Validation of a drooling questionnaire in Indian children with cerebral palsy.
Drooling of saliva is a common problem in children with cerebral palsy. In addition to causing impairment in articulation, drooling also affects socialization, interpersonal relationships and integration into society for these children. There are various methods to assess drooling which measure directly the amount of saliva drooled. However the most convenient and popular method is the use of questionnaires which are mostly western based and need slight modification for the Indian scenario Aim-Validation of a modified questionnaire for the assessment of drooling in children with cerebral palsy. The modified questionnaire was administered to parents of children with cerebral palsy willing to participate in the study. The drooling score was compared with objective tests, namely cotton pad test and drooling quotient. Internal consistency was assessed using the Cronbach's alpha, test retest reliability by Intraclass Correlation and sensitivity analysis by the Receiver operating characteristic curve. The modified questionnaire was found to be easy to administer. The Cronbach's alpha coefficient was between 0.867 and 0.879 which implies a high degree on internal consistency. The intraclass correlation and the test retest reliability was found to be statistically significant with a p value < 0.001 which show that the questionnaire was highly reliable for repeat administration as well as administration by different investigators. The ROC Area was found to be 0.94 with a standard error of 0.02 with a 95% confidence interval of 0.88-0.99, which suggests that the score has great specificity, closer agreement between specificity and sensitivity and excellent precision. Our modified questionnaire was easy to administer, highly reliable and valid with high internal consistency. A score of 24 on the questionnaire was found to be the most sensitive and specific point to discriminate between the mild and severe droolers in children with cerebral palsy.


Evaluation of the psychometric properties of the SF-36 health survey for use among Turkish and Moroccan ethnic minority populations in the Netherlands.
To evaluate the psychometrics of the SF-36 Health Survey among Turkish and Moroccan ethnic minority populations in the Netherlands and to compare the results to those based on the indigenous Dutch population. Data were derived from the Second Dutch National Survey of General Practice. In total, SF-36 data were available for 409 Turkish, 377 Moroccan, and 9,628 Dutch respondents. Language subgroup analyses were performed for the Turkish (n = 162) and Moroccan-Arabic (n = 52) versions of the SF-36. Psychometric properties were evaluated by descriptive statistics, factor analysis, internal consistency estimates, known-group comparisons, and differential item function (DIF) analysis. Low levels of missing SF-36 data were observed across all groups. Confirmatory factor analysis supported the hypothesized SF-36 scale structure for the Dutch and Turkish group, but high correlations between the MH and VT factors were observed in the Moroccan group. All Cronbach's alpha coefficients were above 0.70, except for the Vitality scale in the Moroccan group and the Vitality and Social Functioning scales in the Moroccan-Arabic language subgroup. Known-groups validity was found across samples using age, sex, education, and comorbidity as grouping variables, but not marital status. Some evidence for DIF was found in both ethnic group samples. The results generally support the use of the SF-36 for general population research among Turkish and Moroccan ethnic minorities in the Netherlands. Additional studies are needed to confirm the psychometrics of the questionnaire when used among these populations in other Western European countries.


The Global Longitudinal Study of Osteoporosis in Women (GLOW): rationale and study design.
The Global Longitudinal study of Osteoporosis in Women (GLOW) is a prospective cohort study involving 723 physicians and 60,393 women subjects >or=55 years. The data will provide insights into the management of fracture risk in older women over 5 years, patient experience with prevention and treatment, and distribution of risk among older women on an international basis. Data from cohort studies describing the distribution of osteoporosis-related fractures and risk factors are not directly comparable and do not compare regional differences in patterns of patient management and fracture outcomes. The GLOW is a prospective, multinational, observational cohort study. Practices typical of each region were identified through primary care networks organized for administrative, research, or educational purposes. Noninstitutionalized patients visiting each practice within the previous 2 years were eligible. Self-administered questionnaires were mailed, with 2:1 oversampling of women >or=65 years. Follow-up questionnaires will be sent at 12-month intervals for 5 years. A total of 723 physicians at 17 sites in ten countries agreed to participate. Baseline surveys were mailed (October 2006 to February 2008) to 140,416 subjects. After the exclusion of 3,265 women who were ineligible or had died, 60,393 agreed to participate. GLOW will provide contemporary information on patterns of management of fracture risk in older women over a 5-year period. The collection of data in a similar manner in ten countries will permit comparisons of patient experience with prevention and treatment and provide insights into the distribution of risk among older women on an international basis.


A Subsidiary Cell-Localized Glucose Transporter Promotes Stomatal Conductance and Photosynthesis.
It has long been recognized that stomatal movement modulates CO2 availability and as a consequence the photosynthetic rate of plants, and that this process is feedback-regulated by photoassimilates. However, the genetic components and mechanisms underlying this regulatory loop remain poorly understood, especially in monocot crop species. Here, we report the cloning and functional characterization of a maize (Zea mays) mutant named closed stomata1 (cst1). Map-based cloning of cst1 followed by confirmation with the clustered regularly interspaced short palindromic repeats (CRISPR)/ CRISPR associated protein 9 system identified the causal mutation in a Clade I Sugars Will Eventually be Exported Transporters (SWEET) family gene, which leads to the E81K mutation in the CST1 protein. CST1 encodes a functional glucose transporter expressed in subsidiary cells, and the E81K mutation strongly impairs the oligomerization and glucose transporter activity of CST1. Mutation of CST1 results in reduced stomatal opening, carbon starvation, and early senescence in leaves, suggesting that CST1 functions as a positive regulator of stomatal opening. Moreover, CST1 expression is induced by carbon starvation and suppressed by photoassimilate accumulation. Our study thus defines CST1 as a missing link in the feedback-regulation of stomatal movement and photosynthesis by photoassimilates in maize.


Proteogenomic Analysis of Human Colon Cancer Reveals New Therapeutic Opportunities.
We performed the first proteogenomic study on a prospectively collected colon cancer cohort. Comparative proteomic and phosphoproteomic analysis of paired tumor and normal adjacent tissues produced a catalog of colon cancer-associated proteins and phosphosites, including known and putative new biomarkers, drug targets, and cancer/testis antigens. Proteogenomic integration not only prioritized genomically inferred targets, such as copy-number drivers and mutation-derived neoantigens, but also yielded novel findings. Phosphoproteomics data associated Rb phosphorylation with increased proliferation and decreased apoptosis in colon cancer, which explains why this classical tumor suppressor is amplified in colon tumors and suggests a rationale for targeting Rb phosphorylation in colon cancer. Proteomics identified an association between decreased CD8 T cell infiltration and increased glycolysis in microsatellite instability-high (MSI-H) tumors, suggesting glycolysis as a potential target to overcome the resistance of MSI-H tumors to immune checkpoint blockade. Proteogenomics presents new avenues for biological discoveries and therapeutic development.


Optically modulated electrokinetic manipulation and concentration of colloidal particles near an electrode surface.
We study a recently demonstrated AC electrokinetic technique for manipulation and concentration of colloidal particles on an electrode surface. The technique uses indium tin oxide (ITO)-based parallel-plate electrodes on which highly localized infrared (1064 nm) laser illumination is shone. We show that the highly localized laser illumination leads to a highly nonuniform heating of the electrode substrate, which in turn drives an electrothermal microvortex resulting in a rapid transport of particles toward the illuminated site. Hundreds of polystyrene particles, with diameters ranging from 2.0 to 0.1 microm, suspended in a low conductivity solution (2.0 mS/m) could be aggregated at selected locations on the electrode by activating the laser illumination at suitable AC frequencies. Subsequent deactivation of the laser illumination causes the particles to scatter, and we explore this dynamical behavior for 1.0 microm particles using Delaunay tessellations and high-speed videography. We establish that drag from the electrothermal microvortex acts against a repulsive force, which decreases with increasing AC frequency, to create stable particle clusters. Moreover, experimentally we show that this particle capturing technique can be characterized by a critical frequency: a frequency at which the captured colloidal particle cluster becomes unstable and particles are carried away into the bulk by the electrothermal microvortex. This critical frequency increases with decreasing particle diameter for similar particles. For 0.1 microm particles, comparison of aggregation at different AC frequencies is achieved by the comparison of fluorescent intensity profiles of the aggregations.


Fludarabine, cytarabine, granulocyte colony-stimulating factor, and idarubicin (FLAG-IDA) for the treatment of children with poor-prognosis acute leukemia: the Hacettepe experience.
Fludarabine, cytarabine, granulocyte colony-stimulating factor (G-CSF), and idarubicin (FLAG-IDA) regimen has been proven to be a potentially useful chemotherapy regimen for relapsed or poor-prognosis childhood leukemia. The aim of the study was to evaluate complete remission (CR) rate, toxicity, and overall survival of children with poor-prognosis acute leukemia who received the FLAG-IDA regimen. Furthermore, the authors investigated the children who achieved CR following FLAG-IDA treatment regarding their eligibility for allogeneic hematopoietic stem cell transplantation (HSCT). Between January 2002 and April 2007, 25 children with poor-prognosis acute leukemia were treated with FLAG-IDA regimen in our center. Of the 25 children (16 AML, 9 ALL) with poor-prognosis acute leukemia, 7 (28.0%) received 1 cycle, 17 (68.0%) received 2 cycles, and 1 (4%) received 3 cycles of FLAG or FLAG-IDA regimen. After 44 cycles of FLAG-IDA or FLAG regimen, 10/25 (40%) children were nonresponders, 15/25 (60.0%) showed CR. Five (20%) of these patients in CR who underwent allogeneic HSCT are still in remission. The remaining 20 (80.0%) children were lost due to infection or relapse of the primary diseases. The overall survival of patients who are still alive and underwent allogeneic HSCT (mean: 40.6 ± 4.7, median: 40, range: 34-46 months) was longer than that of patients (mean: 5.5 ± 4.3, median: 4, range: 1-15 months) who did not undergo allogeneic HSCT. The CR rate was quite high in the present study using the FLAG-IDA regimen, and the authors believe this regimen is a possible option prior to allogeneic HSCT in children with poor-prognosis acute leukemia.


