Presentation and validation of the Abbreviated Self Completion Teen-Addiction Severity Index (ASC T-ASI): A preference-based measure for use in health-economic evaluations.
Economic evaluations of new youth mental health interventions require preference-based outcome measures that capture the broad benefits these interventions can have for adolescents. The Abbreviated Self Completion Teen-Addiction Severity Index (ASC T-ASI) was developed to meet the need for such a broader measure. It assesses self reported problems in seven important domains of adolescents' lives, including school performance and family relationships, and is intended for use in economic evaluations of relevant interventions. The aim of the current study was to present the ASC T-ASI and examine its validity as well as its ability to distinguish between adolescents with and without problems associated with substance use and delinquency. The validation study was conducted in a sample of adolescents (n = 167) aged 12-18 years, who received in- or outpatient care in a youth mental health and (enclosed) care facility in the Netherlands. To examine its feasibility, test-retest reliability, and convergent validity, respondents completed the ASC T-ASI, as well as the EQ-5D-3L and SDQ at baseline and after a two-week interval using a counterbalanced method. The ASC T-ASI descriptive system comprises seven domains: substance use, school, work, family, social relationships, justice, and mental health, each expressing self reported problems on a five-point Likert scale (ranging from having 'no problem' to having a 'very large problem'). The majority of respondents (>70%) completed the ASC T-ASI within 10 minutes and appraised the questions as (very) easy and (very) comprehensible. Test-retest reliability was adequate (Kw values 0.26-0.55). Correlations with the supplementary measures were moderate to high (rs 0.30-0.50), suggesting convergent validity. The ASC T-ASI is a promising and valid measure for assessing self reported problems in important domains in adolescents' lives, allowing benefits beyond health and health-related quality of life to be included in economic evaluations of youth mental health interventions. Future studies of the ASC T-ASI should consider the comprehensiveness of its domains and sensitivity to change.


Diversity of the active methanotrophic community in acidic peatlands as assessed by mRNA and SIP-PLFA analyses.
The active methanotroph community was investigated for the first time in heather (Calluna)-covered moorlands and Sphagnum/Eriophorum-covered UK peatlands. Direct extraction of mRNA from these soils facilitated detection of expression of methane monooxygenase genes, which revealed that particulate methane monooxygenase and not soluble methane monooxygenase was probably responsible for CH(4) oxidation in situ, because only pmoA transcripts (encoding a subunit of particulate methane monooxygenase) were readily detectable. Differences in methanotroph community structures were observed between the Calluna-covered moorland and Sphagnum/Eriophorum-covered gully habitats. As with many other Sphagnum-covered peatlands, the Sphagnum/Eriophorum-covered gullies were dominated by Methylocystis. Methylocella and Methylocapsa-related species were also present. Methylobacter-related species were found as demonstrated by the use of a pmoA-based diagnostic microarray. In Calluna-covered moorlands, in addition to Methylocella and Methylocystis, a unique group of peat-associated type I methanotrophs (Gammaproteobacteria) and a group of uncultivated type II methanotrophs (Alphaproteobacteria) were also found. The pmoA sequences of the latter were only distantly related to Methylocapsa and also to the RA-14 group of methanotrophs, which are believed to be involved in oxidation of atmospheric concentrations of CH(4). Soil samples were also labelled with (13)CH(4), and subsequent analysis of the (13)C-labelled phospholipid fatty acids (PLFAs) showed that 16:1 omega 7, 18:1 omega 7 and 18:1 omega 9 were the major labelled PLFAs. The presence of (13)C-labelled 18:1 omega 9, which was not a major PLFA of any extant methanotrophs, indicated the presence of novel methanotrophs in this peatland.


Spontanous periodic breathing is associated with sympathetic hyperreactivity and baroreceptor dysfunction in hypertension.
Intermittent periods of hypoxemia such as during periodic breathing are associated with hypertension and increased sympathetic activity. In patients with sleep apnea syndrome, hypertension is common. Treating apnea improves hypertension and reduces sympathetic outflow. The aim of the present study was to investigate the phenomenon and mechanisms of spontaneous periodic breathing in patients with hypertension. We examined 43 hypertensive patients with untreated hypertension without left-ventricular dysfunction, heart failure or sleep apnea syndrome. Muscle sympathetic nerve activity (MSA), heart rate (HR), blood pressure (BP) and respiration were continuously recorded at rest and during cold-pressor testing. Oxygen and a CO2-enriched gas were used to test central and peripheral chemoreceptors, respectively. Baroreceptor gain was measured using the alpha method. Seven out of 43 patients showed spontaneous periodic breathing while awake. No difference in MSA, HR and BP was seen between patients with and without periodic breathing at rest except the breathing pattern. However, the cold-pressor test caused a larger increase of MSA in patients with periodic breathing (203 +/- 62 vs. 62 +/- 8%, P < 0.0001 by ANOVA), as well as systolic (46 +/- 6 vs. 25 +/- 3 mmHg, P = 0.002) and diastolic BP (26 +/- 5 vs. 12 +/- 1 mmHg, P = 0.004, ANOVA). Baroreceptor gain was markedly higher in patients with periodic breathing. Chemoreceptor sensitivity was comparable. Spontaneous periodic breathing is relatively common in patients with hypertension and is associated with greatly enhanced responses to cold-pressor testing. We suggest increased baroreceptor gain and sympathetic outflow as a cause for the oscillatory respiration pattern via barorespiratory coupling.


A Novel Morphological Marker for the Analysis of Molecular Activities at the Single-cell Level.
For more than a century, hematoxylin and eosin (H&E) staining has been the de facto standard for histological studies. Consequently, the legacy of histological knowledge is largely based on H&E staining. Due to the recent advent of multi-photon excitation microscopy, the observation of live tissue is increasingly being used in many research fields. Adoption of this technique has been further accelerated by the development of genetically encoded biosensors for ions and signaling molecules. However, H&E-based histology has not yet begun to fully utilize in vivo imaging due to the lack of proper morphological markers. Here, we report a genetically encoded fluorescent marker, NuCyM (Nucleus, Cytosol, and Membrane), which is designed to recapitulate H&E staining patterns in vivo. We generated a transgenic mouse line ubiquitously expressing NuCyM by using a ROSA26 bacterial artificial chromosome (BAC) clone. NuCyM evenly marked the plasma membrane, cytoplasm and nucleus in most tissues, yielding H&E staining-like images. In the NuCyM-expressing cells, cell division of a single cell was clearly observed as five basic phases during M phase by three-dimensional imaging. We next crossed NuCyM mice with transgenic mice expressing an ERK biosensor based on the principle of Förster resonance energy transfer (FRET). Using NuCyM, ERK activity in each cell could be extracted from the FRET images. To further accelerate the image analysis, we employed machine learning-based segmentation methods, and thereby automatically quantitated ERK activity in each cell. In conclusion, NuCyM is a versatile cell morphological marker that enables us to grasp histological information as with H&E staining.Key words: in vivo imaging, histology, machine learning, molecular activity.


Constitutive activation of a phosphoinositidase C-linked G protein in murine fibroblasts decreases agonist-stimulated Ca2+ mobilization.
We compared Ca2+ signaling and inositol polyphosphate metabolism in NIH-3T3 cells stably transfected with cDNA encoding either the wild-type G protein G16 alpha subunit or a GTPase-deficient alpha 16 subunit (Q212L-alpha 16). Constitutive activation of phosphoinositidase C (PIC) in cells expressing Q212L-alpha 16 was demonstrated by 1) an increased basal level of [3H]inositol polyphosphates, 2) an enhanced rate of [3H]inositol polyphosphate accumulation in cells treated with 10 mM LiCl, and 3) an increased rate of incorporation of [3H]inositol into cell lipids. Q212L-alpha 16 cells had a diminished cell growth rate. Basal intracellular Ca2+ concentration was equivalent in Fura-2 acetoxymethyl ester-loaded Q212L-alpha 16 cells compared with controls; however, calcium release in Q212L-alpha 16 cells exposed to ionomycin, ATP (a G protein-linked agonist), or platelet-derived growth factor (a tyrosine kinase-linked agonist) was decreased. Permeabilized, 45Ca-loaded Q212L-alpha 16 cells released less 45Ca at each concentration of inositol-1,4,5-trisphosphate than did control cells. Accordingly, the total amount of inositol trisphosphate (IP3) receptor protein was decreased in Q212L-alpha 16 cells relative to controls. These data demonstrate that Q212L-alpha 16 cells maintain physiological levels of cytoplasmic calcium and partially loaded Ca2+ stores in the face of constitutively active PIC. This is accomplished, at least in part, by down-regulation of IP3 receptor number. Thus, diminution in cell growth rate in Q212L-alpha 16 cells seems to be attributable to a combination of at least two effects: a direct effect of PIC activation leading to partial depletion of Ca2+ stores and an indirect, adaptive response resulting in a decreased IP3 receptor number.


Reduced retina microglial activation and improved optic nerve integrity with minocycline treatment in the DBA/2J mouse model of glaucoma.
In the context of the retinal ganglion cell (RGC) axon degeneration in the optic nerve that occurs in glaucoma, microglia become activated, then phagocytic, and redistribute in the optic nerve head. The authors investigated the potential contribution of retinal microglia activation to glaucoma progression in the DBA/2J chronic mouse glaucoma model. The authors treated 6-week-old DBA/2J mice for 25 weeks with minocycline, a tetracycline derivative known to reduce microglia activation and to improve neuronal survival in other models of neurodegenerative disease. They quantified RGC numbers and characterized microglia activation, gliosis, and both axonal integrity and retrograde tracer transport by RGCs in mice systemically treated with minocycline or vehicle only. Minocycline reduced microglial activation and improved RGC axonal transport and integrity, yet it had no effect on the characteristic age-related ocular changes that lead to chronically elevated pressure and did not alter Müller or astrocyte gliosis. Specifically, minocycline increased the fraction of microglia with resting ramified morphology and reduced levels of Iba1 mRNA and protein, a microglia-specific calcium ligand linked to activation. The reduction in microglial activation was coupled to significant improvement in RGC axonal transport, as measured by neuronal retrograde tracing from the superior colliculus. Finally, minocycline treatment significantly decoupled RGC axon loss from increased intraocular pressure. These observations suggest that in glaucoma, retina and optic nerve head microglia activation may be a factor in the early decline in function of the optic nerve and its subsequent degeneration.


