Optimized Photoactivatable Lipid Nanoparticles Enable Red Light Triggered Drug Release.
Encapsulation of small molecule drugs in long-circulating lipid nanoparticles (LNPs) can reduce toxic side effects and enhance accumulation at tumor sites. A fundamental problem, however, is the slow release of encapsulated drugs from these liposomal systems at the disease site resulting in limited therapeutic benefit. Methods to trigger release at specific sites are highly warranted. Here, it is demonstrated that incorporation of ultraviolet (UV-A) or red-light photoswitchable-phosphatidylcholine analogs (AzoPC and redAzoPC) in conventional LNPs generates photoactivatable LNPs (paLNPs) having comparable structural integrity, drug loading capacity, and size distribution to the parent DSPC-cholesterol liposomes. It is shown that 65-70% drug release (doxorubicin) can be induced from these systems by irradiation with pulsed light based on trans-to-cis azobenzene isomerization. In vitro it is confirmed that paLNPs are non-toxic in the dark but convey cytotoxicity upon irradiation in a human cancer cell line. In vivo studies in zebrafish embryos demonstrate prolonged blood circulation and extravasation of paLNPs comparable to clinically approved formulations, with enhanced drug release following irradiation with pulsed light. Conclusively, paLNPs closely mimic the properties of clinically approved LNPs with the added benefit of light-induced drug release making them promising candidates for clinical development.


Legacy motivations & the psychology of intergenerational decisions.
Critical issues in society today involve long time horizons and multiple generations of people. A central challenge in intergenerational decisions is that the interests of present and future generations are not always aligned. The goals and focus of research on intergenerational decisions has been to (1) identify the key features of intergenerational decisions and use those insights to develop experiments that emulate those features, (2) identify the central barriers to intergenerational beneficence, and importantly, (3) identify variables that lead people to act on the behalf of future generations, void of any material or economic incentive to do so. Critically, intergenerational contexts are characterized by the intersection of intertemporal and interpersonal dimensions, which enables the enactment of legacy motivations-a key underlying force in overcoming the many barriers to intergenerational beneficence.


Clonify: unseeded antibody lineage assignment from next-generation sequencing data.
Defining the dynamics and maturation processes of antibody clonal lineages is crucial to understanding the humoral response to infection and immunization. Although individual antibody lineages have been previously analyzed in isolation, these studies provide only a narrow view of the total antibody response. Comprehensive study of antibody lineages has been limited by the lack of an accurate clonal lineage assignment algorithm capable of operating on next-generation sequencing datasets. To address this shortcoming, we developed Clonify, which is able to perform unseeded lineage assignment on very large sets of antibody sequences. Application of Clonify to IgG+ memory repertoires from healthy individuals revealed a surprising lack of influence of large extended lineages on the overall repertoire composition, indicating that this composition is driven less by the order and frequency of pathogen encounters than previously thought. Clonify is freely available at www.github.com/briney/clonify-python.


Postmortem blood free and total morphine concentrations in medical examiner cases.
This purpose of this study was to determine the relationships between postmortem free morphine and total morphine levels in a large series of medical examiner morphine and heroin related deaths. Free morphine, total morphine, and 6-monoacetylmorphine (6-MAM) concentrations were measured by gas chromatography-mass spectrometry (GC-MS) in 87 medical examiner cases over 20 months. The mean total morphine concentration, mean free morphine concentration, and mean percent free morphine for all cases were: 2.3 mg/L (SD 5.2 mg/L), 0.5 mg/L (SD 1.6 mg/L), and 19.4% (SD 22.8%); respectively. Regression analyses showed weak correlations between total and free morphine concentrations over the entire concentration range (0 to 36.6 m/L, r = 0.603, n = 91) and over a subset concentration range of 0 to 1.0 mg/L (r = 0.369, n = 54). Twenty-three out of 56 (41%) tested positive for 6-MAM, indicative heroin abuse cases. Lower total and free morphine concentrations and a higher percent free morphine were found in individuals with detectable 6-MAM. Comparing blood concentrations for cases with and without detectable 6-MAM demonstrated mean total morphine concentrations of 0.9 mg/L versus 2.1 mg/L (p = 0.05), mean free morphine concentrations of 0.3 mg/L versus 0.4 mg/L (p = 0.21), and mean percent free morphine of 34.7% versus 13.7% (p < 0.003), respectively. Our findings demonstrate higher free to total morphine ratios in individuals with detectable 6-MAM than in individuals without 6-MAM. The database established in this study may assist medical examiners in the evaluation of postmortem blood opiates regarding the cause of death in opiate related ingestion cases.


A chip-scale, telecommunications-band frequency conversion interface for quantum emitters.
We describe a chip-scale, telecommunications-band frequency conversion interface designed for low-noise operation at wavelengths desirable for common single photon emitters. Four-wave-mixing Bragg scattering in silicon nitride waveguides is used to demonstrate frequency upconversion and downconversion between the 980 nm and 1550 nm wavelength regions, with signal-to-background levels > 10 and conversion efficiency of ≈ -60 dB at low continuous wave input pump powers (< 50 mW). Finite element simulations and the split-step Fourier method indicate that increased input powers of ≈ 10 W (produced by amplified nanosecond pulses, for example) will result in a conversion efficiency > 25 % in existing geometries. Finally, we present waveguide designs that can be used to connect shorter wavelength (637 nm to 852 nm) quantum emitters with 1550 nm.


Summer freezing resistance decreased in high-elevation plants exposed to experimental warming in the central Chilean Andes.
Alpine habitats have been proposed as particularly sensitive to climate change. Shorter snow cover could expose high-elevation plants to very low temperatures, increasing their risk of suffering damage by freezing, hence decreasing their population viability. In addition, a longer and warmer growing season could affect the hardening process on these species. Thus, understanding the ability of these species to withstand freezing events under warmer conditions is essential for predicting how alpine species may respond to future climate changes. Here we assessed the freezing resistance of 11 species from the central Chilean Andes by determining their low temperature damage (LT(50)) and freezing point (FP) after experimental warming in the field. Plants were exposed during two growing seasons to a passive increase in the air temperature using open top chambers (OTCs). OTCs increased by ca. 3 K the mean air and soil daytime temperatures, but had smaller effects on freezing temperatures. Leaf temperature of the different species was on average 5.5 K warmer inside OTCs at midday. While LT(50) of control plants ranged from -9.9 to -22.4, that of warmed plants ranged from -7.4 to -17.3 degrees C. Overall, high-Andean species growing inside OTCs increased their LT(50) ca. 4 K, indicating that warming decreased their ability to survive severe freezing events. Moreover, plants inside OTCs increased the FP ca. 2 K in some studied species, indicating that warming altered processes of ice crystal formation. Resistance of very low temperatures is a key feature of high-elevation species; our results suggest that current climate warming trends will seriously threaten the survival of high-elevation plants by decreasing their ability to withstand severe freezing events.


Are Men's Perceptions of Sexually Dimorphic Vocal Characteristics Related to Their Testosterone Levels?
Feminine physical characteristics in women are positively correlated with markers of their mate quality. Previous research on men's judgments of women's facial attractiveness suggests that men show stronger preferences for feminine characteristics in women's faces when their own testosterone levels are relatively high. Such results could reflect stronger preferences for high quality mates when mating motivation is strong and/or following success in male-male competition. Given these findings, the current study investigated whether a similar effect of testosterone occurs for men's preferences for feminine characteristics in women's voices. Men's preferences for feminized versus masculinized versions of women's and men's voices were assessed in five weekly test sessions and saliva samples were collected in each test session. Analyses showed no relationship between men's voice preferences and their testosterone levels. Men's tendency to perceive masculinized men's and women's voices as more dominant was also unrelated to their testosterone levels. Together, the results of the current study suggest that testosterone-linked changes in responses to sexually dimorphic characteristics previously reported for men's perceptions of faces do not occur for men's perceptions of voices.


Self-Propelled Water Drops on Bare Glass Substrates in Air: Fast, Controllable, and Easy Transport Powered by Surfactants.
Self-propelled drops are capable of motion without external intervention. As such, they constitute attractive entities for fundamental investigations in active soft matter, hydrodynamics, and surface sciences, as well as promising systems for autonomous microfluidic operations. In contrast with most of the examples relying on organic drops or specifically treated substrates, here we describe the first system of nonreactive water drops in air that can propel themselves on a commercially available ordinary glass substrate that was used as received. This is achieved by exploiting the dynamic adsorption behavior of common n-alkyltrimethylammonium bromide (CnTAB) surfactants added to the drop. We precisely analyze the drop motion for a broad series of surfactants carrying n = 6 to 18 carbon atoms in their tail and establish how the motion characteristics (speed, probability of motion) are tuned by both the hydrophobicity and the concentration of the surfactant. We show that motion occurs regardless of the n value but only in a specific concentration range with a maximum speed at around one tenth of the critical micelle concentration (CMC/10) for most of the tested surfactants. Surfactants of intermediate hydrophobicity are shown to be the best candidates to power drops that can move at a high speed (1-10 cm s-1), the optimal performance being reached with [C12TAB] = 800 μM. We propose a mechanism where the motion originates from the anisotropic wettability of the substrate created by the electrostatic adsorption of surfactants beneath the moving drop. Simply drawing lines with a marker pen allows us to create guiding paths for drop motion and to achieve operations such as complex trajectory control, programmed drop fusion, drop refilling, as well as drop moving vertically against gravity. This work revisits the role of surfactants in dynamic wetting and self-propelled motion as well as brings an original strategy to build the future of microfluidics with lower-cost, simpler, and more autonomous portable devices that could be made available to everyone and everywhere.


Cytotoxic T cells in HIV2 seropositive Gambians. Identification of a virus-specific MHC-restricted peptide epitope.
A preliminary study of gag-specific, MHC-restricted CD8+ CTL has been performed in nine Gambian patients infected with HIV2. Such CTL were present in at least 55% of patients in fresh peripheral blood mononuclear cells without the requirement for in vitro restimulation. We have identified a nonamer peptide from HIV2 gag that is recognized by CD8+ HLA-B53 CTL using an amino acid sequence motif predicted from analysis of endogenous peptides eluted from HLA-B53 molecules. This peptide, from an HIV2/SIV conserved sequence, has previously been reported to be recognized by CTL from non-human primates vaccinated with recombinant vaccinia virus expressing the gag protein of SIV or infected with SIV virus. HLA-B53-restricted, HIV2 gag-specific CTL did not recognize target cells expressing HIV1 gag proteins, indicating that no cellular cross protection to HIV1 could be expected in this case.


