Antimicrobial effects of Melaleuca alternifolia (tea tree) essential oil against biofilm-forming multidrug-resistant cystic fibrosis-associated Pseudomonas aeruginosa as a single agent and in combination with commonly nebulized antibiotics.

Broth microdilution assays were used to determine minimum inhibitory concentrations (MICs) and fractional inhibitory concentration indices (FICIs) of tea tree oil (TTO), tobramycin, colistin and aztreonam (ATM) against clinical cystic fibrosis-associated Pseudomonas aeruginosa (CFPA) isolates (n = 20). The minimum biofilm eradication concentration (MBEC) and fractional biofilm eradication concentration index (FBECI) were also determined using a similar microbroth dilution checkerboard assay, with biofilms formed using the MBEC device® . TTO was effective at lower concentrations against multidrug-resistant (MDR) CFPA isolates (n = 3) in a biofilm compared to in a planktonic state (MBEC 18·7-fold lower than MIC). CFPA within biofilm was less susceptible to ATM, colistin and tobramycin compared to planktonic cells (MBEC 6·3-fold, 9·3-fold, and 2·1-fold higher than MIC respectively). All combinations of essential oil and antibiotic showed indifferent relationships (FICI 0·52-1·72) when tested against planktonic MDR CFPA isolates (n = 5). Against CFPA isolates (n = 3) in biofilm, combinations of TTO/aztreonam and TTO/colistin showed indifferent relationships (mean FBECI 0·85 and 0·60 respectively), whereas TTO/tobramycin showed a synergistic relationship (mean FBECI 0·42). The antibiofilm properties of TTO and the synergistic relationship seen between TTO and tobramycin against CFPA in vitro make inhaled TTO a promising candidate as a potential therapeutic agent.

An economic evaluation of the randomized controlled trial of topical corticosteroid and home-based narrowband ultraviolet B for active and limited vitiligo (the HI-Light Vitiligo Trial).

BACKGROUND: Economic evidence for vitiligo treatments is absent. OBJECTIVES: To determine the cost-effectiveness of (i) handheld narrowband ultraviolet B (NB-UVB) and (ii) a combination of topical corticosteroid (TCS) and NB-UVB compared with TCS alone for localized vitiligo. METHODS: Cost-effectiveness analysis alongside a pragmatic, three-arm, placebo-controlled randomized controlled trial with 9 months' treatment. In total 517 adults and children (aged ≥ 5 years) with active vitiligo affecting < 10% of skin were recruited from secondary care and the community and were randomized 1: 1: 1 to receive TCS, NB-UVB or both. Cost per successful treatment (measured on the Vitiligo Noticeability Scale) was estimated. Secondary cost-utility analyses measured quality-adjusted life-years using the EuroQol 5 Dimensions 5 Levels for those aged ≥ 11 years and the Child Health Utility 9D for those aged 5 to < 18 years. The trial was registered with number ISRCTN17160087 on 8 January 2015. RESULTS: The mean ± SD cost per participant was £775 ± 83·7 for NB-UVB, £813 ± 111.4 for combination treatment and £600 ± 96·2 for TCS. In analyses adjusted for age and target patch location, the incremental difference in cost for combination treatment compared with TCS was £211 (95% confidence interval 188-235), corresponding to a risk difference of 10·9% (number needed to treat = 9). The incremental cost was £1932 per successful treatment. The incremental difference in cost for NB-UVB compared with TCS was £173 (95% confidence interval 151-196), with a risk difference of 5·2% (number needed to treat = 19). The incremental cost was £3336 per successful treatment. CONCLUSIONS: Combination treatment, compared with TCS alone, has a lower incremental cost per additional successful treatment than NB-UVB only. Combination treatment would be considered cost-effective if decision makers are willing to pay £1932 per additional treatment success.

Randomized controlled trial of topical corticosteroid and home-based narrowband ultraviolet B for active and limited vitiligo: results of the HI-Light Vitiligo Trial.