Parental Sense of Coherence in the first 2 years of life is not related to parental and child diurnal cortisol rhythm or proxies of anthroposophic lifestyle.
Sense of Coherence (SOC) is hypothesized to have direct physiological consequences on endocrine and immunological processes. In this study, we compare parental SOC scores from pregnancy in groups of infants and parents representing different lifestyles (anthroposophic, partly anthroposophic and nonanthroposophic). We also analyse whether these could predict cortisol levels of the parents and their infants at 6-24 months postpartum. Parental SOC-13 was collected during the third trimester of pregnancy from a birth cohort of families with different lifestyles. Salivary samples were collected from the whole family when the child was 6 months (n = 210), 12 (n = 178) and 24 months of age (n = 149), and cortisol levels were analysed with radioimmunoassay technique. Sense of Coherence scores did not differ between the three lifestyle groups, and there were no correlations between SOC scores and salivary cortisol concentrations in separate analyses of mothers, fathers and children at any sampling age or at any sampling time during the day (morning, afternoon, bedtime). Sense of Coherence scores did not vary in parents with different lifestyles and were not associated with salivary cortisol levels in parents or in children.


Group sequential tests for treatment effect on survival and cumulative incidence at a fixed time point.
Medical research frequently involves comparing an event time of interest between treatment groups. Rather than comparing the entire survival or cumulative incidence curves, it is sometimes preferable to evaluate these probabilities at a fixed point in time. Performing a covariate adjusted analysis can improve efficiency, even in randomized clinical trials, but no currently available group sequential test for fixed point analysis provides this adjustment. This paper introduces covariate adjusted group sequential pointwise comparisons of survival and cumulative incidence probabilities. Their test statistics have an asymptotic distribution with independent increments, permitting use of common stopping boundary specification methods. These tests are demonstrated through a redesign of BMT CTN 0402, a clinical trial that evaluated a prophylactic treatment for adverse outcomes following blood and marrow transplantation. A simulation study demonstrates that these tests maintain the type I error rate and power at nominal levels under a variety of settings involving influential covariates.


Striae Distensae: Preventative and Therapeutic Modalities to Improve Aesthetic Appearance.
Striae distensae (SD) are aesthetically troublesome to patients and therapeutically challenging. Herein, the authors comprehensively review the literature pertaining to the history, pathogenesis, clinical presentation, clinical rating scales, and laboratory, imaging, and histologic features of SD. A review of PubMed, MEDLINE, Scopus, Embase, and Google scholar was conducted, including literature published from 1773 to August 6, 2016. The authors identified 68 articles that met inclusion and exclusion criteria. There are few randomized controlled trials evaluating the long-term efficacy and safety of various topical and energy-based devices. Based on clinical and anecdotal experience, both nonablative and ablative fractionated lasers have shown modest SD improvement compared with other treatment modalities (including Excimer laser, CuBr laser, pulsed dye laser, and 1,064-nm Nd:YAG laser). In the authors' experience, 1,540-nm nonablative fractionated laser is a worthy first-line modality for the treatment of SD. Future researchers may consider greater focus on enhanced study design, including larger, long-term split-body, or split-SD head-to-head randomized comparative trials with objective outcome measures and end points, such as biopsy and molecular studies demonstrating increased collagen and elastic fibers that correlate to clinical improvement.


[Relationship between body mass index and self-perception among university students].
To assess the relationship between body mass index and self-perception of body image. A study was carried out in a sample comprising 106 female and male university students aged 18 years or more in Ribeirão Preto, Southeastern Brazil, in 2003. The Contour Drawing Rating Scale and Visual Analogue Scale were used to evaluate body image perception; the former was applied using two different psychometric methods. A body image questionnaire was used to assess the subjective component of body image. Subjects were classified according to body mass index. Statistical analyses were performed through variance analysis and Newman-Keuls post-hoc test. Most normal weight or overweight women (87%) overestimated their body size while obese women and all men (73%), regardless of their BMI, underestimated their body size. The differences of body image perception between men and women were statistically significant as well as the overall dissatisfaction with their perceived body size, revealed by a desired lower body mass index. Overweight women were more concerned and uncomfortable with their own body. Both men and women had a distorted self-perception of body image, underestimating or overestimating it. The study results suggest dissatisfaction of subjects with their body image as they desire to have leaner bodies.


Hypoxia and Hyperoxia Affect Serum Angiogenic Regulators in T2DM Men during Cycling.
Exercise-induced transient increases in pro-angiogenic regulators can promote angiogenesis.This pilot study aims to analyze the potential of exercise to positively affect angiogenic regulators in patients with type 2 diabetes mellitus (T2DM), who often exhibit abnormal angiogenesis, under different environmental conditions. 9 overweight/obese men with uncomplicated T2DM (8 took anti-diabetic drugs) performed submaximal cycling for 40 min in normoxia (≈21 vol%O2), hypoxia (≈14 vol%O2) and during alternating hypoxia/hyperoxia (≈14 vol%O2/≈30 vol%O2, 5-min intervals) (3×3 crossover design). Serum pro-angiogenic vascular endothelial growth factor (VEGF), matrix metalloproteinase (MMP)-2, MMP-9 and anti-angiogenic endostatin were quantified using enzyme-linked immunosorbent assay (ELISA) kits. Non-parametric statistical tests (Wilcoxon, Friedman analysis of variance) were applied. VEGF increased significantly from pre- to post-exercise with hypoxia and hypoxia/hyperoxia. MMP-2 increased significantly in all experimental runs, while MMP-9 only increased significantly with hypoxia and hypoxia/hyperoxia. Endostatin increased significantly with normoxia and hypoxia. However, the magnitude of changes did not differ significantly between conditions. Capillary blood lactate was significantly lower following cycling with hypoxia/hyperoxia than with hypoxia alone. Although differences in subjective ratings of perceived exertion failed significance, 7 subjects were less exerted with hypoxia/hyperoxia than with hypoxia. Submaximal cycling with hypoxia or alternating hypoxia/hyperoxia may induce a more reliable up-regulation of pro-angiogenic regulators compared with normoxia, while hypoxia/hyperoxia may be better tolerated than hypoxia alone.


Relationship between age, molecular markers, and response to sulphadoxine-pyrimethamine treatment in Kampala, Uganda.
Sulphadoxine/pyrimethamine (SP) has become the first-line treatment of uncomplicated malaria in a number of African countries. Molecular surveillance of resistance-mediating mutations in Plasmodium falciparum dihydrofolate reductase (dhfr) and dihydropteroate synthase (dhps) has been proposed as a means of predicting SP treatment outcomes, but optimal methods of surveillance in different populations have not been well established. To investigate the relationship between molecular markers of SP resistance, host immunity, and response to therapy, we evaluated the association between the presence of five key dhfr and dhps mutations at enrollment and clinical outcome in children and adults treated with SP for uncomplicated malaria in Kampala, Uganda. Clinical treatment failure was 11% at 14 days, increasing to 30% at 28 days, after excluding new infections. Outcomes varied markedly based on the number of dhfr and dhps mutations and on the age of treated subjects. All infections with less than two dhfr/dhps mutations were successfully treated. Treatment failure associated with any two, three, or four dhfr/dhps mutations occurred in nine of 24 (38%) children up to 5 years, but not in older patients (0/20). In the presence of all five mutations, treatment failure occurred equally in children aged 5 years or younger [7/16 (44%)] and in older patients [8/16 (50%)]. Our results showed that age, a surrogate marker of antimalarial immunity, had a major impact on the relationship between polymorphisms in SP target enzymes and treatment outcomes. The use of molecular markers of SP resistance to predict treatment failure rates should take age into account.


Initial ball flight characteristics of curve and instep kicks in elite women's football.
Initial ball flight characteristics of curve and instep kicks were investigated. Fifteen international female footballers performed curve and instep kicks from a distance of 20 m from goal and at a 1 m2 target. Seventeen Vicon cameras tracked three-dimensional coordinates of four reflective markers adhered to the ball. Ball flight characteristics were quantified, and the coordinates of the ball relative to the target center were recorded. The lateral launch angle and the angle of the spin axis relative to the horizontal best predicted the horizontal placement of the ball relative to the target. The vertical launch angle, antero-posterior velocity and amount of backspin best predicted the vertical coordinate. Regression models demonstrated how carefully controlled the flight characteristics must be with launch angles constrained within 3° to hit the target. Curve kicks were characterized by significantly greater lateral and vertical launch angles, increased sidespin and spin about the antero-posterior axis, and a more vertical spin axis. This information is beneficial for coaches in training players to achieve the characteristics required to score a goal and avoid a defensive wall. For example, if players consistently kick above or below the target, these findings identify the variables that will help rectify that error.


Implementation of an EMR System for a Comprehensive Dental Service within a Large Regional Hospital Network: Challenges and Opportunities Presented by the Introduction of new Technology.
The development of new information technology has significant effects on the health care system, and its implementation and the associated change management process can bring some positive changes and gains in understanding, but there are challenges with making the transition. These benefits and challenges are explored in the context of a hospital based dental department. Additionally, the concept of the integration of oral health to overall systemic health is explored in context with an Electronic Medical Records system implementation, and the American Dental Association's recent recognition of dental anesthesiology as a clinical subspecialty. Qualitative survey of attending dental faculty members of the department, who represent a broad range of dental specialties and experience in private practice, hospital based practice, teaching, and public health practice. The faculty survey yielded some consistent themes, ranging from enhanced information to make better diagnoses, to challenges in transitioning to EMR, as well as concerns about data security and too much time and effort in front of a computer screen. A brief summary of the history of the stand-alone development of dentistry is given, which contributed to the separate development of dental EMRs from hospital EMRs. The various modalities of clinical care provided by the Department of Dentistry at Advocate Illinois Masonic Medical Center, Chicago, IL are presented to give a scope of the areas of need a successful EMR solution must meet in a hospital based dental setting. Public health aspects are included in the discussion. Macro level health data sets (ie NHANES, state level datasets) have the potential to be expanded to include more thorough data, combining medical health data and oral health data in the same datasets.


Spatial beam shaping of ultrashort laser pulses: theory and experiment.
We studied the spatial intensity profile of an ultrashort laser pulse passing through a laser beam shaping system, which uses diffractive optical elements to reshape a Gaussian beam profile into a flat-topped distribution. Both dispersion and nonlinear self-phase modulation are included in the theoretical model. Our calculation shows that this system works well for ultrashort pulses (approximately 100 fs) when the pulse peak intensity is less than 5 x 10(11) W/cm2. Experimental results are presented for 136 fs pulses at 800 nm wavelength from a Ti:sapphire laser with a 6 nJ pulse energy. We also studied the effects of lateral misalignment, beam-size deviation, and defocusing on the energy fluence profile.