On-the-fly ab initio semiclassical dynamics: identifying degrees of freedom essential for emission spectra of oligothiophenes.
Vibrationally resolved spectra provide a stringent test of the accuracy of theoretical calculations. We combine the thawed Gaussian approximation (TGA) with an on-the-fly ab initio (OTF-AI) scheme to calculate the vibrationally resolved emission spectra of oligothiophenes with up to five rings. The efficiency of the OTF-AI-TGA permits treating all vibrational degrees of freedom on an equal footing even in pentathiophene with 105 vibrational degrees of freedom, thus obviating the need for the global harmonic approximation, popular for large systems. Besides reproducing almost perfectly the experimental emission spectra, in order to provide a deeper insight into the associated physical and chemical processes, we also develop a novel systematic approach to assess the importance and coupling between individual vibrational degrees of freedom during the dynamics. This allows us to explain how the vibrational line shapes of the oligothiophenes change with increasing number of rings. Furthermore, we observe the dynamical interplay between the quinoid and aromatic characters of individual rings in the oligothiophene chain during the dynamics and confirm that the quinoid character prevails in the center of the chain.


Comparison of adults who stutter with and without social anxiety disorder.
Social anxiety disorder is a debilitating anxiety disorder associated with significant life impairment. The purpose of the present study is to evaluate overall functioning for adults who stutter with and without a diagnosis of social anxiety disorder. Participants were 275 adults who stuttered (18-80 years), including 219 males (79.6%) and 56 females (20.4%), who were enrolled to commence speech treatment for stuttering. Comparisons were made between participants diagnosed with social anxiety disorder (n = 82, 29.8%) and those without that diagnosis (n = 193, 70.2%). Although the socially anxious group was significantly younger than the non-socially anxious group, no other demographic differences were found. When compared to the non-socially anxious group, the socially anxious group did not demonstrate significantly higher self-reported stuttering severity or percentage of syllables stuttered. Yet the socially anxious group reported more speech dissatisfaction and avoidance of speaking situations, significantly more psychological problems, and a greater negative impact of stuttering. Significant differences in speech and psychological variables between groups suggest that, despite not demonstrating more severe stuttering, socially anxious adults who stutter demonstrate more psychological difficulties and have a more negative view of their speech. The present findings suggest that the demographic status of adults who stutter is not worse for those with social anxiety disorder. These findings pertain to a clinical sample, and cannot be generalized to the wider population of adults who stutter from the general community. Further research is needed to understand the longer-term impact of social anxiety disorder for those who stutter.


A randomized trial of two e-learning strategies for teaching substance abuse management skills to physicians.
To compare the educational effectiveness of two virtual patient (VP)-based e-learning strategies, versus no training, in improving physicians' substance abuse management knowledge, attitudes, self-reported behaviors, and decision making. The 2011-2012 study was a posttest-only, three-arm, randomized controlled trial in 90 resident and 30 faculty physicians from five adult medicine primary care training programs. The intervention was one of two 2-hour VP-based e-learning programs, designed by national experts to teach structured screening, brief interventions, referral, and treatment skills. One used traditional problem solving with feedback (unworked example), and the other incorporated an expert demonstration first, followed by problem solving with feedback (worked example). The main outcome measure was performance on the Physicians' Competence in Substance Abuse Test (P-CSAT, maximum score = 315), a self-administered, previously validated measure of physicians' competence in managing substance abuse. The survey was completed at the outset of the study and two months later. Overall P-CSAT scores were virtually identical (202-211, P > .05) between both intervention groups and the no-training control group at both times. Average faculty P-CSAT scores (221.9, 224.6) were significantly higher (P < .01) than resident scores (203.7, 202.5) at both times. This study did not provide evidence that a brief, worked example, VP-based e-learning program or a traditional, unworked, VP-based e-learning program was superior to no training in improving physicians' substance abuse management skills. The study did provide additional evidence that the P-CSAT distinguishes between physicians who should possess different levels of substance abuse management skills.


Clinical recommendation radical trachelectomy for fertility preservation in patients with early-stage cervical cancer.
Radical trachelectomy (RT) is a fertility-sparing procedure with the aim to provide adequate oncological safety to patients with cervical cancer while preserving their fertility. In the current review, indications, development of the procedure, technical aspects, preoperative and postoperative management, and oncological, fertility, and obstetric outcomes are discussed and studied with respect to whether the procedure is performed abdominally or vaginally. Complications of RT, staging, and more conservative alternatives to RT are discussed as well. A systematic MEDLINE search was performed, which yielded 218 articles, of which 75 were selected for further analysis based on the number of patients and the quality of the study. Strict morphologic criteria should be applied to the candidates to maintain oncological safety. When limited to a tumor less than 2 cm in diameter, the overall recurrence rate after vaginal RT is 3% to 6% and the death rate is 2% to 5%. Data on fertility and obstetric outcome are mostly based on the results of patients who underwent vaginal RT. More data are needed to be able to draw the same conclusions for abdominal RT. Fertility seems not to be decreased, but the risk for premature delivery is 2 to 3 times higher compared to women with an intact cervix. In locally advanced cervical tumors with a diameter larger than 2 cm, neoadjuvant chemotherapy followed by RT may be offered after explaining the experimental nature to the patient. In conclusion, RT is an oncologically safe technique in women with early invasive cancer. The rate of term pregnancies still needs improvement. Fertility-preserving treatment of women with tumors larger than 2 cm in diameter can be done by combining neoadjuvant chemotherapy and trachelectomy; however, experience is still limited.


Responses of plant species diversity and soil physical-chemical-microbial properties to Phragmites australis invasion along a density gradient.
The invasion of ecosystems by strongly colonising plants such as Phragmites australis is viewed as one of the greatest threats to plant diversity and soil properties. This study compared a range of diversity measures including soil properties and mycorrhizal potential under different degrees of Phragmites density among three populations in coastal wetland, Victoria, Australia. Species richness, evenness and Shanon-Wiener index had significantly higher values in low degree of Phragmites density in all populations. Higher densities had the lowest diversity, with Shannon-Wiener index = 0 and Simpson's index = 1 indicating its mono-specificity. Significant alterations in soil properties associated with different degrees of Phragmites density were noticed. These had interactive effects (population × density) on water content, dehydrogenase activity, microbial biomass (C, N and P) but not on pH, electrical conductivity, phenolics, organic carbon, and spore density. Furthermore, the study elucidated decrease of competitive abilities of native plants, by interfering with formation of mycorrhizal associations and biomass. Overall, our results suggest that significant ecological alterations in vegetation and soil variables (including mycorrhizal potential) were strongly dependent on Phragmites density. Such changes may lead to an important role in process of Phragmites invasion through disruption of functional relationships amongst those variables.


Architecture of the RNA polymerase-Spt4/5 complex and basis of universal transcription processivity.
Related RNA polymerases (RNAPs) carry out cellular gene transcription in all three kingdoms of life. The universal conservation of the transcription machinery extends to a single RNAP-associated factor, Spt5 (or NusG in bacteria), which renders RNAP processive and may have arisen early to permit evolution of long genes. Spt5 associates with Spt4 to form the Spt4/5 heterodimer. Here, we present the crystal structure of archaeal Spt4/5 bound to the RNAP clamp domain, which forms one side of the RNAP active centre cleft. The structure revealed a conserved Spt5-RNAP interface and enabled modelling of complexes of Spt4/5 counterparts with RNAPs from all kingdoms of life, and of the complete yeast RNAP II elongation complex with bound Spt4/5. The N-terminal NGN domain of Spt5/NusG closes the RNAP active centre cleft to lock nucleic acids and render the elongation complex stable and processive. The C-terminal KOW1 domain is mobile, but its location is restricted to a region between the RNAP clamp and wall above the RNA exit tunnel, where it may interact with RNA and/or other factors.


Common variants in P2RY11 are associated with narcolepsy.
Growing evidence supports the hypothesis that narcolepsy with cataplexy is an autoimmune disease. We here report genome-wide association analyses for narcolepsy with replication and fine mapping across three ethnic groups (3,406 individuals of European ancestry, 2,414 Asians and 302 African Americans). We identify a SNP in the 3' untranslated region of P2RY11, the purinergic receptor subtype P2Y₁₁ gene, which is associated with narcolepsy (rs2305795, combined P = 6.1 × 10⁻¹⁰, odds ratio = 1.28, 95% CI 1.19-1.39, n = 5689). The disease-associated allele is correlated with reduced expression of P2RY11 in CD8(+) T lymphocytes (339% reduced, P = 0.003) and natural killer (NK) cells (P = 0.031), but not in other peripheral blood mononuclear cell types. The low expression variant is also associated with reduced P2RY11-mediated resistance to ATP-induced cell death in T lymphocytes (P = 0.0007) and natural killer cells (P = 0.001). These results identify P2RY11 as an important regulator of immune-cell survival, with possible implications in narcolepsy and other autoimmune diseases.


Complex phenotype of mice lacking occludin, a component of tight junction strands.
Occludin is an integral membrane protein with four transmembrane domains that is exclusively localized at tight junction (TJ) strands. Here, we describe the generation and analysis of mice carrying a null mutation in the occludin gene. Occludin -/- mice were born with no gross phenotype in the expected Mendelian ratios, but they showed significant postnatal growth retardation. Occludin -/- males produced no litters with wild-type females, whereas occludin -/- females produced litters normally when mated with wild-type males but did not suckle them. In occludin -/- mice, TJs themselves did not appear to be affected morphologically, and the barrier function of intestinal epithelium was normal as far as examined electrophysiologically. However, histological abnormalities were found in several tissues, i.e., chronic inflammation and hyperplasia of the gastric epithelium, calcification in the brain, testicular atrophy, loss of cytoplasmic granules in striated duct cells of the salivary gland, and thinning of the compact bone. These phenotypes suggested that the functions of TJs as well as occludin are more complex than previously supposed.


Oxygen on Ganymede: laboratory studies.
To test proposals for the origin of oxygen absorption bands in the visible reflectance spectrum of Ganymede, the reflectance of condensed films of pure oxygen (O2) and O2-water mixtures and the evolution of O2 from the films as a function of temperature were determined. Absorption band shapes and positions for oxygen at 26 kelvin were similar to those reported for Ganymede, whereas those for the mixtures were slightly shifted. The band intensity dropped by more than two orders of magnitude when the ice mixture was warmed to 100 kelvin, although about 20 percent of the O2 remained trapped in the ice, which suggested that at these temperatures O2 molecules dissolve in the ice rather than aggregate in clusters or bubbles. The experiments suggest that the absorption bands in Ganymede's spectrum were not produced in the relatively warm surface of the satellite but in a much colder source. Solid O2 may exist in a cold subsurface layer or in an atmospheric haze.


