On the way to rotaxane-based molecular motors: studies in molecular mobility and topological chirality.
ATP synthase represents a machine at the molecular level which couples the rotation of an axle in a wheel with the endergonic production of ATP, the general source of chemical energy in the cell. The natural system prototypically bears all features of a macroscopic motor: a rotor within a stator held by a membrane and fueled by a difference in chemical potential in the form of a proton gradient combined with a machine for ATP production. The assembly of axle and wheel to a rotor device reminds one very much of a rotaxane. In this Account, we discuss some important features of motors and their (potential) realization in simpler artificial model systems, that is, the molecular mobility of mechanically bound molecules, the importance of chirality for unidirectional motion, the sources of energy for driving the rotation, and the potential of using membranes and surfaces for ordering a large number of devices to achieve macroscopic effects.


Giardia lamblia RNA polymerase II: amanitin-resistant transcription.
Giardia lamblia is an early branching eukaryote, and although distinctly eukaryotic in its cell and molecular biology, transcription and translation in G. lamblia demonstrate important differences from these processes in higher eukaryotes. The cyclic octapeptide amanitin is a relatively selective inhibitor of eukaryotic RNA polymerase II (RNAP II) and is commonly used to study RNAP II transcription. Therefore, we measured the sensitivity of G. lamblia RNAP II transcription to alpha-amanitin and found that unlike most other eukaryotes, RNAP II transcription in Giardia is resistant to 1 mg/ml amanitin. In contrast, 50 microg/ml amanitin inhibits 85% of RNAP III transcription activity using leucyl-tRNA as a template. To better understand transcription in G. lamblia, we identified 10 of the 12 known eukaryotic rpb subunits, including all 10 subunits that are required for viability in Saccharomyces cerevisiae. The amanitin motif (amanitin binding site) of Rpb1 from G. lamblia has amino acid substitutions at six highly conserved sites that have been associated with amanitin resistance in other organisms. These observations of amanitin resistance of Giardia RNA polymerase II support previous proposals of the mechanism of amanitin resistance in other organisms and provide a molecular framework for the development of novel drugs with selective activity against G. lamblia.


Inhibiting ex-vivo Th17 responses in Ankylosing Spondylitis by targeting Janus kinases.
Treatment options for Ankylosing Spondylitis (AS) are still limited. The T helper cell 17 (Th17) pathway has emerged as a major driver of disease pathogenesis and a good treatment target. Janus kinases (JAK) are key transducers of cytokine signals in Th17 cells and therefore promising targets for the treatment of AS. Here we investigate the therapeutic potential of four different JAK inhibitors on cells derived from AS patients and healthy controls, cultured in-vitro under Th17-promoting conditions. Levels of IL-17A, IL-17F, IL-22, GM-CSF and IFNγ were assessed by ELISA and inhibitory effects were investigated with Phosphoflow. JAK1/2/3 and TYK2 were silenced in CD4+ T cells with siRNA and effects analyzed by ELISA (IL-17A, IL-17F and IL-22), Western Blot, qPCR and Phosphoflow. In-vitro inhibition of CD4+ T lymphocyte production of multiple Th17 cytokines (IL-17A, IL-17F and IL-22) was achieved with JAK inhibitors of differing specificity, as well as by silencing of JAK1-3 and Tyk2, without impacting on cell viability or proliferation. Our preclinical data suggest JAK inhibitors as promising candidates for therapeutic trials in AS, since they can inhibit multiple Th17 cytokines simultaneously. Improved targeting of TYK2 or other JAK isoforms may confer tailored effects on Th17 responses in AS.


The effect of cryotherapy on intraarticular temperature and postoperative care after anterior cruciate ligament reconstruction.
The objective of this study was to elucidate how cryotherapy after anterior cruciate ligament reconstruction affects intraarticular temperature and clinical results. A prospective and randomized study was performed on 21 knees of 21 patients. The ligament reconstruction was performed by single-incision arthroscopy using autogenous hamstring tendon. On completion of the surgery, thermosensors were implanted in the suprapatellar pouch and the intracondylar notch, and the intraarticular temperature was monitored while the joint was cooled. Cooling was performed in one group at 5 degrees C (N = 7) and in another at 10 degrees C (N = 7), for 48 hours. A control group (N = 7) did not undergo cryotherapy. The cooled groups showed three temperature phases: a low-temperature phase immediately after the ligament reconstruction, followed by a temperature-rising phase and a thermostatic phase. The control group had no low-temperature phase and immediately entered a thermostatic phase. During the low-temperature phase in the treated groups, the temperature of the suprapatellar pouch and of the intercondylar notch were significantly lower than the body temperature. The pain score and the number of times an analgesic had to be administered were both significantly lower in the 10 degrees C group than in the control group. Blood loss was significantly less in the 5 degrees C group than in the control group.


Metabotropic purinergic receptors in lipid membrane microdomains.
There is broad evidence that association of transmembrane receptors and signalling molecules with lipid rafts/caveolae provides an enriched environment for protein-protein interactions necessary for signal transduction, and a mechanism for the modulation of neurotransmitter and/or growth factor receptor function. Several receptors translocate into submembrane compartments after ligand binding, while others move in the opposite direction. The role of such a dynamic localization and functional facilitation is signalling modulation and receptor desensitization or internalization. Purine and pyrimidine nucleotides have been viewed as primordial precursors in the evolution of all forms of intercellular communication, and they are now regarded as fundamental extracellular signalling molecules. They propagate the purinergic signalling by binding to ionotropic and metabotropic receptors expressed on the plasma membrane of almost all cell types, tissues and organs. Here, we have illustrated the localization in lipid rafts/caveolae of G protein-coupled P1 receptors for adenosine and P2Y receptors for nucleoside tri- and di-phosphates. We have highlighted that microdomain partitioning of these purinergic GPCRs is cell-specific, as is the overall expression levels of these same receptors. Moreover, we have described that disruption of submembrane compartments can shift the purinergic receptors from raft/caveolar to non-raft/non-caveolar fractions, and then abolish their ability to activate lipid signalling pathways and to integrate with additional lipid-controlled signalling events. This modulates the biological response to purinergic ligands and most of all indicates that the topology of the various purinergic components at the cell surface not only organizes the signal transduction machinery, but also controls the final cellular response.


Biology of VO2 max: looking under the physiology lamp.
In this review, we argue that several key features of maximal oxygen uptake (VO2 max) should underpin discussions about the biological and reductionist determinants of its interindividual variability: (i) training-induced increases in VO2 max are largely facilitated by expansion of red blood cell volume and an associated improvement in stroke volume, which also adapts independent of changes in red blood cell volume. These general concepts are also informed by cross-sectional studies in athletes that have very high values for VO2 max. Therefore, (ii) variations in VO2 max improvements with exercise training are also likely related to variations in these physiological determinants. (iii) All previously untrained individuals will respond to endurance exercise training in terms of improvements in VO2 max provided the stimulus exceeds a certain volume and/or intensity. Thus, genetic analysis and/or reductionist studies performed to understand or predict such variations might focus specifically on DNA variants or other molecular phenomena of relevance to these physiological pathways.


Silencing of aphid genes by feeding on stable transgenic Arabidopsis thaliana.
Aphids are economically important pests that predominantly feed from the plant phloem. Genome, transcriptome, and proteome data are being generated for these insects, and predicted secreted proteins in aphid saliva have been identified. These secreted proteins are candidate effectors that may modulate plant processes and aid aphid colonization of plants. The next step is to develop post-genomics strategies to study the functions of identified aphid genes. One such strategy is to express aphid effector genes in planta to assess whether aphid effectors alter plant development and aphid survival and fecundity. A second strategy is to knock down the expression of aphid target genes by RNA interference (RNAi). In this chapter, we describe how to knock down aphid gene expression using plant-mediated RNAi. This strategy is useful for assessing the contribution of aphid effectors to aphid colonization of plants.


Nucleotide sequence analysis and DNA hybridization studies of the ant(4')-IIa gene from Pseudomonas aeruginosa.
The ant(4')-IIa gene was previously cloned from Pseudomonas aeruginosa on a 1.6-kb DNA fragment (G. A. Jacoby, M. J. Blaser, P. Santanam, H. Hächler, F. H. Kayser, R. S. Hare, and G. H. Miller, Antimicrob. Agents Chemother. 34:2381-2386, 1990). In the current study, the ant(4')-IIa gene was localized by gamma-delta mutagenesis. A region of approximately 600 nucleotides which contained the ant(4')-IIa gene was identified, and DNA sequence analysis revealed two overlapping open reading frames (ORFs) within this region. Northern (RNA) blot analysis demonstrated expression of both ORFs in P. aeruginosa; therefore, site-directed mutagenesis was used to identify the ORF which encodes the ant(4')-IIa gene. No homology was found between ant(4')-IIa and ant(4')-Ia DNA sequences. Hybridization experiments confirmed that the ant(4')-Ia probe hybridized only to gram-positive presumptive ANT(4')-I strains and that the ant(4')-IIa probe hybridized only to gram-negative strains presumed to carry ANT(4')-II. Seven gram-negative strains which had been classified as having ANT(4')-II resistance profiles did not hybridize with probes for either ant(4')-Ia or ant(4')-IIa, suggesting that at least one additional ant(4') gene may exist. The predicted amino-terminal sequences of the ANT(4')-Ia and ANT(4')-IIa proteins showed significant sequence similarity between residues 38 and 63 of the ANT(4')-Ia protein and residues 26 and 51 of the ANT(4')-IIa protein.


Pseudo-random binary sequence phase modulation for narrow linewidth, kilowatt, monolithic fiber amplifiers.
We report on pseudo random binary sequence (PRBS) phase modulation for narrow-linewidth, kilowatt-class, monolithic (all-fiber) amplifiers. Stimulated Brillouin scattering (SBS) threshold enhancement factors for different patterns of PRBS modulated fiber amplifiers were experimentally analyzed and agreed well with the theoretical predictions. We also examined seeding of the SBS process by phase modulated signals when the effective linewidth is on the same order as the Brillouin shift frequency. Here ~30% variations in SBS power thresholds were observed from small tunings of the modulation frequency. In addition, a 3 GHz PRBS modulated, 1.17 kW fiber amplifier was demonstrated. Near diffraction-limited beam quality was achieved (M2 = 1.2) with an optical pump efficiency of 83%. Overall, the improved SBS suppression and narrow linewidth achieved through PRBS modulation can have a significant impact on the beam combining of kilowatt class fiber lasers.