Total variation regularization for nonlinear fluorescence tomography with an augmented Lagrangian splitting approach.
Fluorescence tomography is an imaging modality that seeks to reconstruct the distribution of fluorescent dyes inside a highly scattering sample from light measurements on the boundary. Using common inversion methods with L(2) penalties typically leads to smooth reconstructions, which degrades the obtainable resolution. The use of total variation (TV) regularization for the inverse model is investigated. To solve the inverse problem efficiently, an augmented Lagrange method is utilized that allows separating the Gauss-Newton minimization from the TV minimization. Results on noisy simulation data provide evidence that the reconstructed inclusions are much better localized and that their half-width measure decreases by at least 25% compared to ordinary L(2) reconstructions.


18β-Glycyrrhetinic Acid Inhibits Osteoclastogenesis In Vivo and In Vitro by Blocking RANKL-Mediated RANK-TRAF6 Interactions and NF-κB and MAPK Signaling Pathways.
Bone metabolism is determined by a delicate balance between bone resorption by osteoclasts and bone formation by osteoblasts. The imbalance due to over-activated osteoclasts plays an important role in various diseases. Activation of NF-κB and MAPK signaling pathways by receptor activator of nuclear factor -κB ligand (RANKL) is vital for osteoclastogenesis. Here, we for the first time explored the effects of 18β-glycyrrhetinic acid (18β-GA), a pentacyclic triterpenoid found in the Glycyrrhiza glabra L roots, on RANKL-induced osteoclastogenesis, osteoclast functions and signaling pathways in vitro and in vivo. In bone marrow monocytes (BMMs) and RAW264.7 cells, 18β-GA inhibited osteoclastogenesis, decreased expression of TRAP, cathepsin K, CTR and MMP-9, blocked actin ring formation and compromised osteoclasts functions in a dose-dependent manner at an early stage with minimal effects on osteogenic and adipogenic differentiation of bone marrow mesenchymal stem cells (BMSCs). For underlying molecular mechanisms, 18β-GA inhibited RANKL-induced phosphorylation of p65, p50, and IκB, blocked p65 nuclear translocation and decreased the DNA-binding activity of NF-κB. Besides, 18β-GA inhibited the activation of the MAPK pathways. Co-immunoprecipitation showed that 18β-GA treatment blocked RANK-TRAF6 association at an upstream site. In vivo, 18β-GA treatment inhibited ovariectomy-induced osteoclastogenesis and reduced bone loss in mice. Overall, our results demonstrated that 18β-GA inhibited RANKL-induced osteoclastogenesis by inhibiting RANK expression in preosteoclasts and blocking the binding of RANK and TRAF6 which lead to the inhibition of NF-κB and MAPK signaling pathways. 18β-GA is a promising novel candidate in the treatment of osteoclast-related diseases such as postmenopausal osteoporosis.


Ventricular tachyarrhythmias after cardiac arrest in public versus at home.
The incidence of ventricular fibrillation or pulseless ventricular tachycardia as the first recorded rhythm after out-of-hospital cardiac arrest has unexpectedly declined. The success of bystander-deployed automated external defibrillators (AEDs) in public settings suggests that this may be the more common initial rhythm when out-of-hospital cardiac arrest occurs in public. We conducted a study to determine whether the location of the arrest, the type of arrhythmia, and the probability of survival are associated. Between 2005 and 2007, we conducted a prospective cohort study of out-of-hospital cardiac arrest in adults in 10 North American communities. We assessed the frequencies of ventricular fibrillation or pulseless ventricular tachycardia and of survival to hospital discharge for arrests at home as compared with arrests in public. Of 12,930 evaluated out-of-hospital cardiac arrests, 2042 occurred in public and 9564 at home. For cardiac arrests at home, the incidence of ventricular fibrillation or pulseless ventricular tachycardia was 25% when the arrest was witnessed by emergency-medical-services (EMS) personnel, 35% when it was witnessed by a bystander, and 36% when a bystander applied an AED. For cardiac arrests in public, the corresponding rates were 38%, 60%, and 79%. The adjusted odds ratio for initial ventricular fibrillation or pulseless ventricular tachycardia in public versus at home was 2.28 (95% confidence interval [CI], 1.96 to 2.66; P < 0.001) for bystander-witnessed arrests and 4.48 (95% CI, 2.23 to 8.97; P<0.001) for arrests in which bystanders applied AEDs. The rate of survival to hospital discharge was 34% for arrests in public settings with AEDs applied by bystanders versus 12% for arrests at home (adjusted odds ratio, 2.49; 95% CI, 1.03 to 5.99; P = 0.04). Regardless of whether out-of-hospital cardiac arrests are witnessed by EMS personnel or bystanders and whether AEDs are applied by bystanders, the proportion of arrests with initial ventricular fibrillation or pulseless ventricular tachycardia is much greater in public settings than at home. The incremental value of resuscitation strategies, such as the ready availability of an AED, may be related to the place where the arrest occurs.


Proteasome inhibition preserves longitudinal growth of denervated muscle and prevents neonatal neuromuscular contractures.
Muscle contractures are a prominent and disabling feature of many neuromuscular disorders, including the 2 most common forms of childhood neurologic dysfunction: neonatal brachial plexus injury (NBPI) and cerebral palsy. There are currently no treatment strategies to directly alter the contracture pathology, as the pathogenesis of these contractures is unknown. We previously showed in a mouse model of NBPI that contractures result from impaired longitudinal muscle growth. Current presumed explanations for growth impairment in contractures focus on the dysregulation of muscle stem cells, which differentiate and fuse to existing myofibers during growth, as this process has classically been thought to control muscle growth during the neonatal period. Here, we demonstrate in a mouse model of NBPI that denervation does not prevent myonuclear accretion and that reduction in myonuclear number has no effect on functional muscle length or contracture development, providing definitive evidence that altered myonuclear accretion is not a driver of neuromuscular contractures. In contrast, we observed elevated levels of protein degradation in NBPI muscle, and we demonstrate that contractures can be pharmacologically prevented with the proteasome inhibitor bortezomib. These studies provide what we believe is the first strategy to prevent neuromuscular contractures by correcting the underlying deficit in longitudinal muscle growth.


Dynamics of the IL-33/ST2 network in the progression of human colorectal adenoma to sporadic colorectal cancer.
Most sporadic colorectal cancers (CRCs) develop from preformed adenomas. Cytokines are involved in the transition from adenoma to CRC. Interleukin-33 (IL-33) is a newly discovered proinflammatory cytokine belonging to the IL-1 cytokine family and involved in the development of chronic inflammation and cancer. The aim of this study was to evaluate the dynamics of the IL-33/ST2 axis during the sequence of progression from normal colorectum to adenoma to carcinoma and to investigate the association of IL-33 and ST2 expression with clinicopathological parameters and prognosis. The results demonstrated that the levels of IL-33 and ST2 in adenomas (n = 50), determined by real-time PCR, were significantly higher than those of normal controls (n = 30); the levels of both IL-33/ST mRNA in CRCs (n = 50) were higher than in normal controls but lower than in adenomas. Further analysis revealed that the expression level of ST2 in CRCs was associated with tumor/node/metastasis (TNM) stage. The log-rank test showed that neither the IL-33 nor the ST2 expression level was correlated with overall survival in patients with CRC. The increased expression of IL-33/ST2 in adenomas and CRC tissues was confirmed by immunohistochemistry and was observed in both the tumor stromal cells and adenomatous/cancerous cells. Notably, increased densities of IL-33-positive and ST2-positive microvessels were found in the stroma of adenomas and CRCs. In conclusion, increased expression of the IL-33/ST2 axis along the colorectal adenoma-carcinoma sequence might be involved in the neoplastic transformation via the participation of this axis in the regulation of angiogenesis.


Decoupling of receptor and downstream signals in the Akt pathway by its low-pass filter characteristics.
In cellular signal transduction, the information in an external stimulus is encoded in temporal patterns in the activities of signaling molecules; for example, pulses of a stimulus may produce an increasing response or may produce pulsatile responses in the signaling molecules. Here, we show how the Akt pathway, which is involved in cell growth, specifically transmits temporal information contained in upstream signals to downstream effectors. We modeled the epidermal growth factor (EGF)-dependent Akt pathway in PC12 cells on the basis of experimental results. We obtained counterintuitive results indicating that the sizes of the peak amplitudes of receptor and downstream effector phosphorylation were decoupled; weak, sustained EGF receptor (EGFR) phosphorylation, rather than strong, transient phosphorylation, strongly induced phosphorylation of the ribosomal protein S6, a molecule downstream of Akt. Using frequency response analysis, we found that a three-component Akt pathway exhibited the property of a low-pass filter and that this property could explain decoupling of the peak amplitudes of receptor phosphorylation and that of downstream effectors. Furthermore, we found that lapatinib, an EGFR inhibitor used as an anticancer drug, converted strong, transient Akt phosphorylation into weak, sustained Akt phosphorylation, and, because of the low-pass filter characteristics of the Akt pathway, this led to stronger S6 phosphorylation than occurred in the absence of the inhibitor. Thus, an EGFR inhibitor can potentially act as a downstream activator of some effectors.


Why seeking help from teammates is a blessing and a curse: a theory of help seeking and individual creativity in team contexts.
Research has not explored the extent to which seeking help from teammates positively relates to a person's own creativity. This question is important to explore as help seeking is commonly enacted in organizations and may come with reciprocation costs that may also diminish creativity. Results based on 291 employees in a single division of a large multinational organization revealed that seeking help predicted creativity and mediated the relationship between intrinsic motivation and creativity. However, help seekers also incurred reciprocation costs in that they tended to give more help to teammates, and giving help to teammates was negatively related to creativity. In general, giving higher levels of help attenuated the positive relationship between help seeking and creativity. We also tested an integrated model to show that help giving moderated the mediated relationship between intrinsic motivation and creativity via help seeking, such that higher levels of help giving attenuated this mediated effect. We discuss theoretical and practical implications recommending additional research regarding the interpersonal creative process in team contexts.