The function of a TiO2 compact layer in dye-sensitized solar cells incorporating "planar" organic dyes.
We present a device based study into the operation of liquid electrolyte dye-sensitized solar cells (DSSC's) using organic dyes. We find that, for these systems, it is entirely necessary to employ a compact TiO2 layer between the transparent fluorine doped SnO2 (FTO) anode and the electrolyte in order to reduce charge recombination losses. By incorporation of a compact layer, the device efficiency can be increased by over 160% under simulated full sun illumination and more than doubled at lower light intensities. This is strong evidence that the more widely employed ruthenium based sensitizers act as to "insulate" the anode against recombination losses and that many planar organic dyes employed in DSSC's could greatly benefit from the use of a compact TiO2 blocking layer. This is in strong contrast to DSSC's sensitized with ruthenium based systems, where the introduction of compact TiO2 has only marginal effects on conversion efficiencies.


Quantitative and functional alterations of 6-sulfo LacNac dendritic cells in multiple myeloma.
Multiple myeloma (MM) results from expansion of abnormal plasma cells in the bone marrow (BM). Previous studies have shown that monocytes play a crucial role in MM pathophysiology. A 6-sulfo LacNAc-expressing population of dendritic cells (Slan-DCs) that overlaps with intermediate and non-classical monocytes in terms of phenotype has been described. Slan-DCs represent a circulating and tissue proinflammatory myeloid population which has been shown to play a role in different cancer contexts, and which exhibits a remarkable plasticity. Herein, we studied Slan-DCs from the BM and blood of MM patients. We performed quantitative and functional analyses of these cells from 54 patients with newly diagnosed, symptomatic MM, 21 patients with MGUS and 24 responding MM patients. We found that circulating Slan-DCs were significantly decreased in MM patients as compared to those of healthy donors or patients with MGUS, while CD14+CD16+ intermediate monocytes accumulate in the BM. Moreover, after activation with TLR7/8 ligand R848, IL-12-producing Slan-DCs from the BM or peripheral blood from MM patients were decreased as compared with healthy donors. We show that MM cell lines or MM cells isolated from patients at diagnosis were able to inhibit the production of IL-12 by Slan-DCs, as well as to shift the phenotype of Slan-DCs towards an intermediate monocyte-like phenotype. Finally, Slan-DCs that have been cultured with MM cells reduced their capacity to induce T cell proliferation and Th1 polarization. We conclude that Slan-DCs represent previously unrecognized players in MM development and may represent a therapeutic target.


Challenges and proposed improvements for reviewing symptoms and catheter use to identify National Healthcare Safety Network catheter-associated urinary tract infections.
Retrospective medical record review is used to categorize urinary tract infections (UTIs) as symptomatic, catheter-associated, and/or healthcare-associated to generate National Healthcare Safety Network (NHSN) surveillance and claims data. We assessed how often patients with UTI diagnoses in claims data had a catheter in place, had documented symptoms, or met the NHSN criteria for catheter-associated UTI (CAUTI). Two physicians retrospectively reviewed medical records for 294 randomly selected patients hospitalized with UTI as a secondary diagnosis, discharged between October 2008 and September 2009 from the University of Michigan. We applied a modification of recent NHSN criteria to estimate how often UTIs in claims data may be an NHSN CAUTI. The 294 patients included 193 women (66%). The mean patient age was 63 years, and the median length of hospital stay was 7.5 days. Catheter use was noted for 216 of 294 postadmission records (74%), including 126 (43%) with a Foley catheter. NHSN symptoms were noted in 113 records (38%); 62 (21%) had symptoms other than fever. Of 136 hospitalizations meeting urine culture criteria, 17 (5.8%) met the criteria for a potential NHSN CAUTI. Retrospective medical record review to identify symptoms and catheter use is complicated and resource-intensive. Requiring standard documentation of symptoms and catheter status when ordering urine cultures could simplify and improve CAUTI surveillance and its fidelity as a hospital quality indicator.


Chronic mTOR inhibition in mice with rapamycin alters T, B, myeloid, and innate lymphoid cells and gut flora and prolongs life of immune-deficient mice.
The mammalian (mechanistic) target of rapamycin (mTOR) regulates critical immune processes that remain incompletely defined. Interest in mTOR inhibitor drugs is heightened by recent demonstrations that the mTOR inhibitor rapamycin extends lifespan and healthspan in mice. Rapamycin or related analogues (rapalogues) also mitigate age-related debilities including increasing antigen-specific immunity, improving vaccine responses in elderly humans, and treating cancers and autoimmunity, suggesting important new clinical applications. Nonetheless, immune toxicity concerns for long-term mTOR inhibition, particularly immunosuppression, persist. Although mTOR is pivotal to fundamental, important immune pathways, little is reported on immune effects of mTOR inhibition in lifespan or healthspan extension, or with chronic mTOR inhibitor use. We comprehensively analyzed immune effects of rapamycin as used in lifespan extension studies. Gene expression profiling found many and novel changes in genes affecting differentiation, function, homeostasis, exhaustion, cell death, and inflammation in distinct T- and B-lymphocyte and myeloid cell subpopulations. Immune functions relevant to aging and inflammation, and to cancer and infections, and innate lymphoid cell effects were validated in vitro and in vivo. Rapamycin markedly prolonged lifespan and healthspan in cancer- and infection-prone mice supporting disease mitigation as a mechanism for mTOR suppression-mediated longevity extension. It modestly altered gut metagenomes, and some metagenomic effects were linked to immune outcomes. Our data show novel mTOR inhibitor immune effects meriting further studies in relation to longevity and healthspan extension.


The pathogenesis of the bone disease of multiple myeloma.
Multiple myeloma is a fatal hematologic malignancy associated with clonal expansion of malignant plasma cells within the bone marrow and the development of a destructive osteolytic bone disease. The principal cellular mechanisms involved in the development of myeloma bone disease are an increase in osteoclastic bone resorption, and a reduction in bone formation. Myeloma cells are found in close association with sites of active bone resorption, and the interactions between myeloma cells and other cells within the specialized bone marrow microenvironment are essential, both for tumor growth and the development of myeloma bone disease. This review discusses the many different factors which have been implicated in myeloma bone disease, including the evidence for their role in myeloma and subsequent therapeutic implications.


Poverty, Dietary Intake, Intestinal Parasites, and Nutritional Status among School-Age Children in the Rural Philippines.
Intestinal helminths are endemic throughout the Philippines; however, there is limited evidence with respect to their prevalence, intensity, and impact on children's nutritional status. A cross-sectional survey was carried out on 693 children from five rural villages in Northern Samar, the Philippines. Data on dietary intake, nutritional status, and intestinal parasites were collected. Infection with Schistosoma japonicum, Ascaris lumbricoides, Trichuris trichiura, and hookworm was evident in 20.1, 54.4, 71.4, and 25.3% of the children. The majority (84.7%) was infected with one or more helminth species, with about one-quarter of the sample (24.7%) infected with three or more. About half (49.2%, n = 341) of the children were stunted and 27.8% (n = 193) were wasted. A lower prevalence of normal height-for-age (48.3%) appeared in those with polyparasitism, while the prevalence of stunted children increased with infection (46.7% monoparasitism and 51.7% polyparasitism). There was a decreasing trend between infection intensity and the mean values of HAZ and BAZ identified for T. trichiura or hookworm infections. Stunted children were more likely to be male (AOR = 1.58; 95% CI: 1.05⁻2.39; p = 0.028), older in age (10⁻14 years) (AOR = 1.93; 95% CI: 1.29⁻2.88; p = 0.001), and living in poorer households with palm leaves/nipa roof (AOR = 1.85; 95% CI: 1.14⁻3.01; p = 0.013). Intestinal parasitic treatment needs to be combined with nutrient supplements and health education in order to interrupt the parasite life cycle and achieve sustainable control.


Ethyleneglycol tungsten complexes of calix[6 and 8]arenes: synthesis, characterization and ROP of ε-caprolactone.
By varying the reaction conditions, the reaction of [W(eg)3] (eg = 1,2-ethanediolato) with p-tert-butylcalix[n]areneHn (n = 6 or 8) in refluxing toluene affords, following work-up, a number of products which have been fully characterized. From the reaction of p-tert-butylcalix[6]areneH6 with one or two equivalents of [W(eg)3], only the oxo-bridged complex {[W(eg)]2(μ-O)p-tert-butylcalix[6]arene} (1) could be isolated, whereas the use of four equivalents of [W(eg)3], in the presence of molecular sieves, afforded {[W(eg)2]2p-tert-butylcalix[6]areneH2}·2MeCN (2); molecules of 2 pack in bi-layers. Under similar conditions, use of one or two equivalents of [W(eg)3] and p-tert-butylcalix[8]areneH8 afforded {[W(eg)]2p-tert-butylcalix[8]arene}·MeCN (3) in which each tungsten centre was bound by four calixarene oxygens. By contrast, the small orange prisms resulting from the use of four equivalents of [W(eg)3] and p-tert-butylcalix[8]areneH8 were shown by synchrotron radiation to be a mixture of two isomers 4a/4b·3.5MeCN). In the major isomer {1,2-[W(eg)2]2p-tert-butylcalix[8]areneH4} (4a), two tungsten centres bind to neighbouring sets of phenolate oxygens, whereas in the minor isomer {1,3-[W(eg)2]2p-tert-butylcalix[8]areneH4} (4b), there is a protonated phenolic group between the two pairs of phenolate oxygens bound to tungsten; the major : minor ratio is about 83 : 17. Use of p-tert-butyltetrahomodioxacalix[6]areneH6 with two equivalents of [W(eg)3] resulted in the isolation of {[WO(eg)]2p-tert-butyltetrahomodioxacalix[6]areneH2} (5·0.83toluene·MeCN), in which each dimethyleneoxa bridge is bound to an oxotungsten(vi) centre. Complexes 1-5, together with the known complex [W(eg)p-tert-butylcalix[4]arene] (6), have been screened for their ability to ring open polymerize (ROP) ε-caprolactone; for 1, 2 and 5, 6 conversion rates were good (>88%) at 110 °C over 12 or 24 h, whereas the calix[8]arene complexes 3 and 4 under the same conditions were inactive.