[Childhood anemia in populations residing at different geographical altitudes of Arequipa, Peru: A descriptive and retrospective study].
To identify the characteristics of anemia in children living at different geographical altitude in the Arequipa Region of Peru. We did a descriptive and retrospective study. The population consisted of 106 499 children under five years of age living in the Arequipa Region at different geographical altitude and receiving care by the Ministry of Health of Arequipa. Of these, 32 454 had anemia (30.5%). Socio-demographic data related to age, province, and beneficiary of the Comprehensive Health System program, home visits, and growth and development were obtained, in addition to data regarding anemia such as frequency, hemoglobin, and severity. The results were categorized by the geographical altitude of origin of children under 0 to 59 months of age and hemoglobin levels according to geographical altitude. Of the sample studied, the frequency of anemia at different geographical altitudes of Arequipa in children under five for the 2017 to 2019 period was: 18.7% at 0 to 999 meters above sea level; 29.6% at 1000 to 1999 meters; 31.6% at 2000 to 2999 meters; 42.9% at 3000 to 3999 meters; and 54.4% at 4000 to 4999 meters. There was a higher prevalence of anemia when the geographic altitude correction factor was used. A significant statistical relationship (p < 0.05) was observed between the corrected hemoglobin according to the World Health Organization parameters and the altitude at which children with anemia live: the higher the geographical altitude, the greater the anemia in children. Supplementation programs should be included during home visits and at the time of evaluation of children's growth and development.


[Inhibitory mechanism of melanin synthesis by glutathione].
Glutathione dose dependently inhibited melanin synthesis in the reaction of tyrosinase and L-DOPA. The inhibition of melanin synthesis was recovered by increasing the concentration of L-DOPA, but not recovered by increasing tyrosinase. Glutathione inhibited the binding between tyrosinase and L-DOPA. Although the synthesized melanin was aggregated within 1 h, the aggregation was inhibited by the addition of glutathione. These results indicate that glutathione inhibits the synthesis and agglutination of melanin by interrupting the function of L-DOPA.


HOXB13 is a susceptibility gene for prostate cancer: results from the International Consortium for Prostate Cancer Genetics (ICPCG).
Prostate cancer has a strong familial component but uncovering the molecular basis for inherited susceptibility for this disease has been challenging. Recently, a rare, recurrent mutation (G84E) in HOXB13 was reported to be associated with prostate cancer risk. Confirmation and characterization of this finding is necessary to potentially translate this information to the clinic. To examine this finding in a large international sample of prostate cancer families, we genotyped this mutation and 14 other SNPs in or flanking HOXB13 in 2,443 prostate cancer families recruited by the International Consortium for Prostate Cancer Genetics (ICPCG). At least one mutation carrier was found in 112 prostate cancer families (4.6 %), all of European descent. Within carrier families, the G84E mutation was more common in men with a diagnosis of prostate cancer (194 of 382, 51 %) than those without (42 of 137, 30 %), P = 9.9 × 10(-8) [odds ratio 4.42 (95 % confidence interval 2.56-7.64)]. A family-based association test found G84E to be significantly over-transmitted from parents to affected offspring (P = 6.5 × 10(-6)). Analysis of markers flanking the G84E mutation indicates that it resides in the same haplotype in 95 % of carriers, consistent with a founder effect. Clinical characteristics of cancers in mutation carriers included features of high-risk disease. These findings demonstrate that the HOXB13 G84E mutation is present in ~5 % of prostate cancer families, predominantly of European descent, and confirm its association with prostate cancer risk. While future studies are needed to more fully define the clinical utility of this observation, this allele and others like it could form the basis for early, targeted screening of men at elevated risk for this common, clinically heterogeneous cancer.


Dynamical mechanism for sharp orientation tuning in an integrate-and-fire model of a cortical hypercolumn.
Orientation tuning in a ring of pulse-coupled integrate-and-fire (IF) neurons is analyzed in terms of spontaneous pattern formation. It is shown how the ring bifurcates from a synchronous state to a non-phase-locked state whose spike trains are characterized by clustered but irregular fluctuations of the interspike intervals (ISIs). The separation of these clusters in phase space results in a localized peak of activity as measured by the time-averaged firing rate of the neurons. This generates a sharp orientation tuning curve that can lock to a slowly rotating, weakly tuned external stimulus. Under certain conditions, the peak can slowly rotate even to a fixed external stimulus. The ring also exhibits hysteresis due to the subcritical nature of the bifurcation to sharp orientation tuning. Such behavior is shown to be consistent with a corresponding analog version of the IF model in the limit of slow synaptic interactions. For fast synapses, the deterministic fluctuations of the ISIs associated with the tuning curve can support a coefficient of variation of order unity.


Targeting BTK with ibrutinib in relapsed or refractory mantle-cell lymphoma.
Bruton's tyrosine kinase (BTK) is a mediator of the B-cell-receptor signaling pathway implicated in the pathogenesis of B-cell cancers. In a phase 1 study, ibrutinib, a BTK inhibitor, showed antitumor activity in several types of non-Hodgkin's lymphoma, including mantle-cell lymphoma. In this phase 2 study, we investigated oral ibrutinib, at a daily dose of 560 mg, in 111 patients with relapsed or refractory mantle-cell lymphoma. Patients were enrolled into two groups: those who had previously received at least 2 cycles of bortezomib therapy and those who had received less than 2 complete cycles of bortezomib or had received no prior bortezomib therapy. The primary end point was the overall response rate. Secondary end points were duration of response, progression-free survival, overall survival, and safety. The median age was 68 years, and 86% of patients had intermediate-risk or high-risk mantle-cell lymphoma according to clinical prognostic factors. Patients had received a median of three prior therapies. The most common treatment-related adverse events were mild or moderate diarrhea, fatigue, and nausea. Grade 3 or higher hematologic events were infrequent and included neutropenia (in 16% of patients), thrombocytopenia (in 11%), and anemia (in 10%). A response rate of 68% (75 patients) was observed, with a complete response rate of 21% and a partial response rate of 47%; prior treatment with bortezomib had no effect on the response rate. With an estimated median follow-up of 15.3 months, the estimated median response duration was 17.5 months (95% confidence interval [CI], 15.8 to not reached), the estimated median progression-free survival was 13.9 months (95% CI, 7.0 to not reached), and the median overall survival was not reached. The estimated rate of overall survival was 58% at 18 months. Ibrutinib shows durable single-agent efficacy in relapsed or refractory mantle-cell lymphoma. (Funded by Pharmacyclics and others; ClinicalTrials.gov number, NCT01236391.)


Income-related inequities: Cross-sectional analyses of the use of medicare services in British Columbia in 1992 and 2002.
The primary demonstration of the principle of income-related equity in Canada is the provision of health care services based on need rather than ability to pay. Despite this principle, Canada, along with other OECD countries, exhibits income-related variations in the use of health care services. This paper extends previous analyses to include surgical day care, assesses changes in income-related equity between 1992 and 2002 in British Columbia and tests the feasibility of using administrative data for general equity analyses. Data derive from the BC Linked Health Database and from a custom tabulation of income tax filer data provided by Statistics Canada. Cross-sectional analyses measure inequity in the probability and conditional use of services using concentration indices, which summarize health care services use for individuals ranked by income, after standardization for age, sex, region of residence and need for health care services. Small but systematic relationships were found between income and use of health care services for all types of services, with the exception of visits to general practitioners (GPs). Lower income is associated with greater conditional use of GPs and greater use of acute inpatient care. Higher income is associated with the greater use of specialist and surgical day care services; the latter inequity was found to grow substantially over time. Deviations from equity deserve further investigation, especially because the use of day care surgery is continually expanding. For example, an understanding of the reasons for differential admission rates to acute and day surgery might provide insight as to whether community-based services could help shift some acute care use among lower income groups to surgical day care. It is possible to use administrative data to monitor income-related equity, and future research should take advantage of this possibility.


Allogeneic stem cell transplantation for adult Philadelphia chromosome-negative acute lymphocytic leukemia: comparable survival rates but different risk factors between related and unrelated transplantation in first complete remission.
To identify factors to improve the outcomes of related and unrelated allogeneic stem cell transplantations (allo-SCT) for Philadelphia chromosome-negative acute lymphocytic leukemia (Ph(-) ALL) in the first complete remission (CR1), we retrospectively analyzed 1139 Ph(-) ALL patients using the registry data, particularly the details of 641 patients transplanted in CR1. Overall survival was significantly superior among patients transplanted in CR1, but no significant difference was observed between related and unrelated allo-SCTs (related vs unrelated: 65% vs 62% at 4 years, respectively; P = .19). Among patients transplanted in CR1, relapse rates were significantly higher in related allo-SCT compared with unrelated allo-SCT, and multivariate analysis demonstrated that less than 6 months from diagnosis to allo-SCT alone was associated with relapse. On the other hand, nonrelapse mortality (NRM) was significantly higher in unrelated allo-SCT compared with related allo-SCT, and multivariate analysis demonstrated that 10 months or longer from diagnosis to allo-SCT, human leukocyte antigen mismatch, and abnormal karyotype were associated with NRM. In conclusion, our study showed comparable survival rates but different relapse rates, NRM rates, and risk factors between related and unrelated allo-SCTs. After a close consideration of these factors, the outcome of allo-SCT for adult Ph(-) ALL in CR1 could be improved.


Treatment of lupus nephritis with abatacept: the Abatacept and Cyclophosphamide Combination Efficacy and Safety Study.
To assess the efficacy and safety of a 24-week course of abatacept in the treatment of active lupus nephritis and to assess the potential of abatacept to induce "clinical tolerance," defined as sustained clinical quiescence of lupus nephritis after discontinuation of immunosuppressive therapy. Patients with active lupus nephritis (n = 134) were enrolled in a randomized, double-blind phase II add-on trial in which they received either abatacept or placebo in conjunction with the Euro-Lupus Nephritis Trial regimen of low-dose cyclophosphamide (CYC) followed by azathioprine (AZA). The primary efficacy outcome was the frequency of complete response at week 24. Thereafter, patients who met either complete or partial response criteria continued blinded treatment through week 52. During this phase of the study, subjects in the abatacept treatment group in whom a complete response was achieved at week 24 discontinued immunosuppressive therapy other than prednisone (10 mg/day). There were no statistically significant differences between groups with respect to the primary outcome or any of the secondary outcomes, including measures of safety. A complete response was achieved in 33% of the subjects in the treatment group and in 31% of the subjects in the control group at week 24. Fifty percent of the subjects in the treatment group who met complete response criteria and therefore discontinued immunosuppressive therapy at week 24 maintained their complete response status through week 52. The addition of abatacept to a regimen of CYC followed by AZA did not improve the outcome of lupus nephritis at either 24 or 52 weeks. No worrisome safety signals were encountered.