Interplay of nanoscale domain purity and size on charge transport and recombination dynamics in polymer solar cells.
Charge transport and bimolecular recombination dynamics were correlated with nanomorphology in polymer solar cells. The morphology of poly(diketopyrrolopyrrole-terthiophene) (PDPP3T) and phenyl-C61-butyric acid methyl ester (PC60BM) blend films was modified using different solvent additives namely 1-chloronaphthalene (CN), 1,8-diiodooctane (DIO) and 1,8-octanedithiol (ODT) and their role on steady state and transient optoelectronic properties was investigated. The energy filtered transmission electron microscopy (EFTEM) images showed that additives (e.g. CN and DIO) improved the domain purity which leads to significantly higher short circuit current densities (Jsc). However when the cells were processed with the ODT additive, the fill factor (FF) and open circuit voltage (Voc) decreased dramatically. Films processed with the ODT additive showed a smaller domain size but were more connected compared to films processed using CN and DIO additives. Transient photocurrent analysis indicates faster charge collection in the case of CN and DIO processed solar cells and the slowest charge collection in ODT processed solar cells. Interestingly devices processed with the ODT additive also showed the longest charge carrier recombination lifetime and lowest bimolecular recombination coefficient. This is attributed to the smaller donor domains that are connected with each other to provide a more interconnected and efficient charge transport matrix but longer pathways in ODT films. Such a matrix helped the charge to escape from the donor-acceptor interfaces and thus reduces the bimolecular recombination, while the longer pathway increases the charge collection time. Further insight is provided into the selection of processing conditions to achieve an ideal active layer morphology consisting of domains with higher polymer purity and optimal size that lead to higher Jsc and FF.


Common data elements for pediatric traumatic brain injury: recommendations from the working group on demographics and clinical assessment.
The Common Data Elements (CDEs) initiative is a National Institutes of Health (NIH) interagency effort to standardize naming, definitions, and data structure for clinical research variables. Comparisons of the results of clinical studies of neurological disorders have been hampered by variability in data coding, definitions, and procedures for sample collection. The CDE project objective is to enable comparison of future clinical trials results in major neurological disorders, including traumatic brain injury (TBI), stroke, multiple sclerosis, and epilepsy. As part of this effort, recommendations for CDEs for research on TBI were developed through a 2009 multi-agency initiative. Following the initial recommendations of the Working Group on Demographics and Clinical Assessment, a separate workgroup developed recommendations on the coding of clinical and demographic variables specific to pediatric TBI studies for subjects younger than 18 years. This article summarizes the selection of measures by the Pediatric TBI Demographics and Clinical Assessment Working Group. The variables are grouped into modules which are grouped into categories. For consistency with other CDE working groups, each variable was classified by priority (core, supplemental, and emerging). Templates were produced to summarize coding formats, guide selection of data points, and provide procedural recommendations. This proposed standardization, together with the products of the other pediatric TBI working groups in imaging, biomarkers, and outcome assessment, will facilitate multi-center studies, comparison of results across studies, and high-quality meta-analyses of individual patient data.


Developmental biology and the redirection or replacement of cells.
The aim of developmental biology is to understand how an egg converts itself into a complete organism through the processes of cell differentiation, morphogenesis and size regulation. The principles that have emerged over recent decades include the constancy of the genome in nearly all cells of an individual, the existence of stem cells in many organs and the overwhelming importance of signalling between cells for the determination of their fate. These and other characteristics of development are discussed here in relation to the prospect of achieving cell and tissue correction or replacement with the help of nuclear transplantation and signalling factors. Nuclear transplantation offers a one-step procedure for generating multipotent embryo cells from the cells of an adult tissue such as skin. It should be possible to proliferate the resulting cells as can be done for mouse embryonic stem cells. Embryo cells can be made to differentiate in many directions by exposing them to various agents or to different concentrations of a single factor such as the transforming growth factor beta class signalling molecule activin. The possibility of a cancerous condition being acquired during these experimental manipulations can be guarded against by transfecting cells with a conditional suicide gene. Thus it may be possible to generate replacement cells or tissues from an adult human for transplantation back to the original donor, without the disadvantage of any genetic incompatibility.


Development of the Italian version of the Pain Catastrophising Scale (PCS-I): cross-cultural adaptation, factor analysis, reliability, validity and sensitivity to change.
The aim of this study was to create an Italian version of the Pain Catastrophising Scale (PCS-I) and evaluate its psychometric properties in a sample with chronic low back pain. The PCS was culturally adapted in accordance with international standards. The psychometric testing included factor analysis, reliability by internal consistency (Cronbach's alpha) and test-retest repeatability (intraclass coefficient correlations), and concurrent validity by comparing the PCS-I with a numerical rating scale (NRS), the Tampa Scale of Kinesiophobia (TSK), the Roland Morris Disability Questionnaire (RMDQ), the Hospital Anxiety and Depression Scale (HADS) and the Positive Affect and Negative Affect Scale (PANAS) (Pearson's correlation). It took 4 months to develop an agreed version of the PCS-I, which was satisfactorily administered to 180 subjects with chronic low back pain. Factor analysis revealed a three-factor 13-item solution (68% of explained variance). The questionnaire was internally consistent with one exception (α = 0.92 as a whole; 0.89 for Helplessness, 0.87 for Rumination and 0.56 for Magnification subscales) and showed a high degree of test-retest reliability (ICC = 0.842). Concurrent validity showed moderate correlations with the NRS (r = 0.44), TSK (r = 0.59), RMDQ (r = 0.45), HADS (Anxiety: r = 0.57; Depression r = 0.46) and PANAS (Negative Affect r = 0.54). The minimum detectable change was 10.45. The subscales were also psychometrically analysed. The successfully translated Italian version of the PCS has good psychometric properties replicating those of other versions.


Breast milk-mediated transfer of an antigen induces tolerance and protection from allergic asthma.
Allergic asthma is a chronic disease characterized by airway obstruction in response to allergen exposure. It results from an inappropriate T helper type 2 response to environmental airborne antigens and affects 300 million individuals. Its prevalence has increased markedly in recent decades, most probably as a result of changes in environmental factors. Exposure to environmental antigens during infancy is crucial to the development of asthma. Epidemiological studies on the relationship between breastfeeding and allergic diseases have reached conflicting results. Here, we have investigated whether the exposure of lactating mice to an airborne allergen affects asthma development in progeny. We found that airborne antigens were efficiently transferred from the mother to the neonate through milk and that tolerance induction did not require the transfer of immunoglobulins. Breastfeeding-induced tolerance relied on the presence of transforming growth factor (TGF)-beta during lactation, was mediated by regulatory CD4+ T lymphocytes and depended on TGF-beta signaling in T cells. In conclusion, breast milk-mediated transfer of an antigen to the neonate resulted in oral tolerance induction leading to antigen-specific protection from allergic airway disease. This study may pave the way for the design of new strategies to prevent the development of allergic diseases.


De-identifying Swedish clinical text - refinement of a gold standard and experiments with Conditional random fields.
In order to perform research on the information contained in Electronic Patient Records (EPRs), access to the data itself is needed. This is often very difficult due to confidentiality regulations. The data sets need to be fully de-identified before they can be distributed to researchers. De-identification is a difficult task where the definitions of annotation classes are not self-evident. We present work on the creation of two refined variants of a manually annotated Gold standard for de-identification, one created automatically, and one created through discussions among the annotators. The data is a subset from the Stockholm EPR Corpus, a data set available within our research group. These are used for the training and evaluation of an automatic system based on the Conditional Random Fields algorithm. Evaluating with four-fold cross-validation on sets of around 4-6 000 annotation instances, we obtained very promising results for both Gold Standards: F-score around 0.80 for a number of experiments, with higher results for certain annotation classes. Moreover, 49 false positives that were verified true positives were found by the system but missed by the annotators. Our intention is to make this Gold standard, The Stockholm EPR PHI Corpus, available to other research groups in the future. Despite being slightly more time-consuming we believe the manual consensus gold standard is the most valuable for further research. We also propose a set of annotation classes to be used for similar de-identification tasks.


Repression of hTERT transcription by the introduction of chromosome 3 into human oral squamous cell carcinoma.
Telomerase is a ribonucleoprotein enzyme that maintains telomere length. Telomerase activity is primarily attributed to the expression of telomerase reverse transcriptase (TERT). It has been reported that introduction of an intact human chromosome 3 into the human oral squamous cell carcinoma cell line HSC3 suppresses the tumorigenicity of these cells. However, the mechanisms that regulate tumorigenicity have not been elucidated. To determine whether this reduction in tumorigenicity was accompanied by a reduction in telomerase activity, we investigated the transcriptional activation of TERT in HSC3 microcell hybrid clones with an introduced human chromosome 3 (HSC3#3). HSC#3 cells showed inhibition of hTERT transcription compared to that of the parental HSC3 cells. Furthermore, cell fusion experiments showed that hybrids of HSC3 cells and cells of the RCC23 renal carcinoma cell line, which also exhibits suppression of TERT transcription by the introduction of human chromosome 3, also displayed suppressed TERT transcription. These results suggested that human chromosome 3 may carry functionally distinct, additional TERT repressor genes.


Clinical outcomes of Bryan cervical disc arthroplasty a prospective, randomized, controlled, single site trial with 48-month follow-up.
Prospective, randomized, controlled. Level 1 evidence. To report functional outcomes at 48 months follow-up on prospectively randomized patients to either the Bryan cervical disc prosthesis or anterior cervical discectomy and fusion (ACDF) at a single site. Surgical treatment of cervical disc pathology can involve discectomy and fusion (ACDF), the gold standard technique. The safety and effectiveness of this procedure has been established and demonstrated in the literature, however, limitations have evolved and alternatives such as disc replacement are being investigated. Intervertebral disc replacement is designed to preserve motion, both at affected and adjacent levels avoiding limitations of fusion such as adjacent level degeneration. New onset degenerative changes and possible recurring neurologic symptoms may be deferred or eliminated with cervical disc replacement. A recent multicenter trial with 24 months follow-up has shown the Bryan disc to compare favorably with ACDF. Continued follow-up is needed to further evaluate and compare functional outcomes in both these cohorts. A total of 47 patients were enrolled at our site as part of an ongoing multicenter prospectively randomized study investigating ACDF versus Bryan cervical disc prosthesis. Functional outcomes are now reported at 48 months follow-up for our cohort of participants. Neck disability index score (NDI), VAS neck and arm and SF-36 both physical and mental as well as complications and reoperations will be reported. Functional outcome data collected at routine follow-up for 48-months has favorably demonstrated improved functional outcomes for NDI, neck/arm pain VAS scores, and the SF-36 physical/mental health component scores for the Bryan arthroplasty and ACDF cohorts. The NDI scores for the Bryan arthroplasty preoperatively was 51 and at 48 months 10. For ACDF preoperative NDI score was also 51 and at 48 months 16.7. At 48 months NDI success, measured by >or=15 points NDI improvement demonstrated a 93.3% success for Bryan arthroplasty and an 82.4% success for ACDF. VAS neck pain scores for the Bryan arthroplasty preoperatively was 76.2 and at 48 months was 13.6. VAS neck pain scores for ACDF preoperatively was 80.6 and at 48 months was 28.1. Arm Pain scores were also measured and for the Bryan arthroplasty preoperatively measured 78.8 and at 48 months 10.8. For ACDF arm pain scores preoperatively measured 77.1 and at 48 months 21.7. These outcomes have not been associated with any degradation of outcome measures from 2 to 4 years. During the 48 months of follow-up at our institution we also report 6 secondary surgeries in our control group (ACDF) and only 1 in our investigational group (Bryan). Of the 6 surgeries in the control group performed, 3 or 12% to date were for adjacent level degenerative disease and 1 or 4% for remote level degenerative disc disease. The remaining 2 surgeries were performed on the same patient for a pseudarthrosis. In the investigational group there was only 1 secondary surgery performed to date for adjacent level disease 5%. At 48 months, cervical arthroplasty with the Bryan cervical disc prosthesis continues to compare favorably to ACDF at our institution. There has been no degradation of functional outcomes from 24 to 48 months for NDI, VAS of neck and arm, and SF-36. There has been a lower incidence of secondary surgeries for the Bryan arthroplasty cohort to date.