[Clinical observation on infantile cerebral palsy treated with quick meridian needling therapy plus scalp acupuncture].
To assess the efficacy of quick meridian needling therapy plus scalp acupuncture on infantile cerebral palsy (CP). One hundred and forty-seven cases of CP were randomly divided into a quick meridian needling therapy plus scalp acupuncture group (group A), a conventional acupuncture group (group B) and a scalp acupuncture group (group C), 49 cases in each one. In group A, quick needling was applied to the Conception Vessel, Governor Vessel, Hand-Yangming, Hand-Jueyin, Foot-Yangming and Foot-Taiyin meridians distributed on four limbs and trunk. One pricking point was 10 mm far from the other one. In scalp acupuncture, motor area, equilibrium area, sensory area, tremor-control area, foot-motor-sensory area, speech No. 2 area, speech No. 3 area, Baihui (GV 20), Sishencong (EX-HN 1), etc. were selected. The needles were stimulated with rotating manipulation and remained for 30-60 min. In group B, the conventional acupuncture was adopted mainly at Dazhui (GV 14), Shenzhu (GV 12), Fengfu (GV 16) and others. In group C, the scalp acupuncture was used and the points selected were same as those in group A for scalp acupuncture treatment. The scores of Gross Motor Function Measure (GMFM) were observed before and after treatment for children. The clinical efficacy of each group was evaluated. By follow-up for 12 months, the condition of independent walking was observed. The total effective rate in group A was 79.6% (39/49), which was superior to that of group B [49.0% (24/49)] and group C [51.0% (25/ 49)] respectively (both P < 0.05). After treatment, GMFM scores of children were all improved significantly in 3 groups (P < 0.001, P < 0.05), of which, the improvement extent in group A was superior to that of other two groups (both P < 0.05). It was found after follow-up for 1 year that 31 cases could walk independently in group A, which was more than group B (17 cases) and group C (16 cases). The quick meridian needling therapy plus scalp acupuncture can improve significantly limb motor function of children with cerebral palsy and its therapeutic effect is superior to conventional acupuncture and simple scalp acupuncture.


IgG B memory cell subpopulations: differences in susceptibility to stimulation by TI-1 and TI-2 antigens.
Spleen cells from mice primed with the thymus dependent (TD) antigen trinitrophenyl keyhole limpet hemocyanin several months earlier can be stimulated in vitro to produce an IgG anti-hapten response to TD as well as thymus independent (TI) forms of the hapten. Selective killing of TD or TI-2 responding B cells can be accomplished with the corresponding antigen by bromouridine deoxyribose (BUdR) and light treatment without affecting the other population. In contrast, we show here that selective killing with TI-1 antigens does not occur. Rather, the TI-1 antigens, TNP-Brucella abortus or TNP-lipopolysaccharide, eliminate both TD and TI-2 responding IgG memory B cells. All TNP-responding B cells are similarly eliminated if cultures are challenged simultaneously with TD and TI-2 antigens before BUdR and light but not when they are challenged with either a TD or TI-2 antigen separately. We conclude that IgG memory B cell precursors stimulated to produce anti-TNP by TD or TI-2 forms of the hapten are defined by only two functionally distinct subpopulations and that TI-1 antigens can stimulate both of these populations at least to divide.


Liquid crystalline bilayers self-assembled from rod-coil diblock copolymers.
The structure and phase behaviour of bilayer membranes self-assembled from rod-coil diblock copolymers are studied using the self-consistent field theory, focusing on the occurrence and relative stability of liquid crystalline phases induced by the geometric shape and orientational interaction of the rod-blocks. A variety of liquid crystalline bilayers, corresponding to the smectic phases in bulk systems, are predicted to occur as equilibrium phases of the system. The ordered morphologies and phase behaviour of the system are analyzed. Phase diagrams of the self-assembled bilayers are constructed. The theoretical results provide an understanding of the formation mechanisms of these intricate phases.


Tiny videos: a large data set for nonparametric video retrieval and frame classification.
In this paper, we present a large database of over 50,000 user-labeled videos collected from YouTube. We develop a compact representation called "tiny videos" that achieves high video compression rates while retaining the overall visual appearance of the video as it varies over time. We show that frame sampling using affinity propagation-an exemplar-based clustering algorithm-achieves the best trade-off between compression and video recall. We use this large collection of user-labeled videos in conjunction with simple data mining techniques to perform related video retrieval, as well as classification of images and video frames. The classification results achieved by tiny videos are compared with the tiny images framework [24] for a variety of recognition tasks. The tiny images data set consists of 80 million images collected from the Internet. These are the largest labeled research data sets of videos and images available to date. We show that tiny videos are better suited for classifying scenery and sports activities, while tiny images perform better at recognizing objects. Furthermore, we demonstrate that combining the tiny images and tiny videos data sets improves classification precision in a wider range of categories.


Tuberculosis among health-care workers in low- and middle-income countries: a systematic review.
The risk of transmission of Mycobacterium tuberculosis from patients to health-care workers (HCWs) is a neglected problem in many low- and middle-income countries (LMICs). Most health-care facilities in these countries lack resources to prevent nosocomial transmission of tuberculosis (TB). We conducted a systematic review to summarize the evidence on the incidence and prevalence of latent TB infection (LTBI) and disease among HCWs in LMICs, and to evaluate the impact of various preventive strategies that have been attempted. To identify relevant studies, we searched electronic databases and journals, and contacted experts in the field. We identified 42 articles, consisting of 51 studies, and extracted data on incidence, prevalence, and risk factors for LTBI and disease among HCWs. The prevalence of LTBI among HCWs was, on average, 54% (range 33% to 79%). Estimates of the annual risk of LTBI ranged from 0.5% to 14.3%, and the annual incidence of TB disease in HCWs ranged from 69 to 5,780 per 100,000. The attributable risk for TB disease in HCWs, compared to the risk in the general population, ranged from 25 to 5,361 per 100,000 per year. A higher risk of acquiring TB disease was associated with certain work locations (inpatient TB facility, laboratory, internal medicine, and emergency facilities) and occupational categories (radiology technicians, patient attendants, nurses, ward attendants, paramedics, and clinical officers). In summary, our review demonstrates that TB is a significant occupational problem among HCWs in LMICs. Available evidence reinforces the need to design and implement simple, effective, and affordable TB infection-control programs in health-care facilities in these countries.


Replication and robustness in developmental research.
Replications and robustness checks are key elements of the scientific method and a staple in many disciplines. However, leading journals in developmental psychology rarely include explicit replications of prior research conducted by different investigators, and few require authors to establish in their articles or online appendices that their key results are robust across estimation methods, data sets, and demographic subgroups. This article makes the case for prioritizing both explicit replications and, especially, within-study robustness checks in developmental psychology. It provides evidence on variation in effect sizes in developmental studies and documents strikingly different replication and robustness-checking practices in a sample of journals in developmental psychology and a sister behavioral science-applied economics. Our goal is not to show that any one behavioral science has a monopoly on best practices, but rather to show how journals from a related discipline address vital concerns of replication and generalizability shared by all social and behavioral sciences. We provide recommendations for promoting graduate training in replication and robustness-checking methods and for editorial policies that encourage these practices. Although some of our recommendations may shift the form and substance of developmental research articles, we argue that they would generate considerable scientific benefits for the field. (PsycINFO Database Record (c) 2014 APA, all rights reserved).


Proviral DNA integration and transcriptional patterns of equine infectious anemia virus during persistent and cytopathic infections.
The structure and integration patterns of equine infectious anemia virus (EIAV) proviral DNA and the patterns of viral transcription were examined in persistent and cytopathic infections of cultured cells. The results of Southern blot analyses indicated that, in persistently infected cells, about 30% of the EIAV provirus exists as randomly integrated DNA, while the remaining 70% is equally divided between unintegrated linear and closed circular forms. The cytopathic infection, in contrast, is characterized by levels of integrated provirus ranging from 65 to more than 90% of the total proviral DNA, depending on the extent of cytopathology exhibited by the virus strain employed. In both persistent and cytopathic infections, extensive Northern (RNA) blot analyses have revealed the presence of two major virus-specific transcripts, an 8.2-kilobase (kb) full-length genomic mRNA and a 3.5-kb single-spliced mRNA. A low-abundance 1.5-kb mRNA, presumably formed by a double-splicing event of the full-length RNA, was also detected in the cytopathic EIAV infection. The two major viral transcripts are present in approximately equal quantities in persistently infected cells, while the cytopathic infection reveals nearly a 30-fold higher level of viral transcripts in which the 3.5-kb species constitutes over 75% of the total viral mRNA. The relatively high proportion of proviral DNA integration and the simple pattern of viral transcription observed during EIAV infections appeared to be different from the generally observed patterns of predominantly unintegrated proviral DNA and multi-spliced viral mRNAs in cells infected with other lentiviruses such as visna virus or human immunodeficiency virus type 1. Moreover, the data suggested that the cytopathology of EIAV may be correlated in part with the degree of proviral DNA integration and levels of viral mRNA in infected cells, particularly that of the spliced 3.5-kb mRNA.


The effects of excretions/secretions of Ostertagia circumcincta on ovine abomasal tissues in vitro.
Products excreted/secreted by Ostertagia circumcincta stimulated the in vitro release of pepsinogen from intact abomasal mucosal sheets and caused the contraction of strips of abomasal smooth muscle, also in vitro. However, responses occurred only when tissues had been derived from animals that were assumed to have experienced prior exposure to the parasite. The overall median responses for pepsinogen secretion in response to ES, expressed as the ratio of stimulated secretion to basal secretion, were 1.8 for previously exposed animals and 0.9 for parasite-naive animals. For the smooth muscle from the previously exposed animals, the overall median response to ES, expressed as a percentage of the maximal response to carbachol in the same tissues, was 27.0. No responses were seen in muscle from any parasite-naive animal. These results suggest that the responses obtained were hypersensitivity reactions to antigens released by the worms during in vitro culture, and occurring in tissues from animals sensitised by exposure to O. circumcincta in the natural environment.