Optimization and profile calculation of ODE models using second order adjoint sensitivity analysis.
Parameter estimation methods for ordinary differential equation (ODE) models of biological processes can exploit gradients and Hessians of objective functions to achieve convergence and computational efficiency. However, the computational complexity of established methods to evaluate the Hessian scales linearly with the number of state variables and quadratically with the number of parameters. This limits their application to low-dimensional problems. We introduce second order adjoint sensitivity analysis for the computation of Hessians and a hybrid optimization-integration-based approach for profile likelihood computation. Second order adjoint sensitivity analysis scales linearly with the number of parameters and state variables. The Hessians are effectively exploited by the proposed profile likelihood computation approach. We evaluate our approaches on published biological models with real measurement data. Our study reveals an improved computational efficiency and robustness of optimization compared to established approaches, when using Hessians computed with adjoint sensitivity analysis. The hybrid computation method was more than 2-fold faster than the best competitor. Thus, the proposed methods and implemented algorithms allow for the improvement of parameter estimation for medium and large scale ODE models. The algorithms for second order adjoint sensitivity analysis are implemented in the Advanced MATLAB Interface to CVODES and IDAS (AMICI, https://github.com/ICB-DCM/AMICI/). The algorithm for hybrid profile likelihood computation is implemented in the parameter estimation toolbox (PESTO, https://github.com/ICB-DCM/PESTO/). Both toolboxes are freely available under the BSD license. Supplementary data are available at Bioinformatics online.


Immunization strategies for epidemic processes in time-varying contact networks.
Spreading processes represent a very efficient tool to investigate the structural properties of networks and the relative importance of their constituents, and have been widely used to this aim in static networks. Here we consider simple disease spreading processes on empirical time-varying networks of contacts between individuals, and compare the effect of several immunization strategies on these processes. An immunization strategy is defined as the choice of a set of nodes (individuals) who cannot catch nor transmit the disease. This choice is performed according to a certain ranking of the nodes of the contact network. We consider various ranking strategies, focusing in particular on the role of the training window during which the nodes' properties are measured in the time-varying network: longer training windows correspond to a larger amount of information collected and could be expected to result in better performances of the immunization strategies. We find instead an unexpected saturation in the efficiency of strategies based on nodes' characteristics when the length of the training window is increased, showing that a limited amount of information on the contact patterns is sufficient to design efficient immunization strategies. This finding is balanced by the large variations of the contact patterns, which strongly alter the importance of nodes from one period to the next and therefore significantly limit the efficiency of any strategy based on an importance ranking of nodes. We also observe that the efficiency of strategies that include an element of randomness and are based on temporally local information do not perform as well but are largely independent on the amount of information available.


The Responses of Medical General Practitioners to Unreasonable Patient Demand for Antibiotics--A Study of Medical Ethics Using Immersive Virtual Reality.
Dealing with insistent patient demand for antibiotics is an all too common part of a General Practitioner's daily routine. This study explores the extent to which portable Immersive Virtual Reality technology can help us gain an accurate understanding of the factors that influence a doctor's response to the ethical challenge underlying such tenacious requests for antibiotics (given the threat posed by growing anti-bacterial resistance worldwide). It also considers the potential of such technology to train doctors to face such dilemmas. Twelve experienced GPs and nine trainees were confronted with an increasingly angry demand by a woman to prescribe antibiotics to her mother in the face of inconclusive evidence that such antibiotic prescription is necessary. The daughter and mother were virtual characters displayed in immersive virtual reality. The specific purposes of the study were twofold: first, whether experienced GPs would be more resistant to patient demands than the trainees, and second, to investigate whether medical doctors would take the virtual situation seriously. Eight out of the 9 trainees prescribed the antibiotics, whereas 7 out of the 12 GPs did so. On the basis of a Bayesian analysis, these results yield reasonable statistical evidence in favor of the notion that experienced GPs are more likely to withstand the pressure to prescribe antibiotics than trainee doctors, thus answering our first question positively. As for the second question, a post experience questionnaire assessing the participants' level of presence (together with participants' feedback and body language) suggested that overall participants did tend towards the illusion of being in the consultation room depicted in the virtual reality and that the virtual consultation taking place was really happening.


Predicting the length of stay for neonates using heart-rate Markov models.
Recently, there has been an increase in preterm newborn's survival and as a result longer length of stay (LOS) in neonatal intensive care units (NICU) have been observed. Conseqyently, there has been an increased interest in accurate prediction of the LOS. Most of the existing techniques are based on physiological parameters i.e., scores such as (SNAP-II or SNAPPE-II). In this paper we propose to predict the length of stay using heart-rate measurements combined with physiological scores. We propose to model the heart-rate using Markov chain model and estimate transition probabilities using maximum likelihood estimator and the patient population from Neonatal Intensive Care Unit at McMaster Hospital. We then derive the maximum likelihood estimators of LOS using physiological measurements and transition probabilities. Training and test data sets were used to verify the proposed linear and nonlinear estimators.


Self-help interventions for depressive disorders and depressive symptoms: a systematic review.
Research suggests that depressive disorders exist on a continuum, with subthreshold symptoms causing considerable population burden and increasing individual risk of developing major depressive disorder. An alternative strategy to professional treatment of subthreshold depression is population promotion of effective self-help interventions that can be easily applied by an individual without professional guidance. The evidence for self-help interventions for depressive symptoms is reviewed in the present work, with the aim of identifying promising interventions that could inform future health promotion campaigns or stimulate further research. A literature search for randomised controlled trials investigating self-help interventions for depressive disorders or depressive symptoms was performed using PubMed, PsycINFO and the Cochrane Database of Systematic Reviews. Reference lists and citations of included studies were also checked. Studies were grouped into those involving participants with depressive disorders or a high level of depressive symptoms, or non-clinically depressed participants not selected for depression. A number of exclusion criteria were applied, including trials with small sample sizes and where the intervention was adjunctive to antidepressants or psychotherapy. The majority of interventions searched had no relevant evidence to review. Of the 38 interventions reviewed, the ones with the best evidence of efficacy in depressive disorders were S-adenosylmethionine, St John's wort, bibliotherapy, computerised interventions, distraction, relaxation training, exercise, pleasant activities, sleep deprivation, and light therapy. A number of other interventions showed promise but had received less research attention. Research in non-clinical samples indicated immediate beneficial effects on depressed mood for distraction, exercise, humour, music, negative air ionisation, and singing; while potential for helpful longer-term effects was found for autogenic training, light therapy, omega 3 fatty acids, pets, and prayer. Many of the trials were poor quality and may not generalize to self-help without professional guidance. A number of self-help interventions have promising evidence for reducing subthreshold depressive symptoms. Other forms of evidence such as expert consensus may be more appropriate for interventions that are not feasible to evaluate in randomised controlled trials. There needs to be evaluation of whether promotion to the public of effective self-help strategies for subthreshold depressive symptoms could delay or prevent onset of depressive illness, reduce functional impairment, and prevent progression to other undesirable outcomes such as harmful use of substances.


Recent and historical recombination in the admixed Norwegian Red cattle breed.
Comparison of recent patterns of recombination derived from linkage maps to historical patterns of recombination from linkage disequilibrium (LD) could help identify genomic regions affected by strong artificial selection, appearing as reduced recent recombination. Norwegian Red cattle (NRF) make an interesting case study for investigating these patterns as it is an admixed breed with an extensively recorded pedigree. NRF have been under strong artificial selection for traits such as milk and meat production, fertility and health. While measures of LD is also crucial for determining the number of markers required for association mapping studies, estimates of recombination rate can be used to assess quality of genomic assemblies. A dataset containing more than 17,000 genome-wide distributed SNPs and 2600 animals was used to assess recombination rates and LD in NRF. Although low LD measured by r2 was observed in NRF relative to some of the breeds from which this breed originates, reports from breeds other than those assessed in this study have described more rapid decline in r2 at short distances than what was found in NRF. Rate of decline in r2 for NRF suggested that to obtain an expected r2 between markers and a causal polymorphism of at least 0.5 for genome-wide association studies, approximately one SNP every 15 kb or a total of 200,000 SNPs would be required. For well known quantitative trait loci (QTLs) for milk production traits on Bos Taurus chromosomes 1, 6 and 20, map length based on historic recombination was greater than map length based on recent recombination in NRF. Further, positions for 130 previously unpositioned contigs from assembly of the bovine genome sequence (Btau_4.0) found using comparative sequence analysis were validated by linkage analysis, and 28% of these positions corresponded to extreme values of population recombination rate. While LD is reduced in NRF compared to some of the breeds from which this admixed breed originated, it is elevated over short distances compared to some other cattle breeds. Genomic regions in NRF where map length based on historic recombination was greater than map length based on recent recombination coincided with some well known QTL regions for milk production traits. Linkage analysis in combination with comparative sequence analysis and detection of regions with extreme values of population recombination rate proved to be valuable for detecting problematic regions in the Btau_4.0 genome assembly.


Low-socioeconomic-status enrollees in high-deductible plans reduced high-severity emergency care.
One-third of US workers now have high-deductible health plans, and those numbers are expected to grow in 2014 as implementation of the Affordable Care Act continues. There is concern that high-deductible health plans might cause enrollees of low socioeconomic status to forgo emergency care as a result of burdensome out-of-pocket costs. We analyzed emergency department (ED) visits and hospitalizations over two years among enrollees insured in high-deductible plans through small employers in Massachusetts. We found that plan members of low socioeconomic status experienced 25-30 percent reductions in high-severity ED visits over both years, while hospitalizations declined by 23 percent in year 1 but rose again in year 2. Similar trends were not found among high-deductible plan members of high socioeconomic status. Our findings suggest that plan members of low socioeconomic status at small firms responded inappropriately to high-deductible plans and that initial reductions in high-severity ED visits might have increased the need for subsequent hospitalizations. Policy makers and employers should consider proactive strategies to educate high-deductible plan members about their benefit structures or identify members at higher risk of avoiding needed care. They should also consider implementing means-based deductibles.