Silicon graphene Bragg gratings.
We propose the use of interleaved graphene sections on top of a silicon waveguide to implement tunable Bragg gratings. The filter central wavelength and bandwidth can be controlled changing the chemical potential of the graphene sections. Apodization techniques are also presented.


Automated analysis of FDG PET as a tool for single-subject probabilistic prediction and detection of Alzheimer's disease dementia.
To introduce, evaluate and validate a voxel-based analysis method of ¹⁸F-FDG PET imaging for determining the probability of Alzheimer's disease (AD) in a particular individual. The subject groups for model derivation comprised 80 healthy subjects (HS), 36 patients with mild cognitive impairment (MCI) who converted to AD dementia within 18 months, 85 non-converter MCI patients who did not convert within 24 months, and 67 AD dementia patients with baseline FDG PET scan were recruited from the AD Neuroimaging Initiative (ADNI) database. Additionally, baseline FDG PET scans from 20 HS, 27 MCI and 21 AD dementia patients from our institutional cohort were included for model validation. The analysis technique was designed on the basis of the AD-related hypometabolic convergence index adapted for our laboratory-specific context (AD-PET index), and combined in a multivariable model with age and gender for AD dementia detection (AD score). A logistic regression analysis of different cortical PET indexes and clinical variables was applied to search for relevant predictive factors to include in the multivariable model for the prediction of MCI conversion to AD dementia (AD-Conv score). The resultant scores were stratified into sixtiles for probabilistic diagnosis. The area under the receiver operating characteristic curve (AUC) for the AD score detecting AD dementia in the ADNI database was 0.879, and the observed probability of AD dementia in the six defined groups ranged from 8% to 100% in a monotonic trend. For predicting MCI conversion to AD dementia, only the posterior cingulate index, Mini-Mental State Examination (MMSE) score and apolipoprotein E4 genotype (ApoE4) exhibited significant independent effects in the univariable and multivariable models. When only the latter two clinical variables were included in the model, the AUC was 0.742 (95% CI 0.646 - 0.838), but this increased to 0.804 (95% CI 0.714 - 0.894, bootstrap p=0.027) with the addition of the posterior cingulate index (AD-Conv score). Baseline clinical diagnosis of MCI showed 29.7% of converters after 18 months. The observed probability of conversion in relation to baseline AD-Conv score was 75% in the high probability group (sixtile 6), 34% in the medium probability group (merged sixtiles 4 and 5), 20% in the low probability group (sixtile 3) and 7.5% in the very low probability group (merged sixtiles 1 and 2). In the validation population, the AD score reached an AUC of 0.948 (95% CI 0.625 - 0.969) and the AD-Conv score reached 0.968 (95% CI 0.908 - 1.000), with AD patients and MCI converters included in the highest probability categories. Posterior cingulate hypometabolism, when combined in a multivariable model with age and gender as well as MMSE score and ApoE4 data, improved the determination of the likelihood of patients with MCI converting to AD dementia compared with clinical variables alone. The probabilistic model described here provides a new tool that may aid in the clinical diagnosis of AD and MCI conversion.


Single-Cell Analysis Reveals a Close Relationship between Differentiating Dopamine and Subthalamic Nucleus Neuronal Lineages.
Stem cell engineering and grafting of mesencephalic dopamine (mesDA) neurons is a promising strategy for brain repair in Parkinson's disease (PD). Refinement of differentiation protocols to optimize this approach will require deeper understanding of mesDA neuron development. Here, we studied this process using transcriptome-wide single-cell RNA sequencing of mouse neural progenitors expressing the mesDA neuron determinant Lmx1a. This approach resolved the differentiation of mesDA and neighboring neuronal lineages and revealed a remarkably close relationship between developing mesDA and subthalamic nucleus (STN) neurons, while also highlighting a distinct transcription factor set that can distinguish between them. While previous hESC mesDA differentiation protocols have relied on markers that are shared between the two lineages, we found that application of these highlighted markers can help to refine current stem cell engineering protocols, increasing the proportion of appropriately patterned mesDA progenitors. Our results, therefore, have important implications for cell replacement therapy in PD.


Multiplexed labeling of genomic loci with dCas9 and engineered sgRNAs using CRISPRainbow.
A lack of techniques to image multiple genomic loci in living cells has limited our ability to investigate chromosome dynamics. Here we describe CRISPRainbow, a system for labeling DNA in living cells based on nuclease-dead (d) Cas9 combined with engineered single guide RNA (sgRNA) scaffolds that bind sets of fluorescent proteins. We demonstrate simultaneous imaging of up to six chromosomal loci in individual live cells and document large differences in the dynamic properties of different chromosomal loci.


Intracranial hemorrhage is much more common after carotid stenting than after endarterectomy: evidence from the National Inpatient Sample.
Intracranial hemorrhage (ICH) is a rare and devastating complication of carotid revascularization. We sought to determine the prevalence of, type of, and risk factors associated with ICH among recipients of carotid endarterectomy (CEA) and carotid angioplasty and stenting (CAS) within the National Inpatient Sample (NIS). Postoperative cases of ICH after CEA (International Classification of Disease 9th edition [ICD-9]: 38.12) or CAS (ICD-9: 00.63) were retrieved from the 2001 to 2008 NIS. Clinical presentation (asymptomatic versus symptomatic), discharge status, in-hospital mortality, demographics, and hospital characteristics were extracted from NIS data. Charlson indices of comorbidity were determined based on ICD-9 and clinical classification software codes. Multivariate regression was used to determine the impact of revascularization procedure type and symptom status on adverse outcomes, including ICH, in-hospital mortality, and unfavorable discharge status. Among 57,663,486 NIS hospital admissions, 215,012 CEA and 13,884 CAS procedures were performed. Symptomatic presentations represented the minority of CEA (N=10,049; 5%) and CAS cases (N=1251; 10%). ICH occurred significantly more frequently after CAS than CEA in both symptomatic (4.4% versus 0.8%; P<0.0001) and asymptomatic presentations (0.5% versus 0.06%; P<0.0001). Multivariate regression suggested that symptomatic presentations (versus asymptomatic) and CAS procedures (versus CEA) were both independently predictive of 6-fold to 7-fold increases in the frequency of postoperative ICH. ICH was independently predictive in a 30-fold increased risk of mortality before discharge. CAS procedures are associated with elevated adverse outcomes, including ICH, in-hospital death, and unfavorable discharges, especially among symptomatic presentations.


Impact of phase on collision between vortex solitons in three-dimensional cubic-quintic complex Ginzburg-Landau equation.
We present a systematic analysis for three generic collisional outcomes between stable dissipative vortices with intrinsic vorticity S = 0, 1, or 2 upon variation of relative phase in the three-dimensional (3D) cubic-quintic complex Ginzburg-Landau equation. The first type outcome is merger of the vortices into a single one, of which velocity can be effectively controlled by relative phase. With the increase of the collision momentum, the following is creation of an extra vortex, and its velocity also increases with growth of relative phase. However, at largest collision momentum, the variety of relative phase cannot change the third type collisional outcomes, quasielastic interaction. In addition, the dynamic range of the outcome of creating an extra vortex decreases with the reduction of cubic-gain. The above features have potential applications in optical switching and logic gates based on interaction of optical solitons.


Isolated, well-defined organovanadium(iii) on silica: single-site catalyst for hydrogenation of alkenes and alkynes.
A well-defined, isolated, single-site organovanadium(iii) catalyst on SiO2 [(SiO2)V(Mes)(THF)] was synthesized via surface organometallic chemistry, and fully characterized using a combination of analytical and spectroscopic techniques (EA, ICP, 1H NMR, TGA-MS, EPR, XPS, DR-UV/Vis, UV-Raman, DRIFTS, XAS). The catalyst exhibits unprecedented reactivity in liquid- and gas-phase alkene/alkyne hydrogenation. Kinetic poisoning experiments revealed that 100% of the V sites are active for hydrogenation.


In vitro anti-listerial activities of crude n-hexane and aqueous extracts of Garcinia kola (heckel) seeds.
We assessed the anti-Listerial activities of crude n-hexane and aqueous extracts of Garcinia kola seeds against a panel of 42 Listeria isolates previously isolated from wastewater effluents in the Eastern Cape Province of South Africa and belonging to Listeria monocytogenes, Listeria grayi and Listeria ivanovii species. The n-hexane fraction was active against 45% of the test bacteria with zones of inhibition ranging between 8-17 mm, while the aqueous fraction was active against 29% with zones of inhibition ranging between 8-11 mm. The minimum inhibitory concentrations (MIC) were within the ranges of 0.079-0.625 mg/mL for the n-hexane extract and 10 to >10 mg/mL for the aqueous extract. The rate of kill experiment carried out for the n-hexane extract only, revealed complete elimination of the initial bacterial population for L. grayi (LAL 15) at 3× and 4× MIC after 90 and 60 min; L. monocytogenes (LAL 8) at 3× and 4× MIC after 60 and 15 min; L. ivanovii (LEL 18) at 3× and 4× MIC after 120 and 15 min; L. ivanovii (LEL 30) at 2, 3 and 4× MIC values after 105, 90 and 15 min exposure time respectively. The rate of kill activities were time- and concentration-dependant and the extract proved to be bactericidal as it achieved a more than 3log(10) decrease in viable cell counts after 2 h exposure time for all of the four test organisms at 3× and 4× MIC values. The results therefore show the potential presence of anti-Listerial compounds in Garcinia kola seeds that can be exploited in effective anti-Listerial chemotherapy.