Respiratory syncytial virus is present in the neonatal intensive care unit.
Nosocomial transmission of respiratory syncytial virus (RSV) occurs in children within the neonatal intensive care unit (NICU). During peak community RSV transmission, three swabs were collected from the nose, hand and personal clothing of visitors and health care workers (HCW) in NICU once every week for eight weeks. Nasal swabs were collected from every third neonate and from any neonate clinically suspected of having a respiratory infection. Environmental sampling of high touch areas was done once during the study period. All swabs were tested for RSV using real time RT-PCR. There were 173 (519 total) and 109 (327 total) swabs, each of nose, hand and dress from 84 HCWs and 80 visitors respectively and 81 nasal swabs from 55 neonates collected. Thirty five environmental swabs from surfaces of the beds, side tables, counter tops, chairs, tables and computers were collected. Overall 1% of nasal swabs from each of HCWs, visitors and neonates, 4% of dress specimens from visitors and 9% of environmental swabs were positive for RSV-RNA. The results suggest that though the risk for RSV in the NICU remains low, personnel clothing are contaminated with RSV-RNA and may have a role in transmission.


The assessment of quality of life in acute cough with the Leicester Cough Questionnaire (LCQ-acute).
Acute cough has a significant impact on physical and psychosocial health and is associated with an impaired quality of life (QOL). The Leicester Cough Questionnaire (LCQ) is a validated cough-related health status questionnaire designed for patients with chronic cough. The purpose of this study was to validate the LCQ for the assessment of health related QOL in patients with acute cough and determine the clinical minimal important difference (MID). 10 subjects with cough due to acute upper respiratory tract infection underwent focused interviews to investigate the face validity of the LCQ. The LCQ was also evaluated by a multidisciplinary team. 30 subjects completed the revised LCQ-acute and a cough visual analogue score (VAS: 0-100 mm) within one week of onset of cough and again <2 weeks later and at resolution of cough. The concurrent validity, internal reliability, repeatability and responsiveness of the LCQ-acute were also assessed. Patients also completed a Global Rating of Change Questionnaire that assessed the change in cough severity between visits. The MID was calculated as the change in LCQ-acute score for patients responding to GRCQ category representing the smallest change in health status that patients found worthwhile. Health status was severely impaired at baseline affecting all domains; median (interquartile range) total LCQ-acute score 13.0 (3.4). All subjects found the LCQ-acute questionnaire acceptable for assessing their cough. Internal reliability of the LCQ-acute was good for all domains and total score, Cronbach's α coefficients >0.9. There was a significant correlation between LCQ-acute and VAS (ρ = -0.48, p = 0.007). The LCQ-acute and its domains were highly responsive to change; effect sizes 1.7-2.3. The MID for total LCQ and VAS were 2.5 and 13 mm respectively. The LCQ-acute is a brief, simple and valid instrument to assess cough specific health related QOL in patients with acute cough. It is a highly responsive tool suggesting that it will be particularly useful to assess the effect of antitussive therapy.


Lack of association between worsening renal function and mortality in heart failure patients treated with nesiritide.
Nesiritide is approved for the treatment of decompensated heart failure. Because nesiritide may exacerbate renal dysfunction, it has been claimed that there may be a link between worsening renal function caused by nesiritide and increased mortality. We analyzed our inpatient experience with nesiritide to determine if changes in renal function were associated with worsened mortality. A retrospective study of 167 patients [65% male, median age 70 years, left ventricular ejection fraction (LVEF) 30%, glomerular filtration rate (GFR) 36 mL/min/1.73 m(2), serum creatinine (sCr)1.8 mg/dL] treated with standard dose nesiritide was undertaken for the period September 2001-March 2005. Mortality was 12.5% at 1 month and 51% at 24 months (median follow-up was 5.4 months [interquartile ranges (IQR) 1.3, 18.6]. Higher mortality was associated with lower pre-infusion GFR (hazard ratio for a 10 mL/min/1.72 m(2) decrease=1.22, p=0.004). Pre-to-post nesiritide changes in GFR (median 0.0, IQR-5.8, 8.4; p=0.51) and creatinine (median 0.0, IQR -0.3, 0.3; p=0.91), however, were not significant and not detected to be associated with worsened mortality by Cox proportional hazards (p=0.46 and p=0.40, respectively). While we found that nesiritide infusion in decompensated heart failure patients worsened renal function (decrease in GFR) in 29% of patients, changes in renal function could not be related to an incremental worsening of mortality. Our findings do not support an association of worsening renal function and worsening mortality after nesiritide.


First Report of Didymella bryoniae Causing Gummy Stem Blight of Chayote in Taiwan.
Chayote (Sechium edule (Jacq.) Swartz, Cucurbitaceae), originally native to Mexico, is an important vegetable known as "dragon-whisker vegetable" and is cultivated for its shoots in Ji-an, Hualien County in eastern Taiwan. In June 2010, 70 to 80% of the chayote plants grown in Ji-an developed necrotic spots on stems, leaves, and fruits. The disease was severe during the warm and rainy season from June to August. The symptoms on stems, leaves, and fruits were water-soaked lesions that eventually dried up, cracked, and produced perithecia on necrotic tissues. A single ascospore was isolated from perithecia harvested from diseased stems and cultured on potato dextrose agar (PDA) at 25°C for 1 month. Colonies of three isolates (SE5, SE6, and SE7) were white to olivaceous green bearing unicellular conidia measuring 2 to 5 × 3 to 10 μm, which is consistent with the morphological characteristics of Didymella bryoniae (Auersw.) Rehm (anamorph Phoma cucurbitacearum (Fr.:Fr.) Sacc.) (1,2,3). DNA of SE5, SE6, and SE7 isolates were obtained using microwave-based method (4). The internal transcribed spacer (ITS) rDNA (GenBank accessions AB714984, AB714985, and AB714986), PCR-amplified using primers ITS1 and ITS4, had 98 to 99% nucleotide sequence identity with D. bryoniae (GenBank Accession Nos. GU045304 and GU592001). A pathogenicity test was conducted in a greenhouse with temperature ranging from 20 to 30°C. Three-day-old mycelial plugs (5 × 5 mm) of the three isolates were placed on the needle-pricked wounds of stems and leaves of 36 4-month-old potted chayote plants wrapped in plastic bags to maintain 100% relative humidity for 2 days. Six days after inoculation, water-soaked lesions formed on the stems and leaves. Controls inoculated with sterile water had no symptoms. The fungus reisolated from the lesions of diseased stems and leaves had morphological characteristics of D. bryoniae. Based on the results of morphology, molecular data, and pathogenicity tests, we reported for the first time to our knowledge that gummy stem blight of chayote is caused by D. bryoniae in Taiwan. References: (1) J. W. Huang and W. R. Hsieh. Plant Prot. Bull. 27:325, 1985. (2) A. P. Keinath et al. Phytopathology 85:364, 1995. (3) E. Punithalingam and P. Holliday. P. 332 in: Descriptions of Pathogenic Fungi and Bacteria. CMI, Kew, Surrey, UK, 1972. (4) S. R. Tendulkar et al. Biotechnol. Lett. 22:1941, 2003.


Elevated inflammatory markers in response to prolonged sleep restriction are associated with increased pain experience in healthy volunteers.
Sleep disturbances, pain, and inflammation co-occur in various medical conditions, but their interrelationships are poorly understood. We investigated the effects of reduced sleep duration (by approximately 50%) to 4 h/night across 10 days, on peripherally circulating inflammatory mediators. In addition, we tested the prediction that degree of inflammation is quantitatively related to the extent to which pain is increased in response to prolonged sleep restriction. Randomized, 16 day controlled in-laboratory study conducted in GCRC. Eighteen volunteers were randomly assigned to either 12 days of sleeping 8 h/night or 4 h/night. Participants rated mood and pain symptoms throughout experimental days. Urine was collected and blood was drawn frequently on the baseline day and after the 10th experimental day for 25 hours. Levels of plasma interleukin (IL)-6, serum C-reactive protein (CRP), plasma soluble tumor necrosis factor receptor p55 (sTNF-R p55), urinary levels of prostaglandin (PG) metabolites D2 and E2, subjective assessment of pain and tiredness-fatigue. IL-6 levels were elevated in the 4-h sleep condition over the 8-h sleep condition (P <0.05). CRP levels showed the same trend as IL-6, but did not differ significantly between groups (P = 0.11). Levels of sTNF-R p55 were unchanged in both groups. PG E2 and 11beta-F2alpha metabolite increased in 4-h sleepers, but did not differ significantly from the 8-h sleepers. Elevated IL-6 levels were strongly associated with increased pain ratings in response to sleep restriction (r = 0.67, P <0.01), and this association could not be explained by elevations in tiredness-fatigue. Insufficient sleep quantity may facilitate and/or exacerbate pain through elevations of IL-6. In disorders where sleep disturbances are common, insufficient sleep quantity itself may establish and maintain its co-occurrence with pain and increased inflammation.


Failure of miltefosine treatment for visceral leishmaniasis in children and men in South-East Asia.
High frequency of relapse in miltefosine-treated visceral leishmaniasis (VL) patients in India and Nepal followed up for twelve months. To identify epidemiological and clinical risk factors for relapse of VL in patients recently treated with standard dosing of miltefosine in India and Nepal. Prospective observational study in three Primary Health Centers and one reference center in Muzaffarpur district, Bihar, India; and two zonal hospitals and a university hospital in South-east Nepal; records of all consenting patients diagnosed with VL and treated with miltefosine according to the current treatment guidelines of the Kala azar elimination program between 2009 and 2011. We compared the clinical records of 78 cases of relapse with those of 775 patients who had no record of subsequent relapse. Relapse was 2 times more common amongst male patients (IRR 2.14, 95% CI 1.27-3.61), and 2 to 3 times more frequent in the age groups below 15 compared to the over 25 year olds (age 10 to 14: IRR 2.53; 95% CI 1.37-4.65 and Age 2 to 9: IRR 3.19; 95% CI 1.77-5.77). History of earlier VL episodes, or specific clinical features at time of diagnosis such as duration of symptoms or spleen size were no predictors of relapse. Young age and male gender were associated with increased risk of VL relapse after miltefosine, suggesting that the mechanism of relapse is mainly host-related i.e. immunological factors and/or drug exposure (pharmacokinetics). The observed decrease in efficacy of miltefosine may be explained by the inclusion of younger patients compared to the earlier clinical trials, rather than by a decreased susceptibility of the parasite to miltefosine. Our findings highlight the importance of proper clinical trials in children, including pharmacokinetics, to determine the safety, efficacy, drug exposure and therapeutic response of new drugs in this age group.