Comparison of monitoring performance of bioreactance versus esophageal Doppler in pediatric patients.
Cardiac output (CO) monitoring and goal-directed therapy during major abdominal surgery is currently used to decrease postoperative complications. However, few monitors are currently available for pediatric patients. Nicom(®) is a noninvasive CO monitoring technique based on the bioreactance principle (analysis of frequency variations of a delivered oscillating current traversing the thoracic cavity). Nicom(®) may be a useful monitor for pediatric patients. Pediatric patients undergoing major abdominal surgery under general anesthesia with cardiac monitoring by transesophageal Doppler (TED) were included. Continuously recorded hemodynamic variables obtained from both bioreactance and TED were compared. Data were analyzed using the Bland-Altman method. A total of 113 pairs of cardiac index (CI) measurments from 16 patients were analyzed. Mean age was 59 months (95% CI: 42-75) and mean weight was 17 kg (95% CI: 15-20). In the overall population, Bland-Altman analysis revealed a bias of 0.4 L/min/m(2), precision of 1.55 L/min/m(2), limits of agreement of -1.1 to 1.9 L/min/m(2) and a percentage error of 47%. For children weighing >15 kg, results were: Bias 0.51 L/min/m(2), precision 1.17 L/min/m(2), limits of agreement -0.64 to 1.66 L/min/m(2) and percentage error 34%. Simultaneous CI estimations made by bioreactance and TED showed high percentage of errors that is not clinically acceptable. Bioreactance cannot be considered suitable for monitoring pediatric patients.


Distribution pattern of surgically treated symptomatic prolapsed lumbar and sacral intervertebral discs in males.
The pattern of distribution of surgically treated symptomatic prolapsed lumbar and sacral intervertebral discs has been published, though scantily, especially in males. We decided to look at our own series, compare and contrast ours with some of those published. We treated 88 locations of this lesion in 68 males. The clinical features were those of lower back pains, with or without radiation into the lower extremities, sensory loss and paresis of the limbs. There was a case of loss of urinary bladder and ano-rectal control. All lesions were confirmed through cauda-equinograms and treated under general anaesthesia in knee-chest position (MECCA position). The patients were followed up for 3-6 months post-operatively. There were 88 locations in 68 males of 21-70 years of age, with 29 prolapses occurring during the age range 31-40 years, while 54 locations were on the left and 48 at L4/5. The procedures were well tolerated by all patients and there were no post-operative complications. This lesion in our series occurred mostly on the left, at the L4/5 level and peaked at 31-40 years age range. The predictability of occurrence of this disease, using side, level and age is still not feasible in males from our series.


Disrupted neural variability during propofol-induced sedation and unconsciousness.
Variability quenching is a widespread neural phenomenon in which trial-to-trial variability (TTV) of neural activity is reduced by repeated presentations of a sensory stimulus. However, its neural mechanism and functional significance remain poorly understood. Recurrent network dynamics are suggested as a candidate mechanism of TTV, and they play a key role in consciousness. We thus asked whether the variability-quenching phenomenon is related to the level of consciousness. We hypothesized that TTV reduction would be compromised during reduced level of consciousness by propofol anesthetics. We recorded functional magnetic resonance imaging signals of resting-state and stimulus-induced activities in three conditions: wakefulness, sedation, and unconsciousness (i.e., deep anesthesia). We measured the average (trial-to-trial mean, TTM) and variability (TTV) of auditory stimulus-induced activity under the three conditions. We also examined another form of neural variability (temporal variability, TV), which quantifies the overall dynamic range of ongoing neural activity across time, during both the resting-state and the task. We found that (a) TTM deceased gradually from wakefulness through sedation to anesthesia, (b) stimulus-induced TTV reduction normally seen during wakefulness was abolished during both sedation and anesthesia, and (c) TV increased in the task state as compared to resting-state during both wakefulness and sedation, but not anesthesia. Together, our results reveal distinct effects of propofol on the two forms of neural variability (TTV and TV). They imply that the anesthetic disrupts recurrent network dynamics, thus prevents the stabilization of cortical activity states. These findings shed new light on the temporal dynamics of neuronal variability and its alteration during anesthetic-induced unconsciousness.


Systematic meta-analyses of Alzheimer disease genetic association studies: the AlzGene database.
The past decade has witnessed hundreds of reports declaring or refuting genetic association with putative Alzheimer disease susceptibility genes. This wealth of information has become increasingly difficult to follow, much less interpret. We have created a publicly available, continuously updated database that comprehensively catalogs all genetic association studies in the field of Alzheimer disease (http://www.alzgene.org). We performed systematic meta-analyses for each polymorphism with available genotype data in at least three case-control samples. In addition to identifying the epsilon4 allele of APOE and related effects, we pinpointed over a dozen potential Alzheimer disease susceptibility genes (ACE, CHRNB2, CST3, ESR1, GAPDHS, IDE, MTHFR, NCSTN, PRNP, PSEN1, TF, TFAM and TNF) with statistically significant allelic summary odds ratios (ranging from 1.11-1.38 for risk alleles and 0.92-0.67 for protective alleles). Our database provides a powerful tool for deciphering the genetics of Alzheimer disease, and it serves as a potential model for tracking the most viable gene candidates in other genetically complex diseases.


UWB Channel Impulse Responses for Positioning in Complex Environments: A Detailed Feature Analysis.
Radio signal-based positioning in environments with complex propagation paths is a challenging task for classical positioning methods. For example, in a typical industrial environment, objects such as machines and workpieces cause reflections, diffractions, and absorptions, which are not taken into account by classical lateration methods and may lead to erroneous positions. Only a few data-driven methods developed in recent years can deal with these irregularities in the propagation paths or use them as additional information for positioning. These methods exploit the channel impulse responses (CIR) that are detected by ultra-wideband radio systems for positioning. These CIRs embed the signal properties of the underlying propagation paths that represent the environment. This article describes a feature-based localization approach that exploits machine-learning to derive characteristic information of the CIR signal for positioning. The approach is complete without highly time-synchronized receiver or arrival times. Various features were investigated based on signal propagation models for complex environments. These features were then assessed qualitatively based on their spatial relationship to objects and their contribution to a more accurate position estimation. Three datasets collected in environments of varying degrees of complexity were analyzed. The evaluation of the experiments showed that a clear relationship between the features and the environment indicates that features in complex propagation environments improve positional accuracy. A quantitative assessment of the features was made based on a hierarchical classification of stratified regions within the environment. Classification accuracies of over 90% could be achieved for region sizes of about 0.1 m 2 . An application-driven evaluation was made to distinguish between different screwing processes on a car door based on CIR measures. While in a static environment, even with a single infrastructure tag, nearly error-free classification could be achieved, the accuracy of changes in the environment decreases rapidly. To adapt to changes in the environment, the models were retrained with a small amount of CIR data. This increased performance considerably. The proposed approach results in highly accurate classification, even with a reduced infrastructure of one or two tags, and is easily adaptable to new environments. In addition, the approach does not require calibration or synchronization of the positioning system or the installation of a reference system.


Reaction of bromine and chlorine with phenolic compounds and natural organic matter extracts--Electrophilic aromatic substitution and oxidation.
Phenolic compounds are known structural moieties of natural organic matter (NOM), and their reactivity is a key parameter for understanding the reactivity of NOM and the disinfection by-product formation during oxidative water treatment. In this study, species-specific and/or apparent second order rate constants and mechanisms for the reactions of bromine and chlorine have been determined for various phenolic compounds (phenol, resorcinol, catechol, hydroquinone, phloroglucinol, bisphenol A, p-hydroxybenzoic acid, gallic acid, hesperetin and tannic acid) and flavone. The reactivity of bromine with phenolic compounds is very high, with apparent second order rate constants at pH 7 in the range of 10(4) to 10(7) M(-1) s(-1). The highest value was recorded for the reaction between HOBr and the fully deprotonated resorcinol (k = 2.1 × 10(9) M(-1) s(-1)). The reactivity of phenolic compounds is enhanced by the activating character of the phenolic substituents, e.g. further hydroxyl groups. With the data set from this study, the ratio between the species-specific rate constants for the reactions of chlorine versus bromine with phenolic compounds was confirmed to be about 3000. Phenolic compounds react with bromine or chlorine either by oxidation (electron transfer, ET) or electrophilic aromatic substitution (EAS) processes. The dominant process mainly depends on the relative position of the hydroxyl substituents and the possibility of quinone formation. While phenol, p-hydroxybenzoic acid and bisphenol A undergo EAS, hydroquinone, catechol, gallic acid and tannic acid, with hydroxyl substituents in ortho or para positions, react with bromine by ET leading to quantitative formation of the corresponding quinones. Some compounds (e.g. phloroglucinol) show both partial oxidation and partial electrophilic aromatic substitution and the ratio observed for the pathways depends on the pH. For the reaction of six NOM extracts with bromine, electrophilic aromatic substitution accounted for only 20% of the reaction, and for one NOM extract (Pony Lake fulvic acid) it accounted for <10%. This shows that for natural organic matter samples, oxidation (ET) is far more important than bromine incorporation (EAS).


Female contraception over 40.
The majority of women 40-49 years of age need an effective method of contraception because the decline in fertility with age is an insufficient protection against unwanted pregnancy. Although pregnancy is less likely after the age of 40 years, the clinical and social consequences of an unexpected pregnancy are potentially detrimental. No contraceptive method is contraindicated by advanced reproductive age alone; thus there is a need to discuss the effectiveness, risks and non-contraceptive benefits of all family planning methods for women in this age group. MEDLINE searches were done by topic (epidemiology, age and reproduction, sexual function, delayed childbearing and specific contraceptive methods). The topic summaries were presented to the Workshop Group and omissions or disagreements were resolved by discussion. The decline in fecundity in the fifth decade is insufficient for contraceptive purposes. Thus a family planning method is needed. Sterilization is by far the most common method in several countries. Copper intrauterine devices and hormone intrauterine systems have similar effectiveness, with fewer than 1% failures in the first year of typical use. Special considerations in this age group include the frequency of menstrual irregularity, sexual problems and the possibility of menopausal symptoms, all of which may respond to hormonal methods of contraception. Women should be advised to continue with a contraceptive method until they have reached the menopause with its natural state of sterility.