BACKGROUND: Evidence for the effectiveness of vitiligo treatments is limited. OBJECTIVES: To determine the effectiveness of (i) handheld narrowband UVB (NB-UVB) and (ii) a combination of potent topical corticosteroid (TCS) and NB-UVB, compared with TCS alone, for localized vitiligo. METHODS: A pragmatic, three-arm, placebo-controlled randomized controlled trial (9-month treatment, 12-month follow-up). Adults and children, recruited from secondary care and the community, aged ≥ 5 years and with active vitiligo affecting < 10% of skin, were randomized 1 : 1 : 1 to receive TCS (mometasone furoate 0·1% ointment + dummy NB-UVB), NB-UVB (NB-UVB + placebo TCS) or a combination (TCS + NB-UVB). TCS was applied once daily on alternating weeks; NB-UVB was administered on alternate days in escalating doses, adjusted for erythema. The primary outcome was treatment success at 9 months at a target patch assessed using the participant-reported Vitiligo Noticeability Scale, with multiple imputation for missing data. The trial was registered with number ISRCTN17160087 on 8 January 2015. RESULTS: In total 517 participants were randomized to TCS (n = 173), NB-UVB (n = 169) and combination (n = 175). Primary outcome data were available for 370 (72%) participants. The proportions with target patch treatment success were 17% (TCS), 22% (NB-UVB) and 27% (combination). Combination treatment was superior to TCS: adjusted between-group difference 10·9% (95% confidence interval 1·0%-20·9%; P = 0·032; number needed to treat = 10). NB-UVB alone was not superior to TCS: adjusted between-group difference 5·2% (95% CI - 4·4% to 14·9%; P = 0·29; number needed to treat = 19). Participants using interventions with ≥ 75% expected adherence were more likely to achieve treatment success, but the effects were lost once treatment stopped. Localized grade 3 or 4 erythema was reported in 62 (12%) participants (including three with dummy light). Skin thinning was reported in 13 (2·5%) participants (including one with placebo ointment). CONCLUSIONS: Combination treatment with home-based handheld NB-UVB plus TCS is likely to be superior to TCS alone for treatment of localized vitiligo. Combination treatment was relatively safe and well tolerated but was successful in only around one-quarter of participants.

Assessing child quality of life impairments following pediatric burn injuries: Rasch analysis of the children's dermatology life quality index.

AIMS: There is a need for a brief, validated measure of quality of life (QOL) for children to monitor their adjustment to burn injuries. We aimed to apply a Rasch analysis to an existing measure of QOL from the dermatology literature to a clinical sample of pediatric burn patients. METHODS: The Children's Dermatology Life Quality Index (CDLQI) was administered to pediatric burn patients (N = 253) during a standard clinic visit. Rasch analysis was used to examine psychometric properties of this measure with a burn sample. RESULTS: The CDLQI showed an adequate fit to the Rasch model. Test difficulty is .61 logits greater than person ability. Results of item reliability and separation analyses were sufficiently strong and indicated a unidimensional latent trait. Person reliability (.74) and separation analyses (1.64) were moderate. Finally, the CDLQI was able to moderately separate the group of respondents into low and high levels of QOL impairments related to burn injuries. CONCLUSION: The Rasch model demonstrated that the CDLQI is a reliable and valid scale that adequately measures QOL impairments in children following burn injuries.

Targeting palmitoyl acyltransferase ZDHHC21 improves gut epithelial barrier dysfunction resulting from burn-induced systemic inflammation.