Isolation and expression of a complementary DNA that confers multidrug resistance.
The emergence and outgrowth of a population of tumour cells resistant to multiple drugs is a major problem in the chemotherapeutic treatment of cancer. We have used highly drug-resistant cell lines developed in vitro to study the molecular basis of multidrug resistance. In these cell lines high levels of resistance are frequently associated with amplification and overexpression of a small group of genes termed mdr or gp170. Direct evaluation of the role of these genes in multidrug resistance has awaited the isolation of a member of this gene family in a biologically active form. Here we report the isolation of DNA clones complementary to the cellular messenger RNA transcripts of mdr genes and show that high-level expression of a full-length complementary DNA clone in an otherwise drug-sensitive cell confers a complete multidrug-resistant phenotype. Our results demonstrate that overexpression of a single member of the mdr group is sufficient to confer drug resistance. Furthermore, because the cDNA was isolated from a drug-sensitive cell, mutations in the primary sequence of mdr are not required to produce a multidrug-resistance phenotype.


Max-independent functions of Myc in Drosophila melanogaster.
Myc proteins are powerful proto-oncoproteins and important promoters of growth and proliferation during normal development. They are thought to exercise their effects upon binding to their partner protein Max, and their activities are largely antagonized by complexes of Max with Mnt or an Mxd family protein. Although the biological functions of Myc, Mxd and Mnt have been intensively studied, comparatively little is known about the in vivo role of Max. Here we generate Max loss-of-function and reduction-of-function mutations in Drosophila melanogaster to address the contribution of Max to Myc-dependent growth control. We find that many biological activities of Myc do not, or only partly, require the association with Max--for example, the control of endoreplication and cell competition-and that a Myc mutant that does not interact with Max retains substantial biological activity. We further show that Myc can control RNA polymerase III independently of Max, which explains some of Myc's observed biological activities. These studies show the ability of Myc to function independently of Max in vivo and thus change the current model of Max network function.


A Review Of Innovative International Financing Mechanisms To Address Noncommunicable Diseases.
Noncommunicable diseases have become prevalent in low- and middle-income countries. A key question that remains unresolved is how to support the development of systems to prevent and treat noncommunicable disease through international financing mechanisms. We conducted a review of articles and grey literature published from 2000 through 2014 on innovative financing models proposed or used for other disease control efforts. We found that the greatest available evidence supported pooled funding models, where funding from multiple groups is combined for a specific investment, with such models previously deployed in vaccine and infectious disease funding areas. Robust evidence also supported the viability of international transactions taxes or levies placed on specific transactions to fund investments in drug procurement and supply, and of the front-loading of development aid through bond sales, particularly to stabilize funding and subsidize drug procurement. Far less compelling evidence was available to support diaspora bonds or debt reduction programs as mechanisms to aid low- and middle-income countries' health systems in financing noncommunicable disease prevention and care services.


Pelvic fracture in multiple trauma: A 67-case series.
Severe pelvic trauma remains associated with elevated mortality, largely due to hemorrhagic shock. The main study objective was to test for correlation between fracture type and mortality. The secondary objective was to assess the efficacy in terms of mortality of multidisciplinary management following a decision-tree in multiple trauma victims admitted to a level 1 trauma center. Between July 2011 and July 2013, 534 severe trauma patients were included in a single-center continuous prospective observational study. All patients with hemorrhagic shock received early treatment by pelvic binder. Patients with active bleeding on full-body CT or persisting hemorrhagic shock underwent arteriography with or without embolization. Pelvic trauma was graded on the Tile classification. The principle end-point was mortality. Median age was 40 years (range, 26-48 years), with a 79% male/female sex ratio. Thirty-two of the 67 patients with pelvic trauma (48%) were in hemorrhagic shock at admission. Median injury severity score (ISS) was 36 (range, 24-43). On the Tile classification, 22 patients (33%) were grade A, 33 (49%) grade B and 12 (18%) grade C. Overall mortality was 19%, and 42% in case of hemorrhagic shock. Mortality was significantly higher with Tile C than A or B (58% vs. 9.1% and 12.1%, respectively; P=0.001). Vertical shear fracture (Tile C) was associated with greater mortality from hemorrhagic shock. IV, case series.


The development of cortical sensitivity to visual word forms.
The ability to extract visual word forms quickly and efficiently is essential for using reading as a tool for learning. We describe the first longitudinal fMRI study to chart individual changes in cortical sensitivity to written words as reading develops. We conducted four annual measurements of brain function and reading skills in a heterogeneous group of children, initially 7-12 years old. The results show age-related increase in children's cortical sensitivity to word visibility in posterior left occipito-temporal sulcus (LOTS), nearby the anatomical location of the visual word form area. Moreover, the rate of increase in LOTS word sensitivity specifically correlates with the rate of improvement in sight word efficiency, a measure of speeded overt word reading. Other cortical regions, including V1, posterior parietal cortex, and the right homologue of LOTS, did not demonstrate such developmental changes. These results provide developmental support for the hypothesis that LOTS is part of the cortical circuitry that extracts visual word forms quickly and efficiently and highlight the importance of developing cortical sensitivity to word visibility in reading acquisition.


Valproic acid enhances protein L-isoaspartyl methyltransferase expression by stimulating extracellular signal-regulated kinase signaling pathway.
Proteins are susceptible to various non-enzymatic post-translational modifications occurring during aging and in certain pathological states. The protein L-isoaspartyl methyltransferase (PIMT) is an enzyme that recognizes and repairs the abnormal L-isoaspartyl residues in proteins. Recently, we reported that PIMT expression was stimulated by the anti-epileptic drug valproic acid and that this was mediated through the glycogen synthase kinase-3 (GSK-3)/beta-catenin pathway. In this study, to gain further insights into which of the signaling pathways activated by valproic acid regulate PIMT abundance, astrocytoma U-87 MG and neuroblastoma SH-SY5Y cells were treated with this drug to investigate the possible involvement of the extracellular-regulated kinase (ERK) pathway in PIMT induction. Valproic acid increased ERK1/2 phosphorylation on Thr202/Tyr204 and Thr185/Tyr187, respectively. Pharmacological inhibitors against the kinases Src, c-Raf, MEK1/2 and ERK1/2 abolished the ERK1/2 phosphorylation stimulated by valproic acid, thus preventing PIMT induction by the drug. Furthermore, MEK1/2 inhibition with U0126 blocked the higher phosphorylation of RSK-1 on Thr359/Ser363 and of GSK-3beta on Ser9 as well as the increased expression of RSK-1, beta-catenin and PIMT upon treatment with valproic acid. RSK-1 knockdown by interfering RNA abrogated the increased expression of RSK-1, beta-catenin and PIMT as well as the induced phosphorylation of RSK-1 and GSK-3beta due to valproic acid. Thus, our findings demonstrated that PIMT up-regulation by valproic acid required the activation of the ERK signaling pathway including RSK-1 the latter being responsible for inactivating GSK-3 and subsequently leading to beta-catenin stabilization.


Validation of the Mass-Extraction-Window for Quantitative Methods Using Liquid Chromatography High Resolution Mass Spectrometry.
A paradigm shift is underway in the field of quantitative liquid chromatography-mass spectrometry (LC-MS) analysis thanks to the arrival of recent high-resolution mass spectrometers (HRMS). The capability of HRMS to perform sensitive and reliable quantifications of a large variety of analytes in HR-full scan mode is showing that it is now realistic to perform quantitative and qualitative analysis with the same instrument. Moreover, HR-full scan acquisition offers a global view of sample extracts and allows retrospective investigations as virtually all ionized compounds are detected with a high sensitivity. In time, the versatility of HRMS together with the increasing need for relative quantification of hundreds of endogenous metabolites should promote a shift from triple-quadrupole MS to HRMS. However, a current "pitfall" in quantitative LC-HRMS analysis is the lack of HRMS-specific guidance for validated quantitative analyses. Indeed, false positive and false negative HRMS detections are rare, albeit possible, if inadequate parameters are used. Here, we investigated two key parameters for the validation of LC-HRMS quantitative analyses: the mass accuracy (MA) and the mass-extraction-window (MEW) that is used to construct the extracted-ion-chromatograms. We propose MA-parameters, graphs, and equations to calculate rational MEW width for the validation of quantitative LC-HRMS methods. MA measurements were performed on four different LC-HRMS platforms. Experimentally determined MEW values ranged between 5.6 and 16.5 ppm and depended on the HRMS platform, its working environment, the calibration procedure, and the analyte considered. The proposed procedure provides a fit-for-purpose MEW determination and prevents false detections.


Inhibition of nociceptive responses after systemic administration of amidated kyotorphin.
Kyotorphin (KTP; L-Tyr-L-Arg), an endogenous neuropeptide, is potently analgesic when delivered directly to the central nervous system. Its weak analgesic effects after systemic administration have been explained by inability to cross the blood-brain barrier (BBB) and detract from the possible clinical use of KTP as an analgesic. In this study, we aimed to increase the lipophilicity of KTP by amidation and to evaluate the analgesic efficacy of a new KTP derivative (KTP-amide - KTP-NH(2) ). We synthesized KTP-NH(2) . This peptide was given systemically to assess its ability to cross the BBB. A wide range of pain models, including acute, sustained and chronic inflammatory and neuropathic pain, were used to characterize analgesic efficacies of KTP-NH(2) . Binding to opioid receptors and toxicity were also measured. KTP-NH(2) , unlike its precursor KTP, was lipophilic and highly analgesic following systemic administration in several acute and chronic pain models, without inducing toxic effects or affecting motor responses and blood pressure. Binding to opioid receptors was minimal. KTP-NH(2) inhibited nociceptive responses of spinal neurons. Its analgesic effects were prevented by intrathecal or i.p. administration of naloxone. Amidation allowed KTP to show good analgesic ability after systemic delivery in acute and chronic pain models. The indirect opioid-mediated actions of KTP-NH(2) may explain why this compound retained its analgesic effects although the usual side effects of opioids were absent, which is a desired feature in next-generation pain medications.