Combined β-adrenergic and corticosteroid receptor activation regulates AMPA receptor function in hippocampal neurons.
Shortly after stress, limbic neurons are exposed to high levels of noradrenaline and corticosterone. These hormones are necessary for optimal behavioural adaptation. Behavioural effects critically depend on noradrenaline acting via β-adrenergic receptors, but these effects are strongly modulated by corticosterone, indicating putative interactions between the two hormones. Since both noradrenaline and corticosterone are known to quickly affect properties of AMPA-type glutamate receptors (AMPAR), we here examined - in hippocampal neurons - three parameters which give insight in the functionality of AMPARs: phosphorylation, surface expression and spontaneous synaptic transmission. In homogenates of adult hippocampal slices, application of corticosterone (30 nM for 15 min) by itself did not affect phosphorylation of the AMPAR GluA1 subunit at S845 or S831. Co-application of the β-adrenergic receptor agonist isoproterenol (10 µM) largely increased S845 (but not S831) phosphorylation. Corticosterone also did not change GluA1 and GluA2 surface expression in hippocampal primary cultures. However, combined administration of corticosterone and 1 µM isoproterenol - which by itself was ineffective - enhanced surface expression. Interestingly, 10 µM isoproterenol alone enhanced GluA1 surface expression, but this was decreased by corticosterone. Finally, in hippocampal primary cultures, the inter-event interval of miniature excitatory postsynaptic currents (mEPSCs) was decreased by the combination of 1 µM isoproterenol and corticosterone (which were ineffective by themselves) while the same combination did not affect the amplitude. We conclude that AMPAR phosphorylation, surface expression and mEPSC inter-event interval respond most strongly to a combination of corticosterone and β-adrenergic receptors. These combined hormonal effects on glutamate transmission might contribute to their memory-enhancing effects.


TAPA-1, the target of an antiproliferative antibody, defines a new family of transmembrane proteins.
A murine monoclonal antibody was identified by its ability to induce a reversible antiproliferative effect on a human lymphoma cell line. Immunoprecipitation studies revealed that the antibody reacted with a 26-kilodalton cell surface protein (TAPA-1). A diverse group of human cell lines, including hematolymphoid, neuroectodermal, and mesenchymal cells, expressed the TAPA-1 protein. Many of the lymphoid cell lines, in particular those derived from large cell lymphomas, were susceptible to the antiproliferative effects of the antibody. TAPA-1 may therefore play an important role in the regulation of lymphoma cell growth. A cDNA clone coding for TAPA-1 was isolated by using the monoclonal antibody to screen an expression library in COS cells. Analysis of the deduced amino acid sequence indicated that the protein is highly hydrophobic and that it contains four putative transmembrane domains and a potential N-myristoylation site. TAPA-1 showed strong homology with the CD37 leukocyte antigen and with the ME491 melanoma-associated antigen, both of which have been implicated in the regulation of cell growth.


Immune checkpoint inhibitors in malignancies with mismatch repair deficiency: a review of the state of the current knowledge.
The use of immune checkpoint inhibitors to treat malignant tumors with microsatellite instability is an emerging new modality. This is based on the observations that these tumors may have a high mutation rate-thus a potential source of tumor-specific neoantigens-and harbor infiltrating cytotoxic T cells in response, suggesting that they may be particularly susceptible to immune checkpoint therapy. PUBMED and ASCO library were systematically reviewed to identify all relevant data that involved the use of immune checkpoint inhibitors in the treatment of cancers with microsatellite instability. The manual search retrieved a total of 3 relevant articles and 1 abstract published between 2015 and 2016. A total of 61 patients with colorectal, 3 with ampullary/cholangiocarcinoma, 2 with endometrial carcinomas, 3 with small bowel cancers, 2 with glioblastoma multiforme, and 1 with bladder cancer with reported efficacy results were reviewed. All the patients had stage IV cancer and were treated with immune checkpoint inhibitors until progression of disease or intolerable side effects emerged. The range of objective response rate was 25-71%. Responses were also durable with progression-free survival at 20 weeks of around 67-78% and to 46% at 1 year. The use of immune checkpoint inhibitors is effective in cancers that express microsatellite instability.


The reinfection threshold.
Thresholds in transmission are responsible for critical changes in infectious disease epidemiology. The epidemic threshold indicates whether infection invades a totally susceptible population. The reinfection threshold indicates whether self-sustained transmission occurs in a population that has developed a degree of partial immunity to the pathogen (by previous infection or vaccination). In models that combine susceptible and partially immune individuals, the reinfection threshold is technically not a bifurcation of equilibria as correctly pointed out by Breban and Blower. However, we show that a branch of equilibria to a reinfection submodel bifurcates from the disease-free equilibrium as transmission crosses this threshold. Consequently, the full model indicates that levels of infection increase by two orders of magnitude and the effect of mass vaccination becomes negligible as transmission increases across the reinfection threshold.


Electrochemically Scalable Production of Fluorine-Modified Graphene for Flexible and High-Energy Ionogel-Based Microsupercapacitors.
Scalable production of high-quality heteroatom-modified graphene is critical for microscale supercapacitors but remains a great challenge. Herein, we demonstrate a scalable, single-step electrochemical exfoliation of graphite into highly solution-processable fluorine-modified graphene (FG), achieved in an aqueous fluorine-containing neutral electrolyte, for flexible and high-energy-density ionogel-based microsupercapacitors (FG-MSCs). The electrochemically exfoliated FG nanosheets are characterized by atomic thinness, large lateral size (up to 12 μm), a high yield of >70% with ≤3 layers, and a fluorine doping of 3 at%, allowing for large-scale production of FG-MSCs. Our ionogel-based FG-MSCs deliver high energy density of 56 mWh cm-3, by far outperforming the most reported MSCs. Furthermore, the all-solid-state microdevices offer exceptional cyclability with ∼93% after 5000 cycles, robust mechanical flexibility with 100% of capacitance retention bended at 180°, and outstanding serial and parallel integration without the requirement of metal-based interconnects for high-voltage and high-capacitance output. Therefore, these FG-MSCs represent remarkable potential for electronics.


Genetic contributions to the development of ADHD subtypes from childhood to adolescence.
Little is known about how genes influence the development of symptoms included in the DSM-IV subtypes of attention-deficit/hyperactivity disorder (ADHD) from childhood to adolescence. The aim of this study was to examine genetic influences contributing to the development of hyperactive-impulsive symptoms and inattentive symptoms of ADHD from childhood to adolescence. The sample included all 1,480 twin pairs born in Sweden between May 1985 and December 1986. Parents responded to mailed questionnaires on three occasions, when the twins were 8 to 9, 13 to 14, and 16 to 17 years old. The authors used dimensional scales of hyperactivity-impulsivity and inattention derived from a checklist of items based on the DSM symptoms of ADHD. Symptoms of hyperactivity-impulsivity declined with increasing age, whereas there was no decline in symptoms of inattention. Persistent genetic influences explain between 45% and 90% of the total genetic variance in hyperactivity-impulsivity and inattention across age. Persistent genetic variance was primarily operating across subtypes, even though persistent subtype-specific influences were also significant. The finding of persistent cross-subtype (i.e., combined) and persistent subtype-specific genetic influences (i.e., primarily hyperactive-impulsive and primarily inattentive) are in line with a genetic basis for the DSM-IV classification of ADHD subtypes.


Deuterium content of snow cores from Sierra Nevada area.
The relative deuterium content was measured on 37 snow cores collected in April 1969 in the Sierra Nevada. The deuterium content varies inversely with altitude of collection (approximately 40 per mil per 1000 meters) but is unrelated to latitude. The altitude relationship is particularly well defined west of the crest of the range but is not well defined east of the crest. However, samples from east of the crest tend to be depleted by about 10 to 15 per mil relative to samples collected at the same elevation west of the crest. We propose that the deuterium content of snow cores, collected so as to include the total winter's precipitation, can be used as a climatic indicator to compare the climate of one winter with that of another.


Effects of ribose as an ergogenic aid.
The amount of adenosine triphosphate (ATP) stored in the muscle available for immediate use is limited, and once used, must be resynthesized in the muscle. Ribose, a naturally occurring pentose sugar, helps resynthesize ATP for use in muscles. There have been claims that ribose supplements increase ATP levels and improve performance. Other studies have provided mixed results on the effectiveness of ribose as an ergogenic aid at high doses. None of these studies have compared the impact of the recommended dose of ribose on athletes and nonathletes under exercise conditions that are most conducive for effectiveness. The purpose of this study was to evaluate the effectiveness of ribose as an ergogenic aid at the dose recommended for supplements currently on the market during an exercise trial to maximize its efficacy. Male subjects (n = 11) performed 2 trials 1 week apart. Each trial consisted of three 30-second Wingate tests with a 2-minute recovery between each test. Trials were counterbalanced, with 1 trial being performed with 625 mg of ribose and the other with a placebo. Peak power, mean power, and percent decrease in power were recorded during each Wingate test. Repeated-measures analysis of variance (p > 0.05) found no significant differences between ribose and placebo. These results suggest that ribose had no effect on performance when taken orally, at the dose suggested by the distributor.


Strengthening E-cadherin adhesion via antibody mediated binding stabilization.
E-cadherins (Ecads) are a crucial cell-cell adhesion protein with tumor suppression properties. Ecad adhesion can be enhanced by the monoclonal antibody 66E8, which has potential applications in inhibiting cancer metastasis. However, the biophysical mechanisms underlying 66E8 mediated adhesion strengthening are unknown. Here, we use molecular dynamics simulations, site directed mutagenesis and single molecule atomic force microscopy experiments to demonstrate that 66E8 strengthens Ecad binding by stabilizing the primary Ecad adhesive conformation: the strand-swap dimer. By forming electrostatic interactions with Ecad, 66E8 stabilizes the swapped β-strand and its hydrophobic pocket and impedes Ecad conformational changes, which are necessary for rupture of the strand-swap dimer. Our findings identify fundamental mechanistic principles for strengthening of Ecad binding using monoclonal antibodies.


