Novel AM60-SiO2 Nanocomposite Produced via Ultrasound-Assisted Casting; Production and Characterization.
There has been growing interest in developing new materials with higher strength-to-weight ratios. Therefore, AM60 magnesium alloy reinforced with SiO2 nanoparticles was synthesized using ultrasound-casting method for the first time, in this study. We introduced 1 and 2 wt.% of SiO2 nanoparticles into the samples. Introduction of nanoparticles led to the grain size drop in MS2 (AM60 + 2 wt.% SiO2) samples. In addition, this increased the hardness of samples from 34.8 Vickers hardness (HV) in M (AM60) to 51.5 HV in MS2, and increased the compressive strength of MS2. Improvement of the mechanical properties can be attributed to a combination of Orowan, Hall-Petch and load-bearing mechanisms. However, ductility of the composites decreased with fracture strains being 0.41, 0.39 and 0.37, respectively, for samples M, MS1 and MS2. Fracture surfaces showed shear fracture in both composite samples with microcracks and a more brittle fracture in MS2.


A computational model of bidirectional axonal growth in micro-tissue engineered neuronal networks (micro-TENNs).
Micro-Tissue Engineered Neural Networks (Micro-TENNs) are living three-dimensional constructs designed to replicate the neuroanatomy of white matter pathways in the brain and are being developed as implantable micro-tissue for axon tract reconstruction, or as anatomically-relevant in vitro experimental platforms. Micro-TENNs are composed of discrete neuronal aggregates connected by bundles of long-projecting axonal tracts within miniature tubular hydrogels. In order to help design and optimize micro-TENN performance, we have created a new computational model including geometric and functional properties. The model is built upon the three-dimensional diffusion equation and incorporates large-scale uni- and bi-directional growth that simulates realistic neuron morphologies. The model captures unique features of 3D axonal tract development that are not apparent in planar outgrowth and may be insightful for how white matter pathways form during brain development. The processes of axonal outgrowth, branching, turning and aggregation/bundling from each neuron are described through functions built on concentration equations and growth time distributed across the growth segments. Once developed we conducted multiple parametric studies to explore the applicability of the method and conducted preliminary validation via comparisons to experimentally grown micro-TENNs for a range of growth conditions. Using this framework, the model can be applied to study micro-TENN growth processes and functional characteristics using spiking network or compartmental network modeling. This model may be applied to improve our understanding of axonal tract development and functionality, as well as to optimize the fabrication of implantable tissue engineered brain pathways for nervous system reconstruction and/or modulation.


Molecular design of upconversion nanoparticles for gene delivery.
Due to their large anti-Stokes shifts, sharp emission spectra and long excited-state lifetimes, upconversion nanoparticles (UCNPs) have attracted an increasing amount of research interests, and have shown great potential for enhancing the practical utility of gene therapy, whose versatility has been limited by existing gene delivery technologies that are basically mono-functional in nature. Despite this, up to now in-depth analysis of the development of UCNPs for gene delivery has been scant in the literature, even though there has been an upsurge of reviews on the chemistry of UCNPs and their applications in bioimaging and drug delivery. To fill this gap, this review aims to present the latest advances in the development and applications of UCNPs as gene carriers. Prior to describing the prominent works published in the field, a critical view on the properties, chemistry and molecular design of UCNPs for gene delivery is provided. With a synopsis of the recent advances in UCNP-mediated gene delivery, challenges and opportunities could be illuminated for clinical translation of works in this nascent field of research.


Comparison of nerve transfers and nerve grafting for traumatic upper plexus palsy: a systematic review and analysis.
In treating patients with brachial plexus injury, there are no comparative data on the outcomes of nerve grafts or nerve transfers for isolated upper trunk or C5-C6-C7 root injuries. The purpose of our study was to compare, with systematic review, the outcomes for modern intraplexal nerve transfers for shoulder and elbow function with autogenous nerve grafting for upper brachial plexus traumatic injuries. PubMed, EMBASE, and the Cochrane Central Register of Controlled Trials were searched for studies in which patients had surgery for traumatic upper brachial plexus palsy within one year of injury and with a minimum follow-up of twelve months. Strength and shoulder and elbow motion were assessed as outcome measures. The Fisher exact test and Mann-Whitney U test were used to compare outcomes, with an alpha level of 0.05. Thirty-one studies met the inclusion criteria. Two hundred and forty-seven (83%) and 286 (96%) of 299 patients with nerve transfers achieved elbow flexion strength of grade M4 or greater and M3 or greater, respectively, compared with thirty-two (56%) and forty-seven (82%) of fifty-seven patients with nerve grafts (p < 0.05). Forty (74%) of fifty-four patients with dual nerve transfers for shoulder function had shoulder abduction strength of grade M4 or greater compared with twenty (35%) of fifty-seven patients with nerve transfer to a single nerve and thirteen (46%) of twenty-eight patients with nerve grafts (p < 0.05). The average shoulder abduction and external rotation was 122° (range, 45° to 170°) and 108° (range, 60° to 140°) after dual nerve transfers and 50° (range, 0° to 100°) and 45° (range, 0° to 140°) in patients with nerve transfers to a single nerve. In patients with demonstrated complete traumatic upper brachial plexus injuries of C5-C6, the pooled international data strongly favors dual nerve transfer over traditional nerve grafting for restoration of improved shoulder and elbow function. These data may be helpful to surgeons considering intraoperative options, particularly in cases in which the native nerve root or trunk may appear less than optimal, or when long nerve grafts are contemplated.


Interleukin 35 Delays Hindlimb Ischemia-Induced Angiogenesis Through Regulating ROS-Extracellular Matrix but Spares Later Regenerative Angiogenesis.
Interleukin (IL) 35 is a novel immunosuppressive heterodimeric cytokine in IL-12 family. Whether and how IL-35 regulates ischemia-induced angiogenesis in peripheral artery diseases are unrevealed. To fill this important knowledge gap, we used loss-of-function, gain-of-function, omics data analysis, RNA-Seq, in vivo and in vitro experiments, and we have made the following significant findings: i) IL-35 and its receptor subunit IL-12RB2, but not IL-6ST, are induced in the muscle after hindlimb ischemia (HLI); ii) HLI-induced angiogenesis is improved in Il12rb2-/- mice, in ApoE-/-/Il12rb2-/- mice compared to WT and ApoE-/- controls, respectively, where hyperlipidemia inhibits angiogenesis in vivo and in vitro; iii) IL-35 cytokine injection as a gain-of-function approach delays blood perfusion recovery at day 14 after HLI; iv) IL-35 spares regenerative angiogenesis at the late phase of HLI recovery after day 14 of HLI; v) Transcriptome analysis of endothelial cells (ECs) at 14 days post-HLI reveals a disturbed extracellular matrix re-organization in IL-35-injected mice; vi) IL-35 downregulates three reactive oxygen species (ROS) promoters and upregulates one ROS attenuator, which may functionally mediate IL-35 upregulation of anti-angiogenic extracellular matrix proteins in ECs; and vii) IL-35 inhibits human microvascular EC migration and tube formation in vitro mainly through upregulating anti-angiogenic extracellular matrix-remodeling proteins. These findings provide a novel insight on the future therapeutic potential of IL-35 in suppressing ischemia/inflammation-triggered inflammatory angiogenesis at early phase but sparing regenerative angiogenesis at late phase.


Evaluation of patient reporting of adverse drug reactions to the UK 'Yellow Card Scheme': literature review, descriptive and qualitative analyses, and questionnaire surveys.
The monitoring of adverse drug reactions (ADRs) through pharmacovigilance is vital to patient safety. Spontaneous reporting of ADRs is one method of pharmacovigilance, and in the UK this is undertaken through the Yellow Card Scheme (YCS). Yellow Card reports are submitted to the Medicines and Healthcare products Regulatory Agency (MHRA) by post, telephone or via the internet. The MHRA electronically records and reviews information submitted so that important safety issues can be detected. While previous studies have shown differences between patient and health-care professional (HCP) reports for the types of drugs and reactions reported, relatively little is known about the pharmacovigilance impact of patient reports. There have also been few studies on the views and experiences of patients/consumers on the reporting of suspected ADRs. To evaluate the pharmacovigilance impact of patient reporting of ADRs by analysing reports of suspected ADRs from the UK YCS and comparing reports from patients and HCPs. To elicit the views and experiences of patients and the public about patient reporting of ADRs. (1) Literature review and survey of international experiences of consumer reporting of ADRs; (2) descriptive analysis of Yellow Card reports; (3) signal generation analysis of Yellow Card reports; (4) qualitative analysis of Yellow Card reports; (5) questionnaire survey of patients reporting on Yellow Cards; (6) qualitative analysis of telephone interviews with patient reporters to the scheme; (7) qualitative analysis of focus groups and usability testing of the patient YCS; and (8) national omnibus telephone survey of public awareness of the YCS. Patients (n = 5180) and HCPs (n = 20,949) submitting Yellow Card reports from October 2005 to September 2007. Respondents to questionnaire survey (n = 1362). Participants at focus groups and usability testing sessions (n = 40). National omnibus telephone survey (n = 2028). The literature review included studies in English from across the world. All other components included populations from the UK; the omnibus survey was restricted to Great Britain. None. Characteristics of patient reports: types of drug and suspected ADR reported; seriousness of reports; and content of reports. The relative contributions of patient reports and of HCP reports to signal generation. Views and experiences of patient reporters. Views of members of the public about the YCS, including user-friendliness and usability of different ways of patient reporting. Public awareness of the YCS. Suggestions for improving patient reporting to the YCS. Compared with HCPs, patient reports to the YCS contained a higher median number of suspected ADRs per report, and described reactions in more detail. The proportions of reports categorised as 'serious' were similar; the patterns of drugs and reactions reported differed. Patient reports were richer in their descriptions of reactions than those from HCPs, and more often noted the effects of ADRs on patients' lives. Combining patient and HCP reports generated more potential signals than HCP reports alone; some potential signals in the 'HCP-only' data set were lost when combined with patient reports, but fewer than those gained; the addition of patient reports to HCP reports identified 47 new 'serious' reactions not previously included in 'Summaries of Product Characteristics'. Most patient reporters found it fairly easy to make reports, although improvements to the scheme were suggested, including greater publicity and the redesign of web- and paper-based reporting systems. Among members of the public, 8.5% were aware of the YCS in 2009. Patient reporting of suspected ADRs has the potential to add value to pharmacovigilance by reporting types of drugs and reactions different from those reported by HCPs; generating new potential signals; and describing suspected ADRs in enough detail to provide useful information on likely causality and impact on patients' lives. These findings suggest that further promotion of patient reporting to the YCS is justified, along with improvements to existing reporting systems. In order of priority, future work should include further investigation of (1) the pharmacovigilance impact of patient reporting in a longer-term study; (2) the optimum approach to signal generation analysis of patient and HCP reports; (3) the burden of ADRs in terms of impact on patients' lives; (4) the knowledge and attitudes of HCPs towards patient reporting of ADRs; (5) the value of using patient reports of ADRs to help other patients and HCPs who are seeking information on patient experiences of ADRs; and (6) the impact of increasing publicity and/or enhancements to reporting systems on the numbers and types of Yellow Card reports from patients. The National Institute for Health Research Health Technology Assessment programme.