Dietary cholesterol: from physiology to cardiovascular risk.
Dietary cholesterol comes exclusively from animal sources, thus it is naturally present in our diet and tissues. It is an important component of cell membranes and a precursor of bile acids, steroid hormones and vitamin D. Contrary to phytosterols (originated from plants), cholesterol is synthesised in the human body in order to maintain a stable pool when dietary intake is low. Given the necessity for cholesterol, very effective intestinal uptake mechanisms and enterohepatic bile acid and cholesterol reabsorption cycles exist; conversely, phytosterols are poorly absorbed and, indeed, rapidly excreted. Dietary cholesterol content does not significantly influence plasma cholesterol values, which are regulated by different genetic and nutritional factors that influence cholesterol absorption or synthesis. Some subjects are hyper-absorbers and others are hyper-responders, which implies new therapeutic issues. Epidemiological data do not support a link between dietary cholesterol and CVD. Recent biological data concerning the effect of dietary cholesterol on LDL receptor-related protein may explain the complexity of the effect of cholesterol on CVD risk.


Making a haplotype catalog with estimated frequencies based on SNP homozygotes.
Understanding the structure and frequencies of haplotypes is important for associating genetic polymorphisms with a given trait and for inferring the genetic genealogy of alleles in a population. Single nucleotide polymorphism (SNP) haplotypes can be determined without ambiguity when an individual does not have more than one heterozygous site in a given genomic region. Using genome-wide SNP genotypes for 3397 individuals from the Japanese population, we detected SNP homozygotes in the genomic regions of 1955 genes, determined haplotypes, and examined the efficiency of haplotype frequency estimation based on the proportion of SNP homozygotes in the sample. The estimated haplotype frequencies were very similar to the frequencies obtained by two statistical methods, PHASE and SNPHAP. We applied this approach to the genomic regions of 11 351 genes, and the results suggested that the sum of the frequencies of unobserved haplotypes is negligible for an analysis of a 100 kb genomic region with approximately 20 SNPs. Determination of haplotypes from homozygotes using genotype data from thousands of individuals, without a long computation time, appears to be useful for detecting real haplotypes including some low-frequency haplotypes. In addition, the unambiguously determined haplotypes with their estimated frequencies can be used as a catalog of haplotypes for the population, which is useful for the design of genome-wide association studies.


Oxidative stress induced by fluoride in adult mice and their suckling pups.
To assess renal and liver damages in pregnant and lactating mice as well as in their suckling pups, Wistar female mice were given 500 ppm NaF (226 ppm F-) in drinking water from the 15th day of pregnancy until day 14 after delivery. All mice were sacrificed on day 14 after parturition. In the present work, we evaluated the effects of sodium fluoride on histopathological aspects of kidney, antioxidant status, lipid peroxidation levels and on the expression of four stress proteins (namely, the cytosolic heat shock proteins: HSP72, 73, 90 and the reticulum-associated GRP94). Histological studies have shown many abnormalities in mothers and their pups. Biochemical results showed that lipid peroxidation increased in NaF-treated mice, as evidenced by high kidney and liver thiobarbituric acid reactive substance (TBARS) levels. Alteration of the antioxidant system was confirmed by the significant decline of serum total antioxidant status and of superoxide dismutase (SOD) and glutathione peroxidase (GSH-Px) activities in red blood cells. Besides, fluoride treatment induced a decrease in serum levels of non-enzymatic antioxidants such as uric acid and of some oligoelements: zinc and copper, known to be cofactors of superoxide dismutase (SOD-Cu-Zn). Compared to control group, the 72kDa protein was found to be overexpressed in kidney of 14-day-old mice only. HSP90 expression in liver appeared moderately inhibited in mothers, but decreased significantly in their pups. No significant changes were detected in the expression of 94kDa protein in both liver and kidney. Results showed that fluoride given to dams led to an oxidative stress in mothers as well as in offspring able to induce enhanced lipid peroxidation levels and protein conformational changes, as suggested by stress protein (HSP, GRP) expression changes.


Microscopic pathways for stress relaxation in repulsive colloidal glasses.
Residual stresses are well-known companions of all glassy materials. They affect and, in many cases, even strongly modify important material properties like the mechanical response and the optical transparency. The mechanisms through which stresses affect such properties are, in many cases, still under study, and their full understanding can pave the way to a full exploitation of stress as a primary control parameter. It is, for example, known that stresses promote particle mobility at small length scales, e.g., in colloidal glasses, gels, and metallic glasses, but this connection still remains essentially qualitative. Exploiting a preparation protocol that leads to colloidal glasses with an exceptionally directional built-in stress field, we characterize the stress-induced dynamics and show that it can be visualized as a collection of "flickering," mobile regions with linear sizes of the order of ≈20 particle diameters (≈2 μm here) that move cooperatively, displaying an overall stationary but locally ballistic dynamics.


Socioeconomic status and changing inequalities in colorectal cancer? A review of the associations with risk, treatment and outcome.
Upcoming mass screening for colorectal cancer (CRC) makes a review of recent literature on the association with socioeconomic status (SES) relevant, because of marked and contradictory associations with risk, treatment and outcome. The Pubmed database using the MeSH terms 'Neoplasms' or 'Colorectal Neoplasms' and 'Socioeconomic Factors' for articles added between 1995 and 1st October 2009 led to 62 articles. Low SES groups exhibited a higher incidence compared with high SES groups in the US and Canada (range risk ratio (RR) 1.0-1.5), but mostly lower in Europe (RR 0.3-0.9). Treatment, survival and mortality all showed less favourable results for people with a lower socioeconomic status: Patients with a low SES received less often (neo)adjuvant therapy (RR ranging from 0.4 to 0.99), had worse survival rates (hazard ratio (HR) 1.3-1.8) and exhibited generally the highest mortality rates up to 1.6 for colon cancer in Europe and up to 3.1 for rectal cancer. A quite consistent trend was observed favouring individuals with a high SES compared to those with a low SES that still remains in terms of treatment, survival and thus also mortality. We did not find evidence that the low/high SES gradients for treatment chosen and outcome are decreasing. To meet increasing inequalities in mortality from CRC in Europe for people with a low SES and to make mass screening successful, a high participation rate needs to be realised of low SES people in the soon starting screening program.


Unraveling the mechanism of recognition of the 3' splice site of the adenovirus major late promoter intron by the alternative splicing factor PUF60.
Pre-mRNA splicing is critical for achieving required amounts of a transcript at a given time and for regulating production of encoded protein. A given pre-mRNA may be spliced in many ways, or not at all, giving rise to multiple gene products. Numerous splicing factors are recruited to pre-mRNA splice sites to ensure proper splicing. One such factor, the 60 kDa poly(U)-binding splicing factor (PUF60), is recruited to sites that are not always spliced, but rather function as alternative splice sites. In this study, we characterized the interaction of PUF60 with a splice site from the adenovirus major late promoter (the AdML 3' splice site, AdML3'). We found that the PUF60-AdML3' dissociation constants are in the micromolar range, with the binding affinity predominantly provided by PUF60's two central RNA recognition motifs (RRMs). A 1.95 Å crystal structure of the two PUF60 RRMs in complex with AdML3' revealed a dimeric organization placing two stretches of nucleic acid tracts in opposing directionalities, which can cause looping of nucleic acid and explain how PUF60 affects pre-mRNA geometry to effect splicing. Solution characterization of this complex by light-scattering and UV/Vis spectroscopy suggested a potential 2:1 (PUF602:AdML3') stoichiometry, consistent with the crystal structure. This work defines the sequence specificity of the alternative splicing factor PUF60 at the pre-mRNA 3' splice site. Our observations suggest that control of pre-mRNA directionality is important in the early stage of spliceosome assembly, and advance our understanding of the molecular mechanism by which alternative and constitutive splicing factors differentiate among 3' splice sites.


Intraocular in vivo imaging of activated T-lymphocytes expressing green-fluorescent protein after stimulation with endotoxin.
Intravital microscopy allows imaging of specific cell populations in vivo. The value of this technique is well established, but would be enhanced if one could distinguish functional states of cells in vivo. Interleukin-2 (IL-2) is expressed upon stimulation of T-cells and is a commonly used marker for T-cell activation. This study tests the use of enhanced green fluorescent protein (GFP) as a reporter gene for interleukin-2 (IL-2) expression in vivo. Characterization of mice that have the GFP gene under the control of IL-2 regulatory sequences has previously been published. Uveitis was induced by injection of E. coli endotoxin into the vitreous of these IL-2/GFPki transgenic mice. Four hours later, 3 microg of recombinant mouse IL-2 was injected into the anterior chambers of one group of mice. In vivo imaging of infiltrating cells in the iris stroma was performed with fluorescence microscopy at 6, 24, 48, and 72 h after endotoxin injection. The absolute number of fluorescent cells per mm2 was evaluated. Eyes with endotoxin-induced uveitis had cells that expressed GFP and were identifiable by intravital microscopy. The fluorescent cells were exclusively seen in the subset of cells that had infiltrated the iris stroma or arrested along the vascular endothelium. The number of GFP-positive infiltrating cells in the iris increased from undetectable at baseline to 0.5 cells/mm2 at 6 h and 1.3 cells/mm2 at 72 h. The animals that received endotoxin as well as IL-2 tended to have more GFP-positive cells at the 48-h and 72-h time points, but these differences were not statistically significant GFP is commonly used as a reporter gene for in vitro expression assays. The results presented here document that transgenic mice with GFP under the control of IL-2 regulatory elements can be used with intravital microscopy for in vivo expression assays that allow detection of activated T-cells at multiple time points within the same animal. This provides a novel method for temporal and spatial studies on the state of cell activation in inflammatory responses.


Does paracetamol (acetaminophen) reduce the pain of osteoarthritis? A meta-analysis of randomised controlled trials.
To assess the best available evidence for efficacy of paracetamol (acetaminophen) in the treatment of osteoarthritis (OA). Systematic review and meta-analysis of randomised controlled trials (RCTs). Medline, Embase, Scientific Citation Index, CINAHL, Cochrane Library, and conference abstracts in the past 2 years from the British Society for Rheumatology, the European League Against Rheumatism, the American College of Rheumatology, and the Osteoarthritis Research Society International. 10 RCTs including 1712 patients with either symptomatic OA of the knee (6 trials) or hip/knee (3 trials) or multiple joints (1 trial). (a). effect size (ES) for pain, stiffness, and functional scores from baseline to end point; (b). rate ratio (RR) and number needed to treat for clinical response rate and patient preference for treatment. Paracetamol was effective in relieving pain due to OA (ES = 0.21, 95% confidence interval (CI) 0.02 to 0.41). Non-steroidal anti-inflammatory drugs (NSAIDs) were better than paracetamol for pain relief (ES = 0.20, 95% CI 0.10 to 0.30). Clinical response rate was higher with NSAIDs than with paracetamol (RR = 1.24, 95% CI 1.08 to 1.41), and the number of patients who preferred NSAIDs was more than twice the number of those preferring paracetamol (RR = 2.46, 95% CI 1.51 to 4.12). NSAIDs were associated with more frequent gastrointestinal discomfort than paracetamol (RR = 1.35, 95% CI 1.05 to 1.75). Paracetamol is an effective agent for pain relief due to OA. Although safer, it is less effective than NSAIDs. For safety reasons paracetamol should be the first line treatment, with NSAIDs reserved for those who do not respond.