Management of patients with multiple myeloma in the era of COVID-19 pandemic: a consensus paper from the European Myeloma Network (EMN).
Patients with multiple myeloma (MM) seem to be at increased risk for more severe COVID-19 infection and associated complications due to their immunocompromised state, the older age and comorbidities. The European Myeloma Network has provided an expert consensus statement in order to guide therapeutic decisions in the era of the COVID-19 pandemic. Patient education for personal hygiene and social distancing measures, along with treatment individualization, telemedicine and continuous surveillance for early diagnosis of COVID-19 are essential. In countries or local communities where COVID-19 infection is widely spread, MM patients should have a PCR test of nasopharyngeal swab for SARS-CoV-2 before hospital admission, starting a new treatment line, cell apheresis or ASCT in order to avoid ward or community spread and infections. Oral agent-based regimens should be considered, especially for the elderly and frail patients with standard risk disease, whereas de-intensified regimens for dexamethasone, bortezomib, carfilzomib and daratumumab should be used based on patient risk and response. Treatment initiation should not be postponed for patients with end organ damage, myeloma emergencies and aggressive relapses. Autologous (and especially allogeneic) transplantation should be delayed and extended induction should be administered, especially in standard risk patients and those with adequate MM response to induction. Watchful waiting should be considered for standard risk relapsed patients with low tumor burden, and slow biochemical relapses. The conduction of clinical trials should continue with appropriate adaptations to the current circumstances. Patients with MM and symptomatic COVID-19 disease should interrupt anti-myeloma treatment until recovery. For patients with positive PCR test for SARS-CoV-2, but with no symptoms for COVID-19, a 14-day quarantine should be considered if myeloma-related events allow the delay of treatment. The need for surveillance for drug interactions due to polypharmacy is highlighted. The participation in international COVID-19 cancer registries is greatly encouraged.


Complementary and alternative medicine in VA specialized PTSD treatment programs.
This survey documented the provision of complementary and alternative medicine (CAM) treatments in U.S. Department of Veterans Affairs (VA) specialized posttraumatic stress disorder (PTSD) treatment programs. Program coordinators or designated staff from 125 of 170 VA specialized PTSD treatment programs completed and returned surveys between September 2010 and March 2011, indicating which of 32 CAM treatments were offered in that program. Ninety-six percent of programs reported use of at least one CAM treatment. Eighty-eight percent offered CAMs other than those that are commonly part of conventional PTSD treatments (guided imagery, progressive muscle relaxation, and stress management-relaxation therapies). The widespread use of CAM treatments in VA PTSD programs presents an opportunity for researchers to assess the effect of CAM on mental health service use and PTSD symptoms among veterans. Future research should assess the effectiveness of CAM treatments and develop methods to tailor these treatments to veterans with PTSD.


A methodology to quantify alterations in human upper limb movement during co-manipulation with an exoskeleton.
While a large number of robotic exoskeletons have been designed by research teams for rehabilitation, it remains rather difficult to analyse their ability to finely interact with a human limb: no performance indicators or general methodology to characterize this capacity really exist. This is particularly regretful at a time when robotics are becoming a recognized rehabilitation method and when complex problems such as 3-D movement rehabilitation and joint rotation coordination are being addressed. The aim of this paper is to propose a general methodology to evaluate, through a reduced set of simple indicators, the ability of an exoskeleton to interact finely and in a controlled way with a human. The method involves measurement and recording of positions and forces during 3-D point to point tasks. It is applied to a 4 degrees-of-freedom limb exoskeleton by way of example.


Genetic Variation of Lactobacillus delbrueckii subsp. lactis Bacteriophages Isolated from Cheese Processing Plants in Finland.
The genomes of four Lactobacillus delbrueckii subsp. lactis bacteriophages were characterized by restriction endonuclease mapping, Southern hybridization, and heteroduplex analysis. The phages were isolated from different cheese processing plants in Finland between 1950 and 1972. All four phages had a small isometric head and a long noncontractile tail. Two different types of genome (double-stranded DNA) organization existed among the different phages, the pac type and the cos type, corresponding to alternative types of phage DNA packaging. Three phages belonged to the pac type, and a fourth was a cos-type phage. The pac-type phages were genetically closely related. In the genomes of the pac-type phages, three putative insertion/deletions (0.7 to 0.8 kb, 1.0 kb, and 1.5 kb) and one other region (0.9 kb) containing clustered base substitutions were discovered and localized. At the phenotype level, three main differences were observed among the pac-type phages. These concerned two minor structural proteins and the efficiency of phage DNA packaging. The genomes of the pac-type phages showed only weak homology with that of the cos-type phage. Phage-related DNA, probably a defective prophage, was located in the chromosome of the host strain sensitive to the cos-type phage. This DNA exhibited homology under stringent conditions to the pac-type phages.


Multiple Geographical Origins of Environmental Sex Determination enhanced the diversification of Darwin's Favourite Orchids.
Environmental sex determination (ESD) - a change in sexual function during an individual life span driven by environmental cues - is an exceedingly rare sexual system among angiosperms. Because ESD can directly affect reproduction success, it could influence diversification rate as compared with lineages that have alternative reproductive systems. Here we test this hypothesis using a solid phylogenetic framework of Neotropical Catasetinae, the angiosperm lineage richest in taxa with ESD. We assess whether gains of ESD are associated with higher diversification rates compared to lineages with alternative systems while considering additional traits known to positively affect diversification rates in orchids. We found that ESD has evolved asynchronously three times during the last ~5 Myr. Lineages with ESD have consistently higher diversification rates than related lineages with other sexual systems. Habitat fragmentation due to mega-wetlands extinction, and climate instability are suggested as the driving forces for ESD evolution.


Computational pharmacokinetics of solute penetration into human intervertebral discs - effects of endplate permeability, solute molecular weight and disc size.
A finite element model is developed to predict the penetration time-history of three different solutes into the human lumbar disc following intravenous injection. Antibiotics are routinely administered intravenously in spinal surgery to prevent disc infection. Successful prophylaxis requires antibiotics to reach adequate inhibitory levels. Here, the transient diffusion of cephazolin is investigated over 10h post-injection in a human disc model subject to reported concentrations in the blood stream as the prescribed boundary sources. Post-injection variation of cephazolin concentrations in the disc adjacent to supply sources closely followed the decay curve in the blood stream and fell sharply with time. Much lower concentrations were computed in the inner annulus and nucleus; much of the disc (80% at 1h and 49% at 4h) experienced concentrations below required inhibitory level of 1mg/L in agreement with measurements. Changes in endplate permeability, disc size, and solute molecular weight had profound effects on concentration profiles at all times and regions, especially in the disc centre, demonstrating their crucial roles on the adequate delivery of drugs. Larger solutes markedly slow transport into the disc. The failure to reach critical therapeutic levels in the central disc regions, especially when endplates calcify and in larger discs, raises concerns and calls for caution in attempts to extrapolate findings of studies on animals with much smaller and non degenerate discs to the human discs. The current study also demonstrates the capability of computational models in predicting the transport of intravenously injected solutes into the disc.


Dynamics of the endoplasmic reticulum and other membranous organelles in growth cones of cultured neurons.
The fluorescent lipophilic dye 3,3'-dihexyloxacarbocyanine iodide [DiOC6(3)] was used to examine the distribution of membrane-bound organelles in growth cones of cultured rat sympathetic neurons. Within chemically fixed growth cones, intense DiOC6(3) fluorescence was localized predominately to the base or central region of growth cones. However, in most growth cones several thin DiOC6(3)-fluorescent processes radiated from the base into the periphery, and double fluorescence imaging of single growth cones indicated that these processes were highly colocalized (approximately 79%) with microtubules. The distribution of DiOC6(3) fluorescence in living growth cones was examined using low light-level fluorescence video microscopy. We observed thin fluorescent processes within the periphery of growth cones to undergo length excursions (extension/retraction) and to change orientation (move laterally). During growth cone advance, processes became progressively thicker and were gradually engulfed by the advancing fluorescent mass. When growth cones were viewed with video-enhanced differential interference contrast microscopy, the position of the fluorescent processes correlated with thickened extensions of central-type cytoplasm through which vesiclelike organelle transport often occurred. These observations indicate several features concerning the organization and movement of membranous organelles (MOs) in growth cones: (1) MOs are highly compartmentalized, the majority being localized to the growth cone base; (2) MOs advance into the periphery along distinct pathways probably associated with microtubules; (3) one or more thin continuous MOs, which most likely represent a thin tubular component of the endoplasmic reticulum, generally precedes advance of vesiclelike MOs along individual transport pathways; and (4) transport pathways with their associated MOs are spatially and temporally dynamic.


Biochemical findings in sarcoidosis.
The majority of patients with sarcoidosis in this large series have had a number of biochemical investigations performed. Abnormal calcium metabolism was demonstrated in 40% of the patients but permanent renal damage due to nephrocalcinosis as a result of persistent derangement of calcium metabolism was rare. Raised immunoglobulin levels were seen. Half the white and two-thirds of the West Indian patients had elevated IgG levels. Abnormal immunoglobulin levels carried no obvious diagnostic or prognostic significance. Raised alkaline phosphatase levels reflected space-occupying hepatic granulomas and occurred in 23% of patients. Serum angiotensin converting enzyme (SACE) was elevated in half the patients. The highest SACE activity was found in patients with severe parenchymal lung infiltration due to sarcoidosis, and the lowest levels in those with inactive disease or after successful management with steroid drugs. SACE levels were not significantly elevated in four other granulomatous conditions: Crohn's disease, primary biliary cirrhosis, Hodgkin's disease, and active tuberculosis.


Development of a repository of individual participant data from randomized controlled trials of therapists delivered interventions for low back pain.
Individual patient data (IPD) meta-analysis of existing randomized controlled trials (RCTs) is a promising approach to achieving sufficient statistical power to identify sub-groups. We created a repository of IPD from multiple low back pain (LBP) RCTs to facilitate a study of treatment moderators. Due to sparse heterogeneous data, the repository needed to be robust and flexible to accommodate millions of data points prior to any subsequent analysis. We systematically identified RCTs of therapist delivered intervention for inclusion to the repository. Some were obtained through project publicity. We requested both individual items and aggregate scores of all baseline characteristics and outcomes for all available time points. The repository is made up of a hybrid database: entity-attribute-value and relational database which is capable of storing sparse heterogeneous datasets. We developed a bespoke software program to extract, transform and upload the shared data. There were 20 datasets with more than 3 million data points from 9328 participants. All trials collected covariates and outcomes data at baseline and follow-ups. The bespoke standardized repository is flexible to accommodate millions of data points without compromising data integrity. Data are easily retrieved for analysis using standard statistical programs. The bespoke hybrid repository is complex to implement and to query but its flexibility in supporting datasets with varying sets of responses and outcomes with different data types is a worthy trade off. The large standardized LBP dataset is also an important resource useable by other LBP researchers. A flexible adaptive database for pain studies that can easily be expanded for future researchers to map, transform and upload their data in a safe and secure environment. The data are standardized and harmonized which will facilitate future requests from other researchers for secondary analyses.