Isolation and characterization of a new subtype of Borna disease virus.
Borna disease virus (BDV), the causative agent of severe meningoencephalitis in a wide variety of animal species, has been considered to be genetically invariable and to form a single type within the genus Bornavirus of the family Bornaviridae. BDV infections are of particular interest, because for the first time a virus infection appears to be linked to human psychiatric disorders. We now describe a new subtype of BDV isolated from a horse which was euthanatized due to severe, incurable neurological disease. The nucleotide sequence of this new strain, named No/98, differs from the reference strains by more than 15%, and the subtype is difficult to detect by standard reverse transcriptase PCR protocols. The nucleotide exchanges of the novel BDV isolate have surprisingly little effect on the primary structures of most viral proteins, with the notable exception of the X protein (p10), which is only 81% identical to its counterpart in reference strains. Our data indicate that the genome of BDV is far more variable than previously assumed and that naturally occurring subtypes may escape detection by currently used diagnostic assays.


Pseudonocardia eucalypti sp. nov., an endophytic actinobacterium with a unique knobby spore surface, isolated from roots of a native Australian eucalyptus tree.
A novel strain, designated EUM 374(T), was isolated from the root of a native Australian eucalyptus tree, Eucalyptus microcarpa, and subjected to a range of morphological, phylogenetic and chemotaxonomic analyses. The strain was Gram-reaction-positive with well-developed aerial mycelia, which fragmented into rod-shaped spores that had unique knobby protrusions on the spore surface. Substrate mycelia were not present in the media used. Strain EUM 374(T) grew as a film on the surface of static liquid culture medium but did not grow under shaking conditions. Phylogenetic evaluation based on 16S rRNA gene sequences identified the new isolate as belonging to the family Pseudonocardiaceae with sequence similarities of 96.1 and 96.3 % to Pseudonocardia acaciae GMKU095(T) and Pseudonocardia spinosispora LM 141(T), respectively, and 93-96 % sequence similarity to other members of the genus Pseudonocardia. The results of comprehensive phylogenetic analyses, including physiological and biochemical tests, differentiated strain EUM 374(T) from related members of the genus Pseudonocardia. Based on the phenotypic, phylogenetic and chemotaxonomic evidence, strain EUM 374(T) represents a novel species of the genus Pseudonocardia, for which the name Pseudonocardia eucalypti sp. nov. is proposed. The type strain is EUM 374(T) ( = DSM 45351(T) = ACM 5285(T)).


Captopril potentiates chronotropic baroreflex responses to carotid stimuli in humans.
Angiotensin II is a potent vasoconstrictor and has profound effects on autonomic neural mechanisms in experimental animals. Human carotid baroreflex control of arterial pressure and heart period was examined before and after acutely decreasing angiotensin II levels by administering 50 mg of oral captopril, an angiotensin converting enzyme inhibitor. Carotid baroreceptor stimuli were delivered by a neck chamber worn by 14 normotensive volunteers. Four subjects received placebo. Arterial pressure responses to carotid distention and tachycardia in response to carotid compression were not changed in captopril or placebo groups; however, there was an augmented bradycardic response to carotid stretch in captopril-treated subjects. These results indicate that captopril has an asymmetrical effect on carotid baroreflex function and suggest that enhanced baroreflex mediated bradycardia is due to a reduction in central nervous system angiotensin II levels by captopril, which augments vagal-cardiac responses to carotid stimuli.


Sero-prevalence and risk factors for leptospirosis in abattoir workers in New Zealand.
Leptospirosis is an important occupational disease in New Zealand. The objectives of this study were to determine risk factors for sero-prevalence of leptospiral antibodies in abattoir workers. Sera were collected from 567 abattoir workers and tested by microscopic agglutination for Leptospira interrogans sv. Pomona and Leptospira borgpetersenii sv. Hardjobovis. Association between prevalence and risk factors were determined by species specific multivariable analysis. Eleven percent of workers had antibodies against Hardjobovis or/and Pomona. Workers from the four sheep abattoirs had an average sero-prevalence of 10%-31%, from the two deer abattoirs 17%-19% and the two beef abattoirs 5%. The strongest risk factor for sero-positivity in sheep and deer abattoirs was work position. In sheep abattoirs, prevalence was highest at stunning and hide removal, followed by removal of the bladder and kidneys. Wearing personal protective equipment such as gloves and facemasks did not appear to protect against infection. Home slaughtering, farming or hunting were not significantly associated with sero-prevalence. There is substantial risk of exposure to leptospires in sheep and deer abattoirs in New Zealand and a persisting, but lower risk, in beef abattoirs. Interventions, such as animal vaccination, appear necessary to control leptospirosis as an occupational disease in New Zealand.


Indirect determination of dissolved silicate in surface water using energy-dispersive X-ray fluorescence spectrometry.
An ultrasound-assisted methodology for the determination of dissolved silicate in water has been developed by combining the miniaturized ion-associated based preconcentration method with energy dispersive X-ray fluorescence spectrometry (EDXRF). The method has been devised for the determination of silicate through molybdenum in the ion-associated complex between 12-molybdosilicate and crystal violet. Because silicate is determined indirectly via molybdenum fluorescent radiation the difficulties resulting from low fluorescence yield and low energy of silicon radiation were successfully overcome. A good ratio of silicon to molybdenum (1 to 41) and a sensitive Kα line of molybdenum make possible the determination of low concentration of silicon in the form of dissolved silicate. Under optimized conditions, good linearity, up to 5 μg of silicon in the form of silicate (r = 0.9990) and a detection limit of 9 ng mL(-1) were achieved. The total RSD for the EDXRF determination of silicate, followed by precipitation of the ion-associated complex and its dissolution in a microdrop of isoamyl alcohol, was 6.5%. The enrichment factor was equal to 142. The developed method was used to determine the dissolved silica content in surface waters. The accuracy and repeatability of the proposed procedure were checked by the standard addition method and compared to the results obtained using the ICP-OES technique. The recovery (in the 93-97% range) was satisfactory and indicated the usefulness of the developed procedure.


Staphylococcus aureus skin colonization in atopic dermatitis patients.
A study was conducted to compare the Staphylococcus aureus skin colonization of 21 patients with atopic dermatitis (AD) and 22 healthy controls. It was found that the total aerobe count (total CFU/cm2), the S. aureus fraction thereof and the S. aureus carrier frequency were significantly higher in apparently normal skin of AD patients than in healthy individuals. In addition, compared to normal skin of patients S. aureus density was 100 to 1,000 times higher in the 3 different kinds of lesional skin (dermatitic, lichenified and impetiginized sites). 190 S. aureus strains isolated from the skin of AD patients were tested for sensitivity to 5 topically used antibiotics and the results reported. Besides the biological consequences for the person affected by AD this severe colonization with S. aureus is of epidemiological importance. Several outbreaks of S. aureus infections by dispersal from dermatitic skin have been described. Therefore some preventive and therapeutic aspects are discussed.


Functional MRI of mild traumatic brain injury (mTBI): progress and perspectives from the first decade of studies.
Mild traumatic brain injury (mTBI) represents the great majority of traumatic brain injuries, and is a common medical problem affecting cognitive and vocational functioning as well as quality of life in some individuals. Functional MRI (fMRI) is an important research method for investigating the neuroanatomic substrates of cognitive disorders and their treatment. Surprisingly, however, relatively little research has utilized fMRI to examine alterations in brain functioning after mTBI. This article provides a critical overview of the published fMRI research on mTBI to date. These topics include examination of frontal lobe/executive functions such as working memory, as well as episodic memory and resting state/functional connectivity. mTBI has also been investigated in military populations where studies have focused on effects of blast injury and comorbid conditions such as post-traumatic stress disorder and major depressive disorder. Finally, we address fMRI evaluations of response to behavioral or pharmacological challenges and interventions targeting cognitive and behavioral sequelae of mTBI. The review concludes with identification and discussion of gaps in current knowledge and future directions for fMRI studies of mTBI. The authors conclude that fMRI in combination with related methods can be expected to play an increasing role in research related to studies of pathophysiological mechanisms of the sequelae of mTBI as well as in diagnosis and treatment monitoring.


Optimization of spot-size converter for low polarization dependent loss of waveguide photodetector.
We present an optimization of spot-size converter (SSC) of waveguide photodetector (PD) for small polarization dependent loss (PDL). Beam-propagation method simulation gives responsivity for each polarization and SSC structure. From the calculated responsivity data, optimum structure of SSC is determined that can be implemented with a sufficient process tolerance. We confirm the optimization by measuring PDL of waveguide PD designed according to the structure obtained through the simulation.


Water quality trends in the Delaware River Basin (USA) from 1980 to 2005.
In 1940, the tidal Delaware River was "one of the most grossly polluted areas in the United States." During the 1950s, water quality was so poor along the river at Philadelphia that zero oxygen levels prevented migration of American shad leading to near extirpation of the species. Since then, water quality in the Delaware Basin has improved with implementation of the 1961 Delaware River Basin Compact and 1970s Federal Clean Water Act Amendments. At 15 gages along the Delaware River and major tributaries between 1980 and 2005, water quality for dissolved oxygen, phosphorus, nitrogen, and sediment improved at 39%, remained constant at 51%, and degraded at 10% of the stations. Since 1980, improved water-quality stations outnumbered degraded stations by a 4 to 1 margin. Water quality remains good in the nontidal river above Trenton and, while improved, remains fair to poor for phosphorus and nitrogen in the tidal estuary near Philadelphia and in the Lehigh and Schuylkill tributaries. Water quality is good in heavily forested watersheds (>50%) and poor in highly cultivated watersheds. Water quality recovery in the Delaware Basin is coincident with implementation of environmental laws enacted in the 1960s and 1970s and is congruent with return of striped bass, shad, blue crab, and bald eagle populations.