A complex role for the progesterone receptor in the response to vascular injury.
Clinical studies of hormone replacement therapy to prevent cardiovascular diseases have heightened interest in the cardiovascular effects of progestins. However, the role of the progesterone receptor (PR) in vascular biology has not been studied in vivo. We studied ovariectomized female PR knockout (PRKO) mice and their wild-type (WT) littermates using the mouse carotid artery injury model. Placebo-treated PRKO mice showed significantly greater vascular medial hypertrophy and vascular smooth muscle cell (VSMC) proliferation in response to vascular injury than did WT mice. Progesterone had no significant effect in the PRKO mice, but worsened the response to injury in WT mice. VSMCs cultured from PRKO mouse aortae were markedly hyperproliferative, and their growth was not affected by progesterone. In contrast to the in vivo findings, progesterone inhibited proliferation of WT-derived VSMCs. Furthermore, reintroduction of PR into PRKO-derived VSMCs using adenoviral methods restored progesterone-mediated inhibition of proliferation to these cells. This effect was reversed by the PR antagonist, RU 486. Thus, the effects of PR and progesterone differ markedly between cultured VSMCs and intact blood vessels. These data demonstrate a direct role for the PR in regulating the response to vascular injury and VSMC proliferation.


Probability of sepsis after infection consultations in primary care in the United Kingdom in 2002-2017: Population-based cohort study and decision analytic model.
Efforts to reduce unnecessary antibiotic prescribing have coincided with increasing awareness of sepsis. We aimed to estimate the probability of sepsis following infection consultations in primary care when antibiotics were or were not prescribed. We conducted a cohort study including all registered patients at 706 general practices in the United Kingdom Clinical Practice Research Datalink, with 66.2 million person-years of follow-up from 2002 to 2017. There were 35,244 first episodes of sepsis (17,886, 51%, female; median age 71 years, interquartile range 57-82 years). Consultations for respiratory tract infection (RTI), skin or urinary tract infection (UTI), and antibiotic prescriptions were exposures. A Bayesian decision tree was used to estimate the probability (95% uncertainty intervals [UIs]) of sepsis following an infection consultation. Age, gender, and frailty were evaluated as association modifiers. The probability of sepsis was lower if an antibiotic was prescribed, but the number of antibiotic prescriptions required to prevent one episode of sepsis (number needed to treat [NNT]) decreased with age. At 0-4 years old, the NNT was 29,773 (95% UI 18,458-71,091) in boys and 27,014 (16,739-65,709) in girls; over 85 years old, NNT was 262 (236-293) in men and 385 (352-421) in women. Frailty was associated with greater risk of sepsis and lower NNT. For severely frail patients aged 55-64 years, the NNT was 247 (156-459) in men and 343 (234-556) in women. At all ages, the probability of sepsis was greatest for UTI, followed by skin infection, followed by RTI. At 65-74 years, the NNT following RTI was 1,257 (1,112-1,434) in men and 2,278 (1,966-2,686) in women; the NNT following skin infection was 503 (398-646) in men and 784 (602-1,051) in women; following UTI, the NNT was 121 (102-145) in men and 284 (241-342) in women. NNT values were generally smaller for the period from 2014 to 2017, when sepsis was diagnosed more frequently. Lack of random allocation to antibiotic therapy might have biased estimates; patients may sometimes experience sepsis or receive antibiotic prescriptions without these being recorded in primary care; recording of sepsis has increased over the study period. These stratified estimates of risk help to identify groups in which antibiotic prescribing may be more safely reduced. Risks of sepsis and benefits of antibiotics are more substantial among older adults, persons with more advanced frailty, or following UTIs.


In vivo protection of nigral dopamine neurons by lentiviral gene transfer of the novel GDNF-family member neublastin/artemin.
The glial cell line-derived neurotrophic factor (GDNF)-family of neurotrophic factors consisted until recently of three members, GDNF, neurturin, and persephin. We describe here the cloning of a new GDNF-family member, neublastin (NBN), identical to artemin (ART), recently published (Baloh et al., 1998). Addition of NBN/ART to cultures of fetal mesencephalic dopamine (DA) neurons increased the number of surviving tyrosine hydroxylase (TH)-immunoreactive neurons by approximately 70%, similar to the maximal effect obtained with GDNF. To investigate the neuroprotective effects in vivo, lentiviral vectors carrying the cDNA for NBN/ART or GDNF were injected into the striatum and ventral midbrain. Three weeks after an intrastriatal 6-hydroxydopamine lesion only about 20% of the nigral DA neurons were left in the control group, while 80-90% of the DA neurons remained in the NBN/ART and GDNF treatment groups, and the striatal TH-immunoreactive innervation was partly spared. We conclude that NBN/ART, similarly to GDNF, is a potent neuroprotective factor for the nigrostriatal DA neurons in vivo.


The relationship between economic growth and carbon emissions in G-7 countries: evidence from time-varying parameters with a long history.
This paper re-investigates the time-varying impacts of economic growth on carbon emissions in the G-7 countries over a long history. In doing so, the historical data spanning the period from the 1800s to 2010 (as constructed) for each country is examined using the time-varying cointegration and bootstrap-rolling window estimation approach. Unlike the previous environmental Kuznets curve (EKC) studies, using this methodology gives us avenue to detect more than one, two, or more turning points for the economic growth-carbon emissions nexus. The empirical findings show that the nexus between economic growth and carbon emission seems over a long history to be M-shaped for Canada and the UK; N-shaped for France; inverted N-shaped for Germany; and inverted M-shaped (W-shaped) for Italy, Japan, and the USA. In addition, the possible validity of EKC hypothesis is examined for both the pre-1973 and post-1973 sub-periods. Based on this investigation, we found that an inverted U-shaped is confirmed only for the pre-1973 period in France, Italy, and the USA. These empirical evidences provide new insights to policy makers to improve environmental quality using economic growth as an economic tool for the long run by observing changes in the environmental impact of this growth from year to year.


Meaning out of chaos: a way to understand chronic pain.
Pain is a multidimensional phenomenon lying at the intersection between biology and culture. The modern understanding of pain takes into account emotional, psychological, socio-political and existential aspects of pain as well as physiological, anatomical factors. Our aim in this study was to deepen the understanding of psychosocial, existential aspects of pain and to discuss how clinicians can better understand and treat patients with chronic pain. A focus group was formed consisting of a researcher and a group of clinicians (n = 3) with various backgrounds working at a specialized pain clinic. The group met once a month during a 6-month period. Questions concerning the life-world of the pain patient as well as inquiries into the conditions for 'the good clinical encounter' were investigated. The results of this study consist of a systematization of the data (focus group meetings) collected and analysed in a collaborative effort between the researcher and group participants. The findings are presented in terms of themes. The main metaphor used to describe the path from the seeking of medical help to successful rehabilitation was order out of chaos. Ordering chaos was a process moving from diagnosis through a phase of heightened self-awareness towards responsibility-taking on the part of the patient. Related themes presented, illustrated and discussed in the paper concern problems of linkage, the role of flexibility and creativity in the healing process and the kind of clinical encounter conducive for the journey from chaos to the creation of new meaning.


Pilot study to evaluate a novel three-dimensional wound measurement device.
As the burden of diabetes continues to grow and treatment standards require careful tracking of wound progress, clinicians increasingly need to rely on technological improvements in wound measurement technologies to track the progress of their treatments. This study aims to determine the accuracy of a new three-dimensional wound measurement (3DWM) device against laser-assisted wound measurement (LAWM) devices and traditional methods of wound measurement. Using several wound models, we demonstrate that the 3DWM device measures wound area, depth and volume similarly to the other methods tested. This is especially apparent when changes in wound measurements were compared between the two devices. Differences between the two technologies were apparent when analysing wound measurement time and measurement repeatability. There was a significantly lower incidence of error in measurements between the 3DWM device and the LAWM device. Finally, the measurement time was significantly faster with the 3DWM device compared to the LAWM device. Together, these data demonstrate that the 3DWM device provides an accurate and reproducible method for measuring changes in wound healing similar to other available technologies. Further, the use of the 3DWM device provides a faster and more consistent measurement, which is critical for clinical application and use.


Filament condition-specific response elements control the expression of NRG1 and UME6, key transcriptional regulators of morphology and virulence in Candida albicans.
Candida albicans is the most frequently isolated human fungal pathogen and can cause a range of mucosal and systemic infections in immunocompromised individuals. Morphogenesis, the ability to undergo a reversible transition from budding yeast to elongated filaments, is an essential virulence trait. The yeast-to-filament transition is associated with expression of genes specifically important for filamentation as well as other virulence-related processes, and is controlled, in part, by the key transcriptional regulators Nrg1 and Ume6. Both of these regulators are themselves controlled at the transcriptional level by filament-inducing environmental cues, although little is known about how this process occurs. In order to address this question and determine whether environmental signals regulate transcription of UME6 and NRG1 via distinct and/or common promoter elements, we performed promoter deletion analyses. Strains bearing promoter deletion constructs were induced to form filaments in YEPD plus 10% serum at 37°C, Spider medium (nitrogen and carbon starvation) and/or Lee's medium pH 6.8 (neutral pH) and reporter gene expression was measured. In the NRG1 promoter we identified several distinct condition-specific response elements for YEPD plus 10% serum at 37°C and Spider medium. In the UME6 promoter we also identified response elements for YEPD plus 10% serum at 37°C. While a few of these elements are distinct, others overlap with those which respond to Lee's pH 6.8 medium. Consistent with UME6 possessing a very long 5' UTR, many response elements in the UME6 promoter are located significantly upstream from the coding sequence. Our data indicate that certain distinct condition-specific elements can control expression of C. albicans UME6 and NRG1 in response to key filament-inducing environmental cues. Because C. albicans encounters a variety of host microenvironments during infection, our results suggest that UME6 and NRG1 expression can be differentially modulated by multiple signaling pathways to control filamentation and virulence in vivo.