Biomechanical evaluation of the suture anchors used in open-door laminoplasty: a cadaveric study.
A cadaveric study. To determine whether the use of suture anchors is warranted in cervical laminoplasty. The use of suture anchors to stabilize elevated laminae has been popularized in laminoplasty. However, the validity of using suture anchors in laminoplasty has not been determined. Six intact fresh frozen cadavers were used. Open-door laminoplasty with a hinge on the cadaver's left side was performed on levels C3-C7. Elevated laminae were stabilized by suture anchors equipped with strain gauges, which were placed on C3, C5, and C7 left lateral masses. After surgery, the cervical spine was manually loaded passively, and the mechanical loads on each suture anchor during each motion were measured. Finally, the incision was opened again, and the failure loads of the suture anchors were also measured. After cervical loading, all elevated laminae were confirmed to be intact without dislodgement or failure of the suture anchors. The loads during left rotation and left bending were significantly higher than those during the respective motion to the right at all levels, except in rotation at C3. The loads on the C5 anchors in flexion and left rotation and on the C7 anchors in extension were relatively high. The maximum load obtained in the present study was 14.9 N, which was one order of magnitude lower than the mean failure load of the suture anchors (131.7 N). Biomechanical laterality was demonstrated, reflecting the asymmetrical nature of open-door laminoplasty. The maximum load on the suture anchors was much lower than the failure load and was consistent with the stability of the suture anchors encountered in clinical cases. This may support the validity of using suture anchors in laminoplasty, although the loads during active motion may be higher than our results. N/A.


Molecular and Biochemical Differences of the Tandem and Cold-Adapted PET Hydrolases Ple628 and Ple629, Isolated From a Marine Microbial Consortium.
Polybutylene adipate terephthalate (PBAT) is a biodegradable alternative to polyethylene and can be broadly used in various applications. These polymers can be degraded by hydrolases of terrestrial and aquatic origin. In a previous study, we identified tandem PETase-like hydrolases (Ples) from the marine microbial consortium I1 that were highly expressed when a PBAT blend was supplied as the only carbon source. In this study, the tandem Ples, Ple628 and Ple629, were recombinantly expressed and characterized. Both enzymes are mesophilic and active on a wide range of oligomers. The activities of the Ples differed greatly when model substrates, PBAT-modified polymers or PET nanoparticles were supplied. Ple629 was always more active than Ple628. Crystal structures of Ple628 and Ple629 revealed a structural similarity to other PETases and can be classified as member of the PETases IIa subclass, α/β hydrolase superfamily. Our results show that the predicted functions of Ple628 and Ple629 agree with the bioinformatic predictions, and these enzymes play a significant role in the plastic degradation by the consortium.


Calcification response of a key phytoplankton family to millennial-scale environmental change.
Coccolithophores are single-celled photosynthesizing marine algae, responsible for half of the calcification in the surface ocean, and exert a strong influence on the distribution of carbon among global reservoirs, and thus Earth's climate. Calcification in the surface ocean decreases the buffering capacity of seawater for CO2, whilst photosynthetic carbon fixation has the opposite effect. Experiments in culture have suggested that coccolithophore calcification decreases under high CO2 concentrations ([CO2(aq)]) constituting a negative feedback. However, the extent to which these results are representative of natural populations, and of the response over more than a few hundred generations is unclear. Here we describe and apply a novel rationale for size-normalizing the mass of the calcite plates produced by the most abundant family of coccolithophores, the Noëlaerhabdaceae. On average, ancient populations subjected to coupled gradual increases in [CO2(aq)] and temperature over a few million generations in a natural environment become relatively more highly calcified, implying a positive climatic feedback. We hypothesize that this is the result of selection manifest in natural populations over millennial timescales, so has necessarily eluded laboratory experiments.


Patterns of spatial and temporal distribution of humpback whales at the southern limit of the Southeast Pacific breeding area.
Understanding the patterns of spatial and temporal distribution in threshold habitats of highly migratory and endangered species is important for understanding their habitat requirements and recovery trends. Herein, we present new data about the distribution of humpback whales (Megaptera novaeangliae) in neritic waters off the northern coast of Peru: an area that constitutes a transitional path from cold, upwelling waters to warm equatorial waters where the breeding habitat is located. Data was collected during four consecutive austral winter/spring seasons from 2010 to 2013, using whale-watching boats as platforms for research. A total of 1048 whales distributed between 487 groups were sighted. The spatial distribution of humpbacks resembled the characteristic segregation of whale groups according to their size/age class and social context in breeding habitats; mother and calf pairs were present in very shallow waters close to the coast, while dyads, trios or more whales were widely distributed from shallow to moderate depths over the continental shelf break. Sea surface temperatures (range: 18.2-25.9°C) in coastal waters were slightly colder than those closer to the oceanic realm, likely due to the influence of cold upwelled waters from the Humboldt Current system. Our results provide new evidence of the southward extension of the breeding region of humpback whales in the Southeast Pacific. Integrating this information with the knowledge from the rest of the breeding region and foraging grounds would enhance our current understanding of population dynamics and recovery trends of this species.


Influence of alpha-adrenergic blockade on the catecholamine response to exercise at 4,300 meters.
This investigation examined the influence of alpha-adrenergic blockade on plasma and urinary catecholamine responses to both exercise and high-altitude exposure. Sixteen nonsmoking, eumenorrheic women (age 23.2 +/- 1.4 years, 68.7 +/- 1.0 kg) were studied at sea level and during 12 days of high-altitude exposure (4,300 m). Subjects received either alpha-blockade (prazosin 3 mg/d) or a placebo in a double-blinded, randomized fashion. Resting plasma and 24-hour urine samples were collected periodically throughout the duration of the study. Further, subjects participated in submaximal exercise tests (50 minutes at 50% sea level maximum oxygen consumption [Vo2max]) at Sea level and on days 1 and 12 at altitude. Urinary norepinephrine (NE) excretion rates increased significantly over time at altitude, with blocked subjects having greater values compared to controls. Plasma NE levels increased significantly with chronic altitude exposure compared to sea level and acute hypoxia both at rest and during exercise. NE levels at rest were greater for blocked compared to control subjects during all conditions. Urinary and plasma epinephrine (EPI) levels increased dramatically, with acute altitude exposure returning to sea level values by day 12 of altitude exposure. EPI levels were greater for blocked compared to placebo both at rest and during exercise for all conditions studied. Changes in alpha-adrenergic activity over time at altitude were associated with select metabolic and physiologic adjustments. The presence of alpha-blockade significantly affected these responses during chronic altitude exposure. It was concluded that: (1) alpha-adrenergic blockade elicited a potentiated sympathoadrenal response to the stress of both exercise as well as high-altitude exposure, and (2) the sympathetics, via alpha-adrenergic stimulation, contribute to a number of key adaptations associated with acclimatization to high altitude.


Visualizing the actin cytoskeleton in living plant cells using a photo-convertible mEos::FABD-mTn fluorescent fusion protein.
The actin cytoskeleton responds quickly to diverse stimuli and plays numerous roles in cellular signalling, organelle motility and subcellular compartmentation during plant growth and development. Molecular and cell biological tools that can facilitate visualization of actin organization and dynamics in a minimally invasive manner are essential for understanding this fundamental component of the living cell. A novel, monomeric (m) Eos-fluorescent protein derived from the coral Lobophyllia hemprichii was assessed for its green to red photo-convertibility in plant cells by creating mEosFP-cytosolic. mEosFP was fused to the F-(filamentous)-Actin Binding Domain of the mammalian Talin gene to create mEosFP::FABDmTalin. Photo-conversion, visualization and colour quantification protocols were developed for EosFP targeted to the F-actin cytoskeleton. Rapid photo-conversion in the entire cell or in a region of interest was easily achieved upon illumination with an approximately 400 nm wavelength light beam using an epi-fluorescent microscope. Dual color imaging after photo-conversion was carried out using a confocal laser-scanning microscope. Time-lapse imaging revealed that although photo-conversion of single mEosFP molecules can be rapid in terms of live-cell imaging it involves a progressive enrichment of red fluorescent molecules over green species. The fluorescence of photo-converted cells thus progresses through intermediate shades ranging from green to red. The time taken for complete conversion to red fluorescence depends on protein expression level within a cell and the quality of the focusing lens used to deliver the illuminating beam. Three easily applicable methods for obtaining information on fluorescent intensity and colour are provided as a means of ensuring experimental repeatability and data quantification, when using mEosFP and similar photo-convertible proteins. The mEosFP::FABD-mTn probe retains all the imaging qualities associated with the well tested GFP::mTn probe while allowing for non-invasive, regional photo-conversion that allows colour based discrimination within a living cell. Whereas a number of precautions should be exercised in dealing with photo-convertible probes, mEosFP::FABD-mTn is a versatile live imaging tool for dissecting the organization and activity of the actin cytoskeleton in plants.


Glioblastoma stem cells.
Glioblastomas are highly malignant primary brain tumors with one of the worst survival rates among all human cancers. With a more profound understanding of the cellular and molecular mechanisms of tumor initiation and acquired resistance to conventional radio- and chemotherapy, novel therapeutic targets might be discovered to optimize therapeutic approaches. In this regard, the identification of a small cellular subpopulation, called glioblastoma stem cell or stem-like cells or glioma-initiating cells or brain tumor propagating cells, has gained attention. In this article, we briefly summarize the current state of knowledge about this tumor cell population and discuss future directions for basic and clinical research.


What is known about tobacco industry efforts to influence tobacco tax? A systematic review of empirical studies.
To systematically review studies of tobacco industry efforts to influence tobacco tax policies. Searches were conducted between 1 October 2009 and 31 March 2010 in 14 databases/websites, in relevant bibliographies and via experts. Studies were included if they focused on industry efforts to influence tobacco tax policies, drew on empirical evidence, were in English and concerned the period 1985-2010. In total, 36 studies met these criteria. Two reviewers undertook data extraction and critical appraisal. A random selection of 15 studies (42%) was subject to second review. Evidence was assessed thematically to identify distinct tobacco industry aims, arguments and tactics. A total of 34 studies examined industry efforts to influence tax levels. They suggest the tobacco industry works hard to prevent significant increases and particularly dislikes taxes 'earmarked' for tobacco control. Key arguments to counter increases are that tobacco taxes are socially regressive, unfair and lead to increased levels of illicit trade and negative economic impacts. For earmarked taxes, the industry also frequently tries to raise concerns about revenue allocation. Assessing industry arguments against established evidence demonstrates most are unsupported. Key industry tactics include: establishing 'front groups', securing credible allies, direct lobbying and publicity campaigns. Only seven studies examined efforts to influence tax structures. They suggest company preferences vary and tactics centre on direct lobbying. The tobacco industry has historically tried to keep tobacco taxes low using consistent tactics and misleading arguments. Further research is required to explore efforts to influence tax structures, excise policies beyond the USA and recent policies.