Regulation of Teladorsagia circumcincta and Trichostrongylus colubriformis worm populations by grazing sheep with differing resistance status.
In an experiment lasting 4 years, changes in the Teladorsagia circumcincta and Trichostrongylus colubriformis populations were compared in lambs and adult sheep with differing resistance statuses. Two flocks of 30 rams (resistant R and susceptible S) grazed separate pastures and 8 rams were slaughtered in the middle and at the end of each grazing season. Five groups of tracer lambs were added each year to estimate the pasture infectivity and were killed for worm counts. The availability of animals with differing resistance statuses (rams and tracer lambs) and differing levels of infection made it possible to investigate the number, size and fecundity of worms of these two species. The inflammatory response was measured in the rams by counting the globule leukocytes, mast cells and eosinophils in the fundic, pyloric and intestinal mucosa. In the tracer lambs, the daily egg production by the female worms of both species was negatively correlated with the worm burden. Worm length accounted for 60 and 70% of the variation in the number of eggs in utero for T. circumcincta and T. colubriformis respectively. Worm length was closely associated with the resistance status of the host; there were greater differences between lambs, and S and R rams for T. colubriformis. T. circumcincta worm lengths were not affected by the worm number. Globule leukocyte counts were related to the worm burdens, and mast cell counts to worm length in the R and S rams. The number, size and fecundity of the worms may well be regulated by similar mechanisms in both species, but T. colubriformis seemed to be more intensively regulated than T. circumcincta. This finding could be useful in devising more effective methods of parasite control.


Changing attitudes to cardiopulmonary resuscitation in older people: a 15-year follow-up study.
while it is well established that individual patient preferences regarding cardiopulmonary resuscitation (CPR) may change with time, the stability of population preferences, especially during periods of social and economic change, has received little attention. to elicit the resuscitation preferences of older Irish inpatients and to compare the results with an identical study conducted 15 years earlier. one hundred and fifty older medical inpatients awaiting discharge in a university teaching hospital or a district general hospital subjects were asked about resuscitation preferences. Results were compared to those elicited from a hundred subjects in 1992. most patients (94%) felt it was a good idea for doctors to discuss CPR routinely with patients, compared with 39% in 1992. In their current health, 6% in 2007 and 76% in 1992 would refuse CPR. The independent predictors of refusal of CPR in current health on logistic regression were age and year of assessment. In the final model, those aged 75-84 years [OR 2.77 (95% CI 1.25-6.13), P = 0.02] and 85 years or more [OR 15.19 (4.26-54.15), P < 0.0001] were more likely than those aged 65-74 years (reference group) to refuse CPR. Those questioned in 2007 [OR 0.04 (0.02-0.81), P < 0.0001] were less likely than those questioned in 1992 (reference group) to refuse CPR. there has been a significant shift in the attitudes of older Irish inpatients over 15 years towards favouring greater patient participation in decision making and an increased desire for resuscitation.


Electroencephalographic and clinical features of cerebral malaria.
Seizures are a prominent feature of childhood cerebral malaria, and are associated with an increased risk of death and neurological sequelae. We present the electroencephalographic (EEG) findings from a detailed clinical and electrophysiological study. Children with cerebral malaria had EEGs recorded within six hours of admission, and at 12 hourly intervals until recovery of consciousness. Ten deeply comatose children underwent intracranial pressure monitoring. Children were not mechanically ventilated, which made it possible to directly correlate the clinical and EEG findings. Of 65 children aged 9 months and above, 40 had one or more seizures, and 18 had an episode of status epilepticus. Most seizures were partial motor, and spike wave activity consistently arose from the posterior temporo-parietal region, a border zone area lying between territories supplied by the carotid and vertebrobasilar circulations. Fifteen children had seizures that were clinically subtle or electrographic. Clinical seizures were associated with an abrupt rise in intracranial pressure. Fifty children recovered fully, seven died, and eight had persistent neurological sequelae. Initial EEG recordings of very slow frequency, or with background asymmetry, burst suppression, or interictal discharges, were associated with an adverse outcome. Serial EEG recording has uncovered a range of clinical, subtle, and electrographic seizures complicating childhood cerebral malaria, and has emphasised their importance in the pathogenesis of coma. Further work is required to determine the most appropriate regimen for the prophylaxis and treatment of seizures in cerebral malaria, in order to improve outcome.


Long-term acclimation to elevated pCO2 alters carbon metabolism and reduces growth in the Antarctic diatom Nitzschia lecointei.
Increasing atmospheric CO2 levels are driving changes in the seawater carbonate system, resulting in higher pCO2 and reduced pH (ocean acidification). Many studies on marine organisms have focused on short-term physiological responses to increased pCO2, and few on slow-growing polar organisms with a relative low adaptation potential. In order to recognize the consequences of climate change in biological systems, acclimation and adaptation to new environments are crucial to address. In this study, physiological responses to long-term acclimation (194 days, approx. 60 asexual generations) of three pCO2 levels (280, 390 and 960 µatm) were investigated in the psychrophilic sea ice diatom Nitzschia lecointei. After 147 days, a small reduction in growth was detected at 960 µatm pCO2. Previous short-term experiments have failed to detect altered growth in N. lecointei at high pCO2, which illustrates the importance of experimental duration in studies of climate change. In addition, carbon metabolism was significantly affected by the long-term treatments, resulting in higher cellular release of dissolved organic carbon (DOC). In turn, the release of labile organic carbon stimulated bacterial productivity in this system. We conclude that long-term acclimation to ocean acidification is important for N. lecointei and that carbon overconsumption and DOC exudation may increase in a high-CO2 world.


Melanopsin regulates visual processing in the mouse retina.
The discovery of melanopsin-dependent inner retinal photoreceptors in mammals has precipitated a fundamental reassessment of such non-image forming (NIF) light responses as circadian photoentrainment and the pupil light reflex. By contrast, it remains unclear whether these new photoreceptors also play a role in classical image-forming vision. The retinal ganglion cells that subserve inner retinal photoreception (ipRGCs) project overwhelmingly to brain areas involved in NIF responses, indicating that, in terms of central signaling, their predominant function is non-image forming. However, ipRGCs also exhibit intraretinal communication via gap junction coupling, which could allow them to modulate classical visual pathways within this tissue. Here, we explore this second possibility by using melanopsin knockout (Opn4-/-) mice to examine the role of inner retinal photoreceptors in diurnal regulation of retinal function. By using electroretinography in wild-type mice, we describe diurnal rhythms in both the amplitude and speed of the retinal cone pathway that are a function of both prior light exposure and circadian phase. Unexpectedly, loss of the melanopsin gene abolishes circadian control of these parameters, causing significant attenuation of the diurnal variation in cone vision. Our results demonstrate for the first time a melanopsin-dependent regulation of visual processing within the retina, revealing an important function for inner retinal photoreceptors in optimizing classical visual pathways according to time of day.


One-pot synthesis of binary metal organic frameworks (HKUST-1 and UiO-66) for enhanced adsorptive removal of water contaminants.
In this study, binary metal organic frameworks (MOFs) with HKUST-1 and UiO-66 have been synthesized in a one-pot process. The synthesized MOFs were characterized by Fourier transform infrared (FTIR) spectroscopy, X-ray diffraction (XRD), scanning electron microscopy (SEM), N2 adsorption, and thermogravimetric analysis (TGA). The meso-porosity and thermal stability of the binary MOFs were higher than those of single HKUST-1 or UiO-66. The synthesized MOF hybrids were then tested for adsorptive removal of methylene blue (MB) from wastewater in terms of kinetic and isothermal adsorption as compared to a commercially available activated carbon (AC). All the synthesized MOFs showed significant removal of MB under a wide range of pH. The adsorption capacities of HKUST-1 are higher than UiO-66 and commercial AC while the binary MOFs presented an even higher adsorption capacity than single MOFs. This is the first time that binary HKUST-1 and UiO-66 MOFs have been successfully synthesized and demonstrated enhanced adsorptive removal of contaminants.


Assembly of GABAA receptor subunits: role of the delta subunit.
GABAA receptor channels (GABARs) composed of different combinations of rat alpha 1, beta 1, gamma 2L, and delta subunits were expressed transiently in mouse fibroblast cells (L929 cells). Whole-cell recordings were obtained from transfected cells to determine which combinations of GABAR subunits formed functional receptor channels, and to compare the electrophysiological and pharmacological characteristics of GABAR channels expressed in the presence and absence of the delta subunit. Only alpha 1 beta 1 gamma 2L, alpha 1 beta 1 gamma 2L delta, and alpha 1 beta 1 delta subunit combinations assembled to form functional GABAR channels and the presence of the delta-subunit slowed the rate of acute desensitization of GABA-evoked current during GABA application and the rate of recovery of GABA-evoked current following GABA application. These three different GABAR channel isoforms also showed distinct pharmacological profiles with differential sensitivity to block by zinc. Zinc was a potent blocker of alpha 1 beta 1 delta GABAR channels, a moderate-strength blocker of alpha 1 beta 1 gamma 2L delta GABAR channels, and did not block the alpha 1 beta 1 gamma 2L GABAR channels. These findings suggest that GABAR isoforms containing the delta subunit constitute a novel GABAR channel with distinct electrophysiological and pharmacological characteristics.


Analogue versus propositional representation in congenitally blind individuals.
Congenitally blind individuals are generally less accurate at mentally manipulating objects than sighted people. However, they often score higher on tests of short- and long-term verbal memory, and it has been suggested that an enhanced propositional representation compensates for inefficiencies in analogue visuospatial representation. Here, congenitally blind, blindfolded, and sighted participants recalled descriptions of relative object locations. In contrast to previous findings, the congenitally blind participants were as accurate as the blindfolded and sighted individuals at remembering the relative locations of objects, but their memory for the verbatim structure of presented descriptions was worse. We propose that, like sighted people, the congenitally blind spontaneously construct and remember analogue representations of object locations and that the performance discrepancies of the blind arise from the process of managing and manipulating these analogue representations.