Top-down proteomics reveals concerted reductions in myofilament and Z-disc protein phosphorylation after acute myocardial infarction.
Heart failure (HF) is a leading cause of morbidity and mortality worldwide and is most often precipitated by myocardial infarction. However, the molecular changes driving cardiac dysfunction immediately after myocardial infarction remain poorly understood. Myofilament proteins, responsible for cardiac contraction and relaxation, play critical roles in signal reception and transduction in HF. Post-translational modifications of myofilament proteins afford a mechanism for the beat-to-beat regulation of cardiac function. Thus it is of paramount importance to gain a comprehensive understanding of post-translational modifications of myofilament proteins involved in regulating early molecular events in the post-infarcted myocardium. We have developed a novel liquid chromatography-mass spectrometry-based top-down proteomics strategy to comprehensively assess the modifications of key cardiac proteins in the myofilament subproteome extracted from a minimal amount of myocardial tissue with high reproducibility and throughput. The entire procedure, including tissue homogenization, myofilament extraction, and on-line LC/MS, takes less than three hours. Notably, enabled by this novel top-down proteomics technology, we discovered a concerted significant reduction in the phosphorylation of three crucial cardiac proteins in acutely infarcted swine myocardium: cardiac troponin I and myosin regulatory light chain of the myofilaments and, unexpectedly, enigma homolog isoform 2 (ENH2) of the Z-disc. Furthermore, top-down MS allowed us to comprehensively sequence these proteins and pinpoint their phosphorylation sites. For the first time, we have characterized the sequence of ENH2 and identified it as a phosphoprotein. ENH2 is localized at the Z-disc, which has been increasingly recognized for its role as a nodal point in cardiac signaling. Thus our proteomics discovery opens up new avenues for the investigation of concerted signaling between myofilament and Z-disc in the early molecular events that contribute to cardiac dysfunction and progression to HF.


XZH-5 inhibits STAT3 phosphorylation and causes apoptosis in human hepatocellular carcinoma cells.
Growing evidence has demonstrated that signal transducer and activator of transcription 3 (STAT3), which is constitutively activated in patients with hepatocellular carcinoma (HCC), plays an important role in HCC development, progression, and prognosis. Interleukin-6 (IL-6) is a multifunctional inflammatory cytokine, which may induce STAT3 activation in a variety of human cancers. In this study, we demonstrated that blocking STAT3 phosphorylation with STAT3 small molecule inhibitors SD-1029 and Stattic caused apoptosis in HCC cells. Then we characterized the inhibitory effects of a novel small molecule XZH-5 on STAT3 phosphorylation in HCC cell lines. XZH-5 reduced constitutive STAT3 phosphorylation at Tyr705 and the expression of STAT3 downstream genes. The inhibition of STAT3 in HCC cells resulted in the induction of apoptosis and reduction of colony forming ability. In addition, XZH-5 also inhibited IL-6-induced STAT3 phosphorylation, nuclear translocation and STAT3 DNA binding activity. In contrast, it had no effect on IFN-γ-induced STAT1 phosphorylation, indicating the more selective effects on STAT3. These results suggested that XZH-5 may serve as a lead compound for further development of STAT3 specific small molecule inhibitors for HCC therapy.


Visualization of Categorical Longitudinal and Times Series Data.
Plotting growth curves is a powerful graphical approach used in exploratory data analysis for continuous longitudinal data. However, plotted growth curves for multiple participants rapidly become uninterpretable with categorical data. Categorical data define specific states (e.g., being single, married, divorced), and these states do not necessarily need to represent any hierarchical order. Thus, a trajectory becomes a sequence of states rather than a continuum. We introduce a horizontal line plot that uses shade or color to differentiate between states on a categorical longitudinal variable for multiple participants. With appropriate sorting, stacking the horizontal lines that represent each participant can reveal important patterns such as the shape of, or heterogeneity in, the trajectories. We illustrate the plotting techniques for large sample sizes, observed groups, the exploration of unobserved latent classes, large numbers of time points such as are found with intensive longitudinal designs or multivariate time series data, individually varying times observation, unique numbers of observations, and missing data. We used the R package longCatEDA to create the illustrations. Illustrative data include both simulated data and alcohol consumption data in adult schizophrenics from the Clinical Antipsychotic Trials of Intervention Effectiveness.


Activation of the DNA damage checkpoint in yeast lacking the histone chaperone anti-silencing function 1.
The packaging of the eukaryotic genome into chromatin is likely to be important for the maintenance of genomic integrity. Chromatin structures are assembled onto newly synthesized DNA by the action of chromatin assembly factors, including anti-silencing function 1 (ASF1). To investigate the role of chromatin structure in the maintenance of genomic integrity, we examined budding yeast lacking the histone chaperone Asf1p. We found that yeast lacking Asf1p accumulate in metaphase of the cell cycle due to activation of the DNA damage checkpoint. Furthermore, yeast lacking Asf1p are highly sensitive to mutations in DNA polymerase alpha and to DNA replicational stresses. Although yeast lacking Asf1p do complete DNA replication, they have greatly elevated rates of DNA damage occurring during DNA replication, as indicated by spontaneous Ddc2p-green fluorescent protein foci. The presence of elevated levels of spontaneous DNA damage in asf1 mutants is due to increased DNA damage, rather than the failure to repair double-strand DNA breaks, because asf1 mutants are fully functional for double-strand DNA repair. Our data indicate that the altered chromatin structure in asf1 mutants leads to elevated rates of spontaneous recombination, mutation, and DNA damage foci formation arising during DNA replication, which in turn activates cell cycle checkpoints that respond to DNA damage.


Evaluation of AAV-DJ vector for retinal gene therapy.
The most common virus vector used in gene therapy research for ophthalmologic diseases is the adeno-associated virus (AAV) vector, which has been used successfully in a number of preclinical and clinical studies. It is important to evaluate novel AAV vectors in animal models for application of clinical gene therapy. The AAV-DJ (type 2/type 8/type 9 chimera) was engineered from shuffling eight different wild-type native viruses. In this study, we investigated the efficiency of gene transfer by AAV-DJ injections into the retina. One microliter of AAV-2-CAGGS-EGFP or AAV-DJ-CAGGS-EGFP vector at a titer of 1.4 × 10e12 vg/ml was injected intravitreally or subretinally in each eye of C57BL/6 mice. We evaluated the transduction characteristics of AAV-2 and -DJ vectors using fluorescence microscopy and electroretinography. The results confirmed that AAV-DJ could deeply transfer gene to photoreceptor layer with intravitreal injection and has an efficient gene transfer to various cell types especially the Mueller cells in the retina. Retinal function was not affected by AAV-DJ infection or ectopic EGFP expression. The AAV-DJ vector efficiently induces the reporter gene in both the inner and outer murine retina without functional toxicity. These data indicated that the AAV-DJ vector is a useful tool for the gene therapy research targeting retinal disorders.


Design and pilot results of a mobile phone weight-loss application for women starting a meal replacement programme.
We developed and tested a mobile phone application (app) to support individuals embarking on a partial meal replacement programme (MRP). Overweight or obese women were randomly allocated to one of two study groups. The intervention group received an MRP Support app. The control group received a static app based on the information available with the MRP. A total of 58 adult women (Support n = 28; Control n = 30) participated in the 8-week trial. Their BMI was 26-43 kg/m2 Usage data suggested that the intervention group were more engaged with using the app throughout the study period. Mixed modelling revealed that the difference in weight loss between the intervention and control groups (estimated mean, EM = 3.2% and 2.2% respectively) was not significant (P = 0.08). Objective data suggested that users of the Support app were more engaged than those using the control app. A total of 1098 prompts (54%) asking people in the intervention group to enter their meals were completed prior to the evening prompt. Women in the intervention group reported a greater increase in positive affect (i.e. mood) than those in the control group (EM = 0.48 and -0.01, respectively) (P = 0.012). At Week 8, those in the control group reported a greater decrease in the effort they were willing to put into staying on the diet than those who received the Support app (EM = -2.8 and -1.4, respectively) (P = 0.024). The Support app could be a useful adjunct to existing MRPs for psychological outcomes.


Evidence that myocardial pertussis toxin substrates are uniquely altered in acute murine Chagas' disease in a manner unrelated to myocardial dysfunction.
In an effort to correlate biochemical characteristics of the beta-adrenergic receptor complex with myocardial function, mouse myocardial GTP-binding proteins, specifically substrates for pertussis toxin (PT), were analysed with regard to the influence of infection with Trypanosoma cruzi, the causative agent of Chagas' cardiomyopathy. Infection was found to decrease in a non-uniform manner the magnitude of ADP-ribosylation in the PT substrates. High detergent concentrations attenuated the infection-associated decrease in PT-dependent ADP-ribosylation. Infection also altered the kinetics of the PT-dependent ADP-ribosylation reaction from a time course wherein maximal PT-dependent ADP-ribosylation occurred after 12 h incubation in control animals to one in which maximal PT-dependent ADP-ribosylation occurred after 3 h incubation and thereafter declined. Immunochemical analysis of the PT-substrates revealed an infection-associated decrease in alpha i1, alpha o, an increase in alpha i2 and no change in alpha i3. Verapamil treatment, which prevents the clinical consequences of infection, did not influence any of the infection-associated changes in PT-dependent ADP-ribosylation of GTP-binding protein substrates or their immunochemical properties. Complementary studies using isolated rat neonatal cardiocytes infected with the parasite further substantiated the finding that the infection-associated decrease in PT-dependent ADP-ribosylation and the associated change in the kinetics of the reaction were properties uniquely associated with the presence of the parasite.