Human placental anticoagulant protein: isolation and characterization.
An anticoagulant protein was purified from the soluble fraction of human placenta by ammonium sulfate precipitation and column chromatography on DEAE-Sepharose, Sephadex G-75, and Mono S (Pharmacia). The yield of the purified protein was approximately 20 mg from one placenta. The purified protein gave a single band by sodium dodecyl sulfate-polyacrylamide gel electrophoresis with a molecular weight of 36,500. This protein prolonged the clotting time of normal plasma when clotting was induced either by brain thromboplastin or by kaolin in the presence of cephalin and Ca2+. It also prolonged the factor Xa induced clotting time of platelet-rich plasma but did not affect thrombin-induced conversion of fibrinogen to fibrin. The purified placental protein completely inhibited the prothrombin activation by reconstituted prothrombinase, a complex of factor Xa-factor Va-phospholipid-Ca2+. The placenta inhibitor had no effect on prothrombin activation when phospholipid was omitted from the above reaction. Also, it neither inhibited the amidolytic activity of factor Xa, nor did it bind to factor Xa. The placenta inhibitor, however, did bind specifically to phospholipid vesicles (20% phosphatidylserine and 80% phosphatidylcholine) in the presence of calcium ions. These results indicate that the placental anticoagulant protein (PAP) inhibits coagulation by binding to phospholipid vesicles. The amino acid sequences of three cyanogen bromide fragments of PAP aligned with those of two distinct regions of lipocortin I and II with a high degree of homology, showing that PAP is a member of the lipocortin family.


[Molecular mechanism of ALS and a possible gene therapy].
We report clinical characteristics of familial amyotrophic lateral sclerosis (FALS) with four different missense point mutations in exons 1, 2, 4, and 5 of the Cu/Zn superoxide dismutase (SOD) gene, that result in amino acid substitutions of cysteine 6 by phenylalanin (C 6 F), histidine 46 by arginine (H46R), leucine 84 by valine (L84V), isoleucine 104 by phenylalanine (I104F), and valine 148 by isoleucine (V148I), in five Japanese families. Although features of progressive neurogenic muscular atrophy was common in patients of these families, patients of each family showed characteristic clinical features. Immunoreactivity for Cu/Zn SOD of the motor neurons was not different between the ALS and controls. In contrast, immunoreactivity for NT was densely detected in motor neurons of ALS while that was not or was only minimally detected in those of controls. Adenovirus-mediated E. coli LacZ gene was transferred and expressed both in the muscle and spinal cord of transgenic mice. These results suggest that familial ALS with different mutations of the Cu/Zn SOD gene showed each clinical characteristics, that nitration of protein-tyrosine residue is upregulated in motor neurons of the spinal cord of ALS, and that there could be a possible future therapy of ALS with exogenous gene transfer.


The challenges of sodium measurements: indirect versus direct ion-selective method.
Diagnosis and treatment of dysnatremia is challenging and further complicated by the pitfalls of different sodium measurement methods. Routinely used sodium measurements are the indirect (plasma/serum) and direct (whole blood) ion-selective electrode (ISE) method, showing discrepant results especially in the setting of acute illness. Few clinicians are aware of the differences between the methods in clinically stable patients or healthy volunteers. Data of 140 patients and 91 healthy volunteers undergoing osmotic stimulation with hypertonic saline infusion were analyzed. Sodium levels were measured simultaneously by indirect and direct ISE method before and at different time points during osmotic stimulation up to a sodium threshold of ≥150 mmol/L. The primary outcome was the difference in sodium levels between the indirect and direct ISE method. 878 sodium measurements were analyzed. Mean (s.d.) sodium levels ranged from 141 mmol/L (2.9) to 151 mmol/L (2.1) by the indirect ISE compared to 140 mmol/L (3) to 149 mmol/L (2.8) by the direct ISE method. The interclass correlation coefficient between the two methods was 0.844 (95% CI: 0.823-0.863). On average, measurements by the indirect ISE were 1.9 mmol/L (95% CI limits: -3.2 to 6.9) higher than those by the direct ISE method (P < 0.001). The tendency of the indirect ISE method resulting in higher levels increased with increasing sodium levels. Intra-individual sodium levels differ significantly between the indirect and direct ISE method also in the absence of acute illness. It is therefore crucial to adhere to the same method in critical situations to avoid false decisions due to measurement differences.


What factors influence the prevalence and accuracy of nursing diagnoses documentation in clinical practice? A systematic literature review.
To identify what determinants influence the prevalence and accuracy of nursing diagnosis documentation in clinical practice. Nursing diagnoses guide and direct nursing care. They are the foundation for goal setting and provide the basis for interventions. The literature mentions several factors that influences nurses' documentation of diagnoses, such as a nurse's level of education, patient's condition and the ward environment. Systematic review. MEDLINE and CINAHL databases were searched using the following headings and keywords: nursing diagnosis, nursing documentation, hospitals, influence, utilisation, quality, implementation and accuracy. The search was limited to articles published between 1995-October 2009. Studies were only selected if they were written in English and were primary studies addressing factors that influence nursing diagnosis documentation. In total, 24 studies were included. Four domains of factors that influence the prevalence and accuracy of diagnoses documentation were found: (1) the nurse as a diagnostician, (2) diagnostic education and resources, (3) complexity of a patient's situation and (4) hospital policy and environment. General factors, which influence decision-making, and nursing documentation and specific factors, which influence the prevalence and accuracy of nursing diagnoses documentation, need to be distinguished. To support nurses in documenting their diagnoses accurately, we recommend taking a comprehensive perspective on factors that influence diagnoses documentation. A conceptual model of determinants that influence nursing diagnoses documentation, as presented in this study, may be helpful as a reference for nurse managers and nurse educators. This review gives hospital management an overview of determinants for possible quality improvements in nursing diagnoses documentation that needs to be undertaken in clinical practice.


Crystal structure of tRNA-guanine transglycosylase: RNA modification by base exchange.
tRNA-guanine transglycosylases (TGT) are enzymes involved in the modification of the anticodon of tRNAs specific for Asn, Asp, His and Tyr, leading to the replacement of guanine-34 at the wobble position by the hypermodified base queuine. In prokaryotes TGT catalyzes the exchange of guanine-34 with the queuine (.)precursor 7-aminomethyl-7-deazaguanine (preQ1). The crystal structure of TGT from Zymomonas mobilis was solved by multiple isomorphous replacement and refined to a crystallographic R-factor of 19% at 1.85 angstrom resolution. The structure consists of an irregular (beta/alpha)8-barrel with a tightly attached C-terminal zinc-containing subdomain. The packing of the subdomain against the barrel is mediated by an alpha-helix, located close to the C-terminus, which displaces the eighth helix of the barrel. The structure of TGT in complex with preQ1 suggests a binding mode for tRNA where the phosphate backbone interacts with the zinc subdomain and the U33G34U35 sequence is recognized by the barrel. This model for tRNA binding is consistent with a base exchange mechanism involving a covalent tRNA-enzyme intermediate. This structure is the first example of a (beta/alpha)-barrel protein interacting specifically with a nucleic acid.


Impact of stress, gender and menstrual cycle on immune system: possible role of nitric oxide.
Stress is a factor found to be involved in the etiology of many diseases. Gender and menstrual cycle phases are other factors affecting the predisposition of individuals for certain diseases. Results from animal and human studies suggest that the distribution of immune system cells may change at different phases of the menstrual cycle. Acute mental stress in humans alters immune variables, too. The increase in the number of natural killer (NK) cells is the most consistent finding among the immune variables, though there are controversies for the other lymphocyte groups. Nitric oxide (NO) as an immune mediator has an unsettled role whether it causes the redistribution of the immune cells, or is an end product of lymphocyte activation. This study was planned to investigate the effect of mental stress on lymphocyte subtypes and the role of NO, for men and women at different phases of the cycle. For this purpose, healthy men (n = 10) and women (n = 10), during the follicular and luteal phases underwent Stroop colour-word interference and cold pressor tests. The immune system responses before and after the tests were determined by cell counts with the flowcytometer. Menstrual cycle phase was ascertained by plasma estrogen and progesterone measurements. Stress response was evaluated by blood pressure (BP) and heart rate (HR) measurements throughout the tests and plasma cortisol and urinary metanephrine and vanillylmandelic acid (VMA) measurements before and after the tests. Plasma and urinary NO determinations were performed before and after the test was completed. All the results were analysed with the appropriate statistical methods. The luteal phase differed from the other groups due to the presence of suppressed immune response to acute stress, including decreased CD4/CD8 ratio and NK cell percentage. On the other hand, acute stress caused a shift from cellular to humoral immunity in men. As indicated by these results, individual reaction towards stress is affected by gender and menstrual cycle phase. NO appears to be a possible effector molecule for these differences.


Fractionated cyclophosphamide, vincristine, liposomal daunorubicin, and dexamethasone plus rituximab and granulocyte-macrophage-colony stimulating factor (GM-CSF) alternating with methotrexate and cytarabine plus rituximab and GM-CSF in patients with Richter syndrome or fludarabine-refractory chronic lymphocytic leukemia.
Therapy for patients with Richter syndrome (RS) or fludarabine-refractory chronic lymphocytic leukemia (CLL) is unsatisfactory. A Phase II study was conducted to evaluate an alternating combination cytotoxic regimen given with rituximab and granulocyte-macrophage-colony stimulating factor (GM-CSF) in these patients. Fludarabine-refractory CLL was defined as failure to respond to most recent prior fludarabine-containing regimen. Patients received up to six cycles of fractionated cyclophosphamide, vincristine, liposomal daunorubicin, and dexamethasone (hyper-CVXD) plus rituximab and GM-CSF alternating with methotrexate and cytarabine plus rituximab and GM-CSF. Response, toxicity, and survival data were compared with data from prior therapy with hyper-CVXD alone in this patient group. Forty-nine patients with RS (n = 30 patients) or refractory CLL (n = 19 patients) were treated on study. Nine patients (18%) achieved a complete remission, and 11 patients achieved a partial remission (22%), for an overall objective response (OR) rate of 41%. With a median follow-up of 7.5 months and a maximum follow-up of 15.2 months, the 12-month failure free survival (FFS) rate was 27%, and the overall survival (OS) rate was 39%. Nine patients (18%) died during the first cycle of therapy, and two patients (4%) died during the second cycle. There were no significant differences between the rates of OR, OS, and FFS in the current study and those obtained with hyper-CVXD alone on a prior study. The study regimen had activity and significant toxicity in patients with RS or fludarabine-refractory CLL. It was not clearly better compared with hyper-CVXD alone in this patient population.