Child-related and parental predictors for thelarche in a general population of girls: the PANIC study.
Obesity has been associated with earlier thelarche, whereas other predictors for it remain unclear. We studied child-related and parental predictors for earlier thelarche in 195 girls aged 6-8 years followed up for 2 years. A physician evaluated breast development by inspection and palpation. Body fat percentage (BF%) was measured by dual-energy X-ray absorptiometry, diet by food records, and physical activity and sedentary time by body movement and heart rate monitors. Parental education, smoking, and alcohol consumption and household income were assessed by questionnaires. Gestational age, birth weight, and maternal prepregnancy BMI were obtained from hospital registers. Predictors for thelarche were examined using logistic regression analysis adjusted for age and follow-up time. The incidence of thelarche during 2 years increased by 11% (OR 1.11, CI 1.06-1.17, p < 0.001) for 1 unit increase in baseline BF%. Girls with a smoking parent had a 2.64 (95% CI 1.21-5.77, p = 0.015) times higher incidence of thelarche than other girls. The associations of lower parental education and higher maternal prepregnancy BMI with the higher incidence of thelarche were largely explained by BF%. Other possible predictors were not associated with thelarche. Higher BF% and exposure to tobacco smoke are independent predictors for earlier thelarche.


Vehicle Modification Prescription: Australian Occupational Therapy Consensus-Based Guidelines.
Vehicle modifications are often essential to ensure that drivers with disabilities can drive independently and safely. Occupational therapy driver assessors and driver rehabilitation specialists routinely prescribe vehicle modifications on the basis of individualized evaluations and support by rehabilitation interventions. After a literature review and consultation with stakeholders, we identified a resource gap in information to support an evidence-based process for vehicle modification prescription. Using an action research approach, we established content validity for draft guideline items and developed consensus-based draft guidelines for vehicle modification prescription that are ready for trialing by occupational therapy driver assessors. We also propose a practice model for vehicle modification prescription that is applicable to international contexts.


In vivo responses to vaccination with Mage-b, GM-CSF and thioglycollate in a highly metastatic mouse breast tumor model, 4T1.
Metastatic breast cancer is an important contributor to morbidity and mortality. Hence, new therapies are needed that target breast cancer metastases. Here, we focus on Mage-b as a possible vaccine target to prevent the development of breast cancer metastases, through activation of Mage-b-specific cytotoxic T lymphocytes (CTL). The syngeneic cell line 4T1, highly expressing Mage-b, was used as a pre-clinical metastatic mouse breast tumor model. BALB/c mice received three preventive intraperitoneal immunizations with Mage-b DNA vaccine mixed with plasmid DNA, secreting granulocyte-macrophage colony stimulating factor (GM-CSF). In addition, antigen-presenting cells were more efficiently recruited to the peritoneal cavity by the injection of thioglycollate broth (TGB), prior to each immunization. Immunization with Mage-b/GM-CSF/TGB significantly reduced the number of metastases by 67% compared to the saline/GM-CSF/TGB and by 69% compared to the vector control/GM-CSF/TGB. Also, tumor growth was significantly reduced by 45% in mice vaccinated with Mage-b/GM-CSF/TGB compared to the saline/ GM-CSF/TGB and by 47% compared to the control vector/ GM-CSF/TGB group. In vivo, the number of CD8 T cells significantly increased in the primary tumors and metastases of mice vaccinated with Mage-b/GM-CSF/TGB compared to the saline/GM-CSF/TGB and the control vector/ GM-CSF/TGB group, while the number of CD4 T cells significantly decreased. The combination of Mage-b, GM-CSF and TGB did not only induce significantly higher levels of IFNgamma in the lymph nodes of vaccinated compared to control mice, but also induced significantly higher expression levels of Fas-ligand (FasL) in the primary tumors (expressing Fas protein constitutively), compared to the control mice. Whether the interaction between Fas and FasL may have contributed to the smaller tumors needs to be further analyzed.


Food marketing expenditures aimed at youth: putting the numbers in context.
In response to concerns about childhood obesity, the Federal Trade Commission (FTC) released two reports documenting food and beverage marketing expenditures to children and adolescents. The recently released 2012 report found an inflation-adjusted 19.5% reduction in marketing expenditures targeted to youth from $2.1 billion in 2006 to $1.8 billion in 2009. The current article highlights features of the FTC's analysis, examines how expenditures relate to youth exposure to food marketing, and assesses changes in the nutritional content of marketed products. Of the $304.0 million decline in expenditures, $117.8 million (38.7%) was from a decline in premium (i.e., restaurant children's meal toys) expenditures rather than direct marketing. Although inflation-adjusted TV expenditures fell by 19.4%, children and teens still see 12-16 TV advertisements (ads)/day for products generally high in saturated fat, sugar, or sodium. In addition, newer digital forms of unhealthy food and beverage marketing to youths are increasing; the FTC reported an inflation-adjusted 50.7% increase in new media marketing expenditures. The self-regulatory Children's Food and Beverage Advertising Initiative (CFBAI) is limited in scope and effectiveness: expenditures increased for many noncovered marketing techniques (i.e., product placement, movie/video, cross-promotion licenses, athletic sponsorship, celebrity fees, events, philanthropy, and other); only two restaurants are members of CFBAI, and nonpremium restaurant marketing expenditures were up by $86.0 million (22.5% inflation-adjusted increase); industry pledges do not protect children aged >11 years, and some marketing appears to have shifted to older children; and, nutritional content remains poor. Continued monitoring of and improvements to food marketing to youth are needed.


Comparison of different methods of calculating CT radiation effective dose in children.
CT radiation dose is a subject of intense interest and concern, especially in children. Effective dose, a summation of whole-body exposure weighted by specific organ sensitivities, is most often used to compute and compare radiation dose; however, there is little standardization, and there are numerous different methods of calculating effective dose. This study compares five such methods in a group of children undergoing routine chest CT and explores their advantages and pitfalls. Patient data from 120 pediatric chest CT examinations were retrospectively used to calculate effective dose: two scanner dose-length product (DLP) methods using published sets of conversion factors by Shrimpton and Deak, the imaging performance and assessment of CT (ImPact) calculator method, the Alessio online calculator, and the Huda method. The Huda method mean effective dose (4.4 ± 2.2 mSv) and Alessio online calculator (5.2 ± 2.8 mSv) yielded higher mean numbers for effective dose than both DLP calculations (Shrimpton, 3.65 ± 1.8 mSv, and Deak, 3.2 ± 1.5 mSv) as well as the ImPact calculator effective dose (3.4 ± 1.7 mSv). Mean differences ranged from 10.2% ± 10.1% lower to 28% ± 37.3% higher than the Shrimpton method (used as the standard for comparison). Differences were more marked at 120 kVp than at 80 or 100 kVp and varied at different ages. Concordance coefficients relative to the Shrimpton DLP method were Deak DLP, 0.907; Alessio online calculator, 0.735; ImPact calculator, 0.926; and Huda, 0.777. Different methods of computing effective dose for pediatric CT produce varying results. The method used must be clearly described to allay confusion about documenting and communicating dose for archiving as well as comparative research purposes.


Bagging Ensemble of Multilayer Perceptrons for Missing Electricity Consumption Data Imputation.
For efficient and effective energy management, accurate energy consumption forecasting is required in energy management systems (EMSs). Recently, several artificial intelligence-based techniques have been proposed for accurate electric load forecasting; moreover, perfect energy consumption data are critical for the prediction. However, owing to diverse reasons, such as device malfunctions and signal transmission errors, missing data are frequently observed in the actual data. Previously, many imputation methods have been proposed to compensate for missing values; however, these methods have achieved limited success in imputing electric energy consumption data because the period of data missing is long and the dependency on historical data is high. In this study, we propose a novel missing-value imputation scheme for electricity consumption data. The proposed scheme uses a bagging ensemble of multilayer perceptrons (MLPs), called softmax ensemble network, wherein the ensemble weight of each MLP is determined by a softmax function. This ensemble network learns electric energy consumption data with explanatory variables and imputes missing values in this data. To evaluate the performance of our scheme, we performed diverse experiments on real electric energy consumption data and confirmed that the proposed scheme can deliver superior performance compared to other imputation methods.


Ubiquitin systems mark pathogen-containing vacuoles as targets for host defense by guanylate binding proteins.
Many microbes create and maintain pathogen-containing vacuoles (PVs) as an intracellular niche permissive for microbial growth and survival. The destruction of PVs by IFNγ-inducible guanylate binding protein (GBP) and immunity-related GTPase (IRG) host proteins is central to a successful immune response directed against numerous PV-resident pathogens. However, the mechanism by which IRGs and GBPs cooperatively detect and destroy PVs is unclear. We find that host cell priming with IFNγ prompts IRG-dependent association of Toxoplasma- and Chlamydia-containing vacuoles with ubiquitin through regulated translocation of the E3 ubiquitin ligase tumor necrosis factor (TNF) receptor associated factor 6 (TRAF6). This initial ubiquitin labeling elicits p62-mediated escort and deposition of GBPs to PVs, thereby conferring cell-autonomous immunity. Hypervirulent strains of Toxoplasma gondii evade this process via specific rhoptry protein kinases that inhibit IRG function, resulting in blockage of downstream PV ubiquitination and GBP delivery. Our results define a ubiquitin-centered mechanism by which host cells deliver GBPs to PVs and explain how hypervirulent parasites evade GBP-mediated immunity.


A calcineurin-independent mechanism of angiogenesis inhibition by a nonimmunosuppressive cyclosporin A analog.
Cyclosporin A (CsA) is a widely used immunosuppressant drug. Its immunosuppressive activity occurs through the inhibition of the protein phosphatase calcineurin via formation of a ternary complex with cyclophilin A (CypA). CsA also inhibits endothelial cell proliferation and angiogenesis. This has been thought to occur through calcineurin inhibition as well. However, CsA is also a potent inhibitor of cyclophilins, a class of prolyl isomerases. Because calcineurin inhibition requires binding, and therefore inhibition of CypA, the relative contributions of calcineurin and cyclophilin inhibition in antiangiogenesis have not been addressed. We have taken a chemical biology approach to explore this question by dissociating the two activities of CsA at the molecular level. We have identified a nonimmunosuppressive analog of CsA that does not inhibit calcineurin but maintains inhibition of endothelial cell proliferation and in vivo angiogenesis. The same analog also maintains inhibition of all cyclophilin isoforms tested. We also show that a second, structurally distinct, cyclophilin inhibitor is sufficient to block endothelial cell proliferation. These results suggest that the inhibition of cyclophilins may play a larger role in the antiangiogenic activity of CsA than previously believed, and that cyclophilins may be potential antiangiogenic drug targets.