Venus fly trap domain of mGluR1 functions as a dominant negative against group I mGluR signaling.
Metabotropic glutamate receptors (mGluRs) form covalently linked homodimers and contain large, N-terminal extracellular ligand binding, "venus fly trap" (VFT) domains. These domains, when expressed separately, are secreted as disulfide linked dimers and can dimerize with full-length receptors. mGluR splice variants have been described that contain only this domain, but the consequences of their interaction on receptor signaling have not been explored. Here it is shown that an mGluR1 mutant containing only the VFT is retained on the cell surface when a full-length receptor is co-expressed. Further, when expressed in rat superior cervical ganglion (SCG) neurons and modulation of native calcium currents is used as an assay for receptor activity, the VFT acts as a dominant negative with respect to mGluR1 signaling. Although full-length mGluR1 and mGluR5 are not known to heterodimerize, the mGluR5 VFT partially occludes mGluR1 signaling and the mGluR1 VFT potently occludes mGluR5 signaling in SCG neurons. In addition, an mGluR1 point mutant, mGluR1 C140G, which cannot covalently dimerize, functions like the wild-type receptor when expressed alone. The C140G mutant is inhibited by the mGluR1 VFT construct but does not retain the mGluR1 VFT on the cell surface, suggesting that the loss of C140 renders the interaction reversible. Finally, a peptide designed to disrupt mGluR1 dimerization reduced signaling through the C140G mutant receptor, but only when applied intracellularly for several hours, indicating that loss of signaling requires disruption of dimerization prior to plasma membrane insertion.


Carvedilol therapy in pediatric patients with dilated cardiomyopathy.
Carvedilol reduces mortality and hospitalization in adults with congestive heart failure. Limited information is available about its use in children. The objective of this study was to determine the dosing, efficacy and side effects of carvedilol for the management of dilated cardiomyopathy in children. Sixteen children with idiopathic dilated cardiomyopathy, aged 7 months to 138 months and with an ejection fraction less than 40%, were treated with carvedilol. The average initial dose was 0.1 mg/kg/day and it was uptitrated to 0.4 mg/kg/day. After six months on carvedilol, there were improvements in clinical scoring system from an average of 2.94 to 2.50 (p<0.05), in mean fractional shortening from 17.2 +/- 6.1% to 22.7 +/- 5.1% (p<0.05), and in ejection fraction from 35.2 +/- 6.8% to 43.1 +/- 11.2% (p<0.05). No side effect was observed during the study period. Two patients died due to serious infection. Carvedilol in addition to standard therapy for dilated cardiomyopathy in children improves cardiac function and symptoms. It is well tolerated, with minimal adverse effects, but close monitoring is necessary.


Cell surface immunoglobulin. VII. Synthesis, shedding, and secretion of immunoglobulin by lymphoid cells of germ-free mice.
Lymphoid cells from the spleen, lymph nodes, and thoracic duct of axenic and control mice were incubated with [(3)H]tyrosine and synthesis and secretion of protein and Ig studied. It was found that only IgM was synthesized by cells from axenic mice whereas cells from control mice also synthesized IgG. Splenocytes from both axenic and control mice had 8S IgM on their surface. Radiolabeled splenocytes from axenic mice were incubated to determine the kinetics of release of (125)I-labeled cell surface IgM and [(3)H]tyrosine-labeled IgM. Cell surface IgM was shed as 8S with an initial half-life of release of 5-8 h whereas [(3)H]tyrosine-labeled Ig was secreted as 19S with an initial half-life of 2-3 h. These findings suggest that two independent pathways are involved. It is suggested that small lymphocytes shed 8S IgM and plasma cells secrete 19S IgM. It was observed that lymphoid cells from axenic mice synthesize a higher proportion of IgM relative to total protein. Electron microscopic examination of splenocytes from such mice revealed a markedly higher proportion of plasma cells and a paucity of lymphoblasts compared to controls. It was suggested, therefore, that axenic mice lack a population of stimulated T cells which can induce a switch from IgM to IgG synthesis and which is capable of suppressing IgM synthesis. Lymphoid cells from axenic mice synthesize and secrete less protein that coprecipitates with antigen-antibody complexes.


Study Protocol: Longitudinal Attention and Temperament Study.
Background: Attention processes may play a central role in shaping trajectories of socioemotional development. Individuals who are clinically anxious or have high levels of trait anxiety sometimes show attention biases to threat. There is emerging evidence that young children also demonstrate a link between attention bias to salient stimuli and broad socioemotional profiles. However, we do not have a systematic and comprehensive assessment of how attention biases, and associated neural and behavioral correlates, emerge and change from infancy through toddlerhood. This paper describes the Longitudinal Attention and Temperament study (LAnTs), which is designed to target these open questions. Method: The current study examines core components of attention across the first 2 years of life, as well as measures of temperament, parental psychosocial functioning, and biological markers of emotion regulation and anxiety risk. The demographically diverse sample (N = 357) was recruited from the area surrounding State College, PA, Harrisburg, PA, and Newark, NJ. Infants and parents are assessed at 4, 8, 12, 18, and 24 months. Assessments include repeated measures of attention bias (via eye-tracking) in both infants and parents, and measures of temperament (reactivity, negative affect), parental traits (e.g., anxiety and depression), biological markers (electrophysiology, EEG, and respiratory sinus arrythmia, RSA), and the environment (geocoding, neighborhood characteristics, perceived stress). Outcomes include temperamental behavioral inhibition, social behavior, early symptom profiles, and cellular aging (e.g., telomere length). Discussion: This multi-method study aims to identify biomarkers and behavioral indicators of attentional and socioemotional trajectories. The current study brought together innovative measurement techniques to capture the earliest mechanisms that may be causally linked to a pervasive set of problem behaviors. The analyses the emerge from the study will address important questions of socioemotional development and help shape future research. Analyses systematically assessing attention bias patterns, as well as socioemotional profiles, will allow us to delineate the time course of any emerging interrelations. Finally, this study is the first to directly assess competing models of the role attention may play in socioemotional development in the first years of life.


The role of synchrony and ambiguity in speech-gesture integration during comprehension.
During face-to-face communication, one does not only hear speech but also see a speaker's communicative hand movements. It has been shown that such hand gestures play an important role in communication where the two modalities influence each other's interpretation. A gesture typically temporally overlaps with coexpressive speech, but the gesture is often initiated before (but not after) the coexpressive speech. The present ERP study investigated what degree of asynchrony in the speech and gesture onsets are optimal for semantic integration of the concurrent gesture and speech. Videos of a person gesturing were combined with speech segments that were either semantically congruent or incongruent with the gesture. Although gesture and speech always overlapped in time, gesture and speech were presented with three different degrees of asynchrony. In the SOA 0 condition, the gesture onset and the speech onset were simultaneous. In the SOA 160 and 360 conditions, speech was delayed by 160 and 360 msec, respectively. ERPs time locked to speech onset showed a significant difference between semantically congruent versus incongruent gesture-speech combinations on the N400 for the SOA 0 and 160 conditions. No significant difference was found for the SOA 360 condition. These results imply that speech and gesture are integrated most efficiently when the differences in onsets do not exceed a certain time span because of the fact that iconic gestures need speech to be disambiguated in a way relevant to the speech context.


An architectural perspective of cell-cycle control at the G1/S phase cell-cycle transition.
A prominent role for the execution of cell cycle and growth regulatory mechanisms within the three-dimensional context of nuclear architecture is becoming increasingly evident. Signaling pathways and regulatory networks that govern activation and suppression of genes controlling proliferation are functionally integrated for the organization and assembly of transcriptional machinery in nuclear microenvironments. The transcriptional activation of histone genes at the G1/S phase transition (S-point) is temporarily, functionally, and spatially distinct from transcriptional mechanisms at the restriction point (R-point). The spatial distinction in R-point versus S-point control is the localization of clustered histone gene loci at cajal bodies, which is modulated during the cell cycle. Histone nuclear factor P (HiNF-P), the principal factor mediating H4 histone gene transcription, is the final link in the signaling cascade that is initiated with growth factor dependent induction of cyclin E/CDK2 kinase activity at the R-point and culminates in the NPAT-mediated activation of histone H4 genes through HiNF-P at the G1/S phase cell-cycle transition.


Ultraviolet photodissociation action spectroscopy of gas-phase protonated quinoline and isoquinoline cations.
The gas-phase photodissociation action spectroscopy of protonated quinoline and isoquinoline cations (quinolineH(+) and isoquinolineH(+)) is investigated at ambient temperature. Both isomers exhibit vibronic detail and wavelength-dependent photoproduct partitioning across two broad bands in the ultraviolet. Photodissociation action spectra are reported spanning 370-285 nm and 250-220 nm and analysed with the aid of electronic structure calculations: TD-DFT (CAM-B3LYP/aug-cc-pVDZ) is used for spectra simulations and CBS-QB3 for dissociation enthalpies. It is shown that the action spectra are afforded predominantly by two-photon excitation. The first band is attributed to both the S1 ← S0 and S2 ← S0 electronic transitions in quinolineH(+), with a S1 ← S0 electronic origin assigned at 27,900 cm(-1). For isoquinolineH(+) the S1 ← S0 transition is observed with an assigned electronic origin at 27,500 cm(-1). A separate higher energy band is observed for both species, corresponding to the S3 ← S0 transition, with origins assigned at 42,100 cm(-1) and 42,500 cm(-1) for quinolineH(+) and isoquinolineH(+), respectively. Franck-Condon absorption simulations provide an explanation for some vibrational structure observed in both bands allowing several normal mode assignments. The nature of the electronic transitions is discussed and it is shown that the excited states active in the reported spectra should be of ππ* character with some degree of charge transfer from the homocycle to the heterocycle.


High specific detection of osteopontin using a three-dimensional copolymer layer support based on electrochemical impedance spectroscopy.
Tumor marker detection is essential for the therapy efficiency of early stage tumors and the evaluation of disease progression. Osteopontin (OPN) is supposed to be closely related to several kinds of tumors. In the present study, we describe a label-free electrochemical detection of OPN based on a specific reaction between OPN and its relevant antibody. An artificial three-dimensional (3D) scaffold structure consisting of 11-mercaptoundecanoic acid/6-mercapto-1-hexanol, dextran amino and synthetic peptides was designed as a substrate for the immobilization of the antibody. This substrate was characterized using cyclic voltammetry, atomic force microscopy and Fourier transform infrared reflection spectroscopy. Antibody immobilization and OPN detection were conducted using electrochemical impedance spectroscopy (EIS). The low limit of detection was 0.17 nM. The concentration of cancer risk (5.77 nM) can be selectively detected with a high EIS signal. The fabricated 3D OPN sensor is proposed for application in clinical analysis.