Clinical studies in burn patients demonstrate a close association between leaky guts and increased incidence or severity of sepsis and other complications. Severe thermal injury triggers intestinal inflammation that contributes to intestinal epithelial hyperpermeability, which exacerbates systemic response leading to multiple organ failure and sepsis. In this study, we identified a significant function of a particular palmitoyl acyltransferase, zinc finger DHHC domain-containing protein-21 (ZDHHC21), in mediating signaling events required for gut hyperpermeability induced by inflammation. Using quantitative PCR, we show that ZDHHC21 mRNA production was enhanced twofold when intestinal epithelial cells were treated with TNF-α-IFN-γ in vitro. In addition, pharmacological targeting of palmitoyl acyltransferases with 2-bromopalmitate (2-BP) showed significant improvement in TNF-α-IFN-γ-mediated epithelial barrier dysfunction by using electric cell-substrate impedance-sensing assays, as well as FITC-labeled dextran permeability assays. Using acyl-biotin exchange assay and click chemistry, we show that TNF-α-IFN-γ treatment of intestinal epithelial cells results in enhanced detection of total palmitoylated proteins and this response is inhibited by 2-BP. Using ZDHHC21-deficient mice or wild-type mice treated with 2-BP, we showed that mice with impaired ZDHHC21 expression or pharmacological inhibition resulted in attenuated intestinal barrier dysfunction caused by thermal injury. Moreover, hematoxylin and eosin staining of the small intestine, as well as transmission electron microscopy, showed that mice with genetic interruption of ZDHHC21 had attenuated villus structure disorganization associated with thermal injury-induced intestinal barrier damage. Taken together, these results suggest an important role of ZDHHC21 in mediating gut hyperpermeability resulting from thermal injury.NEW & NOTEWORTHY Increased mucosal permeability in the gut is one of the major complications following severe burn. Here we report the novel finding that zinc finger DHHC domain-containing protein-21 (ZDHHC21) mediates gut epithelial hyperpermeability resulting from an experimental model of thermal injury. The hyperpermeability response was significantly attenuated with a pharmacological inhibitor of palmitoyl acyltransferases and in mice with genetic ablation of ZDHHC21. These findings suggest that ZDHHC21 may serve as a novel therapeutic target for treating burn-induced intestinal barrier dysfunction.

Interleukin-1ß Mediates ß-Catenin-Driven Downregulation of Claudin-3 and Barrier Dysfunction in Caco2 Cells.

BACKGROUND: IL-1ß is a cytokine involved in mediating epithelial barrier dysfunction in the gut. It is known that IL-1ß mediates activation of non-muscle myosin light chain kinase in epithelial cells, but the precise mechanism by which epithelial barrier dysfunction is induced by IL-1ß is not understood. METHODS AND RESULTS: Using a Caco2 cell model, we show that the expression of the tight junction protein, claudin-3, is transcriptionally downregulated by IL-1ß treatment. In addition, after assessing protein and mRNA expression, and protein localization, we show that inhibition of nmMLCK rescues IL-1ß-mediated decrease in claudin-3 expression as well as junction protein redistribution. Using chromatin immunoprecipitation assays, we also show that ß-catenin targeting of the claudin-3 promoter occurs as a consequence of IL-1ß-mediated epithelial barrier dysfunction, and inhibition of nmMLCK interferes with this interaction. CONCLUSIONS: Taken together, these data represent the first line of evidence demonstrating nmMLCK regulation of claudin-3 expression in response to IL-1ß-treated epithelial cells.

Evaluating the Orbach Mikulincer Mental Pain Scale among Late Adolescent and Early Adult African Americans: A Rasch Analysis.

Although there are only a few psychometric investigations of mental pain measurement in the literature, there are no previous evaluations of mental pain scales among African Americans. The present study examined the Rasch measurement properties of the nine subscales contained in the Orbach-Mikulincer Mental Pain (OMMP) Scale among a sample of older adolescent and young adult African Americans. Results from the analyses suggest that three of the OMMP subscales meet the requirements of the Rasch model and hold promise for use in research and applied settings. Implications for further development and use of the remaining subscales are discussed.

Protein tyrosine kinase 6 mediates TNFα-induced endothelial barrier dysfunction.