Consequences of workplace violence directed at nurses.
The consequences of workplace violence (WPV) are far-reaching, and impact on the nurse, the perpetrator and the organization. However, the authors were unable to identify any research in the literature on nurses' perceptions of the consequences of WPV in non-teaching hospital settings. This study therefore aimed to examine nurses' perspectives of the consequences of WPV, to identify ways to reduce the impact of these incidents. A descriptive, exploratory approach was adopted to collect qualitative survey and interview data from nurses working in several areas of one West Australian non-teaching hospital in 2006. Three themes emerged from the data: nurse, perpetrator and organizational consequences. The sub-themes included nurses accepting that WPV is part of their job; physical and emotional effects; not feeling competent; avoiding patients; organizational costs of WPV; adverse effects of restraint; and disruption to patient care. Participants experienced several negative consequences as a result of WPV. Recommendations for improving the safety of hospitals for staff and patients are made in light of the findings.


Molecular profiling and targeted therapy for advanced thoracic malignancies: a biomarker-derived, multiarm, multihistology phase II basket trial.
We conducted a basket clinical trial to assess the feasibility of such a design strategy and to independently evaluate the effects of multiple targeted agents against specific molecular aberrations in multiple histologic subtypes concurrently. We enrolled patients with advanced non-small-cell lung cancer (NSCLC), small-cell lung cancer, and thymic malignancies who underwent genomic characterization of oncogenic drivers. Patients were enrolled onto a not-otherwise-specified arm and treated with standard-of-care therapies or one of the following five biomarker-matched treatment groups: erlotinib for EGFR mutations; selumetinib for KRAS, NRAS, HRAS, or BRAF mutations; MK2206 for PIK3CA, AKT, or PTEN mutations; lapatinib for ERBB2 mutations or amplifications; and sunitinib for KIT or PDGFRA mutations or amplification. Six hundred forty-seven patients were enrolled, and 88% had their tumors tested for at least one gene. EGFR mutation frequency was 22.1% in NSCLC, and erlotinib achieved a response rate of 60% (95% CI, 32.3% to 83.7%). KRAS mutation frequency was 24.9% in NSCLC, and selumetinib failed to achieve its primary end point, with a response rate of 11% (95% CI, 0% to 48%). Completion of accrual to all other arms was not feasible. In NSCLC, patients with EGFR mutations had the longest median survival (3.51 years; 95% CI, 2.89 to 5.5 years), followed by those with ALK rearrangements (2.94 years; 95% CI, 1.66 to 4.61 years), those with KRAS mutations (2.3 years; 95% CI, 2.3 to 2.17 years), those with other genetic abnormalities (2.17 years; 95% CI, 1.3 to 2.74 years), and those without an actionable mutation (1.85 years; 95% CI, 1.61 to 2.13 years). This basket trial design was not feasible for many of the arms with rare mutations, but it allowed the study of the genetics of less common malignancies.


Skeletal evidence of trepanning on a 5th century skull from Ludbreg, Croatia.
This paper presents a case of skull trepanation from the town of Ludbreg in north-western Croatia. This is the second case of trepanation reported in human skeletal remains from archaeological sites in Croatia. The procedure was performed on an adult male buried in a tomb under tegulae at the "Somođi Garden" site. The archaeological context and radiocarbon analysis date the bone sample to the 5th century AD, i.e. to the Migration Period. The trepanned aperture is located on the left side of the frontal bone and the left parietal bone. The lesion is oval in shape with all three layers of the calvarium breached, leaving dura mater exposed. This neurosurgical intervention was most probably performed by scraping for therapeutic reasons (head injury). The morphology of the lesion strongly indicates that the individual survived the operation and lived for a longer period of time, several years at least.


Complete genome sequencing and genomic characterization of two Escherichia coli strains co-producing MCR-1 and NDM-1 from bloodstream infection.
We previously described the discovery of two Escherichia coli isolates (EC1002 and EC2474) co-harbouring mcr-1 and bla NDM-1 genes, which were recovered from bloodstream infection in China. More importantly, these antibiotic resistance genes were located on different plasmids and signaling the potential spread of pandrug-resistant bacteria. Here, the complete genome sequences of both isolates were determined using Pacbio RS II and Illumina HiSeq2000 systems. The genome of EC1002 consists of a 5,177,501 base pair chromosome and four circular plasmids, while the genome of EC2474 consists of a 5,013,813 base pair chromosome and three plasmids. The plasmid replicon type of pEC1002_NDM and pEC2474_NDM were identified as IncA/C2 and IncF, respectively. The genetic environment of bla NDM-1 in this study was similar to bla NDM-carrying plasmids detected in China, although the overall nucleotide identity and query coverage were variable. The plasmid replicon type of pEC1002_MCR and pEC2474_MCR were identified as IncI2 and IncHI2, respectively. Two different genetic strategies for mcr-1 gene spread were observed in this study and bla NDM-1 genes were also found transferred by two different mobile genetic elements in two plasmids. The findings of this study further support that the diversified transfer mechanisms of bla NDM-1 and mcr-1 present in Enterobacteriaceae.


The influence of the descending pain modulatory system on infant pain-related brain activity.
The descending pain modulatory system (DPMS) constitutes a network of widely distributed brain regions whose integrated function is essential for effective modulation of sensory input to the central nervous system and behavioural responses to pain. Animal studies demonstrate that young rodents have an immature DPMS, but comparable studies have not been conducted in human infants. In Goksan et al. (2015) we used functional MRI (fMRI) to show that pain-related brain activity in newborn infants is similar to that observed in adults. Here, we investigated whether the functional network connectivity strength across the infant DPMS influences the magnitude of this brain activity. FMRI scans were collected while mild mechanical noxious stimulation was applied to the infant's foot. Greater pre-stimulus functional network connectivity across the DPMS was significantly associated with lower noxious-evoked brain activity (p = 0.0004, r = -0.86, n = 13), suggesting that in newborn infants the DPMS may regulate the magnitude of noxious-evoked brain activity.


Mass spectrometry quantification of clusterin in the human brain.
The multifunctional glycoprotein clusterin has been associated with late-onset Alzheimer's disease (AD). Further investigation to define the role of clusterin in AD phenotypes would be aided by the development of techniques to quantify level, potential post-translational modifications, and isoforms of clusterin. We have developed a quantitative technique based on multiple reaction monitoring (MRM) mass spectrometry to measure clusterin in human postmortem brain tissues. A stable isotope-labeled concatenated peptide (QconCAT) bearing selected peptides from clusterin was expressed with an in vitro translation system and purified. This clusterin QconCAT was validated for use as an internal standard for clusterin quantification using MRM mass spectrometry. Measurements were performed on the human postmortem frontal and temporal cortex from control and severe AD cases. During brain tissues processing, 1% SDS was used in the homogenization buffer to preserve potential post-translational modifications of clusterin. However, MRM quantifications in the brain did not suggest phosphorylation of Thr(393), Ser(394), and Ser(396) residues reported for clusterin in serum. MRM quantifications in the frontal cortex demonstrated significantly higher (P < 0.01) level of clusterin in severe AD group (39.1 ± 9.1 pmol/mg tissue protein) in comparison to control group (25.4 ± 4.4 pmol/mg tissue protein). In the temporal cortex, the clusterin levels were not significantly different, 29.0 ± 7.9 pmol/mg tissue protein and 28.0 ± 8.4 pmol/mg tissue protein in control and severe AD groups, respectively. The proposed protocol is a universal quantitative technique to assess expression level of clusterin. It is expected that application of this protocol to quantification of various clusterin isoforms and potential post-translational modifications will be helpful in addressing the role of clusterin in AD.


Hospital characteristics associated with post-discharge suicide of severely depressed patients.
To explore risk factors, such as characteristics of psychiatrist, patient and hospital at index hospitalization, associated with depressed patients who committed suicide within 3 months of discharge using a case-control design. By linking the Taiwanese nationwide mortality database and the National Health Insurance dataset, all hospitalized patients with major depression who committed suicide within a 90-day period post-discharge during the years 2002-2004 were selected as a study cohort (n=85). We randomly selected 425 cases (five for every case in the study cohort) that were matched with the study cohort in terms of age, gender and date of discharge as a control cohort. Cox proportional hazard regression was carried out to compute the adjusted 90-day survival rate after adjusting for other factors. The majority of suicide occurred 30 days after discharge from hospitals, with a mean of 29.9 days. The adjusted hazard for committing suicide after hospital discharge for patients who left on their own initiative was 2.85 times (95% CI=1.387-5.856, p=0.004) greater than for those who were discharged with doctors' approval. Furthermore, the adjusted hazard for patients who were discharged from medical centers was higher than for patients discharged from regional hospitals, by a multiple of 3.38 (95% CI=1.421-8.055, p=0.006). Some patient-specific predictors of post-discharge suicide, such as suicidal ideation prior to admission, recent life events and social relationships with close relatives were not available. We conclude that, in addition to patient characteristics, hospital characteristics also constitute important risk factors for suicide within 90 days of discharge for depressed patients.


Investigation into the optimal prosthetic material for wound healing of abdominal wall defects.
The purpose of this experimental study is to investigate and compare the effects of prosthetic materials used for wound healing of abdominal wall hernias. A total of 60 rats were divided into five equal groups: Group I, control subjected to laparotomy; group II, abdominal wall defect 3×2 cm+polypropylene (PP) mesh; group III, abdominal wall defect 3×2 cm+PP mesh+hyaluronate and carboxymethylcellulose (H-CMC; Seprafilm®); group IV, abdominal wall defect 3×2 cm+polytetrafluoroethylene (PTFE; Composix™); and group V, abdominal wall defect 3×2 cm+polyethylene terephthalate (PET; Dacron®). A total of 14 days after the surgery, rats were sacrificed and the meshes with the surrounding tissue were extracted in block. The breaking strength of the mesh from the fascia was recorded. The healing tissue was examined with the index of histopathology and the hydroxyproline value was analyzed using the Switzer method. Both the breaking strength and histopathological index of the wound healing were significantly improved in groups II and III compared with that in groups IV and V (P<0.001). Hydroxyproline values were the highest in group I (P<0.001). There was also a statistically significant difference between groups II and IV, and group V and the other groups (P<0.001). The present findings demonstrated that PP mesh and PP mesh+H-CMC had a superior breaking strength and improved histopathologic indices compared with PTFE and PET. Furthermore, hydroxyproline values were the lowest in the PET group. In conclusion, wound healing was improved in the PP mesh group and the PP mesh+H-CMC group compared with the PTFE and PET groups according to the present study parameters.