After rehabilitation: meeting the long-term needs of persons with traumatic brain injury.
Many survivors of traumatic brain injury have significant needs for long-term support. The rehabilitation professions, including occupational therapy, have begun to recognize these needs more fully during the last decade. The literature describes various models of supported living that provide housing, support services, and vocational assistance; however, these programs are not widely available due to limitations in public funding and inadequate insurance coverage for long-term care. Occupational therapists can contribute a broad expertise to the provision of supported living services, and the profession is encouraged to provide leadership and advocacy with respect to the long-term care needs of survivors of traumatic brain injury.


Determining thresholds for spatial urban design and transport features that support walking to create healthy and sustainable cities: findings from the IPEN Adult study.
An essential characteristic of a healthy and sustainable city is a physically active population. Effective policies for healthy and sustainable cities require evidence-informed quantitative targets. We aimed to identify the minimum thresholds for urban design and transport features associated with two physical activity criteria: at least 80% probability of engaging in any walking for transport and WHO's target of at least 15% relative reduction in insufficient physical activity through walking. The International Physical Activity and the Environment Network Adult (known as IPEN) study (N=11 615; 14 cities across ten countries) provided data on local urban design and transport features linked to walking. Associations of these features with the probability of engaging in any walking for transport and sufficient physical activity (≥150 min/week) by walking were estimated, and thresholds associated with the physical activity criteria were determined. Curvilinear associations of population, street intersection, and public transport densities with walking were found. Neighbourhoods exceeding around 5700 people per km2, 100 intersections per km2, and 25 public transport stops per km2 were associated with meeting one or both physical activity criteria. Shorter distances to the nearest park were associated with more physical activity. We use the results to suggest specific target values for each feature as benchmarks for progression towards creating healthy and sustainable cities.


Q-switched neodymium-doped Y3Al5O12-based silica fiber laser.
We present pulsed laser operation in a Nd-doped, Y3Al5O12-based silica fiber. The fiber was fabricated using the rod-in-tube technique with a Nd:YAG crystal rod as the core material and a silica tube for the cladding material. A spectroscopy study revealed that the core region had become amorphous in the process of fiber drawing. Q-switched pulsed laser operation was realized at a wavelength of 1058 nm when the fiber was cladding pumped at a wavelength of 808 nm. The laser delivered 38 μJ of energy in 65 ns pulses. The extracted energy was limited due to the multimodal operation of the fiber. Laser slope efficiency in continuous wave operation reached 52%. The spectroscopic properties of the fabricated fiber are discussed and compared to a Nd:YAG crystal and a Nd:Al-doped silica fiber.


Neovascular potential of adipose-derived stromal cells (ASCs) from diabetic patients.
We explored the vascular biology of adipose-derived stromal cells (ASCs) from diabetic patients and applied these cells to a murine ischemic flap model to assess the comparative angiogenic potentials between normal and diabetic human ASCs. ASCs were obtained from diabetic patients (n = 5) and controls (n = 5). Secretion and expression of angiogenic cytokines were measured under normoxic and hypoxic condition in vitro. Conditioned media harvested from ASC cultures were assessed for their ability to stimulate human umbilical vein endothelial cell proliferation and tubulization. The control and diabetic ASCs were injected into the murine ischemic flaps, and the surviving area was measured. Diabetic adipose-derived stromal cells showed a lower level of vascular endothelial growth factor expression and cell proliferation rates than the control cells (p < 0.05). However, vascular endothelial growth factor, hepatocyte growth factor secretion, tubulogenesis, and cell proliferation in diabetic conditioned media were increased in response to hypoxic stimuli (p < 0.05), and it was similar to those of control cells. In an animal study, diabetic and normal ASCs significantly increased flap survival (p < 0.05); however, the functional difference was not found between the two groups. Diabetic ASCs were impaired in their ability to produce vascular endothelial growth factors and to induce cellular proliferation under hypoxic conditions. However, diabetic ASCs showed similar flap salvaging effect compared with controls. These findings may be important in the context of future study of autologous cell-based therapy in diabetic patients.


Type B lactic acidosis secondary to thiamine deficiency in a child with malignancy.
Type B lactic acidosis is an underrecognized clinical entity that must be distinguished from type A (hypoxic) lactic acidosis. We present the case of a 4-year-old boy with medulloblastoma who presented with lactic acidosis in the setting of septic shock. His hyperlactatemia persisted to high levels even after his hemodynamic status improved. After administration of intravenous thiamine, his lactate level rapidly normalized and remained stable. It was determined that his total parenteral nutrition was deficient in vitamins due to a national shortage. Because thiamine is an important cofactor for pyruvate dehydrogenase, he was unable to use glucose through aerobic metabolism pathways. We briefly review type A versus type B lactic acidosis in this case report.


Between a rock and a hard place.
The authors present the case of a patient who presented with acute coronary syndrome requiring percutaneous coronary intervention. He was incidentally diagnosed with a large abdominal abdominal aortic aneurysm which developed acute inflammatory features with rapid sac expansion and imminent rupture. This short report aims to highlight several dilemma that confronted us during the management of this patient.


Urogenital atrophy: prevention and treatment.
Fifteen percent of premenopausal women, 10-40% of postmenopausal women, and 10-25% of women receiving systemic hormone therapy experience urogenital atrophy. The most common symptoms are dryness, burning, pruritus, irritation, and dyspareunia. Estrogen loss, drugs, and chemical sensitivities are causes. Estrogen or hormone replacement therapy (ERT-HRT) is the treatment of choice in postmenopausal women. Dosages prescribed for menopause symptoms or to prevent osteoporosis (and, potentially, other conditions) can restore the vagina to premenopausal physiology and relieve symptoms. Concomitant progestins are necessary for women with an intact uterus to minimize or eliminate estrogen-induced endometrial cancer. Low-dosage oral and vaginal ERT can relieve urogenital atrophy but might not produce systemic effects. Progestins are not necessary with vaginal rings and vaginal tablets. If ERT is given only to treat urogenital atrophy, estrogen creams 1 or 2 times/week may prevent recurrence after symptoms are resolved. Progestins are not required for occasional estrogen cream use. Vaginal moisturizers provide longer relief by changing the fluid content of endothelium and lowering vaginal pH. Vaginal lubricants provide short-term relief. Women with contraindications to ERT-HRT could use lubricants for intercourse-related dryness or moisturizers for more continuous relief. The lay press promotes agrimony, black cohosh, chaste tree, dong quai, witch hazel, and phytoestrogens for vaginal dryness and dyspareunia; however, no evidence exists to support these specific claims. Pharmacists should be actively involved in identifying, preventing, and treating urogenital atrophy.


Regulation of protein turnover by glucose, insulin, and amino acids in adipose tissue.
Protein synthesis and degradation were measured simultaneously in epididymal fat pads of rats by use of the incorporation of [14C]phenylalanine into protein and the sum of net protein breakdown and protein synthesis, respectively. Neither glucose nor insulin altered protein synthesis, but together they promoted this process; pyruvate could be substituted for glucose. Separately, glucose or insulin diminished proteolysis, and these effects were additive. In the presence of glucose and insulin, leucine, alanine, glutamine, glutamate, and aspartate lowered protein degradation to varying degrees but did not alter protein synthesis. Glutamate, but not leucine or alanine, was inhibitory without glucose and insulin present. When aminooxyacetic acid was provided to decrease the rate of transamination of amino acids, the inhibitory effects of leucine, alanine, and aspartate, but not of glutamate, appeared to be diminished. alpha-Ketoglutarate, but neither alpha-ketoisocaproate nor pyruvate, could diminish proteolysis. Inhibition of proteolysis was associated with a higher tissue content of glutamate and a greater production of glutamate and glutamine. These results suggest that glutamate itself may inhibit proteolysis in adipose tissue and mediate, at least in part, the effects of other amino acids.


Interobserver Variability of Quantitative and Qualitative Assessment Using MRI in Malignant Pleural Mesothelioma.
To evaluate the interobserver variability associated with quantitative and qualitative MRI assessments of malignant pleural mesothelioma (MPM). Patients with MPM who underwent uniform-protocol preoperative MRI between 2009 and 2014 were included. The MRI-derived tumor volume was estimated. Unidimensional measurements of maximal pleural thickness (P max) and average pleural thickness (P avg) on axial MR images; maximal fissural thickness (F max); maximal diaphragmatic thickness (D max); and average diaphragmatic thickness (D avg) on sagittal reconstructed images were acquired. Interobserver agreement regarding the American Joint Committee on Cancer (AJCC) tumor stage at each criterion level was assessed by using Cohen κ statistics. Agreement between quantitative measurements was assessed by using Bland-Altman plots and intraclass correlation coefficients (ICCs). The study cohort included 349 patients (median age, 68 years [age range, 30-90 years), 273 (78%) of whom were men and 203 (58%) of whom had epithelioid-subtype tumors. Qualitative assessment performed by using the AJCC staging criteria (eighth edition) was concordant in 31% of cases and yielded considerable disagreement (κ = 0.177). Inspection of the Bland-Altman plots led to decisive agreement between the two reviewers regarding MRI-derived tumor volume (ICC, 0.979). There was also a good degree of agreement between the two reviewers regarding unidimensional measurements of D max (ICC, 0.807), D avg (ICC, 0.823), P max (ICC, 0.787), P avg (ICC, 0.787), and F max (ICC, 0.659). Quantitative assessment can enhance the clinical staging of MPM. Compared with qualitative assessment, quantitative assessment has low interobserver variability and could yield a tumor size criterion that is currently lacking in the AJCC clinical staging of MPM.Supplemental material is available for this article.© RSNA, 2020.


EURADOS intercomparison on emergency radiobioassay.
Nine laboratories participated in an intercomparison exercise organised by the European Radiation Dosimetry Group (EURADOS) for emergency radiobioassay involving four high-risk radionuclides ((239)Pu, (241)Am, (90)Sr and (226)Ra). Diverse methods of analysis were used by the participating laboratories for the in vitro determination of each of the four radionuclides in urine samples. Almost all the methods used are sensitive enough to meet the requirements for emergency radiobioassay derived for this project in reference to the Clinical Decision Guide introduced by the NCRP. Results from most of the methods meet the requirements of ISO 28218 on accuracy in terms of relative bias and relative precision. However, some technical gaps have been identified. For example, some laboratories do not have the ability to assay samples containing (226)Ra, and sample turnaround time would be expected to be much shorter than that reported by many laboratories, as timely results for internal contamination and early decisions on medical intervention are highly desired. Participating laboratories are expected to learn from each other on the methods used to improve the interoperability among these laboratories.