Sphaeranthus indicus Linn.: A phytopharmacological review.
Sphaeranthus indicus Linn. (Asteraceae) is widely used in Ayurvedic system of medicine to treat vitiated conditions of epilepsy, mental illness, hemicrania, jaundice, hepatopathy, diabetes, leprosy, fever, pectoralgia, cough, gastropathy, hernia, hemorrhoids, helminthiasis, dyspepsia and skin diseases. There are reports providing scientific evidences for hypotensive, anxiolytic, neuroleptic, hypolipidemic, immunomodulatory, antioxidant, anti-inflammatory, bronchodialatory, antihyperglycemic and hepatoprotective activities of this plant. A wide range of phytochemical constituents have been isolated from this plant including sesquiterpene lactones, eudesmenolides, flavanoids and essential oil. A comprehensive account of the morphology, phytochemical constituents, ethnobotanical uses and pharmacological activities reported are included in this review for exploring the immense medicinal potential of this plant.


Systematic RNA interference reveals that oncogenic KRAS-driven cancers require TBK1.
The proto-oncogene KRAS is mutated in a wide array of human cancers, most of which are aggressive and respond poorly to standard therapies. Although the identification of specific oncogenes has led to the development of clinically effective, molecularly targeted therapies in some cases, KRAS has remained refractory to this approach. A complementary strategy for targeting KRAS is to identify gene products that, when inhibited, result in cell death only in the presence of an oncogenic allele. Here we have used systematic RNA interference to detect synthetic lethal partners of oncogenic KRAS and found that the non-canonical IkappaB kinase TBK1 was selectively essential in cells that contain mutant KRAS. Suppression of TBK1 induced apoptosis specifically in human cancer cell lines that depend on oncogenic KRAS expression. In these cells, TBK1 activated NF-kappaB anti-apoptotic signals involving c-Rel and BCL-XL (also known as BCL2L1) that were essential for survival, providing mechanistic insights into this synthetic lethal interaction. These observations indicate that TBK1 and NF-kappaB signalling are essential in KRAS mutant tumours, and establish a general approach for the rational identification of co-dependent pathways in cancer.


Therapeutic potential of adult bone marrow-derived mesenchymal stem cells in diseases of the skeleton.
Mesenchymal stem cells (MSCs) are the most popular among the adult stem cells in tissue engineering and regenerative medicine. Since their discovery and functional characterization in the late 1960s and early 1970s, MSCs or MSC-like cells have been obtained from various mesodermal and non-mesodermal tissues, although majority of the therapeutic applications involved bone marrow-derived MSCs. Based on its mesenchymal origin, it was predicted earlier that MSCs only can differentiate into mesengenic lineages like bone, cartilage, fat or muscle. However, varied isolation and cell culturing methods identified subsets of MSCs in the bone marrow which not only differentiated into mesenchymal lineages, but also into ectodermal and endodermal derivatives. Although, true pluripotent status is yet to be established, MSCs have been successfully used in bone and cartilage regeneration in osteoporotic fracture and arthritis, respectively, and in the repair of cardiac tissue following myocardial infarction. Immunosuppressive properties of MSCs extend utility of MSCs to reduce complications of graft versus host disease and rheumatoid arthritis. Homing of MSCs to sites of tissue injury, including tumor, is well established. In addition to their ability in tissue regeneration, MSCs can be genetically engineered ex vivo for delivery of therapeutic molecule(s) to the sites of injury or tumorigenesis as cell therapy vehicles. MSCs tend to lose surface receptors for trafficking and have been reported to develop sarcoma in long-term culture. In this article, we reviewed the current status of MSCs with special emphasis to therapeutic application in bone-related diseases.


Human participants challenges in youth-focused research: perspectives and practices of IRB administrators.
The purpose of this research was to understand institutional review board (IRB) challenges regarding youth-focused research submissions and to present advice from administrators. Semistructured self-report questionnaires were sent via e-mail to administrators identified using published lists of universities and hospitals an Internet searches. Of 183 eligible institutions, 49 responded. One half indicated they never granted parental waivers. Among those considering waivers, decision factors included research risks, survey content, and feasibility. Smoking and substance abuse research among children was generally considered more than minimal risk. These findings are consistent with those from a study conducted by Mammel and Kaplan (1995), which investigated IRB practices concerning protocols involving adolescent participants. IRBs and investigators need to become aware of regulations' flexibility to ensure adequate participant protection. Investigators need to limit jargon and assumptions about participants' understanding of research objectives.


SARS coronavirus unique domain: three-domain molecular architecture in solution and RNA binding.
Nonstructural protein 3 of the severe acute respiratory syndrome (SARS) coronavirus includes a "SARS-unique domain" (SUD) consisting of three globular domains separated by short linker peptide segments. This work reports NMR structure determinations of the C-terminal domain (SUD-C) and a two-domain construct (SUD-MC) containing the middle domain (SUD-M) and the C-terminal domain, and NMR data on the conformational states of the N-terminal domain (SUD-N) and the SUD-NM two-domain construct. Both SUD-N and SUD-NM are monomeric and globular in solution; in SUD-NM, there is high mobility in the two-residue interdomain linking sequence, with no preferred relative orientation of the two domains. SUD-C adopts a frataxin like fold and has structural similarity to DNA-binding domains of DNA-modifying enzymes. The structures of both SUD-M (previously determined) and SUD-C (from the present study) are maintained in SUD-MC, where the two domains are flexibly linked. Gel-shift experiments showed that both SUD-C and SUD-MC bind to single-stranded RNA and recognize purine bases more strongly than pyrimidine bases, whereby SUD-MC binds to a more restricted set of purine-containing RNA sequences than SUD-M. NMR chemical shift perturbation experiments with observations of (15)N-labeled proteins further resulted in delineation of RNA binding sites (i.e., in SUD-M, a positively charged surface area with a pronounced cavity, and in SUD-C, several residues of an anti-parallel beta-sheet). Overall, the present data provide evidence for molecular mechanisms involving the concerted actions of SUD-M and SUD-C, which result in specific RNA binding that might be unique to the SUD and, thus, to the SARS coronavirus.


The Chikungunya epidemic on La Réunion Island in 2005-2006: a cost-of-illness study.
This study was conducted to assess the impact of chikungunya on health costs during the epidemic that occurred on La Réunion in 2005-2006. From data collected from health agencies, the additional costs incurred by chikungunya in terms of consultations, drug consumption and absence from work were determined by a comparison with the expected costs outside the epidemic period. The cost of hospitalization was estimated from data provided by the national hospitalization database for short-term care by considering all hospital stays in which the ICD-10 code A92.0 appeared. A cost-of-illness study was conducted from the perspective of the third-party payer. Direct medical costs per outpatient and inpatient case were evaluated. The costs were estimated in Euros at 2006 values. Additional reimbursements for consultations with general practitioners and drugs were estimated as € 12.4 million (range: € 7.7 million-€ 17.1 million) and € 5 million (€ 1.9 million-€ 8.1 million), respectively, while the cost of hospitalization for chikungunya was estimated to be € 8.5 million (€ 5.8 million-€ 8.7 million). Productivity costs were estimated as € 17.4 million (€ 6 million-€ 28.9 million). The medical cost of the chikungunya epidemic was estimated as € 43.9 million, 60% due to direct medical costs and 40% to indirect costs (€ 26.5 million and € 17.4 million, respectively). The direct medical cost was assessed as € 90 for each outpatient and € 2,000 for each inpatient. The medical management of chikungunya during the epidemic on La Réunion Island was associated with an important economic burden. The estimated cost of the reported disease can be used to evaluate the cost/efficacy and cost/benefit ratios for prevention and control programmes of emerging arboviruses.


Lipoprotein lipase gene expression in THP-1 cells.
Lipoprotein lipase (LPL) mRNA levels are under the control of signals that activate phospholipase C, resulting in activation of protein kinase C (PKC) and mobilization of intracellular Ca2+ in the human monocytic leukemia cell line THP-1. Induction of LPL in THP-1 cells appears to be mediated by PKC since it was affected by both phorbol 12-myristate 13-acetate (PMA) and a diacylglycerol analogue. This induction was blocked by the specific PKC inhibitor H-7. Although Ca2+ mobilization by the ionophore A23187 also induced LPL mRNA, the mechanism is most likely independent of activation of the Ca2+/calmodulin protein kinase. Depletion of cells of PKC made them refractory to induction by A23187, suggesting that Ca2+ mobilization acts by activating PKC. Addition of cycloheximide (CHX) to undifferentiated THP-1 cells resulted in a transient increase in steady-state mRNA levels (3-fold). Sustained superinduction of LPL mRNA occurred when PMA and CHX were added simultaneously. These results suggest that the level of LPL mRNA is regulated either by a labile regulatory protein, which represses transcription of the LPL gene, or by a protein affecting mRNA stability.


A comparison of two computer input devices for uppercase letter matching.
To determine whether the Touch Window or the mouse with an enlarged on-screen arrow was more effective or efficient for an on-screen letter-matching task completed by a boy 9 years of age with spastic quadriplegic cerebral palsy and visual and cognitive deficits. A single-subject research design of 5 treatment phases, A1, B1, A2, B2, and A & B, was used. The total percentage of correct letter matches per treatment session, the total percentage correct per letter, and the amount of time needed to match 5 consecutive letters correctly were evaluated. The range and variability of letters correctly matched per session decreased and trends for correct letter matches accelerated when the participant used the mouse interface. Accuracy with matching 18 of 26 (69%) letters of the alphabet increased when selections were made with the mouse interface. The participant was faster when using the Touch Window to match 5 consecutive letters correctly; however, regardless of the interface device used, letter matching remained slow and tedious. The mouse with an enlarged on-screen arrow and cursor was the more effective interface device with this child. Making a minor adjustment such as increasing the size of the on-screen arrow can make a common piece of equipment accessible to a person with disabilities.