A key event in the progression of systemic inflammation resulting from severe trauma or shock involves microvascular hyperpermeability, which leads to excessive plasma fluid and proteins accumulating in extravascular space resulting in tissue edema. The precise molecular mechanism of the hyperpermeability response is not completely understood. Protein tyrosine kinase 6 (PTK6, also known as breast tumor kinase BRK) is a non-receptor tyrosine kinase related to Src-family proteins. Although it has also been shown that PTK6 participates in regulating epithelial barrier function, the role of PTK6 in endothelial barrier function has not been reported. In this study, we hypothesized that PTK6 is (1) expressed in vascular endothelial cells, and (2) contributes to vascular endothelial hyperpermeability in response to TNFα. Results showed that PTK6 was detected in mouse endothelial cells at the level of protein and mRNA. In addition, PTK6 knockdown attenuated TNFα induced decrease in endothelial barrier function as measured by electric cell-substrate impedance sensing (ECIS) and in vitro transwell albumin-flux assays. Furthermore, we showed that TNFα treatment of endothelial cells increased active PTK6 association with p120-catenin at endothelial cell-cell junctions. Further analysis using immunocytochemistry and immunoprecipitation demonstrated that PTK6 knockdown attenuated TNFα induced VE-cadherin internalization as well as promoting its association with p120-catenin. Our study demonstrates a novel role of PTK6 in mediating endothelial barrier dysfunction.

Wide spectrum of filaggrin-null mutations in atopic dermatitis highlights differences between Singaporean Chinese and European populations.

BACKGROUND: Null mutations in the filaggrin gene (FLG) cause ichthyosis vulgaris (IV) and predispose to atopic dermatitis (AD). Cohort studies in Europe and Japan have reported an FLG mutation carrier frequency of between 14% and 56%, but the prevalent European FLG mutations are rare or absent in Chinese patients with IV and AD. OBJECTIVES: To investigate further the spectrum of FLG-null mutations in Chinese patients and to compare it with that in other populations. METHODS: We conducted comprehensive FLG genetic analysis in a discovery cohort of 92 Singaporean Chinese individuals with IV and/or moderate-to-severe AD. All detected FLG mutations were then screened in a cohort of 425 patients with AD and 440 normal controls. Results In total, 22 FLG-null mutations, of which 14 are novel, were identified in this study; the combined null FLG genotype of 17 mutations detected in cases and controls showed strong association with AD [Fisher's exact test; P = 5·3 × 10⁻9; odds ratio (OR) 3·3], palmar hyperlinearity (Fisher's exact test; P = 9·0 × 10⁻¹5; OR 5·8), keratosis pilaris (Fisher's exact test; P = 0·001; OR 4·7) and with increased severity of AD (permutation test; P = 0·0063). CONCLUSIONS: This study emphasizes the wider genetic landscape of FLG-null mutations in Asia that is slowly emerging.

Statistical modelling of gaze behaviour as categorical time series: what you should watch to save soccer penalties.

Previous research on gaze behaviour in sport has typically reported summary fixation statistics thereby largely ignoring the temporal sequencing of gaze. In the present study on penalty kicking in soccer, our aim was to apply a Markov chain modelling method to eye movement data obtained from goalkeepers. Building on the discrete analysis of gaze employed by Dicks et al. (Atten Percept Psychophys 72(3):706-720, 2010b), we wanted to statistically model the relative probabilities of the goalkeeper's gaze being directed to different locations throughout the penalty taker's approach (Dicks et al. in Atten Percept Psychophys 72(3):706-720, 2010b). Examination of gaze behaviours under in situ and video-simulation task constraints reveals differences in information pickup for perception and action (Attention, Perception and Psychophysics 72(3), 706-720). The probabilities of fixating anatomical locations of the penalty taker were high under simulated movement response conditions. In contrast, when actually required to intercept kicks, the goalkeepers initially favoured watching the penalty taker's head but then rapidly shifted focus directly to the ball for approximately the final second prior to foot-ball contact. The increased spatio-temporal demands of in situ interceptive actions over laboratory-based simulated actions lead to different visual search strategies being used. When eye movement data are modelled as time series, it is possible to discern subtle but important behavioural characteristics that are less apparent with discrete summary statistics alone.

Activation and clinical significance of the unfolded protein response in breast cancer.