Mitochondrial CoQ deficiency is a common driver of mitochondrial oxidants and insulin resistance.
Insulin resistance in muscle, adipocytes and liver is a gateway to a number of metabolic diseases. Here, we show a selective deficiency in mitochondrial coenzyme Q (CoQ) in insulin-resistant adipose and muscle tissue. This defect was observed in a range of in vitro insulin resistance models and adipose tissue from insulin-resistant humans and was concomitant with lower expression of mevalonate/CoQ biosynthesis pathway proteins in most models. Pharmacologic or genetic manipulations that decreased mitochondrial CoQ triggered mitochondrial oxidants and insulin resistance while CoQ supplementation in either insulin-resistant cell models or mice restored normal insulin sensitivity. Specifically, lowering of mitochondrial CoQ caused insulin resistance in adipocytes as a result of increased superoxide/hydrogen peroxide production via complex II. These data suggest that mitochondrial CoQ is a proximal driver of mitochondrial oxidants and insulin resistance, and that mechanisms that restore mitochondrial CoQ may be effective therapeutic targets for treating insulin resistance.


Ursolic-Acid-Enriched Herba Cynomorii Extract Protects against Oxidant Injury in H9c2 Cells and Rat Myocardium by Increasing Mitochondrial ATP Generation Capacity and Enhancing Cellular Glutathione Redox Cycling, Possibly through Mitochondrial Uncoupling.
Mitochondrial decay is considered to be a major contributor to aging-related diseases, including neurodegenerative diseases, cardiovascular disorders, and certain metabolic diseases. Therefore, the maintenance of mitochondrial functional capacity and antioxidant status should play an essential role in preventive health. Herba Cynomorii, which is one of the most potent "Yang-invigorating" Chinese tonic herbs, was found to increase mitochondrial ATP generation capacity (ATP-GC) in rat hearts ex vivo. In the present study, we demonstrated that HCY2, an active fraction of Herba Cynomorii, and its major ingredient ursolic acid (UA) could protect against hypoxia/reoxygenation-induced cell apoptosis in H9c2 cells in vitro and also against ischemia/reperfusion-induced injury in rat hearts ex vivo. The cardioprotection was associated with an increase in ATP-GC and an enhancement of glutathione redox cycling. The results suggest that UA may be one of the active ingredients responsible for the cardioprotection afforded by Herba Cynomorii, and this effect may be mediated, at least in part, by enhancement of mitochondrial functional capacity and antioxidant status, possibly through the induction of mitochondrial uncoupling.


Protective effect of total carotenoid and lycopene intake on the risk of hip fracture: a 17-year follow-up from the Framingham Osteoporosis Study.
In vitro and in vivo studies suggest that carotenoids may inhibit bone resorption, yet no previous study has examined individual carotenoid intake (other than beta-carotene) and the risk of fracture. We evaluated associations of total and individual carotenoid intake (alpha-carotene, beta-carotene, beta-cryptoxanthin, lycopene, lutein + zeaxanthin) with incident hip fracture and nonvertebral osteoporotic fracture. Three hundred seventy men and 576 women (mean age, 75 +/- 5 yr) from the Framingham Osteoporosis Study completed a food frequency questionnaire (FFQ) in 1988-1989 and were followed for hip fracture until 2005 and nonvertebral fracture until 2003. Tertiles of carotenoid intake were created from estimates obtained using the Willett FFQ adjusting for total energy (residual method). HRs were estimated using Cox-proportional hazards regression, adjusting for sex, age, body mass index, height, total energy, calcium and vitamin D intake, physical activity, alcohol, smoking, multivitamin use, and current estrogen use. A total of 100 hip fractures occurred over 17 yr of follow-up. Subjects in the highest tertile of total carotenoid intake had lower risk of hip fracture (p = 0.02). Subjects with higher lycopene intake had lower risk of hip fracture (p =0.01) and nonvertebral fracture (p = 0.02). A weak protective trend was observed for total beta-carotene for hip fracture alone, but associations did not reach statistical significance (p = 0.10). No significant associations were observed with alpha-carotene, beta-cryptoxanthin, or lutein + zeaxanthin. These results suggest a protective role of several carotenoids for bone health in older adults.


Vibrating Mesh Nebulizer Compared With Metered-Dose Inhaler in Mechanically Ventilated Subjects.
The impact of various aerosol delivery devices on patient outcomes during mechanical ventilation is unknown. If one method of delivery results in a higher ventilator-associated pneumonia (VAP) rate than another, multiple patient outcomes may be affected. This study aimed to determine whether there was a difference in VAP occurrence and patient outcomes (days receiving ventilation and in-hospital mortality) between the vibrating mesh nebulizer (AeroNeb Solo) and the metered-dose inhaler (MDI). This retrospective study reviewed medical records for all mechanically ventilated, adult patients with an order for aerosol treatment from August 2011 to August 2013. The hospital converted from MDI to vibrating mesh nebulizers in August 2012, and data were gathered 1 y before/after conversion. Excluded were patients with a tracheostomy, patients who were mechanically ventilated for <24 h, patients who received a combination of nebulizer and MDI treatments, or patients who were re-intubated. Two hundred twenty-eight subjects were included. Forty-eight (21%) received treatment with an MDI, and 180 (79%) were treated with the vibrating mesh nebulizer. Descriptive data did not significantly differ for age or APACHE II (Acute Physiology and Chronic Health Evaluation II) scores between the groups but did for sex (P = .03). Difference in median days receiving ventilation for the MDI (5 d, interquartile range 3.0-8.5 d) and the vibrating mesh nebulizer (6 d, interquartile range 4.0-10.0 d) was not statistically significant. No correlation was found between the use of either device and the primary outcomes of VAP and in-hospital mortality. In multivariable logistic regression analysis, the number of days receiving ventilation increased the odds of VAP (odds ratio [OR] 1.3, 95% CI 1.14-1.49, P < .001) and mortality (OR 1.12, 95% CI 1.04-1.21, P = .002). Higher APACHE II scores increased the odds of mortality (OR 1.05, 95% CI 1.001-1.092, P = .044). We found no association between an MDI or vibrating mesh nebulizer and our primary outcomes, days receiving ventilation, in-hospital mortality, or VAP, in mechanically ventilated subjects.


How Nemo finds home: the neuroecology of dispersal and of population connectivity in larvae of marine fishes.
Nearly all demersal teleost marine fishes have pelagic larval stages lasting from several days to several weeks, during which time they are subject to dispersal. Fish larvae have considerable swimming abilities, and swim in an oriented manner in the sea. Thus, they can influence their dispersal and thereby, the connectivity of their populations. However, the sensory cues marine fish larvae use for orientation in the pelagic environment remain unclear. We review current understanding of these cues and how sensory abilities of larvae develop and are used to achieve orientation with particular emphasis on coral-reef fishes. The use of sound is best understood; it travels well underwater with little attenuation, and is current-independent but location-dependent, so species that primarily utilize sound for orientation will have location-dependent orientation. Larvae of many species and families can hear over a range of ~100-1000 Hz, and can distinguish among sounds. They can localize sources of sounds, but the means by which they do so is unclear. Larvae can hear during much of their pelagic larval phase, and ontogenetically, hearing sensitivity, and frequency range improve dramatically. Species differ in sensitivity to sound and in the rate of improvement in hearing during ontogeny. Due to large differences among-species within families, no significant differences in hearing sensitivity among families have been identified. Thus, distances over which larvae can detect a given sound vary among species and greatly increase ontogenetically. Olfactory cues are current-dependent and location-dependent, so species that primarily utilize olfactory cues will have location-dependent orientation, but must be able to swim upstream to locate sources of odor. Larvae can detect odors (e.g., predators, conspecifics), during most of their pelagic phase, and at least on small scales, can localize sources of odors in shallow water, although whether they can do this in pelagic environments is unknown. Little is known of the ontogeny of olfactory ability or the range over which larvae can localize sources of odors. Imprinting on an odor has been shown in one species of reef-fish. Celestial cues are current- and location-independent, so species that primarily utilize them will have location-independent orientation that can apply over broad scales. Use of sun compass or polarized light for orientation by fish larvae is implied by some behaviors, but has not been proven. Use of neither magnetic fields nor direction of waves for orientation has been shown in marine fish larvae. We highlight research priorities in this area.


Selected In-Season Nutritional Strategies to Enhance Recovery for Team Sport Athletes: A Practical Overview.
Team sport athletes face a variety of nutritional challenges related to recovery during the competitive season. The purpose of this article is to review nutrition strategies related to muscle regeneration, glycogen restoration, fatigue, physical and immune health, and preparation for subsequent training bouts and competitions. Given the limited opportunities to recover between training bouts and games throughout the competitive season, athletes must be deliberate in their recovery strategy. Foundational components of recovery related to protein, carbohydrates, and fluid have been extensively reviewed and accepted. Micronutrients and supplements that may be efficacious for promoting recovery include vitamin D, omega-3 polyunsaturated fatty acids, creatine, collagen/vitamin C, and antioxidants. Curcumin and bromelain may also provide a recovery benefit during the competitive season but future research is warranted prior to incorporating supplemental dosages into the athlete's diet. Air travel poses nutritional challenges related to nutrient timing and quality. Incorporating strategies to consume efficacious micronutrients and ingredients is necessary to support athlete recovery in season.


Sustained Transgene Expression via Hydrogel-Mediated Gene Transfer Results from Multiple Transfection Events.
Sustained delivery of therapeutic genes in vitro and in vivo has a wide range of applications in studying biology and in developing therapies for treating disease or repairing tissue. Nonviral vectors such as cationic polymers still present promising approaches; however, bolus transfection methods with polyethylenimine-based DNA polyplexes suffer from considerable levels of cytotoxicity and short-lived transgene expression levels. Here, we designed and characterized a hyaluronic acid-based porous hydrogel system for nonviral gene delivery by loading with surface-associated DNA polyplexes. With this, we observed tunable, enhanced, and sustained transgene expression over 30 days of cell culture with better cell viability and marked improvements over comparable bolus transfection techniques. Finally, we investigated mechanisms thought to be responsible for the sustained expression profile, finding that multiple transfection events are likely responsible for the observed sustained expression.