Increased barium influx and potassium current in stroke-prone spontaneously hypertensive rats.
Arterial potassium permeability is increased in hypertension. In this study we conducted voltage-clamp experiments to determine whether the whole-cell K+ current is increased in a Ca(2+)-dependent manner in aortic smooth muscle cells from stroke-prone spontaneously hypertensive rats (SHRSP). Aortic cells from Wistar-Kyoto (WKY) rats and SHRSP demonstrated an outward rectifying current elicited by depolarization. The current was carried primarily by K+, because intracellular Cs+ replacement eliminated more than 97% of the current. The current density was higher (P < .05) in SHRSP cells at positive potentials. In the presence of LaCl3 (200 mumol/L) or tetraethylammonium (10 mmol/L), the residual current was similar in WKY and SHRSP cells. Also, the current density did not differ between WKY and SHRSP cells in which the intracellular Ca2+ concentration was clamped at zero. Fura 2 ratio measurement showed similar resting myoplasmic Ca2+ concentration (Ca2+m) in WKY and SHRSP cells (100 +/- 10 versus 117 +/- 9 nmol/L, P = .2). Under low extracellular Na+ conditions, which had a minimal effect on Ca2+m, Ba2+ replacement of Ca2+ caused a continuous and approximately linear increase in the fura 2 ratio, which was twofold faster in SHRSP cells. Because Ca2+ pumps do not transport Ba2+ and Na(+)-Ca2+ exchange was inhibited by low extracellular Na+, this increase reflected unidirectional Ba2+ influx.(ABSTRACT TRUNCATED AT 250 WORDS)


Spinal manipulative therapy for elderly patients with chronic obstructive pulmonary disease: a case series.
The objective of this case series is to report the results of spinal manipulative therapy (SMT) for people with chronic obstructive pulmonary disease (COPD) who were older than 65 years. The study design was a prospective case series. Six patients of a long-term care center who were older than 65 years and having COPD underwent a course of 12 SMT sessions over a 4-week period. Each SMT session consisted of manually applied spinal manipulation and instrument-assisted spinal manipulation delivered by a doctor of chiropractic. Lung function measurements were recorded at baseline and at 2 and 4 weeks. The occurrence and type of any adverse events (AEs) related to SMT were recorded at each SMT session. One male and 5 female patients took part in the study. The average age was 79.1 years (range, 68-89 years). There was a clinically significant increase in forced expiratory volume in the first second after SMT in 4 of the 6 patients at 2 weeks. This was sustained in only 1 patient at 4 weeks. No clinically significant changes were observed for forced vital capacity at 2 or 4 weeks. One hundred forty-four manually applied spinal manipulations and 72 instrument-assisted spinal manipulations were administered during the intervention period. No major or moderate AEs were reported. Only minor AEs were reported after 29% of the intervention sessions, with 1 AE being reported for each patient. All AEs resolved within 48 hours. This case series offers preliminary evidence that SMT may have the potential to benefit lung function in patients with COPD who are older than 65 years.


A solid-phase enzyme-linked immunospot (ELISPOT) assay for enumeration of specific antibody-secreting cells.
A solid-phase enzyme-linked immunosorbent assay (ELISPOT) is described for enumeration of cells secreting specific antibody. Spleen cells from immunized mice are incubated in antigen-coated polystyrene plates. After removal of the cells, bound antibodies are demonstrated by means of an immunoenzyme procedure in which enzyme-substrate reactions are performed in agarose. Dark-brown circular zones (spots), localized in areas of the dish where antibody production has occurred, are enumerated with the naked eye. Spectrophotometric estimation of enzyme-bound activity may be performed by substituting the gel for a liquid buffer, allowing accurate estimation of the total amount of secreted antibody. Versatile, sensitive and very easy to perform, this new assay provides a useful alternative to conventional plaque-forming cell assays.


Online information needs of cancer patients and their organizations.
Increasingly patients, relatives and carers are accessing health information via the internet. However, the health profession and people affected by cancer are becoming concerned with the quality of that information. A European survey was conducted under the auspices of the FP7 European Commission funded Eurocancercoms project during the period September 2010-March 2011. Its aim was to assess current online information needs of people with cancer particularly those who seek information using online social media technologies and the internet more broadly. A literature review was undertaken to gain a greater understanding of health seeking behaviour regarding cancer patients' information needs and patient preferences for accessing different formats and media. This was used to inform the design and validation of online pan-European, multi-lingual questionnaires distributed via patient organizations and via specific Eurocancercoms partner organizations. This paper presents the results of this survey and suggests recommendations to be incorporated into the design of the online platform, ecancerHub, one of the intended outcomes of the Eurocancercoms project following this research. People want a wide variety of easy to find, easy to understand accurate information about cancer and how it is likely to impact on their everyday lives and on those close to them. They differ in the amount and detail of the information they would like and on their ability to identify quality information and understand it sufficiently to base their health-care decisions on. The majority of respondents raised the issue of quality of information and many requested recommendations of websites by the people who usually influence them most, the health professionals involved in their care.


Microcystins affect zooplankton biodiversity in oxbow lakes.
The authors tested the hypothesis that zooplankton diversity and density are affected by the presence of cyanotoxins in the water. The authors focused on 4 oxbow lakes of the Vistula River in southern Poland, which are subjected to mass cyanobacterial development. In 2 of the oxbows (Piekary and Tyniec), microcystins released into the water were found. The highest concentration of microcystins (0.246 μg/L) was observed for microcystins LR. Zooplankton diversity showed a weak response to the presence of microcystins released into the water. The Shannon index (H') of total zooplankton diversity decreased in the Piekary and Tyniec oxbows during periods when the microcystin concentrations were highest. The same trend was noted for diversity of rotifers in both oxbows and for diversity of copepods in Piekary, but not for copepods in Tyniec. No such trends were found for the diversity of cladocerans in any of the oxbows, nor was a relationship found between density of zooplankton and microcystins. Statistical analyses showed that the number of species in individual samples was negatively correlated with the levels of sulfates, phosphates, and ammonia, but the microcystin concentration was positively related to those levels. This points to the complexity of the interactions and synergies among toxins, abiotic factors, and zooplankton biodiversity. In focusing on the problem of cyanotoxins, conservation studies should pay attention to this complexity. Environ Toxicol Chem 2017;36:165-174. © 2016 SETAC.


Simple fabrication of diffraction gratings by two-beam interference method in highly photosensitive hybrid sol-gel films.
Sol-gel hybrid materials containing a large quantity of photoactive molecules exhibited large changes in both refractive index and volume by UV exposure. The materials were used for the fabrication of diffraction gratings using the two-beam interference method. With this technique, we could simply fabricate the diffraction gratings and easily control the grating periods. The diffraction effects and efficiencies of gratings rely heavily on the UV doses and the fabricated diffraction gratings showed a good diffraction performance.


Public Health Laboratory Service IgM antibody capture enzyme linked immunosorbent assay for detecting rubella specific IgM.
A total of 468 sera were selected for the evaluation of the Public Health Laboratory Service's IgM antibody capture enzyme linked immunosorbent assay kit (MACELISA) for detecting rubella specific IgM. The results obtained were compared with those obtained by IgM antibody capture radioimmunoassay (MACRIA). Sera from patients with primary postnatal rubella, congenital rubella, remote rubella, infectious mononucleosis, and recent infection with other agents were included, in addition to sera taken after rubella immunisation and sera containing rheumatoid factor and rubella specific IgG antibody. The assay exhibited a similar ability and comparable specificity to MACRIA for detecting rubella specific IgM antibody. The Public Health Laboratory Service MACELISA can be recommended if, as for all assays that detect rubella specific IgM, all the available clinical and serological data are taken into account when the results are interpreted.


Variables that affect the process and outcome of feedback, relevant for medical training: a meta-review.
Feedback is considered important in medical education. The literature is not clear about the mechanisms that contribute to its effects, which are often small to moderate and at times contradictory. A variety of variables seem to influence the impact of feedback on learning. The aim of this study was to determine which variables influence the process and outcomes of feedback in settings relevant to medical education. A myriad of studies on feedback have been conducted. To determine the most researched variables, we limited our review to meta-analyses and literature reviews published in the period from January 1986 to February 2012. According to our protocol, we first identified features of the feedback process that influence its effects and subsequently variables that influence these features. We used a chronological model of the feedback process to categorise all variables found. A systematic search of ERIC, PsycINFO and MEDLINE yielded 1101 publications, which we reduced to 203, rejecting papers on six exclusion criteria. Of these, 46 met the inclusion criteria. In our four-phase model, we identified 33 variables linked to task performance (e.g. task complexity, task nature) and feedback reception (e.g. self-esteem, goal-setting behaviour) by trainees, and to observation (e.g. focus, intensity) and feedback provision (e.g. form, content) by supervisors that influence the subsequent effects of the feedback process. Variables from all phases influence the feedback process and effects, but variables that influence the quality of the observation and rating of the performance dominate the literature. There is a paucity of studies addressing other, seemingly relevant variables. The larger picture of variables that influence the process and outcome of feedback, relevant for medical education, shows many open spaces. We suggest that targeted studies be carried out to expand our knowledge of these important aspects of feedback in medical education.


Multiple Roles of Ret Signalling During Enteric Neurogenesis.
The majority of the enteric nervous system is formed by vagal neural crest cells which enter the foregut and migrate rostrocaudally to colonise the entire length of the gastrointestinal tract. Absence of enteric ganglia from the distal colon are the hallmark of Hirschsprung disease, a congenital disorder characterised by severe intestinal dysmotility. Mutations in the receptor tyrosine kinase RET have been identified in approximately 50% of familial cases of Hirschsprung disease but the cellular processes misregulated in this condition remain unclear. By lineage tracing neural crest cells in mice homozygous for a knock-in allele of Ret (Ret51/51), we demonstrate that normal activity of this receptor is required in vivo for the migration of enteric nervous system progenitors throughout the gut. In mutant mice, progenitors of enteric neurons fail to colonise the distal colon, indicating that failure of colonisation of the distal intestine is a major contributing factor for the pathogenesis of Hirschsprung disease. Enteric nervous system progenitors in the ganglionic proximal guts of mutant mice are also characterised by reduced proliferation and differentiation. These findings suggest that the functional abnormalities in Hirschsprung disease result from a combination of colonic aganglionosis and deficits in neuronal circuitry of more proximal gut segments. The reduced neurogenesis in the gut of Ret51/51 mutants was reproduced in the multilineage enteric nervous system progenitors isolated from these animals. Correction of the molecular defects of such progenitors fully restored their neurogenic potential in culture. These observations enhance our understanding of the pathogenesis of Hirschsprung disease and highlight potential approaches for its treatment.