INTRODUCTION: The tumour microenvironment is hypoglycaemic, hypoxic and acidotic. This activates a stress signalling pathway: the unfolded protein response (UPR). The UPR is cytoprotective if the stressor is mild, but may initiate apoptosis if severe.Activation of the UPR in breast carcinoma is induced by microenvironmental stress such as glucose and oxygen deprivation, but may also be linked to oestrogen stimulation. It may be clinically significant as it may alter chemosensitivity to doxorubicin. METHODS: 395 human breast adenocarcinomas were immunohistochemically stained for UPR activation markers (glucose-regulated protein (GRP-78 and XBP-1). A model of UPR activation in vitro by glucose deprivation of T47D breast cancer cells was developed to determine how the UPR affects cellular sensitivity to doxorubicin and 5-fluorouracil. Cytotoxicity was assessed using a colorimetric cytotoxicity assay (MTT). The effect of oestrogen stimulation and tamoxifen exposure on UPR activation by T47D cells was determined by western blotting measurement of the key UPR protein, GRP-78. RESULTS: Expression of GRP78 and XBP-1 was demonstrated in 76% and 90% of the breast cancers, respectively, and correlated with oestrogen receptor positivity (P=0.045 and 0.017, respectively). In vitro UPR activation induced resistance to both doxorubicin and 5-flurouracil, (P<0.05). Oestrogen stimulation induced GRP78 and XBP1 over-expression on western blotting. Tamoxifen did not block this response and may induce UPR activation in its own right. CONCLUSIONS: The UPR is activated in the majority of breast cancers and confers resistance to chemotherapy. In vitro oestrogen stimulates UPR induction. UPR activation may contribute to breast cancer chemoresistance and interact with oestrogen response elements.

Hotspots: are some areas of sewer network prone to re-infestation by rats ( Rattus norvegicus ) year after year?

The records of sewer baiting work for three London boroughs were examined to see whether there were locations that exhibited 'rat' (Rattus norvegicus) activity more often than would be expected by chance, a feature we dubbed 'hotspots'. More than 100000 baiting records were checked covering 15 years of the London Borough of Enfield (Enfield) and 5 years each of the London Borough of Barnet (Barnet), and the Royal Borough of Kensington and Chelsea (RBKC). The additional boroughs were included for comparison to see whether any effect observed was confined to Enfield or was a feature that could be found in both inner and outer city locations. Each borough was divided into kilometre squares corresponding with those found both on Ordnance Survey maps and also Thames Water Utilities Asset maps. The number of records per square were logged and then the number of positive records for all the manholes in that square on a year-on-year basis. We examined 350 km2 in Enfield, 377 km2 in Barnet and 66 km2 in RBKC. The data were subjected to a weighted analysis (i.e. allowing for the number of manholes per square and the varying total rat population from year to year). Some areas were shown to exhibit significantly higher amounts of activity than others suggesting that their distribution is not random and that there must be environmental and other factors, which make certain locations attractive to rats. Squares with very high activity, repeating year-on-year, 'hotspots', were found in all three boroughs, suggesting that the phenomena are widespread.

The social context of smoking: the next frontier in tobacco control?

A better understanding of the social context of smoking may help to enhance tobacco control research and practice.

The service architecture of the TeraGyroid experiment.

We describe the service architecture of the successful TeraGyroid experiment. In particular we discuss the use of the open Grid service infrastructure (OGSI) to build the services used during the experiment and illustrate the problems we encountered.

A practical toolkit for computational steering.

Computational steering refers to the real-time interaction of a scientist with their running simulation code. Despite the many benefits associated with computational steering, its uptake to date has been limited. In this paper we discuss the reasons for this and how the computational steering library and associated tools developed as part of the RealityGrid project aim to tackle them. We describe the functionality of the steering library and the use of Grid services in constructing a generic, dynamic architecture for discovering, steering and connecting visualization software to running simulations. The use of on-line visualization for providing feedback to the scientist is described, including the ways in which it may be enhanced through tools such as Chromium and Access Grid. Finally, we illustrate the flexibility of our approach by describing the functionality that has been added to various simulation codes as part of the RealityGrid project.