OTX2 is critical for the maintenance and progression of Shh-independent medulloblastomas.
OTX2 is a developmentally regulated transcription factor involved in early morphogenesis of the central nervous system. This gene is amplified and overexpressed in medulloblastoma cell lines, but the nature and extent of its genetic alterations in primary tumors have not been evaluated. Analysis of a large cohort of primary medulloblastomas revealed frequent focal copy number gain of a region minimally containing OTX2 as a single gene. OTX2 copy number gain was restricted to tumor subtypes that did not express a molecular signature of Wnt or Shh pathway activation. FISH analysis revealed copy number gain in a subset of cells within medulloblastoma samples, suggesting a late event in tumor progression. Gain of OTX2 copy number was associated with the presence of anaplastic histologic features and shorter survival in medulloblastoma patients. In support of a functional role, ectopic OTX2 expression enhanced proliferation and tumorigenicity of immortalized primary cells, whereas OTX2 knockdown in medulloblastoma cells prolonged the survival of animals bearing xenograft tumors. Mechanistic investigations revealed upregulation of MYC as a potential mechanism whereby OTX2 promotes tumor progression. Our findings define OTX2 as an important oncogenic driver in medulloblastoma.


Examining Development Processes for Text Messaging Interventions to Prevent Cardiovascular Disease: Systematic Literature Review.
Interventions delivered by mobile phones have the potential to prevent cardiovascular disease (CVD) by supporting behavior change toward healthier lifestyles and treatment adherence. To allow replication and adaptation of these interventions across settings, it is important to fully understand how they have been developed. However, the development processes of these interventions have not previously been systematically examined. This study aimed to systematically describe and compare the development process of text messaging interventions identified in the Text2PreventCVD systematic review. We extracted data about the development process of the 9 interventions identified in the Text2PreventCVD systematic review. Data extraction, which was guided by frameworks for the development of complex interventions, considered the following development stages: intervention planning, design, development, and pretesting. Following data extraction, we invited the developers of the interventions to contribute to our study by reviewing the accuracy of the extracted data and providing additional data not reported in the available publications. A comprehensive description of the development process was available for 5 interventions. Multiple methodologies were used for the development of each intervention. Intervention planning involved gathering information from stakeholder consultations, literature reviews, examination of relevant theory, and preliminary qualitative research. Intervention design involved the use of behavior change theories and behavior change techniques. Intervention development involved (1) generating message content based on clinical guidelines and expert opinions; (2) conducting literature reviews and primary qualitative research to inform decisions about message frequency, timing, and level of tailoring; and (3) gathering end-user feedback concerning message readability, intervention acceptability, and perceived utility. Intervention pretesting involved pilot studies with samples of 10 to 30 participants receiving messages for a period ranging from 1 to 4 weeks. The development process of the text messaging interventions examined was complex and comprehensive, involving multiple studies to guide decisions about the scope, content, and structure of the interventions. Additional research is needed to establish whether effective messaging systems can be adapted from work already done or whether this level of development is needed for application in other conditions and settings.


Heterozygous splice mutation in PIK3R1 causes human immunodeficiency with lymphoproliferation due to dominant activation of PI3K.
Class IA phosphatidylinositol 3-kinases (PI3K), which generate PIP3 as a signal for cell growth and proliferation, exist as an intracellular complex of a catalytic subunit bound to a regulatory subunit. We and others have previously reported that heterozygous mutations in PIK3CD encoding the p110δ catalytic PI3K subunit cause a unique disorder termed p110δ-activating mutations causing senescent T cells, lymphadenopathy, and immunodeficiency (PASLI) disease. We report four patients from three families with a similar disease who harbor a recently reported heterozygous splice site mutation in PIK3R1, which encodes the p85α, p55α, and p50α regulatory PI3K subunits. These patients suffer from recurrent sinopulmonary infections and lymphoproliferation, exhibit hyperactive PI3K signaling, and have prominent expansion and skewing of peripheral blood CD8(+) T cells toward terminally differentiated senescent effector cells with short telomeres. The PIK3R1 splice site mutation causes skipping of an exon, corresponding to loss of amino acid residues 434-475 in the inter-SH2 domain. The mutant p85α protein is expressed at low levels in patient cells and activates PI3K signaling when overexpressed in T cells from healthy subjects due to qualitative and quantitative binding changes in the p85α-p110δ complex and failure of the C-terminal region to properly inhibit p110δ catalytic activity.


Dimeric c-di-GMP is required for post-translational regulation of alginate production in Pseudomonas aeruginosa.
Pseudomonas aeruginosa is an opportunistic human pathogen that secretes the exopolysaccharide alginate during infection of the respiratory tract of individuals afflicted with cystic fibrosis and chronic obstructive pulmonary disease. Among the proteins required for alginate production, Alg44 has been identified as an inner membrane protein whose bis-(3',5')-cyclic dimeric guanosine monophosphate (c-di-GMP) binding activity post-translationally regulates alginate secretion. In this study, we report the 1.8 Å crystal structure of the cytoplasmic region of Alg44 in complex with dimeric self-intercalated c-di-GMP and characterize its dinucleotide-binding site using mutational analysis. The structure shows that the c-di-GMP binding region of Alg44 adopts a PilZ domain fold with a dimerization mode not previously observed for this family of proteins. Calorimetric binding analysis of residues in the c-di-GMP binding site demonstrate that mutation of Arg-17 and Arg-95 alters the binding stoichiometry between c-di-GMP and Alg44 from 2:1 to 1:1. Introduction of these mutant alleles on the P. aeruginosa chromosome show that the residues required for binding of dimeric c-di-GMP in vitro are also required for efficient alginate production in vivo. These results suggest that the dimeric form of c-di-GMP represents the biologically active signaling molecule needed for the secretion of an important virulence factor produced by P. aeruginosa.


Protocol for a randomised controlled trial for Reducing Arthritis Fatigue by clinical Teams (RAFT) using cognitive-behavioural approaches.
Rheumatoid arthritis (RA) fatigue is distressing, leading to unmanageable physical and cognitive exhaustion impacting on health, leisure and work. Group cognitive-behavioural (CB) therapy delivered by a clinical psychologist demonstrated large improvements in fatigue impact. However, few rheumatology teams include a clinical psychologist, therefore, this study aims to examine whether conventional rheumatology teams can reproduce similar results, potentially widening intervention availability. This is a multicentre, randomised, controlled trial of a group CB intervention for RA fatigue self-management, delivered by local rheumatology clinical teams. 7 centres will each recruit 4 consecutive cohorts of 10-16 patients with RA (fatigue severity ≥ 6/10). After consenting, patients will have baseline assessments, then usual care (fatigue self-management booklet, discussed for 5-6 min), then be randomised into control (no action) or intervention arms. The intervention, Reducing Arthritis Fatigue by clinical Teams (RAFT) will be cofacilitated by two local rheumatology clinicians (eg, nurse/occupational therapist), who will have had brief training in CB approaches, a RAFT manual and materials, and delivered an observed practice course. Groups of 5-8 patients will attend 6 × 2 h sessions (weeks 1-6) and a 1 hr consolidation session (week 14) addressing different self-management topics and behaviours. The primary outcome is fatigue impact (26 weeks); secondary outcomes are fatigue severity, coping and multidimensional impact, quality of life, clinical and mood status (to week 104). Statistical and health economic analyses will follow a predetermined plan to establish whether the intervention is clinically and cost-effective. Effects of teaching CB skills to clinicians will be evaluated qualitatively. Approval was given by an NHS Research Ethics Committee, and participants will provide written informed consent. The copyrighted RAFT package will be freely available. Findings will be submitted to the National Institute for Health and Care Excellence, Clinical Commissioning Groups and all UK rheumatology departments. 52709998; Protocol v3 09.02.2015.


Federal Agency biodefense funding, FY2011-FY2012.
Since 2001, the United States government has spent substantial resources on preparing the nation against a bioterrorist attack. Earlier articles in this series have analyzed civilian biodefense funding by the federal government for fiscal years (FY) 2001 through proposed funding for FY2011. This article updates those figures with budgeted amounts for FY2012, specifically analyzing the budgets and allocations for biodefense at the Departments of Health and Human Services, Defense, Homeland Security, Agriculture, Commerce, and State; the Environmental Protection Agency; and the National Science Foundation. This article also includes an updated assessment of the proportion of biodefense funding provided for programs that address multiple scientific, public health, healthcare, national security, and international security issues in addition to biodefense. The FY2012 federal budget for civilian biodefense totals $6.42 billion. Of that total, $5.78 billion (90%) is budgeted for programs that have both biodefense and nonbiodefense goals and applications, and $637.6 million (10%) is budgeted for programs that have objectives solely related to biodefense.


Is prevention of cancer by sun exposure more than just the effect of vitamin D? A systematic review of epidemiological studies.
The number of studies reporting on the association between sunlight exposure, vitamin D and cancer risk is steadily increasing. We reviewed all published case-control and cohort studies concerning colorectal-, prostate-, breast cancer, non-Hodgkin's lymphoma (NHL) and both sunlight and vitamin D to update our previous review and to verify if the epidemiological evidence is in line with the hypothesis that the possible preventive effect of sunlight on cancer might be mediated not only by vitamin D but also by other pathways. We found that almost all epidemiological studies suggest that chronic (not intermittent) sun exposure is associated with a reduced risk of colorectal-, breast-, prostate cancer and NHL. In colorectal- and to a lesser degree in breast cancer vitamin D levels were found to be inversely associated with cancer risk. In prostate cancer and NHL, however, no associations were found. These findings are discussed and it is concluded that the evidence that sunlight is a protective factor for colorectal-, prostate-, breast cancer and non-Hodgkin's lymphoma is still accumulating. The same conclusion can be drawn concerning high vitamin D levels and the risk of colorectal cancer and possibly breast cancer. Particularly in prostate cancer and NHL other sunlight potentiated and vitamin D independent pathways, such as modulation of the immune system and the circadian rhythm, and the degradation of folic acid might play a role in reduced cancer risk as well.