Simulation Use for Global Away Rotations (SUGAR): preparing residents for emotional challenges abroad--a multicenter study.
Preparation for residents participating in global health (GH) experiences is critical. Active preparatory curricula allowing residents to experience and debrief emotional challenges they may encounter abroad are generally lacking. We sought to evaluate a novel simulation curriculum designed to prepare residents for emotions they may experience in response to challenges abroad. Pediatric GH educators from 7 institutions developed case vignettes incorporating common challenges residents experience abroad. Residents participating in a GH training track or planning to participate in a GH rotation from the 7 institutions were eligible to participate in the simulation curriculum. Participants and trained facilitators completed postsimulation evaluations that were analyzed using descriptive statistics and thematic analysis of written comments to assess the utility of the curriculum, emotions evoked, and changes residents anticipated making to their GH rotation preparation. Fifty-one residents and 16 facilitators completed 160 and 52 evaluations, respectively. Overall, respondents found the simulations useful (mean [SD] resident score 4.49 [0.82] and facilitator score 4.85 [0.36] on a 5-point scale [1 = completely useless, 5 = very useful]). Residents reported strong emotions in 153 (98%) of 156 comments. After the sessions, 131 (96%) of 137 comments reflected anticipated changes to GH rotation preparation plans. Active preparation for GH rotations using simulated cases appears to be a useful tool that can be implemented across a variety of sites with minimal facilitator training or simulation experience. The curriculum successfully elicited powerful emotions in residents and provided an opportunity to debrief these experiences before encountering them abroad.


Resistance to dual blockade of the kinases PI3K and mTOR in KRAS-mutant colorectal cancer models results in combined sensitivity to inhibition of the receptor tyrosine kinase EGFR.
Targeted blockade of aberrantly activated signaling pathways is an attractive therapeutic strategy for solid tumors, but drug resistance is common. KRAS is a frequently mutated gene in human cancer but remains a challenging clinical target. Inhibitors against KRAS signaling mediators, namely, PI3K (phosphatidylinositol 3-kinase) and mTOR (mechanistic target of rapamycin), have limited clinical efficacy as single agents in KRAS-mutant colorectal cancer (CRC). We investigated potential bypass mechanisms to PI3K/mTOR inhibition in KRAS-mutant CRC. Using genetically engineered mouse model cells that had acquired resistance to the dual PI3K/mTOR small-molecule inhibitor PF-04691502, we determined with chemical library screens that inhibitors of the ERBB [epidermal growth factor receptor (EGFR)] family restored the sensitivity to PF-04691502. Although EGFR inhibitors alone have limited efficacy in reducing KRAS-mutant tumors, we found that PF-04691502 induced the abundance, phosphorylation, and activity of EGFR, ERBB2, and ERBB3 through activation of FOXO3a (forkhead box O 3a), a transcription factor inhibited by the PI3K to AKT pathway. PF-04691502 also induced a stem cell-like gene expression signature. KRAS-mutant patient-derived xenografts from mice treated with PF-04691502 had a similar gene expression signature and exhibited increased EGFR activation, suggesting that this drug-induced resistance mechanism may occur in patients. Combination therapy with dacomitinib (a pan-ERBB inhibitor) restored sensitivity to PF-04691502 in drug-resistant cells in culture and induced tumor regression in drug-resistant allografts in mice. Our findings suggest that combining PI3K/mTOR and EGFR inhibitors may improve therapeutic outcome in patients with KRAS-mutant CRC.


American College of Sports Medicine position stand. The female athlete triad.
The female athlete triad (Triad) refers to the interrelationships among energy availability, menstrual function, and bone mineral density, which may have clinical manifestations including eating disorders, functional hypothalamic amenorrhea, and osteoporosis. With proper nutrition, these same relationships promote robust health. Athletes are distributed along a spectrum between health and disease, and those at the pathological end may not exhibit all these clinical conditions simultaneously. Energy availability is defined as dietary energy intake minus exercise energy expenditure. Low energy availability appears to be the factor that impairs reproductive and skeletal health in the Triad, and it may be inadvertent, intentional, or psychopathological. Most effects appear to occur below an energy availability of 30 kcal.kg(-1) of fat-free mass per day. Restrictive eating behaviors practiced by girls and women in sports or physical activities that emphasize leanness are of special concern. For prevention and early intervention, education of athletes, parents, coaches, trainers, judges, and administrators is a priority. Athletes should be assessed for the Triad at the preparticipation physical and/or annual health screening exam, and whenever an athlete presents with any of the Triad's clinical conditions. Sport administrators should also consider rule changes to discourage unhealthy weight loss practices. A multidisciplinary treatment team should include a physician or other health-care professional, a registered dietitian, and, for athletes with eating disorders, a mental health practitioner. Additional valuable team members may include a certified athletic trainer, an exercise physiologist, and the athlete's coach, parents and other family members. The first aim of treatment for any Triad component is to increase energy availability by increasing energy intake and/or reducing exercise energy expenditure. Nutrition counseling and monitoring are sufficient interventions for many athletes, but eating disorders warrant psychotherapy. Athletes with eating disorders should be required to meet established criteria to continue exercising, and their training and competition may need to be modified. No pharmacological agent adequately restores bone loss or corrects metabolic abnormalities that impair health and performance in athletes with functional hypothalamic amenorrhea.


Effects of anesthetic methods on preserving anti-tumor T-helper polarization following hepatectomy.
To investigate the impact of different anesthetic techniques on T-helper (Th) cell subsets in hepatocellular carcinoma (HCC) patients undergoing hepatectomy. Sixty-one HCC patients who received hepatectomies were randomized into an epidural combined general anesthesia (G + E; n = 31) or a general anesthesia (G; n = 30) group. Blood samples were obtained the morning before the operation (d0), and on the second (d2) and seventh (d7) day after the operation. Th cell contents were evaluated using flow cytometry, real-time reverse transcription-polymerase chain reaction and enzyme-linked immunosorbent assay. In all 61 patients, Th1 and Th2 cell frequencies, and interferon-γ (IFN-γ) mRNA expression markedly increased on d2, compared to d0. They recovered slightly on d7, and the Th1/Th2 ratio increased markedly on d7, compared with d2. In contrast, Th17, regulatory T cell (Treg), and interleukin-17 (IL-17) levels and FOXP3 mRNA expression showed no significant change on d2, and then markedly decreased on d7. Similarly, plasma IFN-γ concentration on d2 was much higher than that on d0, and then partly recovered on d7. As compared with the G group, in the G + E group, Th1 cell frequencies and the Th1/Th2 ratio were slightly higher on d2 and significantly higher on d7, while Th2, Th17, and Treg cell frequencies were slightly lower on d2, and significantly lower on d7. Consistently, on d7, IFN-γ mRNA and protein levels and the IFN-γ/IL-4 ratio in the G + E group were higher than those in the G group. In contrast, the IL-17 mRNA level, and IL-17 and transforming growth factor-β₁ concentrations in the G + E group were lower than those in the G group. G + E is superior to G in shifting the Th1/Th2 balance towards Th1, while decreasing Th17 and Treg, potentially benefiting HCC patients by promoting anti-tumor Th polarization.


Angle-and polarization-dependent collective excitation of plasmonic nanoarrays for surface enhanced infrared spectroscopy.
Our recent work has showed that diffractively coupled nanoplasmonic arrays for Fourier transform infrared (FTIR) microspectroscopy can enhance the Amide I protein vibrational stretch by up to 10(5) times as compared to plain substrates. In this work we consider computationally the impact of a microscope objective illumination cone on array performance. We derive an approach for computing angular- and spatially-averaged reflectance for various numerical aperture (NA) objectives. We then use this approach to show that arrays that are perfectly optimized for normal incidence undergo significant response degradation even at modest NAs, whereas arrays that are slightly detuned from the perfect grating condition at normal incidence irradiation exhibit only a slight drop in performance when analyzed with a microscope objective. Our simulation results are in good agreement with microscope measurements of experimentally optimized periodic nanoplasmonic arrays.


VariantClassifier: A hierarchical variant classifier for annotated genomes.
High-throughput DNA sequencing has produced a large number of closed and well annotated genomes. As the focus from whole genome sequencing and assembly moves towards resequencing, variant data is becoming more accessible and large quantities of polymorphisms are being detected. An easy-to-use tool for quickly assessing the potential importance of these discovered variants becomes ever important. Written in Perl, the VariantClassifier receives a list of polymorphisms and genome annotation, and generates a hierarchically-structured classification for each variant. Depending on the available annotation, the VariantClassifier may assign each polymorphism to a large variety of feature types, such as intergenic or genic; upstream promoter region, intronic region, exonic region or downstream transcript region; 5' splice site or 3' splice site; 5' untranslated region (UTR), 3' UTR or coding sequence (CDS); impacted protein domain; substitution, insertion or deletion; synonymous or non-synonymous; conserved or unconserved; and frameshift or amino acid insertion or deletion (indel). If applicable, the truncated or altered protein sequence is also predicted. For organisms with annotation maintained at Ensembl, a software application for downloading the necessary annotation is also provided, although the classifier will function with properly formatted annotation provided through alternative means. We have utilized the VariantClassifier for several projects since its implementation to quickly assess hundreds of thousands of variations on several genomes and have received requests to make the tool publically available. The project website can be found at: http://www.jcvi.org/cms/research/projects/variantclassifier.