The contribution of psychologists to specialist palliative care.

Current approaches to multidisciplinary working in specialist palliative care generally acknowledge the need for psychological care and the contribution of specialist psychological services. Embedded within an integrated approach to care, there has long been a recognition of the psychological needs of patients and their informal caregivers, although there has been less consensus on how these needs should be addressed. Confusion appears to exist regarding the boundaries between psychological support and psychological interventions, and which members of the multidisciplinary team are best qualified to provide them. This article aims to explore the contribution of psychologists and psychological services to specialist palliative care. It offers a conceptual model to help categorize psychological care in hospices and specialist palliative care services. It seeks to help define the boundaries between general psychological support and care that may be offered by nurses and other health care professionals, and the specialist expertise of psychologists.

Platinum carboxylato-pendant-arm macrocycles: structure, redox properties and anti-cancer potential.

In an attempt to generate new platinum compounds that may be effective in the treatment of cancer, as well as having a lower toxicity than traditional platins and being orally viable, we are studing the synthesis and reactivity of platinum complexes of tetraazamacrocycles bearing carboxylato pendant arms. We have synthesized adducts of meso- and rac-5,5,7,12,12,14-hexamethyl-1,4,8,11-tetraazacyclotetradecane-1,7-diacetic acid (L(1)H(2)). The meso-Pt(II)L(1) complex is unstable with respect to disproportionation, forming platinum metal and [meso-Pt(IV)L(1)](2+). The rac-isomer shows less tendency to disproportionate. Cyclic voltammetry suggests that the rac-Pt(II)L(1) complex undergoes two one-electron oxidations. Using bis-triazacyclononanenickel(III), [Ni(III)(tacn)(2)](3+) as an outer-sphere oxidant, the self-exchange rate for the [Pt(II/III)L](0/+) couple has been estimated at 0.034 M(-1) s(-1).

Long-lived emissions from 4'-substituted Pt(trpy)Cl+ complexes bearing aryl groups. Influence of orbital parentage.

Pt(trpy)Cl+, where trpy denotes 2,2':6',2' '-terpyridine, is a versatile binding agent but has a limited photochemistry due to a short excited-state lifetime. However, this work shows that the introduction of aryl substituents at the 4' position of the trpy ligand drastically alters the picture. For the substituents phenyl, p-methoxyphenyl, 1-naphthyl, 2-naphthyl, 9-phenanthrenyl, and 1-pyrenyl, the ligand abbrevations are 4'-Ph-T, 4'-pMeOPh-T, 4'-Npl-T, 4'-Np2-T, 4'-Phe9-T, and 4'-Pyre1-T, respectively. Techniques utilized include electrochemistry as well as absorption and emission spectroscopies. While the lowest energy excited states of Pt(4'-Ph-T)Cl+ and the parent complex Pt(trpy)Cl+ exhibit mainly metal-to-ligand charge-transfer (MLCT) character, the emitting state takes on aryl-to-trpy intraligand charge-transfer (ILCT) character as the substituents become more electron-donating. Studies of Zn(trpy)Cl2, its aryl-substituted analogues, and the free ligands themselves provide information about the relative energies of participating ILCT and intraligand 3pi-pi excited states. Even though the emission energy decreases when larger aryl groups are present, the emission lifetime increases all the way from 85 ns for Pt(4'-Ph-T)Cl+ to 64 micros for Pt(4'-Pyre1-T)Cl+. (Data from deoxygenated, room-temperature dichloromethane solution.) Intraligand character appears to dominate in the case of Pt(4'-Pyre1-T)Cl+, which is unique in the series in that it exhibits singlet and triplet emissions in solution. In aerated solution the complex shows prompt as well as delayed fluorescence. Finally, studies in donor media establish that the introduction of intraligand character inhibits solvent-induced exciplex quenching.