Tying and Tearing in Robotic and Laparoscopic Intracorporeally Hand-Sewn Ileocolic Anastomoses. A Propensity Score-Matched Prospective Study.
The aim of this study was to evaluate the impact of the ergonomics of laparoscopy as well as the lack of tactile feedback in robotic surgery on intracorporeal suturing. This was a prospective cohort study that compared the first 12 consecutive robotic hand-sewn ileocolic anastomoses to matched laparoscopic cases performed by the same surgeon. The endpoints were interrupted suturing and tying time, and the numbers of interrupted sutures placed and torn. Propensity score matching was based on age, body mass index, previous abdominal surgery, and diagnosis. Ileocolic anastomoses were hand-sewn in two layers: running 3-0 polyglycolic acid and interrupted 3-0 silk. Continuous variables were compared using the Student t test, whereas a Chi-square test was used to compare categorical variables. Linear regression and a cumulative sum analysis (CUSUM) were used for quality control. Median time for robotic suturing and tying was significantly shorter than that in matched laparoscopic cases (89.5 sec (IQR 76-103.5) vs. 160 sec (IQR 146-210), p<0.0001). There were fewer torn sutures in laparoscopic suturing (1 (IQR 1-2.5) vs. 0 (IQR 0-0.5), p<0.0026). No differences were found in complication rates (6 vs. 2 p=0.19). CUSUM analysis did not show a significant improvement in the number of torn sutures over time. Although there were more torn sutures with robotic suturing and tying, significantly less time was required than for laparoscopic intracorporeal suturing. This study suggests that torn sutures may be the result of a lack of tactile feedback rather than the presence of a learning curve.


Nonclassical mechanisms of progesterone action in the brain: I. Protein kinase C activation in the hypothalamus of female rats.
The modulation of gene regulation by progesterone (P) and its classical intracellular regulation by progestin receptors in the brain, resulting in alterations in physiology and behavior has been well studied. The mechanisms mediating the short latency effects of P are less well understood. Recent studies have revealed rapid nonclassical signaling action of P involving the activation of intracellular signaling pathways. We explored the involvement of protein kinase C (PKC) in P-induced rapid signaling in the ventromedial nucleus of the hypothalamus (VMN) and preoptic area (POA) of the rat brain. Both the Ca2+-independent (basal) PKC activity representing the activation of PKC by the in vivo treatments and the Ca+2-dependent (total) PKC activity assayed in the presence of exogenous cofactors in vitro were determined. A comparison of the two activities demonstrated the strength and temporal status of PKC regulation by steroid hormones in vivo. P treatment resulted in a rapid increase in basal PKC activity in the VMN but not the POA. Estradiol benzoate priming augmented P-initiated increase in PKC basal activity in both the VMN and POA. These increases were inhibited by intracerebroventricular administration of a PKC inhibitor administered 30 min prior to P. The total PKC activity remained unchanged demonstrating maximal PKC activation within 30 min in the VMN. In contrast, P regulation in the POA significantly attenuated total PKC activity +/- estradiol benzoate priming. These rapid changes in P-initiated PKC activity were not due to changes in PKC protein levels or phosphorylation status.


Cross-resistance between taxanes and new hormonal agents abiraterone and enzalutamide may affect drug sequence choices in metastatic castration-resistant prostate cancer.
Treatment options for patients with metastatic castration-resistant prostate cancer (mCRPC) have expanded in recent years with the introduction of cabazitaxel, abiraterone and enzalutamide. With new systemic therapies available, the optimal treatment sequence of these drugs in mCRPC becomes increasingly important. As shown recently, patients who had previously been treated with abiraterone showed impaired responses to docetaxel, suggesting clinical cross-resistance [1]. In the present study, we aimed to identify cross-resistance between taxanes (docetaxel and cabazitaxel) and the new hormonal agents abiraterone and enzalutamide. As a potential mechanism for cross-resistance, we investigated the effects on androgen receptor (AR) nuclear translocation of these compounds. To identify cross-resistance, we determined the effects of docetaxel, cabazitaxel, abiraterone and enzalutamide on cell viability in prostate cancer cell lines with acquired resistance to abiraterone and enzalutamide. Time-lapse confocal microscopy was used to study the dynamics of AR nuclear translocation. We observed impaired efficacy of docetaxel, cabazitaxel and enzalutamide in the abiraterone-resistant cell line, compared to the non-resistant cell line, providing evidence for in vitro cross-resistance. Impaired efficacy of docetaxel, cabazitaxel and abiraterone was observed in the enzalutamide-resistant cell line. Furthermore, docetaxel and cabazitaxel inhibited AR nuclear translocation, which was also observed for abiraterone and enzalutamide. In conclusion we found substantial preclinical evidence for cross-resistance between the taxanes docetaxel and cabazitaxel, and AR targeting agents abiraterone and enzalutamide. Since these compounds all interfere with AR-signalling, this strongly suggests a common mechanism of action, and thus a potential mechanism for cross-resistance in mCRPC.


Topotecan as salvage therapy for relapsed or refractory primary central nervous system lymphoma.
Treatment for patients with refractory or relapsed primary CNS lymphoma (PCNSL) remains unsatisfactory. Topotecan is an intravenous topoisomerase I inhibitor with good CSF penetration and documented efficacy in patients with relapsed systemic non-Hodgkin's lymphoma. In this study 15 patients with refractory or relapsed PCNSL were treated with intravenous topotecan (1.5 mg/m(2)) for five consecutive days during each 21-day cycle. All 15 patients had measurable, contrast-enhancing tumor on cranial MRI at the time of relapse. Three (20%) patients achieved a complete response after one, three and four cycles, respectively, while three (20%) patients achieved a partial response after two cycles each, for a total response proportion of 40%. Three patients had stable disease at the end of topotecan treatment. Six patients (40%) had progressive disease during treatment. Median overall survival was 981 days (95% CI: 275, NA) and median progression free survival was 60 days (95% CI: 46, 945). Three out of 15 patients had grade 3 thrombocytopenia. Six out of 15 patients had grade 3 neutropenia, while 5/15 patients had grade 4 neutropenia, and 13/15 patients received g-CSF at some point during treatment. There were no deaths directly related to treatment toxicity. Our study shows that topotecan, as a salvage therapy in patients with relapsed or refractory PCNSL, is associated with an overall response proportion of 40% and should be considered in patients who have failed prior methotrexate-based chemotherapy and/or whole brain irradiation. However, progression is frequent and early and most patients required growth factor support due to myelotoxicity.


Morale, stress and coping strategies of staff working in the emergency department: A comparison of two different-sized departments.
Clinical staff in EDs are subject to a range of stressors. The objective of this study was to describe and compare clinical staff perceptions of their ED's working environment across two different Australian EDs. This was a cross-sectional, descriptive, research design that included distribution of three survey tools to clinical staff in two Australian EDs in 2016. Descriptive statistics were reported to characterise workplace stressors, coping styles and the ED environment. These data were compared by hospital and the employee's clinical role (nurse or physician). In total, 146 ED nurses and doctors completed the survey (response rate: 67%). Despite geographical variation, the staff at the two locations had similar demographic profiles in terms of age, sex and years of experience. Staff reported moderate levels of workload and self-realisation but low levels of conflict or nervousness in the workplace. Nurses and physicians reported similar perceptions of the work environment, although nurses reported slightly higher median levels of workload. Staff rated the death or sexual abuse of a child as most stressful, followed by workplace violence and heavy workload. Staff used a large range of coping strategies, and these were similar across both sites. These findings are the first multi-site and multidisciplinary examinations of Australian ED staff perceptions, improving our understanding of staff stressors and coping strategies and highlighting similarities across different EDs. These data support the development and implementation of strategies to improve ED working environments to help ensure professional longevity of ED staff.


TBC1D20 is a Rab1 GTPase-activating protein that mediates hepatitis C virus replication.
Like other viruses, productive hepatitis C virus (HCV) infection depends on certain critical host factors. We have recently shown that an interaction between HCV nonstructural protein NS5A and a host protein, TBC1D20, is necessary for efficient HCV replication. TBC1D20 contains a TBC (Tre-2, Bub2, and Cdc16) domain present in most known Rab GTPase-activating proteins (GAPs). The latter are master regulators of vesicular membrane transport, as they control the activity of membrane-associated Rab proteins. To better understand the role of the NS5A-TBC1D20 interaction in the HCV life cycle, we used a biochemical screen to identify the TBC1D20 Rab substrate. TBC1D20 was found to be the first known GAP for Rab1, which is implicated in the regulation of anterograde traffic between the endoplasmic reticulum and the Golgi complex. Mutation of amino acids implicated in Rab GTPase activation by other TBC domain-containing GAPs abrogated the ability of TBC1D20 to activate Rab1 GTPase. Overexpression of TBC1D20 blocked the transport of exogenous vesicular stomatitis virus G protein from the endoplasmic reticulum, validating the involvement of TBC1D20 in this pathway. Rab1 depletion significantly decreased HCV RNA levels, suggesting a role for Rab1 in HCV replication. These results highlight a novel mechanism by which viruses can hijack host cell machinery and suggest an attractive model whereby the NS5A-TBC1D20 interaction may promote viral membrane-associated RNA replication.