Micromechanical Analysis of the Hyaluronan-Rich Matrix Surrounding the Oocyte Reveals a Uniquely Soft and Elastic Composition.
The cumulus cell-oocyte complex (COC) matrix is an extended coat that forms around the oocyte a few hours before ovulation and plays vital roles in oocyte biology. Here, we analyzed the micromechanical response of mouse COC matrix by colloidal-probe atomic force microscopy. We found that the COC matrix is elastic insofar as it does not flow and its original shape is restored after force release. At the same time, the COC matrix is extremely soft. Specifically, the most compliant parts of in vivo and in vitro expanded COC matrices yielded Young's modulus values of 0.5 ± 0.1 Pa and 1.6 ± 0.3 Pa, respectively, suggesting both high porosity and a large mesh size (≥100 nm). In addition, the elastic modulus increased progressively with indentation. Furthermore, using optical microscopy to correlate these mechanical properties with ultrastructure, we discovered that the COC is surrounded by a thick matrix shell that is essentially devoid of cumulus cells and is enhanced upon COC expansion in vivo. We propose that the pronounced nonlinear elastic behavior of the COC matrix is a consequence of structural heterogeneity and serves important functions in biological processes such as oocyte transport in the oviduct and sperm penetration.


Genomewide linkage scan of schizophrenia in a large multicenter pedigree sample using single nucleotide polymorphisms.
A genomewide linkage scan was carried out in eight clinical samples of informative schizophrenia families. After all quality control checks, the analysis of 707 European-ancestry families included 1615 affected and 1602 unaffected genotyped individuals, and the analysis of all 807 families included 1900 affected and 1839 unaffected individuals. Multipoint linkage analysis with correction for marker-marker linkage disequilibrium was carried out with 5861 single nucleotide polymorphisms (SNPs; Illumina version 4.0 linkage map). Suggestive evidence for linkage (European families) was observed on chromosomes 8p21, 8q24.1, 9q34 and 12q24.1 in nonparametric and/or parametric analyses. In a logistic regression allele-sharing analysis of linkage allowing for intersite heterogeneity, genomewide significant evidence for linkage was observed on chromosome 10p12. Significant heterogeneity was also observed on chromosome 22q11.1. Evidence for linkage across family sets and analyses was most consistent on chromosome 8p21, with a one-LOD support interval that does not include the candidate gene NRG1, suggesting that one or more other susceptibility loci might exist in the region. In this era of genomewide association and deep resequencing studies, consensus linkage regions deserve continued attention, given that linkage signals can be produced by many types of genomic variation, including any combination of multiple common or rare SNPs or copy number variants in a region.


Does the ethical appropriateness of paying donors depend on what body parts they donate?
The idea of paying donors in order to make more human bodily material available for therapy, assisted reproduction, and biomedical research is notoriously controversial. However, while national and international donation policies largely oppose financial incentives they do not treat all parts of the body equally: incentives are allowed in connection to the provision of some parts but not others. Taking off from this observation, I discuss whether body parts differ as regards the ethical legitimacy of incentives and, if so, why. I distinguish two approaches to this issue. On a "principled" approach, some but not all body parts are inherently special in a way that proscribes payment. On a "pragmatic" approach, the appropriateness of payment in relation to a specific part must be determined through an overall assessment of e.g. the implications of payment for the health and welfare of providers, recipients, and third parties, and the quality of providers' consent. I argue that the first approach raises deep and potentially divisive questions about the good life, whereas the second approach invokes currently unsupported empirical assumptions and requires difficult balancing between different values and the interests of different people. This does not mean that any attempt to distinguish between body parts in regard to the appropriateness of payment necessarily fails. However, I conclude, any plausible such attempt should either articulate and defend a specific view of the good life, or gather relevant empirical evidence and apply defensible principles for weighing goods and interests.


Downregulation of uPAR and cathepsin B induces apoptosis via regulation of Bcl-2 and Bax and inhibition of the PI3K/Akt pathway in gliomas.
Glioma is the most commonly diagnosed primary brain tumor and is characterized by invasive and infiltrative behavior. uPAR and cathepsin B are known to be overexpressed in high-grade gliomas and are strongly correlated with invasive cancer phenotypes. In the present study, we observed that simultaneous downregulation of uPAR and cathepsin B induces upregulation of some pro-apoptotic genes and suppression of anti-apoptotic genes in human glioma cells. uPAR and cathepsin B (pCU)-downregulated cells exhibited decreases in the Bcl-2/Bax ratio and initiated the collapse of mitochondrial membrane potential. We also observed that the broad caspase inhibitor, Z-Asp-2, 6-dichlorobenzoylmethylketone rescued pCU-induced apoptosis in U251 cells but not in 5310 cells. Immunoblot analysis of caspase-9 immunoprecipitates for Apaf-1 showed that uPAR and cathepsin B knockdown activated apoptosome complex formation in U251 cells. Downregulation of uPAR and cathepsin B also retarded nuclear translocation and interfered with DNA binding activity of CREB in both U251 and 5310 cells. Further western blotting analysis demonstrated that downregulation of uPAR and cathepsin B significantly decreased expression of the signaling molecules p-PDGFR-β, p-PI3K and p-Akt. An increase in the number of TUNEL-positive cells, increased Bax expression, and decreased Bcl-2 expression in nude mice brain tumor sections and brain tissue lysates confirm our in vitro results. In conclusion, RNAi-mediated downregulation of uPAR and cathepsin B initiates caspase-dependent mitochondrial apoptosis in U251 cells and caspase-independent mitochondrial apoptosis in 5310 cells. Thus, targeting uPAR and cathepsin B-mediated signaling using siRNA may serve as a novel therapeutic strategy for the treatment of gliomas.


The ecology of outcomes: system accountability in children's mental health.
This article provides a conceptual and practical framework called the Ecology of Outcomes. Based on this framework, agencies that serve children and families build and use outcome-oriented information systems to respond to their clients in a more flexible manner. The goal is to improve promising programs by involving stakeholders in outcome identification and in utilization of results. Problems addressed include the emphasis human services place on rules compliance, lack of feedback to program staff to allow for midcourse correction, and lack of input by key stakeholders in the identification of outcomes to be measured. The following components of the framework are described: principles of outcome accountability, prerequisites and building blocks, implementing an outcome information system, and utilizing the results. Key elements of the framework are the integration of outcome information into a service system's decision-making process and the inclusion of client, stakeholder, and provider satisfaction information.


Improvement in immediate memory after 16 weeks of tualang honey (Agro Mas) supplement in healthy postmenopausal women.
The aim of this study was to evaluate the verbal learning and memory performance of postmenopausal women who received tualang honey (Agro Mas) in comparison with women receiving estrogen plus progestin therapy and untreated controls. A total of 102 postmenopausal women were recruited and randomly assigned to three groups: tualang honey (20 g/d)[corrected], estrogen plus progestin therapy (Femoston 1/5), and untreated control. Their verbal learning and memory performances were assessed using the Malay version of the Auditory Verbal Learning Test before and after 16 weeks of intervention. Data were analyzed using the repeated-measures analysis of variance, and a P value of less than 0.05 was considered significant. There were significant differences in the mean scores of total learning as well as the mean scores of trials A1, A5, A6, and A7 between the three groups. There were also significant differences in the overall mean scores of total learning and trials A1 and A5 between both estrogen plus progestin therapy and tualang honey groups when compared with the untreated control group. However, significant differences in the mean score for trials A6 and A7 were only observed between the estrogen plus progestin therapy and untreated control groups. Postmenopausal women who received tualang honey showed improvement in their immediate memory but not in immediate memory after the interference and delayed recall. This is comparable with the improvement seen in women receiving estrogen plus progestin therapy.


Performance of stroke risk scores in older people with atrial fibrillation not taking warfarin: comparative cohort study from BAFTA trial.
To compare the predictive power of the main existing and recently proposed schemes for stratification of risk of stroke in older patients with atrial fibrillation. Comparative cohort study of eight risk stratification scores. Trial of thromboprophylaxis in stroke, the Birmingham Atrial Fibrillation in the Aged (BAFTA) trial. 665 patients aged 75 or over with atrial fibrillation based in the community who were randomised to the BAFTA trial and were not taking warfarin throughout or for part of the study period. Events rates of stroke and thromboembolism. 54 (8%) patients had an ischaemic stroke, four (0.6%) had a systemic embolism, and 13 (2%) had a transient ischaemic attack. The distribution of patients classified into the three risk categories (low, moderate, high) was similar across three of the risk stratification scores (revised CHADS(2), NICE, ACC/AHA/ESC), with most patients categorised as high risk (65-69%, n = 460-457) and the remaining classified as moderate risk. The original CHADS(2) (Congestive heart failure, Hypertension, Age ≥ 75 years, Diabetes, previous Stroke) score identified the lowest number as high risk (27%, n = 180). The incremental risk scores of CHADS(2), Rietbrock modified CHADS(2), and CHA(2)DS(2)-VASc (CHA(2)DS(2)-Vascular disease, Age 65-74 years, Sex) failed to show an increase in risk at the upper range of scores. The predictive accuracy was similar across the tested schemes with C statistic ranging from 0.55 (original CHADS(2)) to 0.62 (Rietbrock modified CHADS(2)), with all except the original CHADS(2) predicting better than chance. Bootstrapped paired comparisons provided no evidence of significant differences between the discriminatory ability of the schemes. Based on this single trial population, current risk stratification schemes in older people with atrial fibrillation have only limited ability to predict the risk of stroke. Given the systematic undertreatment of older people with anticoagulation, and the relative safety of warfarin versus aspirin in those aged over 70, there could be a pragmatic rationale for classifying all patients over 75 as "high risk" until better tools are available.


In situ viability detection assays induce heat-shock protein 70 expression in spermatozoa without affecting the chromatin integrity.
To differentiate dead spermatozoa from viable but immotile spermatozoa, several techniques are being used during ICSI. As processed spermatozoa from poor-quality ejaculate are confronted with a higher risk of experiencing stress on exposure to altered osmotic conditions or chemicals, this study was undertaken to determine the expression of stress response gene Hsp70 and chromatin integrity in spermatozoa subjected to in situ viability assays such as hypo-osmotic swelling (HOS) test, modified hypo-osmotic swelling (M-HOS) test and pentoxifylline in 25 fresh and frozen-thawed asthenozoospermic ejaculates. RT-PCR and immunofluorescence detection of Hsp70 were performed to elucidate the expression and localisation of Hsp70 in spermatozoa, whereas DNA fragmentation analysis was performed by sperm chromatin dispersion assay. Exposure of fresh and frozen-thawed asthenozoospermic spermatozoa to M-HOS and pentoxifylline significantly increased Hsp70 expression as evidenced by increased RNA expression and immunolocalisation of Hsp70 protein in sperm head (P < 0.05-0.001). However, chromatin integrity was not significantly affected in any groups until 6 h of post-exposure time period. Our results suggest that conventional HOS may be preferred for the in situ detection of the viability as there was no immediate stress response and chromatin instability in the exposed spermatozoa.