18F-FDG PET-CT imaging versus bone marrow biopsy in pediatric Hodgkin's lymphoma: a quantitative assessment of marrow uptake and novel insights into clinical implications of marrow involvement.
To evaluate whether positron emission tomography/computed tomography using fluorine-18 fluoro-deoxyglucose (18F-FDG PET-CT) predicts bone marrow involvement (BMI) in pediatric Hodgkin's lymphoma (pHL) with sufficient accuracy to supplant routine staging bone marrow biopsy (BMB), and to assess the clinical importance of marrow disease by comparing the prognosis of stage IV HL with BMI versus that without BMI. Data were retrospectively analyzed for all cases of pHL between July 2010 and June 2015 referred for staging 18F-FDG PET-CT scan and BMB. The reference standard was BMB. Stage IV patients were divided into three groups to compare their progression-free and overall survival: PET+ BMB-, PET+ BMB+, and PET- BMB-. Of the 784 patients, 83.3% were male and 16.7% female, with age ranging from 2 to 18 years (mean 10.3 years). Among the total cases, 104 (13.3%) had BMI; of these, 100 were detected by PET imaging and 58 by BMB. BMB and 18F-FDG PET/CT scans were concordant for BMI detection in 728 patients (93%): positive concordance in 54 and negative in 674. Of the 56 discordant cases, four had a false-negative PET scans and were upstaged by BMB, 46 with focal uptake were PET/CT-positive and BMB-negative (not obtained from active sites), and six with diffuse uptake were false-positive on PET due to paraneoplastic marrow activation. The sensitivity, specificity, PPV, and NPV of PET for identifying BMI was 93.6, 94, 53, and 99.4% respectively. On quantitative assessment, mean iBM-SUVmax of bilateral iliac crests was significantly higher in those with BMI versus those without (p < 0.05). 18F-FDG PET-CT imaging is more sensitive than BMB for BMI detection in pHL staging. BMB should be limited to those with normal marrow uptake in the presence of poor risk factors or those with diffusely increased uptake to exclude marrow involvement in the background of reactive marrow.


Investigation of a Rotavirus Gastroenteritis Outbreak among Immunosuppressed Patients in a Hospital Setting.
Rotavirus (RV) is the most common cause of severe dehydrating diarrhoea in healthy infants and young children. The aims of this study were to investigate a RV outbreak in the pediatric hematology and oncology ward and to examine possible associations between immune status and RV infection. Twenty-eight children (19 boys and 9 girls) who were hospitalized for treatment of hematological malignancy and solid organ tumor during the RV outbreak were enrolled in this study. Fourteen of the 28 patients developed RV gastroenteritis (GE) during the observation period. RV antigen and RV IgG and IgA were measured by enzyme-linked immunosorbent assays. RV G and P types were determined by reverse transcriptase-polymerase chain reaction. Mean duration of RVGE in 14 patients was 13.9 days and mean severity score was 7.4. Two RV strains (G3P [8] and G2P [4]) were mainly circulating in the ward, which might result in the formation of a reassortant G2P [8] strain and mixed infection with G2+3P [8] in the immunocompromised patients. RV antigenemia was detected in 22 of the 28 patients (78.6%). RV-specific IgG titers in acute-phase sera of RVGE group were significantly lower than those in non-RVGE group (P=0.001). Mean age of the patients was significantly lower in RVGE group (5.5 ± 4.6 years) than non RVGE group (10.6 ± 4.5 years) (P=0.015). Our data demonstrate that host factors including age, underlying diseases, and immune status may be associated with the susceptibility of RV infection in immunocompromised patients at the time of the nosocomial infection.


Inhibition of IL-12 production by 1,25-dihydroxyvitamin D3. Involvement of NF-kappaB downregulation in transcriptional repression of the p40 gene.
Interleukin 12 (IL-12), produced by myelomonocytic cells, plays a pivotal role in the development of T helper 1 (Th1) cells, which are involved in the pathogenesis of chronic inflammatory autoimmune disorders. 1,25-Dihydroxyvitamin D3 [1,25(OH)2D3] inhibits IL-12 production by activated macrophages and dendritic cells, thus providing a novel interpretation to its immunosuppressive properties. 1,25(OH)2D3 significantly inhibits mRNA expression for both IL-12 p35 and p40 subunits acting at the transcriptional level. The effect of 1,25(OH)2D3 on p40 promoter activation was analyzed by cotransfecting monocytic RAW264.7 cells with p40 promoter/reporter constructs and expression vectors for vitamin D3 receptor (VDR) and/or retinoid X receptor (RXRalpha). We observed transcriptional repression of the p40 gene by 1,25(OH)2D3, which required coexpression of VDR with RXR and an intact VDR DNA-binding domain. The repressive effect maps to a region in the p40 promoter containing a binding site for NF-kappaB (p40-kappaB). Deletion of the p40-kappaB site abrogates part of the inhibitory effect on the p40 promoter, confirming the functional relevance of this site. Activation of monocytic THP-1 cells in the presence of 1,25(OH)2D3 results in reduced binding to the p40-kappaB site. Thus, 1,25(OH)2D3 may negatively regulate IL-12 production by downregulation of NF-kappaB activation and binding to the p40-kappaB sequence.


The Santa Clara Strength of Religious Faith Questionnaire (SCSORF): a validation study on Iranian Muslim patients undergoing dialysis.
The Santa Clara Strength of Religious Faith Questionnaire (SCSORF) is an often used and validated scale that is uncommonly utilized in culturally diverse populations. The purpose of this research investigation was to adapt the SCSORF for use among Iranian Muslim patients undergoing dialysis and to examine the reliability and validity of the scale among this population. A total of 428 patients (228 females, 200 males, M age = 52.2 years, SD = 10) were selected from five dialysis center in Tehran and Qazvin, Iran. A comprehensive forward-backward translation system was used for cross-cultural translation. Patients completed a baseline questionnaire obtaining demographic and clinical information as well as the SCSORF, the Age Universal Religious Orientation Scale (AUROS), the religious life inventory (RLI), and the Duke University religion index (DUREL). 2 weeks later, patients were asked to complete the SCSORF once again. Reliability of the SCSORF was examined using internal consistency and test-rest reliability. Convergent validity and factor structure using exploratory factor analysis (EFA) and confirmatory factor analysis (CFA) were also examined. Cronbach's α for the single construct of the SCSORF was 0.89 with adequate test-retest reliability measured over a 2 week period. SCSORF scores were significantly correlated with AUROS, RLI and the DUREL. The EFA generated a single factor solution for the SCSORF while these results were confirmed by the CFA in an independent sample. Findings demonstrated that the SCSORF has favorable reliability, convergent validity, and divergent validity among Iranian Muslim patients undergoing dialysis and is recommended for use by clinicians (e.g., nephrologists) to measure strength of religious faith among patients.


Troglitazone amplifies counterregulatory responses to hypoglycemia in nondiabetic subjects.
As insulin sensitizers, thiazolidinediones could affect the hormonal counterregulatory response to hypoglycemia via the modulatory effect of insulin on counterregulation. In addition, recent studies suggest that thiazolidinediones may influence key steps in glucose sensing and glucoregulatory hormone secretion. We therefore evaluated the effects of a short course of troglitazone on counterregulatory hormones in response to mild hypoglycemia in eight lean nondiabetic subjects. Subjects received either troglitazone (400 mg/day) or placebo for 7 days before stepped hypoglycemia clamp studies (5.0, 4.4, 3.9, and 3.3 mmol/L target plasma glucose steps, 50 min each). The glycemic thresholds for secretion of epinephrine (3.77 +/- 0.05 mmol/L) and glucagon (3.83 +/- 0.11 mmol/L) were reset to a higher plasma glucose concentration after troglitazone [4.05 +/- 0.05 mmol/L (P = 0.003) and 4.10 +/- 0.05 mmol/L (P = 0.03), respectively]. In addition, the magnitude of the rise in epinephrine and glucagon concentrations was higher with troglitazone (28% and 11%, respectively; P < 0.05 for both), whereas plasma norepinephrine, GH, and cortisol were comparable in both sets of studies. Endogenous glucose production, measured with [3-(3)H]glucose, rose by 33% (P < 0.05) in the troglitazone studies compared with 17% (P = NS) after placebo. We conclude that thiazolidinediones may induce an amplification of the counterregulatory response to hypoglycemia characterized by a shift in the glycemic threshold for and an increase in the magnitude of glucagon and epinephrine secretion, and subsequent activation of glucose production.


Testing of large optical surfaces with subaperture stitching.
Based on our previously proposed subaperture stitching and localization (SASL) algorithm, we present strategies and a prototype for testing of large optical surfaces with subaperture stitching. First, several strategies are introduced to deal with new problems when applying the SASL algorithm to large surfaces. The uncertainty of the lateral scale of the interferometer is compensated in the same manner as that of the radii of best-fit spheres in the algorithm. Then the coarse-fine stitching strategy is proposed to stitch tens of subapertures efficiently. Second, a prototype for testing of large surfaces with subaperture stitching is developed with a welded structural base. The model of kinematics is built to determine the initial configuration of each subaperture, according to the records of nulling motion. The uncertainty of linear motion is required to be no more than 1 mm, taking advantage of the large range of convergence of the SASL algorithm. Finally we present an experiment to verify the validity of the method and the prototype. A spherical surface is tested and successfully stitched with 37 subapertures.


Defibrillation before EMS arrival in western Sweden.
Bystanders play a vital role in public access defibrillation (PAD) in out-of-hospital cardiac arrest (OHCA). Dual dispatch of first responders (FR) alongside emergency medical services (EMS) can reduce time to first defibrillation. The aim of this study was to describe the use of automated external defibrillators (AEDs) in OHCAs before EMS arrival. All OHCA cases with a shockable rhythm in which an AED was used prior to the arrival of EMS between 2008 and 2015 in western Sweden were eligible for inclusion. Data from the Swedish Register for Cardiopulmonary Resuscitation (SRCR) were used for analysis, on-site bystander and FR defibrillation were compared with EMS defibrillation in the final analysis. Of the reported 6675 cases, 24% suffered ventricular fibrillation (VF), 162 patients (15%) of all VF cases were defibrillated before EMS arrival, 46% with a public AED on site. The proportion of cases defibrillated before EMS arrival increased from 5% in 2008 to 20% in 2015 (p<0.001). During this period, 30-day survival increased in patients with VF from 22% to 28% (p=0.04) and was highest when an AED was used on site (68%), with a median delay of 6.5min from collapse to defibrillation. Adjusted odds ratio for on-site defibrillation versus dispatched defibrillation for 30-day survival was 2.45 (95% CI: 1.02-5.95). The use of AEDs before the arrival of EMS increased over time. This was associated with an increased 30-day survival among patients with VF. Thirty-day survival was highest when an AED was used on site before EMS arrival.