A randomized cross-over study of inhalation of diesel exhaust, hematological indices, and endothelial markers in humans.
Exposure to traffic-related air pollution (TRAP) is considered a trigger for acute cardiovascular events. Diesel Exhaust (DE) is a major contributor to TRAP in the world. We evaluated the effect of DE inhalation on circulating blood cell populations, hematological indices, and systemic inflammatory cytokines in humans using a specialized facility. In a randomized double-blind crossover study balanced to order, 17 metabolic syndrome (MetS) and 15 healthy subjects inhaled filtered air (FA) or DE exposure in two-hour sessions on different days with a minimum 2-week washout period. We collected blood pre-exposure, 7, and 22 hours after exposure initiation and measured the complete blood count and differential. We performed multiplex cytokine assay to measure the changes in the systemic inflammatory cytokines, and endothelial adhesion molecules (n=15). A paired analysis compared the effect of DE and FA exposures for the change from pre-exposure to the subsequent time points. A significant increase in the hematocrit was noted 7 hrs after DE [1.4% (95% CI: 0.9 to 1.9%)] compared to FA exposure [0.5% (95% CI: -0.09 to 1.0%); p=0.008. The hemoglobin levels increased non-significantly at 7 hrs post DE [0.3 gm/dL (95% CI: 0.2 to 0.5 gm/dL)] versus FA exposure [0.2 gm/dL (95% CI: 0 to 0.3 gm/dL)]; p=0.06. Furthermore, the platelet count increased 22 hrs after DE exposure in healthy, but not in MetS subjects [DE: 16.6 (95% CI: 10.2 to 23) thousand platelets/mL versus [FA: 3.4 (95% CI: -9.5 to 16.3) thousand platelets/mL)]; p=0.04. No DE effect was observed for WBC, neutrophils, lymphocytes or erythrocytes. Using the multiplex assay, small borderline significant increases in matrix metalloproteinase-9, interleukins (IL)-1 beta, 6 and 10 occurred 7 hrs post exposure initiation, whereas E-selectin, intercellular adhesion molecule-1, and vascular cell adhesion molecule -1, and myeloperoxidase 22 hrs post exposure. Our results suggest that short-term DE exposure results in hemoconcentration and thrombocytosis, which are important determinants of acute cardiovascular events. Multiplex assay showed a non-significant increase in IL-1β and IL-6 immediately post exposure followed by myeloperoxidase and endothelial activation molecules. Further specific assays in a larger population will improve our understanding of the systemic inflammatory mechanisms following acute exposure to TRAP.


Infrared behavior and fixed points in Landau-gauge QCD.
We investigate the infrared behavior of gluon and ghost propagators in Landau-gauge QCD by means of an exact renormalization group equation. We explain how, in general, the infrared momentum structure of Green functions can be extracted within this approach. An optimization procedure is devised to remove residual regulator dependences. In Landau-gauge QCD this framework is used to determine the infrared leading terms of the propagators. The results support the Kugo-Ojima confinement scenario. Possible extensions are discussed.


Mobile phone radiation inhibits Vigna radiata (mung bean) root growth by inducing oxidative stress.
During the last couple of decades, there has been a tremendous increase in the use of cell phones. It has significantly added to the rapidly increasing EMF smog, an unprecedented type of pollution consisting of radiation in the environment, thereby prompting the scientists to study the effects on humans. However, not many studies have been conducted to explore the effects of cell phone EMFr on growth and biochemical changes in plants. We investigated whether EMFr from cell phones inhibit growth of Vigna radiata (mung bean) through induction of conventional stress responses. Effects of cell phone EMFr (power density: 8.55 microW cm(-2); 900 MHz band width; for 1/2, 1, 2, and 4 h) were determined by measuring the generation of reactive oxygen species (ROS) in terms of malondialdehyde and hydrogen peroxide (H(2)O(2)) content, root oxidizability and changes in levels of antioxidant enzymes. Our results showed that cell phone EMFr significantly inhibited the germination (at > or =2 h), and radicle and plumule growths (> or =1 h) in mung bean in a time-dependent manner. Further, cell phone EMFr enhanced MDA content (indicating lipid peroxidation), and increased H(2)O(2) accumulation and root oxidizability in mung bean roots, thereby inducing oxidative stress and cellular damage. In response to EMFr, there was a significant upregulation in the activities of scavenging enzymes, such as superoxide dismutases, ascorbate peroxidases, guaiacol peroxidases, catalases and glutathione reductases, in mung bean roots. The study concluded that cell phone EMFr inhibit root growth of mung bean by inducing ROS-generated oxidative stress despite increased activities of antioxidant enzymes.


Exogenous BMP7 in aortae of rats with chronic uremia ameliorates expression of profibrotic genes, but does not reverse established vascular calcification.
Hyperphosphatemia and vascular calcification are frequent complications of chronic renal failure and bone morphogenetic protein 7 (BMP7) has been shown to protect against development of vascular calcification in uremia. The present investigation examined the potential reversibility of established uremic vascular calcification by treatment of uremic rats with BMP7. A control model of isogenic transplantation of a calcified aorta from uremic rats into healthy littermates examined whether normalization of the uremic environment reversed vascular calcification. Uremia and vascular calcification were induced in rats by 5/6 nephrectomy, high phosphate diet and alfacalcidol treatment. After 14 weeks severe vascular calcification was present and rats were allocated to BMP7, vehicle or aorta transplantation. BMP7 treatment caused a significant decrease of plasma phosphate to 1.56 ± 0.17 mmol/L vs 2.06 ± 0.34 mmol/L in the vehicle group even in the setting of uremia and high phosphate diet. Uremia and alfacalcidol resulted in an increase in aortic expression of genes related to fibrosis, osteogenic transformation and extracellular matrix calcification, and the BMP7 treatment resulted in a decrease in the expression of profibrotic genes. The total Ca-content of the aorta was however unchanged both in the abdominal aorta: 1.9 ± 0.6 μg/mg tissue in the vehicle group vs 2.2 ± 0.6 μg/mg tissue in the BMP7 group and in the thoracic aorta: 71 ± 27 μg/mg tissue in the vehicle group vs 54 ± 18 μg/mg tissue in the BMP7 group. Likewise, normalization of the uremic environment by aorta transplantation had no effect on the Ca-content of the calcified aorta: 16.3 ± 0.6 μg/mg tissue pre-transplantation vs 15.9 ± 2.3 μg/mg tissue post-transplantation. Aortic expression of genes directly linked to extracellular matrix calcification was not affected by BMP7 treatment, which hypothetically might explain persistent high Ca-content in established vascular calcification. The present results highlight the importance of preventing the development of vascular calcification in chronic kidney disease. Once established, vascular calcification persists even in the setting when hyperphosphatemia or the uremic milieu is abolished.


Monitoring the immune competence of cancer patients to predict outcome.
A new era of cancer immunotherapy has brought not only successful cancer vaccines but also immunomodulators, such as those that target checkpoint blockade in order to induce endogenous host immune responses. However, the immune system of cancer patients can be compromised through multiple means, including immune suppression by the tumor and by prior therapies such as chemotherapy and radiation. Therefore, a comprehensive means of assessing patient immunocompetence would seem helpful for determining whether patients are ready to benefit from immunotherapy, and perhaps even which immunotherapy might be most appropriate for them. Unfortunately, there are no standardized tests for immune competence, nor is there agreement on what to measure and what will be predictive of outcome. In this review, we will discuss the technologies and assays that might be most useful for this purpose. We argue for a comprehensive approach that should maximize the chances of developing predictive biomarkers for eventual clinical use.


Power spectral analysis of heart rate variation in diabetic patients with neuropathic foot ulceration.
To evaluate the relationship between diabetic autonomic neuropathy and diabetic neuropathic foot ulceration, we used power spectral analysis (PSA) of heart rate variation, which provides the accurate simultaneous quantification of parasympathetic and sympathetic activities, to assess autonomic function in diabetic patients. We studied 55 NIDDM patients including 10 diabetic patients without neuropathy, 23 diabetic patients with neuropathy and no history of foot ulceration, and 22 diabetic patients with neuropathic foot ulceration. We performed PSA of 100 R-R intervals at rest and analyzed the results by fast Fourier transformation. The low frequency (LF) power, which reflects sympathetic activity, and the high frequency (HF) power, which reflects parasympathetic (vagal) activity, were inversely correlated with the duration of diabetes and the fasting plasma glucose (FPG) levels. By multiple regression analysis, the FPG remained with significant influence on both LF and HF powers. The LF and HF powers were positively correlated with motor nerve conduction velocity (MCV) and sensory nerve conduction velocity (SCV) in the upper and lower limbs and the coefficient of variation of R-R intervals. The LF and HF powers were significantly reduced in patients with neuropathy and patients with foot ulceration compared with patients without neuropathy. Although the median MCV and SCV were similar between diabetic patients with neuropathy and patients with foot ulceration, both the LF and HF powers were significantly decreased in patients with foot ulceration compared with patients with neuropathy. There was no difference in the value of the LF:HF ratio, an index of sympathovagal balance, among three subgroups. We observed a positive correlation between LF and HF power in all subjects; however, the LF and HF powers were not correlated in the subgroups of patients with foot ulceration. These results showed that diabetic patients with neuropathic foot ulceration have a greater impairment in spectral indexes of autonomic activity obtained by PSA than patients with neuropathy and no history of foot ulceration, whereas no difference was present in nerve conduction velocities.


Sequence and expression analysis of the AMT gene family in poplar.
Ammonium transporters (AMTs) are plasma membrane proteins that exclusively transport ammonium/ammonia. These proteins are encoded by an ancient gene family with many members. The molecular characteristics and evolutionary history of AMTs in woody plants are still poorly understood. We comprehensively evaluated the AMT gene family in the latest release of the Populus trichocarpa genome (version 3.0; Phytozome 9.0), and identified 16 AMT genes. These genes formed four clusters; AMT1 (7 genes), AMT2 (2 genes), AMT3 (2 genes), and AMT4 (5 genes). Evolutionary analyses suggested that the Populus AMT gene family has expanded via whole-genome duplication events. Among the 16 AMT genes, 15 genes are located on 11 chromosomes of Populus. Expression analyses showed that 14 AMT genes were vegetative organs expressed; AMT1;1/1;3/1;6/3;2 and AMT1;1/1;2/2;2/3;1 had high transcript accumulation level in the leaves and roots, respectively and strongly changes under the nitrogen-dependent experiments. The results imply the functional roles of AMT genes in ammonium absorption in poplar.


Shade signals alter the expression of circadian clock genes in newly-formed bioenergy sorghum internodes.
Stem internodes of bioenergy sorghum inbred R.07020 are longer at high plant density (shade) than at low plant density (control). Initially, the youngest newly-formed subapical stem internodes of shade-treated and control plants are comparable in length. However, full-length internodes of shade-treated plants are three times longer than the internodes of the control plants. To identify the early molecular events associated with internode elongation in response to shade, we analyzed the transcriptome of the newly-formed internodes of shade-treated and control plants sampled between 4 and 6 hr after the start of the light period (14 hr light/10 hr dark). Sorghum genes homologous to the Arabidopsis shade marker genes ATHB2 and PIL1 were not differentially expressed. The results indicate that shade signals promote internode elongation indirectly because sorghum internodes are not illuminated and grow while enclosed with leaf sheaths. Sorghum genes homologous to the Arabidopsis morning-phased circadian clock genes LHY, RVE, and LNK were downregulated and evening-phased genes such as TOC1, PRR5, and GI were upregulated in young internodes in response to shade. We hypothesize that a change in the function or patterns of expression of the circadian clock genes is the earliest molecular event associated with internode elongation in response to shade in bioenergy sorghum. Increased expression of CycD1, which promotes cell division, and decreased expression of cell wall-loosening and MBF1-like genes, which promote cell expansion, suggest that shade signals promote internode elongation in bioenergy sorghum in part through increasing cell number by delaying transition from cell division to cell expansion.


