in both directions should be sought at multiple intervals over time. Copyright (C) 2004 John Wiley Sons, Ltd.

Treatment of child anxiety. an exploratory study of the role of maternal anxiety and behaviours in treatment outcome

Anxiety disorders are common among parents of anxious children and have been found to impede child treatment outcomes, yet it is unclear whether it is parental anxiety that needs to be targeted in therapy or associated parental behaviours. Twenty-two children (6-12 years) with a current anxiety disorder and their mothers received cognitive-behavioural treatment (CBT) for child anxiety. In addition, of the 12 mothers who met criteria for a current anxiety disorder, 6 received CBT for their own disorder. Assessments were made of the mother-child interaction. The main findings were: (1) children did less well from treatment where their mothers had a current anxiety disorder; (2) treatment of maternal anxiety disorder did not improve child treatment outcome; and (3) maternal overinvolvement and expression of fear was associated with child treatment outcome. The results suggest that in the context of maternal anxiety disorder, child treatment outcome may be improved by specifically targeting parenting behaviours. Copyright (C) 2008 John Wiley & Sons, Ltd.

Implicit memory and consumer choice: the mediating role of brand familiarity

Two experiments investigated the influence of implicit memory on consumer choice for brands with varying levels of familiarity. Priming was measured using a consideration-choice task, developed by Coates, Butler and Berry (2004). Experiment 1 employed a coupon-rating task at encoding that required participants to meaningfully process individual brand names, to assess whether priming could affect participants' final (preferred) choices for familiar brands. Experiment 2 used this same method to assess the impact of implicit memory on consideration and choice for unknown and leader brands, presented in conjunction with familiar competitors. Significant priming was obtained in both experiments, and was shown to directly influence final choice in the case of familiar and highly familiar leader brands. Moreover, it was shown that a single prior exposure could lead participants to consider buying an unknown, and indeed fictitious, brand. Copyright (c) 2006 John Wiley & Sons, Ltd.

Implicit memory: a prime example for brand consideration and choice

Three experiments investigated the influence of implicit memory for familiar brand names on consumer choice. Priming was measured using modified preference judgment tasks that comprised both brand consideration and choice components. Experiment 1 used a 'complex choice task' where consideration and choice stages were characterized as acting in sequence. Experiment 2 explored a different formulation whereby consideration and choice were assumed to act in parallel, Both experiments demonstrated that priming had an influence on brand consideration but not on final or preferred choice. Finally, Experiment 3 replicated and extended these findings under more realistic conditions where participants actually received some of the products that they selected. Overall, the experiments suggested that for many decisions involving the consideration of familiar brands prior to choice, previous exposure to brand names can increase the likelihood that they will enter the consumers' consideration set. However, the advantage does not appear to extend to choice itself. Copyright (C) 2004 John Wiley Sons, Ltd.

Reverberant auditory environments: the effects of multiple echoes on distraction by 'irrelevant' speech

Two experiments examine the effect on an immediate recall test of simulating a reverberant auditory environment in which auditory distracters in the form of speech are played to the participants (the 'irrelevant sound effect'). An echo-intensive environment simulated by the addition of reverberation to the speech reduced the extent of 'changes in state' in the irrelevant speech stream by smoothing the profile of the waveform. In both experiments, the reverberant auditory environment produced significantly smaller irrelevant sound distraction effects than an echo-free environment. Results are interpreted in terms of changing-state hypothesis, which states that acoustic content of irrelevant sound, rather than phonology or semantics, determines the extent of the irrelevant sound effect (ISE). Copyright (C) 2007 John Wiley & Sons, Ltd.