Prebiotic RNA polymerisation: energetics of nucleotide adsorption and polymerisation on clay mineral surfaces.
We measured the binding energy and bonding parameters between model nucleotide functional groups and model clay mineral surfaces in solutions of acidic pH. We demonstrate that basal surfaces of clay minerals interact most strongly with nucleobases and show that the adsorption of the phosphate group to clay edges could facilitate polymerisation. Our results suggest that Al- and Fe-rich edge sites behave similarly in nucleotide polymerisation through change of the phosphodiester bond strength. We present an internally consistent set of thermodynamic parameters that represent the nucleotide-clay mineral system.


Improved treatment outcomes with (18) F-FDG PET/CT for patients with advanced head and neck squamous cell carcinoma.
The aim of this study was to assess the impact of (18) F-fluoro-2-deoxy-D-glucose ((18) F-FDG) PET/CT on survival for patients with head and neck squamous cell carcinoma correlated with a matched patient cohort. In all, disease in 58 patients was initially staged using (18) F-FDG PET/CT. A case-control analysis was done with 63 patients who disease was staged without (18) F-FDG-PET/CT. Disease-specific survival (DSS) and overall survival (OS) did not show significant differences between both groups. Statistical analysis revealed no difference in DSS and OS between the 2 groups for patients treated by radiochemotherapy (p = .975 and p = .671). In the analysis of survival in patients treated by a combined approach (surgery + radiochemotherapy), a significant difference in favor of patients evaluated by (18) F-FDG PET/CT was found (p = .05 and p = .027). Addition of (18) F-FDG PET/CT in patients treated by surgery and conformal radiochemotherapy improves outcome. This may be due to the more comprehensive topographic orientation of the primary tumor for the surgeon.


Structural characterization of HIV reverse transcriptase: a target for the design of specific virus inhibitors.
The reverse transcriptase (RT) of HIV is an important target for chemotherapy as demonstrated by the effective treatment of AIDS patients with zidovudine, a potent inhibitor of RT. Structural studies of HIV RT were therefore undertaken with a view to designing more effective inhibitors. To obtain sufficient quantities of enzyme for these studies the reverse transcriptase gene of HIV was cloned into a high level expression plasmid yielding reverse transcriptase at a level of 10% of the total Escherichia coli proteins. Monoclonal antibodies to RT were raised in mice and have been used to purify the enzyme by immunoaffinity chromatography. Crystallization of the enzyme has been achieved and studies are underway to determine its three-dimensional structure. In addition, carboxy-terminal truncated mutants were prepared by inserting stop codons into the gene at appropriate sites. The proteins expressed were analysed for RT and RNase H activity and used for mapping RT epitopes. This, together with previous data on site-directed mutagenesis of conserved regions of HIV RT has helped to map some of the structural and functional regions of the enzyme.


Rates and predictors of four-year readmission among late-middle-aged and older substance abuse patients.
This naturalistic study examines treatment, diagnoses and readmission among late-middle-aged and older (age 55+) substance abuse inpatients (N = 16,066) in Department of Veterans Affairs Medical Centers. Over an interval extending from 4 years before to 4 years after an index episode of care, older substance abuse patients used substantial amounts of inpatient and outpatient treatment. Service use was heaviest among patients with a concomitant psychiatric disorder, and there was no evidence that it declined over time in the overall group. Four-year readmission rates in three diagnostic subgroups (alcohol or drug dependence diagnosis only, alcohol or drug psychoses, substance dependence and/or psychoses with one or more psychiatric diagnoses) were very high (57% to 70%); however, they were somewhat lower among patients with less chronic substance abuse problems. Readmission and multiple readmission were predicted by younger age, unmarried status, more prior service use, alcohol psychosis or psychiatric diagnoses, treatment in a psychiatric unit, and shorter hospital stay. These factors may be used at admission and discharge to identify patients at risk for rehospitalization; inpatient treatment and aftercare can be planned accordingly.


Associations between serum 25(OH)D concentrations and prevalent asthma among children living in communities with differing levels of urbanization: a cross-sectional study.
Prior evidence suggests that vitamin D deficiency may increase the risk of asthma and atopy and impair pulmonary function in children. In this cross-sectional analysis nested in a case-control study, we analyzed serum 25(OH)D concentrations in 413 children with asthma and 471 children without asthma living in two geographically adjacent study communities (Pampas and Villa El Salvador). We measured total and antigen-specific IgE levels, pulmonary function, asthma control, and exhaled nitric oxide. Mean 25(OH)D concentrations were 25.2 ng/mL (SD 10.1) in children with asthma and 26.1 ng/mL (SD 13.7) in children without asthma (p = 0.28). Vitamin D deficiency (25(OH)D < 20 ng/ml) was more common in Pampas than in Villa El Salvador (52.7% vs. 10.5%; p < 0.001). In the overall study population, a 10 ng/ml decrease in serum 25(OH)D concentrations was not significantly associated with odds of asthma (OR 1.09, 95% CI: 0.94 to 1.25). However, vitamin D deficiency was associated with a 1.6-fold increase in odds of asthma in the overall cohort (95% CI: 1.14 to 2.25). After stratifying by site, a 10 ng/mL decrease in serum 25(OH)D concentrations was associated with 18% higher odds of having asthma in Pampas (OR = 1.18, 95% CI 1.02 to 1.38), whereas there was no significant association between 25(OH)D concentrations and asthma in Villa El Salvador (OR = 0.95, 95% CI 0.87 to 1.05). Combined data from these geographically adjacent populations suggests a possible threshold for the relationship between 25(OH)D levels and asthma at approximately 27.5 ng/ml. Serum 25(OH)D concentrations were not clearly associated with asthma control, total serum IgE, atopy, or airway inflammation. Serum 25(OH)D concentrations were inversely associated with asthma in one study community with a high prevalence of deficiency. Studies are needed to investigate a possible threshold 25(OH)D concentration after which higher vitamin D levels show no further benefit for asthma.


Analysis of costs borne by families of patients hospitalized for stroke.
Studies focusing on the economic impact of cancer on families have emphasized that costs of chronic disease are substantial for patients and their families. However, little effort has been devoted to measuring the costs of care for families of patients hospitalized with stroke. A total of 215 stroke patients and their families from four teaching hospitals in the Taipei metropolitan area were monitored from the date of the patient's admission to hospital until the date of discharge. The value of labor contributed by families was estimated by assigning the current monetary market rate of providing health aide to the time families spent caring for patients in hospital. Lost earnings of patients and families, expenditure for medical care, and expenses for food, clothes, adult diapers, transportation and other miscellaneous items were determined and summed to arrive at the total family cost of providing care. The average cost of care for one family per inpatient day was NT$4,358.20. A total of 98.6% of the families incurred labor costs, which accounted for about half of family costs for providing care. Hospital bills accounted for almost 19% of total family costs. The income loss for families and patients accounted for about 25% of total family costs. Expenses for food, clothes, transportation, diapers and other illness-related miscellaneous items accounted for about 12% of total family costs. Multiple regression analyses demonstrated that the number of family members involved in giving care and the length of stay are important predictors for the total cost of care. Average total family costs per day increased by 24.3% when an additional family member was involved in providing care. Total family costs increased 2.5% for each hospital day. If direct and indirect nonmedical costs are not included in the total cost calculation for providing hospital care to stroke patients, the economic impact of care on families is likely to be underestimated.


Hidden in plain sight: what remains to be discovered in the eukaryotic proteome?
The first decade of genome sequencing stimulated an explosion in the characterization of unknown proteins. More recently, the pace of functional discovery has slowed, leaving around 20% of the proteins even in well-studied model organisms without informative descriptions of their biological roles. Remarkably, many uncharacterized proteins are conserved from yeasts to human, suggesting that they contribute to fundamental biological processes (BP). To fully understand biological systems in health and disease, we need to account for every part of the system. Unstudied proteins thus represent a collective blind spot that limits the progress of both basic and applied biosciences. We use a simple yet powerful metric based on Gene Ontology BP terms to define characterized and uncharacterized proteins for human, budding yeast and fission yeast. We then identify a set of conserved but unstudied proteins in S. pombe, and classify them based on a combination of orthogonal attributes determined by large-scale experimental and comparative methods. Finally, we explore possible reasons why these proteins remain neglected, and propose courses of action to raise their profile and thereby reap the benefits of completing the catalogue of proteins' biological roles.


Gene signatures of progression and metastasis in renal cell cancer.
To address the progression, metastasis, and clinical heterogeneity of renal cell cancer (RCC). Transcriptional profiling with oligonucleotide microarrays (22,283 genes) was done on 49 RCC tumors, 20 non-RCC renal tumors, and 23 normal kidney samples. Samples were clustered based on gene expression profiles and specific gene sets for each renal tumor type were identified. Gene expression was correlated to disease progression and a metastasis gene signature was derived. Gene signatures were identified for each tumor type with 100% accuracy. Differentially expressed genes during early tumor formation and tumor progression to metastatic RCC were found. Subsets of these genes code for secreted proteins and membrane receptors and are both potential therapeutic or diagnostic targets. A gene pattern ("metastatic signature") derived from primary tumor was very accurate in classifying tumors with and without metastases at the time of surgery. A previously described "global" metastatic signature derived by another group from various non-RCC tumors was validated in RCC. Unlike previous studies, we describe highly accurate and externally validated gene signatures for RCC subtypes and other renal tumors. Interestingly, the gene expression of primary tumors provides us information about the metastatic status in the respective patients and has the potential, if prospectively validated, to enrich the armamentarium of diagnostic tests in RCC. We validated in RCC, for the first time, a previously described metastatic signature and further showed the feasibility of applying a gene signature across different microarray platforms. Transcriptional profiling allows a better appreciation of the molecular and clinical heterogeneity in RCC.